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University of Texas MD Anderson Cancer CenterComparison of Pinnacle and SameSpots for Spot Quantification and Differential Expression in 2-Dimensional Gel Electrophoresis StudiesJeffrey S. Morris Sun, 15 Feb 2009 04:27:12 -0800
2-DE is an important method for proteomics. Accurate spot detection and quantification on the resulting images is a challenging task, but must be done effectively for the technology to fulfill its potential. Traditional analytical methods have significant weaknesses, including spot mismatching and missing data. These problems require time-consuming manual editing to correct, which dramatically decreases throughput and compromises the objectivity and reproducibility of the analysis. To address this issue, we developed Pinnacle, a new method that markedly improves spot detection and quantification precision. Another new method implemented in Progenesis SameSpots, a commercial 2-DE analysis product, has also been touted as an improvement upon traditional approaches. In this study, we compared Pinnacle and SameSpots in spot detection and precision of quantification using two different dilution series, and evaluated the detection of differentially expressed proteins in two differential expression experiments. We found that SameSpots at times had problems with spot delineation, while Pinnacle did not. While both Pinnacle and SameSpots showed marked improvement in precision of spot quantifications over conventional methods, Pinnacle yielded spot quantifications with greater validity and reliability than SameSpots. Pinnacle detected more differentially expressed proteins than SameSpots, which may be a result of its increased precision and improved spot delineation.
Statistical Issues in Proteomic ResearchJeffrey S. Morris Thu, 31 Jul 2008 11:03:24 -0700
Microproteomics: Analysis of protein diversity in small samplesHoward B. Gutstein Fri, 13 Jun 2008 14:38:53 -0700
Proteomics, the large-scale study of protein expression in organisms, offers the potential to evaluate global changes in protein expression and their post-translational modifications that take place in response to normal or pathological stimuli. One challenge has been the requirement for substantial amounts of tissue in order to perform comprehensive proteomic characterization. In heterogeneous tissues, such as brain, this has limited the application of proteomic methodologies. Efforts to adapt standard methods of tissue sampling, protein extraction, arraying, and identification are reviewed, with an emphasis on those appropriate to smaller samples ranging in size from several microliters down to single cells. The effects of miniaturization on these analyses are highlighted using neuroscience-related examples, as are statistical issues unique to the high-dimensional datasets generated by proteomic experiments.
Pinnacle: A Fast, Automatic Method for Detecting and Quantifying Protein Spots in 2-Dimensional Gel Electrophoresis DataJeffrey S. Morris Tue, 04 Dec 2007 09:44:53 -0800
Motivation: One of the key limitations for proteomic studies using 2-dimensional gel electrophoresis (2DE) is the lack of rapid, robust, and reproducible methods for detecting, matching, and quantifying protein spots. The most commonly used approaches involve first detecting spots and drawing spot boundaries on individual gels, then matching spots across gels, and finally quantifying each spot by calculating normalized spot volumes. This approach is time con-suming, error-prone, and frequently requires extensive manual edit-ing, which can unintentionally introduce bias into the results.Results: We introduce a new method for spot detection and quanti-fication called Pinnacle that is automatic, quick, sensitive and spe-cific, and yields spot quantifications that are reliable and precise. This method incorporates a spot definition that is based on simple, straightforward criteria rather than complex arbitrary definitions, and results in no missing data. Using dilution series for validation, we demonstrate Pinnacle outperformed two well-established 2DE analysis packages, proving to be more accurate and yielding smaller CVs. More accurate quantifications may lead to increased power for detecting differentially expressed spots, an idea supported by the results of our group comparison experiment. Our fast, automatic analysis method makes it feasible to conduct very large 2DE-based proteomic studies that are adequately powered to find important protein expression differences.Availability: Matlab code to implement Pinnacle is available from the authors upon request for non-commercial use.
Laser capture sampling and analytical issues in proteomicsHoward Gutstein Tue, 04 Dec 2007 09:35:54 -0800
Proteomics holds the promise of evaluating global changes in protein expression and post-translational modificaiton in response to environmental stimuli. However, difficulties in achieving cellular anatomic resolution and extracting specific types of proteins from cells have limited the efficacy of these techniques. Laser capture microdissection has provided a solution to the problem of anatomical resolution in tissues. New extraction methodologies have expanded the range of proteins identified in subsequent analyses. This review will examine the application of laser capture microdissection to proteomic tissue sampling, and subsequent extraction of these samples for differential expression analysis. Statistical and other quantitative issues important for the analysis of the highly complex datasets generated are also reviewed.
