Mohadese Abdoli; Parisa Fathi Rezaei; Kamran Mansouri
Abstract
The aim of this study was to investigate the cytotoxicity of probiotic mixture on breast cancer cell lines. A mixture of local probiotic bacteria was cultured and the supernatant was lyophilized for performing additional experiments. The antioxidant activity, total phenol content (TPC) and fatty acid ...
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The aim of this study was to investigate the cytotoxicity of probiotic mixture on breast cancer cell lines. A mixture of local probiotic bacteria was cultured and the supernatant was lyophilized for performing additional experiments. The antioxidant activity, total phenol content (TPC) and fatty acid composition of bacterial supernatant (BS) were also measured. The effect of probiotic mixture on cell viability of two breast cancer cell lines and apoptosis induction were investigated using MTT assay and DAPI staining, respectively. The highest antioxidant activity and total phenol content (TPC) of the bacterial supernatant (BS) were detected at 3200 µg/ml. According to the GC–MS analyses, linoleic acid (37.40 %) and oleic acid (26.93 %) were identified as the major fatty acids of the BS. The MTT assay showed that BS has antiproliferative effect on MDA-MB-231 and MCF-7 cells in a time and dose-dependent manner, and also the 3200 μg/ml was determined as IC50. The apoptosis induction was confirmed by morphological characteristics on both cell lines, including nucleus changes, granulation of the nucleus, and the formation of the apoptotic bodies. According to this experiment, cytotoxic activity and apoptosis potential of BS on two breast cancer cell lines including MDA-MB-231 and MCF-7 was shown. BS could be considered as a suitable anticancer agent
Saeedeh Sibevieh; Ensieh Salehghamari; Mohammad Ali Amoozegar; Mohammad Reza Zolfaghari; Mohammad Soleimani; Zohre Nasrollahzadeh; Sara Eftekhari Yazdi
Abstract
With the increased usage of nitrate fertilizers, as it is a stable ion with high solubility in water, its removal is a challenge for human health. Several physicochemical methods have been examined for nitrate removal of water, but nowadays the biological treatment of it from soil and water are widely ...
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With the increased usage of nitrate fertilizers, as it is a stable ion with high solubility in water, its removal is a challenge for human health. Several physicochemical methods have been examined for nitrate removal of water, but nowadays the biological treatment of it from soil and water are widely used because of its high efficiency and low cost. To remove nitrogen from such environments, we studied nitrate reducing halophilic and halotolerant bacteria. A total of 50 strains of halophilic and halotolerant bacteria from different saline and hypersaline environments of Iran, including Incheboron wetland, Aran-Bidgol Lake and Urmia Lake, were screened for nitrate reductase production. Among isolated bacteria, 60% of strains isolated from Urmia Lake and 19% of strains isolated from Incheboron wetland produced nitrate reductase. Nitrate reductase coding genes narG and napA were analyzed for all the nitrate reducing strains. The napA gene was amplified successfully from a gram negative halophilic strain and narG gene was detected in ten halophilic strains. Among nitrate reducing isolates, Kocuria rosea strain R3A34 which has narG gene showed the most nitrate reductase production. The strain R3A34 was selected for optimization of denitrifying activity. Statistical results have shown that the optimum conditions for the production of nitrate reductase were 32 °C, pH 7.0, NaCl 8 % (w/v) and mannitol as carbon source. Furthermore, as these conditions are common in wastewaters, this bacterium can be a proper candidate for bioremediation of wastewaters from nitrate pollutants.
Narges Fazili; Zahra-Soheila Soheili; Saeid Malekzadeh-Shafaroudi; Shahram Samiei; Nasrin Moshtaghi; Abouzar Bagheri
Abstract
Royal jelly (RJ) from queen honeybee larva as a traditional medicine agent has a variety of pharmacological benefits. In the present study, the effect of Royal jelly was investigated on the urinary bladder cancer cell line (HTB-9 5637). To determine the cell viability in different concentrations ...
