Sugar beet (Beta vulgaris L.) is an important industrial crop, the yield of which is strongly affected by numerous diseases caused by fungal pathogens. To the aim of developing transgenic plants resistant to fungi, two transgenic diploid sugar beet genotypes expressing the gene encoding the polygalacturonase inhibiting protein 2 of Phaseolus vulgaris (PvPGIP2) were generated by Agrobacterium tumefaciens-mediated transformation. PGIPs are plant cell wall leucine-rich repeat (LRR) proteins that bind to and inhibit fungal polygalacturonase (PG), thus slowing down the plant cell wall degradation and limiting fungal colonization of the plant tissues. Leaf blade explants carrying the bases of regenerated shoots, a highly regenerative tissue, were used for transformation. PCR screening using specific primers showed the presence of the transgene in more than 40% of the regenerated kanamycin-resistant plants. A transformation rate of 4.4-4.2% (depending on the genotype) was achieved as revealed by agarose diffusion assay of the PvPGIP2 activity in the crude protein extracts of shoot tissues. The intact integration of the transgene cassette into the genome was confirmed by Southern blot analysis. The inhibitory activity against Fusarium phyllophilum polygalacturonase (FpPG) was found at various levels in several transgenic plants. No alterations of growth and development of the transgenic plants were observed.
Abstract In this study, we investigated the design and construct of a chitosan (CA)‐based targeted gene delivery system and evaluated its function. To this end, CA–folic acid/pDNA (CA–FA/pDNA) nanoparticles were prepared in different formulations using the ion gelation method. All the synthesized nanoparticles were characterized using FTIR, TEM, SEM and DLS. Moreover, the effects of molecular weight (MW) of CA, DNA, and CA concentration were inspected on encapsulation efficiency (EE). The results showed that the EE of pDNA was directly proportional with MW of CA and CA concentration but was in an inverse proportion with DNA concentration. In addition, high MW of CA and low MW of CA nanoparticles showed lower and higher pDNA release in all pH ranges, respectively. It is concluded that the N/P ratio increase can cause controlled pDNA release.
Abstract Reduced expression of tumor suppressor miRNAs leads to cancer cell development, so restoring the expression of these miRNAs can be an appropriate treatment option for cancer. Due to the heterogeneity of cancer cells, single-drug therapy often results in drug resistance. Therefore, the combination of chemotherapy with miRNA can be a powerful strategy for cancer treatment. In the current investigation, we researched the synergic effect of miR-34a in combination with doxorubicin (DOX) on cell death of acute T-cell lymphoblastic leukemia (T-ALL) Jurkat cell line, as well as the expression of some genes including Caspase-3, Bcl-2, and p53 which are involved in apoptosis. Our outcomes showed that the combination of miR-34a and doxorubicin remarkably reduced the expression of the Bcl-2 gene, the target gene of miR-34a. According to the results of the MTT assay in cells treated with miR-34a and doxorubicin, the survival rate was significantly decreased compared to the untreated cells. Results of the flow cytometry assay and DAPI staining demonstrated an increased apoptosis rate of Jurkat cells in combination therapy. Moreover, cell cycle arrest was observed at the G2/M phase in cells that were treated with miR-34a/doxorubicin. Most importantly, we showed that the transfection of the Jurkat cells with miR-34a increased the sensitivity of these cells to doxorubicin. Furthermore, the combination of miR-34a and doxorubicin drug effectively increased apoptosis of treated cells. Therefore, this method can be used as an impressive treatment for T-ALL.
Background and Aim: Acute promyelocytic leukemia is a malignancy of myeloid cells that are associated with resistance to apoptosis and differentiation arrest in promyelocytes in the bone marrow.The aim of this study was to investigate the effect of Origanum vulgare aqueous extract on apoptosis induction in an acute promyelocytic leukemia cell line (HL-60). Materials and Methods:The viability of HL-60 cells under treatment with various doses of Origanum vulgare extract was assessed by using a cell cytotoxic assay.Then, IC50 values were determined after 24, 48, and 72 hours.The expression of Caspase8, Caspase9, Bax, Bcl2, and Nrf2 genes were measured by real-time PCR assay and different stages of apoptosis and necrosis in HL-60 cells were investigated using acridine orange/ethidium bromide double staining.Finally, data were introduced into SPSS software and analyzed by one-way ANOVA. Results:The results showed that Origanum vulgare extract decreased HL-60 cell survival and Caspase9, Caspase8, Bax, Bcl2, and Nrf2 gene expressions were changed after 72 hours of treatment with 1/5 of IC50 of Origanum vulgare extract.Also, morphological changes in the nucleus indicated apoptosis induction in HL-60 cells.Conclusion: It seems that the aqueous extract of Origanum vulgare leaves can induce apoptosis in an acute promyelocytic leukemia cell line (HL-60) under experimental conditions.
