RESUMO
Background and purpose: Recently, the use of immunotoxins for targeted cancer therapy has been proposed, to find new anticancer drugs with high efficacy on tumor cells with minimal side effects on normal cells. we designed and compared several arazyme (AraA)-based fusion proteins with different ligands to choose the best-targeted therapy for interleukin 13 receptor alpha 2 (IL13Rα2)-overexpressed cancer cells. For this purpose, IL13Rα2 was selected as a receptor and IL13 and IL13.E13K were evaluated as native and mutant ligands, respectively. In addition, Pep-1 and A2b11 were chosen as the peptide ligands for targeted cancer therapy. Experimental approach: Several bioinformatics servers were used for designing constructs and optimization. The structures of the chimeric proteins were predicted and verified by I-TASSER, Q-Mean, ProSA, Ramachandran plot, and Verify3D program. Physicochemical properties, toxicity, and antigenicity were predicted by ProtParam, ToxinPred, and VaxiJen. HawkDock, LigPlot+, and GROMACS software were used for docking and molecular dynamics simulation of the ligand-receptor interaction. Findings/Results: The in silico results showed AraA-A2b11 has higher values of confidence score and Q-mean score was obtained for high-resolution crystal structures. All chimeric proteins were stable, non-toxic, and non-antigenic. AraA-(A(EAAAK)4ALEA(EAAAK)4A)2-IL13 retained its natural structure and based on ligand-receptor docking and molecular dynamic analysis, the binding ability of AraA-(A(EAAAK)4ALEA(EAAAK)4A)2-IL13 to IL13Rα2 was sufficiently strong. Conclusion and implications: Based on the bioinformatics result AraA-(A(EAAAK)4ALEA(EAAAK)4A)2-IL13 was a stable fusion protein with two separate domains and high affinity with the IL13Rα2 receptor. Therefore, AraA-(A(EAAAK)4ALEA(EAAAK)4A)2-IL13 fusion protein could be a new potent candidate for target cancer therapy.
RESUMO
BACKGROUND AND OBJECTIVES: Tuberculosis (TB) is the oldest known bacterial disease in humans. Due to the rise of morbidity in recent years, early diagnosis of the disease is necessary. MATERIALS AND METHODS: In this study we used Fluorescent Amplification-Based Specific Hybridization (FLASH) PCR to targetIS6110 for rapid detection of M. tuberculosis (MTB). To investigate the important factors influencing the risk of TB, data from patients and their medical records were analyzed. RESULT: The sensitivity and specificity of FLASH-PCR for detecting MTB were determined as 93.33% and 92.5%, respectively. The findings of this study have suggested that removal of the contaminants in FLASH-PCR sign ificantly reduced the detection time, and MTB was much more rapidly detected in the clinical specimens compared to the conventional culture and smear examination. Results of the medical survey showed that the majority of TB patients were males, over 51 years old, smokers, with pulmonary TB and normal chest X-ray (CXR). CONCLUSION: MTB can be rapidly detected inclinical specimens using FLASH-PCR in comparison with culture and smear examination.