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Royal jelly (RJ) from queen honeybee larva as a traditional medicine agent has a variety of pharmacological benefits. In the present study, the effect of Royal jelly was investigated on the urinary bladder cancer cell line (HTB-9 5637). To determine the cell viability in different concentrations of Royal jelly, MTT assay was performed. An in vitro wound healing assay was applied to investigate the effect of RJ on cell migration. The activity and gene expression level of matrix metalloproteinase 2 and 9 was assessed by zymography and Real time PCR respectively. R.J.S at the concentration of 0.7 mg/ml had a significant effect on reducing the proliferation rate of 5637 cells after 72h (p < 0.009). R.J.S significantly decreased cell migration and induced a significant decrease in the transcriptional level of MMP9 after 72h (0.5x; P < 0.049). However R.J.S did not impose any effect on the expression level and activity of matrix metalloproteinase 2. These results indicate the potential of RJ as a promised natural anti-proliferative and anti-metastatic drug in combination with advanced therapy methods for cancer treatment. Royal jelly has the potential to be more focused as an anti-metastatic drug to control tumor growth and can be considered as a more effective alternative to the current chemotherapy drugs.
Ahdiyeh Shahtaghi; Ali Alam Shahnabadi; Kamelia Kohannezhad; Neda Amini; Maria Beihaghi
Abstract
One of the newest diagnostic methods and treatment of cancer is to design new drugs. It is now possible to design a drug with desired properties in theory and evaluate its therapeutic effects through bioinformatics tools. Among the studied drugs; those based on cytokine genes, which increase the body's ...
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One of the newest diagnostic methods and treatment of cancer is to design new drugs. It is now possible to design a drug with desired properties in theory and evaluate its therapeutic effects through bioinformatics tools. Among the studied drugs; those based on cytokine genes, which increase the body's immunity against cancer, are of great interest. Cytokines are small proteins that play an important role in cell signaling and can affect the function and behavior of surrounding cells. CCL21 chemokine is one of the cytokines that possess antitumor properties has potential on chemoa-ttraction of T lymphocytes and dendritic cells. Interleukin 1 beta (IL1β) is a cytokine involving in different cellular activities such as activation of neutrophils, B-Cells and T-Cells. In the present study, we designed a drug-based cytokine gene to activate T cells and B cells by inserting defined CCL21 epitope and IL1β pepteide sequences into a protein construct. Molecular dynamics simulation was performed in Linux space using Gromex software. Results of RMSD, RMSF and the radius of gyration obtained from the simulation showed the stability of both proteins which indicated that there are no significant conformational differences between the commercial CCL21 and recombinant form. The interaction of synthetic construct and human CCL21 with CCR7 receptor was also investigated by HADDOCK software. Obtained results showed that there are no differences between these proteins and recombinant protein has the same structural and conformational characteristics as human commercial CCL21.
Hamid Reza Shojania; Madjid Momeni Moghaddam; Seyed Ebrahim Hossini
Abstract
Wound healing is a complex biological process in which many molecules, including microRNA molecules, play an important role in its regulation. It is well established that reducing mir-155 expression can accelerate wound healing. In this study, the effect of using nanoparticle loaded by vitamin C in wound ...
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Wound healing is a complex biological process in which many molecules, including microRNA molecules, play an important role in its regulation. It is well established that reducing mir-155 expression can accelerate wound healing. In this study, the effect of using nanoparticle loaded by vitamin C in wound healing and mir-133b, collagen I and III expression changes was investigated. In this study, first nanoparticles of albumin protein were produced and then loaded with vitamin C. to evaluate the toxicity and determine the appropriate dose, these nanoparticles were affected on fibroblast cell and cell behavior was investigated. Also in the molecular study, changes in the expression of collagen I and III genes were studied. The results show that nanoparticles containing vitamin C had a positive effect on the expression of collagen I and III compared to the control group. Also we observed a decrease in the expression of mir-133 compared to the control group. Therefore, the results of this study clearly indicate the role of nanoparticles in improving the expression of collagen I and III genes, also the expression of mir-133 gene is reduced, which leads to the activation of collagen genes I and III, which these findings favor the high expression of collagen by fibroblasts, which ultimately have a positive effect on wound healing.
Mohammadreza Nassiri; Azadeh Safarchi; Masoume Vakili-Azghandi; Vinod Gopalan; Mohammad Doosti; Shahrokh Ghovvati; Ahmad Reza Movassaghi
Abstract
p53 is a tumor suppressor protein that plays an essential role in controlling the cell and vascular endothelial growth factor (VEGF) is one of the most strong and specific angiogenic factors. The main objective of this study was to evaluate the impact of p53 and VEGF-C gene expression in the neoplastic ...
