Evaluation of thymidylate synthase RNA expression by polymerase chain reaction

1994 ◽  
Vol 8 (1) ◽  
pp. 67-72 ◽  
Author(s):  
R.M. Elledge ◽  
N. Okuyama ◽  
S.D. Fitzgerald ◽  
S.A.W. Fuqua
Keyword(s):  
Polymerase Chain Reaction ◽  
Thymidylate Synthase ◽  
Rna Expression ◽  
Chain Reaction ◽  
Polymerase Chain

An intensive hands-on course designed to teach molecular biology techniques to physiology graduate students

2002 ◽  
Vol 26 (1) ◽  
pp. 42-49 ◽  
Author(s):  
Andrea D. Weston ◽  
Sasha Stasko ◽  
Gerald M. Kidder
Keyword(s):  
Polymerase Chain Reaction ◽  
Graduate Students ◽  
Molecular Biology ◽  
Historical Perspective ◽  
Rna Expression ◽  
Chain Reaction ◽  
Hands On ◽  
Similar Information ◽  
Polymerase Chain ◽  
Intensive Course

To address a growing need to make research trainees in physiology comfortable with the tools of molecular biology, we have developed a laboratory-intensive course designed for graduate students. This course is offered to a small group of students over a three-week period and is organized such that comprehensive background lectures are coupled with extensive hands-on experience. The course is divided into seven modules, each organized by a faculty member who has particular expertise in the area covered by that module. The modules focus on basic methods such as cDNA subcloning, sequencing, gene transfer, polymerase chain reaction, and protein and RNA expression analysis. Each module begins with a lecture that introduces the technique in detail by providing a historical perspective, describing both the uses and limitations of that technique, and comparing the method with others that yield similar information. Most of the lectures are followed by a laboratory session during which students follow protocols that were carefully designed to avoid pitfalls. Throughout these laboratory sessions, students are given an appreciation of the importance of proper technique and accuracy. Communication among the students, faculty, and the assistant coordinator is focused on when and why each procedure would be used, the importance of each step in the procedure, and approaches to troubleshooting. The course ends with an exam that is designed to test the students’ general understanding of each module and their ability to apply the various techniques to physiological questions.

scholarly journals Altered expression of small leucine-rich proteoglycans (Decorin, Biglycan and Lumican): Plausible diagnostic marker in urothelial carcinoma of bladder

Tumor Biology ◽  
2017 ◽  
Vol 39 (5) ◽  
pp. 101042831769911 ◽  
Author(s):  
Sandeep Appunni ◽  
Vivek Anand ◽  
Madhuram Khandelwal ◽  
Amlesh Seth ◽  
Sandeep Mathur ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Bladder Cancer ◽  
Urothelial Carcinoma ◽  
Messenger Rna ◽  
Quantitative Polymerase Chain Reaction ◽  
Rna Expression ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Lower Expression ◽  
Urothelial Carcinoma Of Bladder

Small leucine-rich proteoglycans are components of extracellular matrix that regulates neoplastic transformation. Among small leucine rich proteoglycans, Decorin, Biglycan and Lumican are most commonly implicated markers, and their expression is well studied in various malignancies. In this novel study, we have collectively evaluated expression of these three molecules in urothelial carcinoma of bladder. Thirty patients of confirmed untreated bladder cancer, 30 healthy controls for blood and 30 controls for adjacent non-tumour tissue were enrolled. Blood was collected from all subjects and tumour/adjacent normal tissue was obtained from the patients. Circulatory levels were estimated by enzyme-linked immunosorbent assay, relative messenger RNA expression by quantitative polymerase chain reaction and protein expression by immunohistochemistry and western-blotting. Circulatory levels of Biglycan (p = 0.0038) and Lumican (p < 0.0001) were significantly elevated, and that of Decorin (p < 0.0001) was significantly reduced in patients as compared with controls. Protein expression by immunohistochemistry and western-blotting showed elevated expression of Lumican and Biglycan and lower expression of Decorin in urothelial carcinoma of bladder. Quantitative polymerase chain reaction for messenger RNA expression from tissue specimens revealed significantly higher expression of Biglycan (p = 0.0008) and Lumican (p = 0.01) and lower expression of Decorin (p < 0.0001) in urothelial carcinoma of bladder. Out of all molecules receiver operating characteristic curve showed that the 0.207 ng/ml cut-off of serum Lumican provided optimum sensitivity (90.0%) and specificity (90.0%). Significant alteration of matrix small leucine-rich proteoglycans in urothelial carcinoma of bladder was observed. Higher expression of Lumican in Bladder cancer patients with the cut-off value of highest optimum sensitivity and specificity shows its importance as a potential non-invasive marker for early detection of UBC following further validation in large patient cohort.

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