The Profile of the Expression of IBMR3 Antigens 3T3 and HT29 Cancer Cell Line after Analysis
Qutaiba Kafi Jassim Alrawi
Issue:
Volume 2, Issue 4, August 2014
Pages:
55-61
Received:
30 June 2014
Accepted:
15 July 2014
Published:
30 July 2014
Abstract: Aims: The aim of this study was to evaluate the effectiveness of specific monoclonal IBMR3 antibodies expression in the cancer cell lines of 3T3 and HT29, in order to recognize specific antigen and make evaluation, using immunoblotting technique which is widely use in this filed. Methods: Protein extracts were extracted from these cancer cell lines and used for western bloat. The Immunobloting were consequently subjected to densitometry analysis using bioimaging machine. This bioimaging process will facilitate to measure the molecular weights, peak height and raw volume of the protein bands for the 3T3 and HT29 cancer cell lines, which helps in diagnosis of any pathogenic antigen. Results: The bands obtained from bioimaging were exposed on the PVDF membrane. In the 3T3 bio imaging process revealed four bands and molecular weight were 299.58, 87.90, 41.67 and 23.54 KDa. However, bioimaging results for HT29 revealed also four bands with molecular weights: 90.11, 41.31, 23.87 and 20.86 KDa. The results of Peak height densitometry for IBMR3 antigen bands for 3T3 were: 1856.985, 551.769, 394.164, and 216.185. However HT29 peak height results were: 281.544, 101.711.202.668, 757.213. The raw volume (amount of protein bands of IBMR3 Ag for 3T3) were: 1460168.75, 206078.47, 161406.89, and 219583.16. However HT29 raw volume results were: 110197.11, 76106.84, 98632.59, and 221395.34. Negative protein staining for 3T3 and HT29 cancer cell line bands were done by using mouse IgM serotype. IgM serotype was not indicated that means no specific antigens for negative control IgM. Conclusion: The bioimaging revealed different results of the expression profile in molecular weight, peak height and raw volume between the two cancer cell lines. The results from this study suggest that the IBMR3 antigens were differentially expressed in 3T3 and HT29 and the molecular weight is higher in the 3T3 than HT29 cancer cell lines. In future it will be beneficial to categorize and study the character analysis of the IBMR3 antigen and its prospective role in cancer cells.
Abstract: Aims: The aim of this study was to evaluate the effectiveness of specific monoclonal IBMR3 antibodies expression in the cancer cell lines of 3T3 and HT29, in order to recognize specific antigen and make evaluation, using immunoblotting technique which is widely use in this filed. Methods: Protein extracts were extracted from these cancer cell lines...
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Relationship between Platelet Aggregatory and Heparin-Induced Thrombocytopenia Type II
Harunobu Iwase,
Hiroko Kariyazono,
Junko Arima,
Hiroyuki Yamamoto,
Kazuo Nakamura
Issue:
Volume 2, Issue 4, August 2014
Pages:
78-86
Received:
21 August 2014
Accepted:
4 September 2014
Published:
20 September 2014
Abstract: Heparin-induced thrombocytopenia type II (HIT) is a significant adverse effect of heparin treatment in either therapeutic or prophylactic doses. HIT is an immune-mediated disorder caused by IgG antibodies that bind to platelet factor 4 (PF4), which becomes an antigenic target when bound to heparin. The possibility of HIT is suspected when patients show a reduction in the platelet count while receiving heparin. However, the diagnosis of HIT may be difficult on the basis of clinical symptoms alone, especially in patients with other diseases that may induce thrombocytopenia. Therefore, confirmation of HIT by using biologic or antigen assays is required. To analyze whether two patients in whom thrombocytopenia was induced after initiating administration of unfractionated heparin during percutaneous transluminal coronary angioplasty or coronary artery bypass grafting surgery had HIT, we measured levels of platelet aggregability, anti-heparin-PF4 complex antibody (anti-HIT antibody), tumor necrosis factor (TNF)-alpha, TNF-receptor 1 (TNF-R1), interleukin (IL)-6, and thrombin-antithrombin III complex (TAT). The two patients were judged to be HIT-positive from the results that platelet aggregation induced with induced by ADP 1 μM as well as collagen 0.2 μg/mL increased after addition of UFH. ELISA results of two patients showing optical density (OD) values greater than 0.40 were regarded as positive. Additionally, the levels of TNF-alpha and TAT in both patients were higher than in the control patients who underwent CABG without thrombocytopenia after heparin therapy initiation. The results suggest that blood coagulation is enhanced and an inflammatory reaction is induced in the endothelial cells of patients with HIT. In conclusion, the combined measurement of platelet aggregation and anti-HIT antibodies is crucial for defining HIT status, and measurement of TNF-alpha and TAT may play a significant role in the practice of anticoagulant therapy.
Abstract: Heparin-induced thrombocytopenia type II (HIT) is a significant adverse effect of heparin treatment in either therapeutic or prophylactic doses. HIT is an immune-mediated disorder caused by IgG antibodies that bind to platelet factor 4 (PF4), which becomes an antigenic target when bound to heparin. The possibility of HIT is suspected when patients ...
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