1Department of Medical Laboratory Sciences, Faculty of Allied Medical Sciences, Al-Ahliyya Amman University, Amman JO 19328, Jordan, a.shraim@ammanu.edu.jo
Dr. Ala'a Shraim
1Department of Medical Laboratory Sciences, Faculty of Allied Medical Sciences, Al-Ahliyya Amman University, Amman JO 19328, Jordan, a.shraim@ammanu.edu.jo
Aptamers are molecules that reveal highly complex and refined molecular recognition properties. These molecules are capable of binding with high affinity and selectivity to targets, ranging from small molecules to whole living cells. Several aptamers have been selected for targeting cellular proteins and they have also used in developing therapeutics and diagnostic strategies. Epithelial cell adhesion molecule (EpCAM) is considered as a cancer stem cell (CSC) biomarker and one of the most promising targets for aptamer selection against CSCs. In this study, we have developed a ssDNA aptamer with high affinity and selectivity of targeting the EpCAM protein extracellular domain. The SELEX technique was applied and the resulted sequences were tested on EpCAM-positive human gastric cancer cell line, KATO III, and the EpCAM-negative mouse embryonic fibroblast, NIH/3T3 cells. Ep1 aptamer was successfully isolated and showed selective binding on EpCAM-positive KATO III cells when compared to EpCAM-negative NIH/3T3 cells, as observed by the flow cytometry and the confocal imaging results. Additionally, the binding of Ep1 to EpCAM protein was assessed using mobility shifting assay and aptamers-protein docking. Furthermore, the binding affinity of Ep1 was measured against EpCAM protein using EpCAM-immobilized on magnetic beads and showed apparent affinity of 118 nM. The results of this study could suggest that Ep1 aptamer can bind specifically to the cellular EpCAM protein, making it an attractive ligand for targeted drug delivery and as an imaging agent for the identification of cancer cells.
Iron oxide nanozymes, which are magnetic nanoparticles with enzymatic properties, are explored, including their robust production techniques and diverse applications in the fields of biomedicine and environmental science.
Background: The national mandatory premarital screening test is based on mean corpuscular volume (MCV) > 80 fL value for the detection of b-thalassemia to provide acceptance for marriage. The objective of this study is to assess the efficacy of MCV as a screening test for b-thalassemia trait in the present population. Methods: This study was conducted on 418 blood samples collected from adult individuals. The diagnosis of b-thalassemia carrier was given to those having HbA2 values equal to or above 3.5%. The diagnostic reliability of different RBC indices and formulas in discriminating cases of b-thalassemia trait were evaluated. Finally, a new index called "Momani" was determined based on MCV, RDW and RBC count. Results: b-thalassemia trait was identified in 10% of the cases. The measured MCV value was significantly lower in b-thalassemia carrier group compared to non-carrier group (p = <0.001). MCV value and RBC count showed a higher diagnostic reliability than other RBC indices. We found that MCV ≤ 74.45 fL is more suitable cutoff value of MCV with 86.2% specificity, 71.4% sensitivity, 36.6% positive predictive value, and 96.4% negative predictive value. Finally, our index "Momani" was found to be useful in predicting carrier and paralleled the performance of Sirdah, Mentzer, and Ehsani indices. Conclusions: MCV<80 is a useful but not a perfect cutoff point for the screening of b-thalassemia carriers from noncarriers. The diagnostic accuracy of MCV can be improved by selecting a new cutoff value. Moreover, "Momani" index shows good discrimination ability in diagnosing b-thalassemia carrier in our population.