Research in Molecular Medicine (RMM)

Investigation of the Impact of Foretinib on AURKA and AURKB Expression in T98 Glioblastoma Cell Line

Mansour Moghimi, Hossein Sadeghi Tafti, Faezeh Namazi, Mansoor Salehi — Mon, 30 Sep 2019 12:07:38 +0000

Background: Gliomas are a common type of the primary brain tumors and account formore than 40% of all central nervous system (CNS) tumors. Glioblastoma (GBM) remainsone of the most fatal human malignancies as it is highly angiogenic. Foretinib is anoral multikinase inhibitor that has been shown to exhibit antitumor activity in previousclinical studies. AURKA and AURKB have been shown to be overexpressed in variouscancers.The purpose of this study was to investigate the effect of foretinib on the expressionof AURKA and AURKB in the T98 cell line.Materials and Methods: In this study, the T98 cell line was selected as an experimentalmodel of glioblastoma. The cultured cells were exposed to different concentrationsof foretinib (5 μM, 10 μM, 20 μM, and 0 μM as control). Following that, we examinedthe changes in the expression of AURKA and AURKB under the influence of foretinibcompared to control using quantitative real-time polymerase chain reaction (qRT-PCR).Results: The expression of AURKA and AURKB were found to be significantly reducedin the foretinib treated group compared to the control. The results demonstrated thatincreasing the concentration of foretinib led to reduction in the expression of both thegenes.Conclusion: These findings indicate that the foretinib can decrease the mRNA levelsof AURKA and AURKB. Thus, we suggest that foretinib may be an effective drug forGBM treatment and can be considered for future studies.

Aqueous Cannabis Extract Prevents Ethylene Glycol-induced Renal Calcium Crystallization

Mon, 30 Sep 2019 12:11:56 +0000

Background: Some plant extracts can prevent kidney stone formation in a dosedependent manner. In our study, we aimed to investigate the protective role of Cannabis sativa aqueous extraction on calcium oxalate formation in ethylene glycolinduced rats. Materials and Methods: To evaluate anti-urolithiasis, variations of the main risk factors (citrate, phosphorous, calcium, and Ph) have been evaluated in 24-hour urine samples of rats one day before the end of the experiment. Rats were divided into 4 groups: group 1 was administered regular drinking water; group 2 was administered 1% ethylene glycol in drinking water, group 3 was administered 100 mg/kg of Cannabis sativa extract for oral administration by gavage along with drinking water containing 1% ethylene glycol; and group 4 was administered 200 mg/kg of Cannabis sativa extract by gavage along with drinking water containing 1% ethylene glycol. Finally, histopathological slides from the kidney were also evaluated. Results: Results showed that 100 mg/kg and 200 mg/kg doses of Cannabis sativa extract significantly reduced the mentioned risk factors in comparison with ethylene glycol-treated rats (p<0.05). Conclusion: Histopathological slides showed reduced calcifications with extract treatment at 200 mg/kg of Cannabis sativa. Thus, this antioxidant herb can prevent kidney stone formation. These findings pave the way for new therapy.

Optimization of Cloning Conditions for High-level Production of Recombinant Mouse Interleukin-2 in Escherichia coli

Mon, 30 Sep 2019 12:15:03 +0000

Backgrounds and objectives: Many proteins have been expressed so far in bacterial host. Due to its simple culture conditions, having a short life cycle, and easily genetic manipulation, E.coli have been regarded as a preferable host to produce recombinant proteins, but protein cloning in bacterial host have many challenges. Therefore, we aimed to review some of these problems by an experience from mice IL-2 recombinant. Methods: cDNA synthesis was performed after RNA extraction of mouse splenocytes. PCR product purification carried out after IL-2 coding sequence amplification and was ligated into the pET-21b (+) vector and transformed into the competent BL21 E.coli. Expression and purification of recombinant mouse IL-2 were done using IPTG inducer and metal affinity chromatography respectively. Results: DNA sequencing confirmed the accuracy of the insertion process. A 23 kDa exogenous protein was observed on the SDS-PAGE. Specificity and concentration of produced mouse recombinant IL-2 protein were confirmed by western blotting and BCA methods. Conclusion: Recombinant IL-2 was produced in BL21 and pET-21b (+) expression system at 24∘C in the soluble form.