Statistical contributions to proteomic researchJeffrey S. Morris Wed, 04 Apr 2007 12:55:09 -0700
Proteomic profiling has the potential to impact the diagnosis, prognosis, and treatment of various diseases. A number of different proteomic technologies are available that allow us to look at many proteins at once, and all of them yield complex data that raise significant quantitative challenges. Inadequate attention to these quantitative issues can prevent these studies from achieving their desired goals, and can even lead to invalid results. In this chapter, we describe various ways the involvement of statisticians or other quantitative scientists in the study team can contribute to the success of proteomic research, and we outline some of the key statistical principles that should guide the experimental design and analysis of such studies.
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The Oral Cancer Foundation - Discusses issues related to these conditions as well as the activities of the organization. Located in Newport Beach, California.
Meta Description: [ Oral cancer information and resources for both the public, health care professionals and care givers. ]
American Oral Cancer Foundation - Dedicated to the early detection through education and screening.
Meta Description: [ The American Oral Cancer Foundation, a nonprofit organization, is committed
to enhancing the quality of lives for all people through the development and
dissemination of programs designed to educate the public, health care
professionals, and health care organizations in order to... ]
ATC: Oral Cancer - Overview summary from the American Thyroid Clinic of Austin, Texas.
Meta Description: [ oral cancer answers ]
Avinash Raina Foundation - Information about squamous cell carcinoma and its impact on a life.
Cancer of the Mouth and Throat - Information from the Calgary Regional Health Authority in Canada.
Dental Oncology Education Program - Texas program aimed to assist dental professionals with the detection, diagnosis and management of oral cancer.
eMedicine Health: Cancer of the Mouth and Throat - Consumer health resource providing information on causes, symptoms and treatment.
Meta Description: [ The oral cavity (mouth) and the upper part of the throat (pharynx) have roles in many important functions, including breathing, talking, chewing, and swallowing. The mouth and upper throat are ... ]
Examination for Oral Cancer - Maryland dentist resource offers instruction for inspection including photographs.
Meta Description: [ Oral Cancer Site includes self exam,links to resources, and information about the new non surgical,highly accurate brush biopsy. ]
Facts About Oral Cancers - Review article by Dr. Gregg Bobier.
Meta Description: [ A review article. This is part of the MedNews.Net Service. ]
Lip and Oral Cavity Cancer - General information with links to specific details of the disease from the U.S. National Cancer Institute.
Meta Description: [ Expert-reviewed information summary about the treatment of lip and oral cavity cancer. ]
MediFocus: Tongue Cancer - Basic information in a fact sheet format.
MedNews: Lip and Oral Cavity Cancer - General information features description, cellular classification, stages, disease overview, and resources.
National Cancer Institute: Lip and Oral Cavity Cancer - Information concerning treatment options, the four stages, recurrent disease, and resources.
Meta Description: [ Expert-reviewed information summary about the treatment of lip and oral cavity cancer. ]
National Institute of Dental and Craniofacial Research - Online resources from this U.S. Federal agency.
Oral Cancer Consortium - Outlines information, screenings and resources related to the member entities as located in the New Jersey and New York areas.
Oral Health Resources: Cancer - General information from the Centers for Disease Control and Prevention.
Raph's Tonsil Cancer - View from a patient including a personal history as well as general information.
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Sangeeta Darvekar Charitable Trust: Oral Cancer - Organization in India dedicated to increasing awareness and aiding patients.
Meta Description: [ A Website Dedicated to Cancer Of Mouth ]
The Extraoral Examination - Demonstration of a less obvious aspect of the search for oral cancer.
Tongue Cancer: A Patient's Story - General advice and information about preparation, tests, treatments and coping.
Meta Description: [ Cancer Patients, Tongue Cancer, Melanoma, Metastatic Melanoma, Lung surgery, Lung Wedge resection, Intussesception, Cancer Treatments, Cancer Tests, Head and Neck Cancer, Living with Cancer ]
University of Manitoba: Oral Self Examination - Inspection for signs of cancer.
Meta Description: [ Oral Cancer Self Examination ]
University of Pittsburgh Cancer Institute: Oral - Gives general information about these conditions and details available services.
What You Need To Know About Oral Cancer - Pamphlet from the National Cancer Institute includes prevention, detection, symptoms, treatment options, clinical trials and support.
Meta Description: [ Information about detection, symptoms, diagnosis, and treatment of oral cancer. NIH Publication No. 03-1574 ]