Numerous diseases caused by fungal pathogens influence the annual production of sugar beet. In order to obtain a plant resistant to fungi, genetic transformation has been applied to the sugar beet. To invade a plant tissue, phytopathogenic fungi produce several cell wall-degrading enzymes (CWDEs); polygalacturonases (PGs) are pathogenicity factors produced at the earlier stages of a fungal infection that depolymerize the homogalacturonan. One of the strategies used by plants to limit the degradation of the cell wall polysaccharides by fungal CWDEs is the production of proteinaceous inhibitors. Against fungal, microbial, and insect PGs, plants produce cell wall-associated polygalacturonase-inhibiting proteins (PGIPs). The overexpression of PGIPs improves the resistance to fungal and bacterial necrotrophs in different plants. In this research, the gene encoding the PGIP1 fused downstream of the leader sequence for secretion in the extracellular environment was isolated from Phaseolus vulgaris and cloned into the expression vector pBI121 for the Agrobacterium-mediated transformation of sugar beet. Modified transformation protocol and selection strategies were developed. In comparison with the preexisting methods, the transformation efficiency was increased and different cultivars were transformed, highlighting the general effectiveness of the method applied. The presence of the transgene and the activity of PvPGIP1 were confirmed by PCR and agarose diffusion assay analyses, respectively, and the present and copy number of the transgene in the T0 plants' genome were demonstrated by Southern blot.
The present study was conducted to identify factors affecting the educational justice in Islamic Azad University. The method of study is applied in terms of objective and it is descriptive-survey in terms of data collection, and quantitative in terms of data type. Interviews and questionnaires were used to collect the initial information. Population of this study included professors of Islamic Azad University, which 40 of them were selected as sample of study using convenient sampling method. Delphi technique based university professors’ view was used to identify the variables affecting educational justice, and factor analysis was used to analyze the data for extracting the indices of questionnaire. Investigations revealed that the impact of each of the factors affecting the educational justice in Islamic Azad University is not equal, so the level of impact of these factors should be assessed and these factors should be ranked accordingly. Identification of the factors affecting educational justice in the university by means of three-branched model allows managers to identify harms in each of the structural, underlying, and behavioral factors in order to implement the educational justice properly. Twenty and six important factors to implement educational justice were identified in the university using a questionnaire and Delphi technique. After identification of the factors, they were classified in three classes. The considered model had appropriate fit, as RMSEA was obtained 0.085 and the ratio of Chi square to degree of freedom (χ2 / df) was obtained 3.7. Based on the model obtained from confirmatory factor analysis using LISREL software, 26 indices of the questionnaire were classified in three factors, including structural, behavioral factors, and underlying factors.
Despite the recent advances in the treatment of other cancers, the 5-year survival rate of pancreatic cancer remains under 9 %. Chemotherapy and surgical resection are the most common therapy methods. The regulatory role of microRNAs in different types of cancer has given them therapeutic importance. miR-612 has been downregulated in colorectal, bladder, liver, and some other types of cancer and could be considered a tumor-suppressor miRNA. 5-FU is one of the most common chemotherapeutic agents used in pancreatic cancer treatment, which is used in multiple drug regimens and combinatorial therapy methods. The aim of this study is the evaluation of miR-612 restoration in the PANC-1 cell line and using the tumor-suppressive effect of it in combination with 5-FU on cell growth and migration. MiR-612 mimic was transfected to PANC-1 cells through electroporation. Following the transfection, expression levels of miR-612 and BAX, BCL-2, Caspase-3, MMP9, and PD-L1 genes were measured by qRT-PCR. MTT assay was used to determine the cytotoxicity of miR-612 and 5-FU on PANC-1 cell viability. To confirm MTT results and to evaluate the quantitative effect of apoptosis induction flow cytometry test was used and in order to confirm apoptosis test results and cell cycle arrest evaluation DAPI staining and cell, cycle tests were conducted, respectively. Finally, to assess the inhibitory effect of miR-612 in combination with 5-FU on migration and growth wound healing and colony formation assays were used, respectively. Results demonstrated that miR-612 alongside 5-FU has an important role in the inhibition of migration and growth and also apoptosis induction in PANC-1 cells and could be considered as a supporting agent of chemotherapy and a novel therapeutic modality in pancreatic cancer treatment.
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