Basil and Dracocephalum kotschyi Alcoholic Extracts Affect BCL2 expression and HepG2 Cell Proliferation

Mansour Moghimi, Sara Rashidian, Farinaz Khosravian, Nasrin Hadi — Mon, 30 Sep 2019 12:16:59 +0000

Background: Liver cancer is the second most common reason for cancer-related death globally, and thus, a major public health challenge. Plant-based drugs are promising therapeutic agents against cancers. Basil and Dracocephalum kotschyi extracts also exhibit therapeutic impact on various cancers. To assess the effects of the extract of Basil and Dracocephalum kotschyi on cell proliferation and BCL2 expression in the HepG2 cell line. Materials and Methods: In this experimental study, the HepG2 cell line was selected for treatment with the alcoholic extracts of Basil and Dracocephalum kotschyi. Cell proliferation was examined in the presence of various concentrations of the Basil and Dracocephalum kotschyi extracts by MTT assay. Moreover, the effect of the alcoholic extracts of Basil and Dracocephalum kotschyi on BCL2 expression was evaluated using quantitative real-time polymerase chain reaction (qRT-PCR). Results: Cell proliferation analyses showed anticancer characteristics of the alcoholic extracts of Basil and Dracocephalum kotschyi. In addition, treatment with the extracts of Basil and Dracocephalum kotschyi led to the decreased expression of BCL2 in HepG2 cell line. Conclusion: These findings indicated that Basil and Dracocephalum kotschyi are promising candidates for future anticancer research.

TEM Gene Detection in Clinical Pseudomonas aeruginosa and Escherichia coli Samples

Elahe Shams, Behnaz Nateghi, Amir Eshaghiyan, Parisa Behshood — Mon, 30 Sep 2019 12:18:49 +0000

Background: Isolation of the TEM beta-lactamase gene from clinical Pseudomonas aeruginosa and Escherichia coli samples provides useful information on the epidemiology of and factors involved in infections caused by these agents as well as their antibiotic resistance patterns. The aim of this study was to evaluate the antibiotic resistance of P. aeruginosa and E. coli isolated from specimens obtained in Isfahan, Iran via detection of the TEM gene. Materials and methods: In this cross-sectional study, 120 P. aeruginosa and 86 E. coli samples isolated from urine and sputum were identified using biochemical methods. Their antimicrobial resistance pattern was investigated using the Kirby-Bauer disc diffusion method. Then, phenotypic detection of extended-spectrum beta-lactamases (ESBL) was performed using a combined disc method. Finally, the TEM gene in isolated samples was examined using polymerase chain reaction (PCR). Results: P. aeruginosa isolates were found to show the highest resistance to tetracycline (97.5%) and amoxicillin (95%) and the highest sensitivity to aztreonam (97.5%) and amikacin (61.66%). 68 P. aeruginosa samples (56.6%) contained a TEM gene. E. coli isolates were found to show the highest resistance to co-trimoxazole (59.34%) and amoxicillin (55.04%), and the highest sensitivity to imipenem (69.66%) and chloramphenicol (61.92%). 62 E. coli samples (72.09%) contained a TEM gene. Conclusions: The alarming spread of ESBL-producing pathogens is a complicating factor in antimicrobial therapies. It is essential to employ diverse strategies for the supervision of the spread of these pathogens.

Acute Beetroot Juice Intake: Hematological, Antioxidant and Lipid Parameters in Female Athletes

Maryam Lotfi, Mohammad Azizi, Worya Tahmasebi, Parviz Bashiri — Mon, 30 Sep 2019 12:20:43 +0000

Background: One of the drinks that are increasingly popular among athletes is beetroot juice. This survey was undertaken to determine the effects of acute beetroot juice consumption on certain hematological parameters, lipid profiles, and total antioxidant capacity in female soccer players. Materials and Methods: This was an applied, semi-experimental study. Female soccer players (n = 30, age = 23.16 ± 0.79 years) were selected randomly and assigned into three groups: experimental (beetroot juice, n = 10), (control (placebo), n = 10) and (mouth rinsing, n = 10). Subjects undertook soccer training for a session (90 min) with consumption of 200 ml juice 2 h before they started. Blood samples were collected and investigated before and after training. Paired sample t-tests were used for comparison within groups, and one-way ANOVA was used for comparison between groups. All statistical analyses were performed at P ≤ 0.05. Results: After a session of using beetroot juice, there were no significant differences in blood indices (levels of hemoglobin, hematocrit, red blood cells, iron, and mean corpuscular volume), lipid profiles (triglycerides, total cholesterol, and high density lipoprotein), and total antioxidant capacity between groups (experimental, control, and mouth rinsing) (P > 0.05), but low density lipoprotein concentrations changed significantly (P < 0.0001). Conclusions: Drinking a dose of beetroot juice did not improve hematological parameters, lipid profiles, and total antioxidant capacity. Therefore, our study of daily consumption of this drink will be pointed out to subsequent researchers.