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p53 is a tumor suppressor protein that plays an essential role in controlling the cell and vascular endothelial growth factor (VEGF) is one of the most strong and specific angiogenic factors. The main objective of this study was to evaluate the impact of p53 and VEGF-C gene expression in the neoplastic and normal mammary gland of canine as an animal model. Elleven benign and malignant specimens and 5 normal specimens were collected. After RNA extraction and cDNA synthesis, relative quantification of p53 and VEGF-C genes were accomplished by Real-time quantitative PCR (RT-qPCR) based on use of β-actin as a reference gene. The relative mRNA expression of the p53 and VEGF-C genes were analyzed by GLM procedure of SAS software v9.2. The results indicated that the VEGF-C and p53 mRNA expression in neoplastic specimens was over-and down-expressed respectively as compared with normal specimens and p53 mRNA expression was significantly negatively associated with VEGF-C (~4 fold) in neoplastic specimens (P <0.01). The findings emphasized that simultaneous evaluation of p53 and VEGF-C expression can be used as tumor biomarker for early diagnosis of malignancy in canine. Furthermore, RT-qPCR is a rapid and sensitive method to for monitoring and investigating of suspicious canine at the beginning stage of malignancy and may provide an alternative explanation for deregulated p53 signalling in breast cancer.
Raheleh Majdani
Abstract
Urinary tract infection, as one of the most prevalent bacterial infections, affecting millions of people yearly worldwide. Due to control of increasingly antibiotic resistant infections, it is needed to introduce the alternative approaches such as phage therapy. In this study, isolation, purification ...
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Urinary tract infection, as one of the most prevalent bacterial infections, affecting millions of people yearly worldwide. Due to control of increasingly antibiotic resistant infections, it is needed to introduce the alternative approaches such as phage therapy. In this study, isolation, purification and enrichment of eight lytic bacteriophages against antibiotic-resistant Escherichia coli strains originated from human urinary tract infections were performed using double layer agar method. Molecular analysis of the bacteriophages was performed using two endonucleases enzymes (EcoRV and XbaI). Then two of eight isolated bacteriophages with the highest host range were more characterized to determine their morphology, one step growth, latent period, burst size and stability in environmental parameters (temperature and pH). Eight isolated bacteriophages showed high genome variation in enzyme digestion process. Two phages with the broadest host range (PEcMa2/17 and PEcMa3/17) showed effective lytic activity against five isolates. In electron microscopy, selected phages were belonged to Siphoviridae and Myoviridae family. Latent period in both of the propagated phages were 15 min and the burst size was estimated to 100 pfu/ ml and 120 pfu/ml in PEcMa2/17 and PEcMa3/17 respectively. Both of the phages showed more than 50% stability at 37 ºC and lower used temperatures and they were survived efficiently in pH=7. The bacteriophages showed strong antibacterial potential against Escherichia coli from UTIs, but as candidates for phage therapy, more characterization such as molecular analysis and experimental assays are needed before therapeutic applications.
Nima Dehdilani; Mohsen Fathi Najafi; Hesam Dehghani
Abstract
To achieve a reliable and persistent expression over time, the transgene should be precisely integrated into the genome safe harbor (GSH) loci. Little attention has been paid to find the safe harbor loci of the chicken (Gallus gallus domesticus) genome. Identification and characterization of GSH loci ...
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To achieve a reliable and persistent expression over time, the transgene should be precisely integrated into the genome safe harbor (GSH) loci. Little attention has been paid to find the safe harbor loci of the chicken (Gallus gallus domesticus) genome. Identification and characterization of GSH loci that allow the persistent and reliable expression of knocked-in genes could be a major area of interest within the field of transgenic technology and central to the development of bioreactor transgenic livestock. Randomly integrated transgenes might be encountered position effects and epigenetic silencing, so unstable phenotypes as well as unreliable and unpredictable expression of knocked-in transgene could be occurred. In contrast to random gene insertion, site-specific gene targeting provides a specific strategy that exploits homologous recombination to insert a transgene of interest into a pre-determined locus, specifically.In this study, using bioinformatics, gene expression atlas, and Hi-C data, the GSH region between drg1 and eif4enif1 genes was predicted in the chicken genome. Contrary to the previous time-consuming and expensive methods, here we introduce a fast and easy-to-use pipeline allowing the prediction of orthologue GSH loci in all organisms, especially in chicken. Then, the standard design of targeting and gRNA/Cas9 vectors is explained in detail.
Sara Yousefi taemeh; Jalil Mehrzad; Hesam Dehghani
Abstract
Primordial germ cells (PGCs) are precursors for mature gametes, which transmit genetic information to the next generation. Due to the importance of PGCs in many fields including developmental biology, genome editing, transgenesis, and conservation of avian genetic resources, a lot of research has focused ...
