Introduction: To the best of our knowledge, there is little information in regards to the cytotoxicity of 1,8- cineole. Insect cells exhibit wide resistance to the lethal effects of chemical compounds. Activation of p53 by acetylation can cause cell toxicity. This study has been carried out in order to investigate whether the use of 1,8-cineole in the Spodoptera frugiperda (SF-9) cell line can induce cytotoxicity by increasing p53 acetylation expression or not. Materials and Methods: SF-9 cell line was cultured in TC100 insect culture medium and treated with 1,8-cineole at concentration of 850.45 μmol/L, based on the half-maximal inhibitory concentration (IC50) index at different times (24, 48, and 72h). The IC50 value was estimated for 1,8-cineole in SF-9. The percentage of alive cells was measured by MTT assay. The ELISA and Bradford protein techniques were used to detect endogenous levels of total and acetylated p53 protein generated in SF-9 cells. Results: The findings of the present study indicated that treatment with 850.45 μmol per liter of 1,8-cineole shows a time-dependent increase in the number of dead cells of SF-9 cell line. The effect of severe toxicity on SF-9 was observed after 72 h of incubation with 1,8-cineole, with approximately 4% of SF-9 cells alive. We observed a significant increase in the level of p53 acetylation up to 48 h in SF-9, indicating that 1,8-cineole resulted in up‑regulation of acetylated P53 and consequently p53 activation in SF-9 cells. Results showed that there is relationship between acetylated p53 protein levels and 1,8-cineole toxicity in SF-9 cell line. Conclusions: 1,8-cineole, may function through common pathways and mediate their cytotoxic effects through targeting p53 and its acetylation in SF-9 cells.
Introduction: Matrix metalloproteinases (MMPs) are a family of zinc-dependent proteinase. Several studies have reported the association between the single nucleotide polymorphisms(C/T) -1562 in the MMP-9 promoter and the risk of cancer. In this study, we decided to carry out a comprehensive meta-analysis to obtain a reasonable result about the association between this polymorphism and the risk of lung cancer. Materials and Methods: A complete literature review was conducted within the databases of ISI Web of Knowledge, google scholar, and PubMed for studies on lung cancer published from 2002 to 2018. A meta-analysis was conducted which included more than 3000 case and control subjects. The pooled Odds Ratio (OR) and 95% Confidence Intervals ( CI) were used for dominant, recessive, and co-dominant MMP-9 genotypes to assess the strength of the association. Results: Analysis indicated no significant association between MMP-9-1562C/T polymorphism and the risk of lung cancer, dominant model; [CT+TT/CC]: OR = 0.972, 95% CI = 0.811–1.164, recessive model [CC+CT/TT] OR= 1.027, 95% CI = 0.651–1.618 and co-dominant model [TT/CC] OR = 0.983, 95% CI = 0.550–1.755. Conclusions: No association was observed between the MMP9 -1562 C/T polymorphism and the incidence of lung cancer. The meta-analysis demonstrated that this polymorphism can't serve as a diagnostic marker for lung cancer.
Introduction: Analysis of genome sequences to search for encoded proteins and motifs is a most widely used technique for the prediction of new drug and vaccine targets. It can effectively leverage computational techniques to deliver effective and pragmatic advantages in the search of new drug and vaccines. Materials and Methods: The diversity and evolution of the SARS-CoV-2 (severe acute respiratory syndrome coronavirus-2) isolates have been examined from different geographical parts of India using phylogenetic tree analysis. A dataset of 172 Indian SARS-CoV-2 genome sequences were collected from a database and a phylogenetic tree was constructed. Results: From the phylogenetic analysis, six different clusters were identified and from each cluster 10 genome sequences were chosen to find open reading frames (ORFs) and common encoded proteins. Four encoded proteins that are common among the 60 genome sequences and correspond to ORF7a protein, Membrane glycoprotein, Surface glycoprotein and Nucleocapsid phosphoproteins were found. Our results highlight that there are six conserved motifs with high frequency of occurrence suggesting a potential of being useful in further studies. Conclusions: The encoded proteins and their detected sequential motifs might be useful for screening potential drugs and vaccine candidates of the SARS-CoV-2 Indian isolates in the current epidemic situation. © 2021 The Author(s).
Introduction: Wide applications in research, clinical and cosmetic industry of human epidermal growth factor (hEGF) made it a research interest target. Its production in different expression systems has shown several limitations. Recombinant expression of hEGF in E. coli is always accompanied by inclusion body formation. The object of this study is to the evaluation of a chromatography-independent approach for the production of EGF in E. coli as soluble form. Materials and Methods: In order to evaluate a chromatogram independent purification approach for recombinant hEGF production in a soluble form, the hEGF gene was fused to an elastin-like protein (ELP) and expressed in E. coli BL21 (DE3) using pET26b expression vector for secretion the product into periplasmic space. Results: Periplasmic protein content analysis confirmed that the recombinant protein is secreted into the periplasm. The purification process was done by using 0.4 M ammonium sulfate in two cycles of inverse phase transition (ITC). After two cycles of purification, purity reached more than 95%. Western blotting analysis with the monoclonal anti-EGF antibody has confirmed the accuracy of EGF. Biological activity of the purified protein was investigated on NIH-3T3 cell line and results indicated EGF-induced proliferation in treated cells. Our results showed periplasmic expression is the proper approach to the production of soluble recombinant hEGF. By using ELP fused to EGF, the purification process was established without applying chromatography which will result in decreasing in final costs. Conclusions: This study introduced a new economic and efficient approach to the production and purification of recombinant hEGF.
Introduction: Understanding the association among various disorders has remarkably improved their diagnosis and therapies. It has been observed that autism and inflammatory bowel diseases cause a sort of inflammation. Exploring the relationship between them will lead to discovering important involved genes and in turn will eventually help discover any possible common therapeutic protocols. The aim of the present study was to determine the correlation between autism spectrum disorders and inflammatory bowel diseases. Materials and Methods: The common genes associated with autism spectrum disorders and inflammatory bowel diseases were retrieved from DisGeNET. SFARI databases and were subjected to an in silico data analysis framework to explore predictive genes and the related pathways. Results: Eleven genes including HLA-DRB1, MTHFR, PON1, IL6, MTOR, SETD2, GSTM1, APC, IFNG, SERPINE1, and MAPK1 regulated by YY1 and IRF1 transcription factors were characterized as discriminating molecules which by further screening were enriched in pathways mostly involved in neutrophil apoptosis, neutrophil homeostasis, chemokine biosynthesis and the regulation of immune system response. Conclusions: According to findings it can be stated that the identified common genes were associated with a wide range of pathogenic mechanisms.
