Background and Objectives: Aspergillus clavatus antimicrobial peptide (AcAMP) is a fungi-derived peptide with a broad spectrum of activity against pathogenic bacteria and fungi. Natural antimicrobial peptides, including AcAMP, have attracted many attentions in the development of new natural antibiotics against pathogenic bacteria, especially multidrug resistant ones. Materials and Methods: In the present study, acamp gene was codon-optimized and chemically synthesized in pUC57 cloning vector, subcloned into pET28a (+) expression vector and transferred into competent Escherichia coli BL21 (DE3) cells. The expression of AcAMP was induced by addition of Isopropyl β- d-1-thiogalactopyranoside (IPTG) and the expressed peptide was purified by Ni-NTA. BALB/c mice were immunized with the purified peptide and the ability of the immunized mice sera for the detection of the native AcAMP secreted by A. clavatus IRAN 142C was examined through ELISA and Western blotting techniques. Results: Both ELISA and Western blotting demonstrated the ability of the sera of the immunized mice to detect the native AcAMP. Conclusion: The results of the present work show that the raised antibody against recombinant AcAMP can be used to detect AcAMP peptide, an issue which paves the way to develop detection kits for the detection of AcAMP-producing organisms, purification of this valuable peptide for further investigations.
Introduction: Chemotherapy, biotherapy, and radiotherapy play a limited but important role in treating breast cancer. For more efficient treatment, combination therapy could be an appropriate option. In this study, radiotherapy using neutron radiation emitted from a 241 Am-Be neutron source, as well as biotherapy using curcumin (80 μM) was combined to investigate the efficiency of treatment towards MCF-7 breast cancer in a 3D culture medium. Methods: MTT, NR uptake assay, NO, GSH assay, catalase, cytochrome c, comet assay, and caspase-3 were used to determine the effect of neutron radiation and also neutron and curcumin combination on the viability of cancer cells. Results: The results of cytotoxicity test showed that neutron irradiation with or without curcumin at 5, 10, 15, and 20 h reduced the survival of tumor cells. Moreover, the rate of apoptosis due to the neutron effect at different irradiation times enhanced with the increasing time. Conclusion: Due to the significant anticancer effect of curcumin in 3D culture, using this molecule before or after neutron therapy is recommended.
Breast cancer is one of the most common types of cancer among women. One of these toxins that inhibits the growth of breast cancer cells in 2D cell culture and has significant anti-tumor effects is Dianthin-30. Therefore, in this manuscript, for the first time, the anti-cancer effect of Dianthin-30 toxin against breast cancer cells (MCF-7) in 3D culture has been studied. Materials and Methods: In order to evaluate the anti-cancer effects and cytotoxicity of the toxin at concentrations of 1.25, 2.5, 5 and 10μg/ml, MTT methods were used and a Neutral red test was used to validate the results of this test. Nitric oxide, Catalase, GSH assays, cytochrome c, Caspase-3 and Comet assay tests were also used to determine the type of mortality in cancer cells.Results: This toxin did not induce nitric oxide production, but at concentrations higher than 5μg/ml increased catalase production compared to the control. However, the level of GSH produced in all of the concentrations was significant compared to the control. In addition, Dianthin-30 increased cytochrome 30 and activation of caspase-3 in the above concentrations, but this effect was not significant compared to the control. The results of alkaline comet test also showed that the rate of induction of apoptosis by toxin was upward compared to the control. Conclusion:The results of this study show that Dianthin-30 has anti-cancer effects and has caused death in breast cancer cells and this toxin probably induced apoptosis in cancer cells more than the non-mitochondrial pathway.
Background and Objectives: Nowadays, infections with antibiotic-resistant bacteria are among the most important causes of mortality worldwide. This has attracted the attention of researchers to seek suitable alternatives for antibiotics. The venom of many toxic species such as arthropods has antibacterial properties. In this study, we investigated antibacterial effects of crude venom of Latrodectus dahli on Escherichia coli, Staphylococcus aureus, and Bacillus subtilis.Methods: Lyophilized crude venom of L. dahli was dissolved in 50 mM Tris-HCl buffer. Protein concentration was determined by the Bradford assay. Then, the bacteria were exposed to different concentrations (31.25-250 ng/mL) of the crude venom. Inhibitory activity of the venom against the bacteria was determined by MTT assay and determining minimum inhibitory concentration (MIC).Results: Results of the MTT assay showed that the crude venom significantly inhibited the growth of E. coli (31.25 and 62.5 ng/mL), S. aureus (at 250 ng/mL) and B. subtilis (at 125 and 250 ng/mL). In the MIC experiment, the crude venom significantly inhibited the growth of E. coli (at concentrations of 31.25 and 62.5ng/mL), S. aureus (at concentrations of 31.25-250 ng/mL) and B. subtilis (at concentrations of 31.25-250ng/mL).Conclusion: The crude venom of L. dahli and its components showed relatively strong antibacterial effects.
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