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Primordial germ cells (PGCs) are precursors for mature gametes, which transmit genetic information to the next generation. Due to the importance of PGCs in many fields including developmental biology, genome editing, transgenesis, and conservation of avian genetic resources, a lot of research has focused on the cultivation of PGCs. Despite considerable progress in the establishment of specified culture media for the expansion of PGCs, a well-defined PGC culture medium has not yet been developed. This might be due to the complexity of the nutritional requirements of PGCs in the culture. Besides the nutritional needs including vitamins, amino acids, salts, carbohydrates, and growth factors, a certain source of energy must be provided to sustain growth and viability. Glutamine is a major energy source for cultured cells, which is commonly added in cell culture media at higher concentrations than other amino acids. However, glutamine is very labile and rapidly degrades in solutions such as culture media. This generates ammonia as a by-product, which is toxic to the cultured cells and can affect cell viability and protein glycosylation. Therefore, the stability of glutamine in culture conditions is another concern for the long-termed culture of PGCs. Here, we study the effect of glutamine stability on PGC culture using glutamine and GlutaMax (a commercial stabilized dipeptide form of glutamine). We found that the addition of GlutaMax in the medium promotes PGC proliferation. This effect might be exerted by minimizing toxic ammonia buildup that results in maximizing cell performance, and also increasing media stability.
Zahra Ghavidel; Madjid Momeni Moghaddam; Toktam Hajar; Eisa Kohan
Abstract
The use of medicinal plants in the treatment of diseases has a long history dating back to the human presence on Earth. Cancer, has always been an incurable disease, and among the various cancers, breast cancer is one of the most important causes of death in women and imposes difficult conditions on ...
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The use of medicinal plants in the treatment of diseases has a long history dating back to the human presence on Earth. Cancer, has always been an incurable disease, and among the various cancers, breast cancer is one of the most important causes of death in women and imposes difficult conditions on patients, although medical and surgical solutions have been proposed for this disease, but it has not been successful in the complete cure of this disease. In recent years, more studies have been done on the effects of Medicinal Plants and scientists are trying to study the control mechanisms of these plants to control the growth of cancer cells. This article focuses on two very important Genes that are of great importance in regulating the growth and proliferation behavior of normal and cancer cells. We have dealt with the effectiveness of their active ingredients. This article is an overview on P53 and MDM2 genes also P53-MDM2 signal beside Plants that exert their medicinal effect through these genes and the result of this article, which is actually a valuable list of these plant families, can be used in research that seeks to control the mentioned doses, thus the process of the effect of plants in Treatment will be seen more clearly.
Mahnaz Ghowsi; Nazli Khajehnasiri; Sajjad Sisakhtnezhad
Abstract
Connexin-43 (Cx-43) plays axial roles in the propagation of action potentials and contractile coupling in heart. Down-regulation of Cx-43 in heart is associated with arrhythmia, dilated cardiomyopathy and heart failure. To date, no studies have examined the effects of androgen deprivation therapy (ADT)-induced ...
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Connexin-43 (Cx-43) plays axial roles in the propagation of action potentials and contractile coupling in heart. Down-regulation of Cx-43 in heart is associated with arrhythmia, dilated cardiomyopathy and heart failure. To date, no studies have examined the effects of androgen deprivation therapy (ADT)-induced hypogonadism on the expression of Cx-43 in heart. This study investigated the effects of testosterone deprivation and its replacement with testosterone on the expression of Cx-43 mRNA and muscle-specific miRNAs miR-206 and miR-1, as two potential regulators of the Cx-43 protein expression in the ventricular tissue. Accordingly, 21 male Wistar rats were divided into three groups: Ι) Normal control, П) ORX-S: castrated rats serving as animal models for ADT and receiving the sesame oil as a solvent of testosterone enanthate for 10 weeks, and Ш) ORX-T: these animals were castrated, receiving testosterone enanthate (25 mg/kg) for 10 weeks. The relative expression of Cx-43 mRNA, miR-206 and miR-1 was determined by qRT-PCR. Cx-43 mRNA was found to be decreased in the ORX-S group. The Cx-43 mRNA was up-regulated after the administration of testosterone enanthate. There were no significant changes in miR-206 and miR-1 levels in the ORX-S and ORX-T groups when compared to the controls. Our results indicated that testosterone should be regarded as an important factor in the regulation of the Cx-43 mRNA expression in heart, and testosterone deprivation may down-regulate the Cx-43 mRNA expression; however, it doesn’t alter miR-1 and miR-206 levels. These results suggest that ADT-induced hypogonadism may put males at risk for cardiac dysfunctions.
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