Different expression systems for production of HCV structural proteins

     Hepatitis C virus (HCV) is an important agent causing chronic liver infection, which often leads to liver cirrhosis and lethal hepatocellular carcinoma (HCC). At present, there is no effective HCV vaccine for prevention of hepatic disease and the standard treatment is neither economical nor fully effective in all the patients. However, vaccination based on structural and nonstructural proteins of HCV has attracted a special interest. Different heterologous systems have been used to generate the recombinant HCV core, E1, and E2 proteins including Escherichia coli, yeast, insects and mammalian cells. Further studies showed that the amounts of HCV recombinant proteins in E. coli are more suitable and un-expensive compared to other systems. It should be considered that this system is not efficient for generation of the glycosylated proteins. Thus, the structure of proteins is an important agent of selection for expression systems. The selection of expression systems will be critical for the use of recombinant proteins as an immunogen. In this mini-review, we briefly describe different expression systems for generation of the HCV recombinant structural proteins applied in vaccine design

Elderberry diet improves memory function and prevents cell death in rat models of Alzheimer’s disease induced by amyloid beta injection

Background: Alzheimer’s disease (AD) is a neurodegenerative disease which adversely affects memory and learning skills. Currently, there is no disease-modifying therapeutic approach for AD. Aim: A growing body of literature suggests elderberry as a promising remedy for neurological disorders. This study investigates the therapeutic effects of the elderberry diet on amyloid beta-induced (Aβ) rat model. Methods:  Initially, Alzheimer’s model was generated by Aβ administration followed by the treatment of elderberry diet. Then, short term spatial memory was assessed. Stereology was also performed for the evaluation of density of neurons and dark neuron in the hippocampus. Results: The findings showed that the administration of the elderberry diet in an Aβ model of Alzheimer’ disease improved memory and learning function and prevented the degeneration of hippocampal neurons. Conclusion: Overall, the findings imply that the elderberry diet attenuates the Aβ-induced degeneration and improves memory performance. As such, the elderberry diet could be used as a therapeutic procedure for Alzheimer due to its neuroprotective effects

Comparison Study on the Effect of Mesenchymal Stem Cells-Conditioned Medium Derived from Adipose and Wharton’s Jelly on Versican Gene Expression in Hypoxia

BACKGROUND: Mesenchymal stem cells enhance tissue repair through paracrine effects following transplantation. The versican protein is one of the important factors contributing to this repair mechanism. Using MSC conditioned medium for cultivating monocytes may increase versican protein production and could be a good alternative for transplantation of MSCs. This study investigates the effect of culture medium conditioned by human MSCs on the expression of the versican gene in PBMCs under hypoxia-mimetic and normoxic conditions. METHODS: The conditioned media used were derived from either adipose tissue or from WJ. Flow cytometry for surface markers (CD105, CD73, and CD90) was used to confirm MSCs. The PBMCs were isolated and cultured with the culture media of the MSC derived from either the adipose tissue or WJ. Desferrioxamine and cobalt chloride (200 and 300 µM final concentrations, respectively) were added to monocytes media to induce hypoxia-mimetic conditions. Western blotting was applied to detect HIF-1α protein and confirm hypoxia-mimetic conditions in PBMC. Versican gene expression was assessed in PBMC using RT-PCR. Western blotting showed that the expression of HIF-1α in PBMC increased significantly (p < 0.01). RESULTS: RT-PCR results demonstrated that the expression of the versican and VEGF genes in PBMC increased significantly (p < 0.01) in hypoxia-mimetic conditions as compared to normoxia. CONCLUSION: Based on the findings in the present study, the secreted factors of MSCs can be replaced by direct use of MSCs to improve damaged tissues

5-Azacytidine Enhancing Expression of E-cadherin in Adenocarcinoma Cell Line

Introduction: In this study, we assessed the expression of E-cadherin in HT29 cell line treated with 5-Azacytidine and colorectal cancer patient in an Iranian population. E-cadherin expression promotes metastasis and prognosis of colorectal cancer (CRC). 5-Azacytidine, a DNA methyl transferase inhibitor, is a clinically used epigenetic drug for treatment of cancer including colorectal cancer, leading to genes activation involved in tumor suppression, especially E-cadherin. Materials and Methods: HT29 cell line treated with 5-Azacitidine and 40 polyps, 20 tumors and 40 adjacent normal tissues samples were enrolled in this study. Using the real-time PCR method, the expression levels of E-cadherin were examined in treated cell line and colorectal cancer tissue. Results: This study proves that 5-Azacytidine induces over expression of E-cadherin in adenocarcinoma cell line, while the expression levels of E-cadherin were not different in tumor and polyp than adjacent normal tissue. Conclusion: To conclude, 5-Azacytidine induces re-expression of E-cadherin in adenocarcinoma cell line. Thus, 5-Azacytidine as demethylation drug activated tumor suppressor gene as E-cadherin

Designing novel construction for cell surface display of protein E on Escherichia coli using non-classical pathway based on Lpp-OmpA

Additional file 2. pNon-OmpA-PE. ab1. Sequencing result of pNon-OmpA-PE

Citral effect in male NMRI mice nonalcoholic steatosis model: assessing biochemical and histological parameters and PPARα gene expression

Citral is a small molecule present in various citrus species, with reported anti-hyperlipidemic and antiinflammation effects. Here, the effect of intraperitoneal (IP) administration of citral is evaluated in a mouse model of non-alcoholic steatosis. Male NMRI mice were divided into the following groups (n = 12): normal control group (NC) receiving a normal diet; high-fat emulsion group (HF) receiving high fat diet for four weeks; positive control group (C+) receiving HF diet for four weeks and then shifted to normal diet with IP-administered silymarin (80 mg/kg) for four weeks; sham group receiving HF diet for four weeks and then shifted to normal diet for four weeks; and EC1, EC2, and EC3 groups receiving HF diet for four weeks and then shifted to normal diet with IP-administered citral doses of 5, 10, and 20 mg/kg, respectively. HF diet resulted in steatohepatitis with impaired lipid profile, high glucose levels and insulin resistance, impaired liver enzymes, antioxidants, adiponectin and leptin levels, decreased PPARα level, and fibrosis in the liver tissue. Upon treatment with citral, improvement in condition was observed in a dose-dependent manner—both at histological level and in the serum of treated animals. and the PPARα level was also increased

Solvent effects on structural and thermochemical properties of p53 tumor-suppressor gene: a molecular modeling approach in drug design

The p53 tumor-suppressor protein is a cellular phosphoprotein and a negative regulator of cell growth. Most p53 mutations occur in exons 5–8 within the DNA-binding domain. Therefore, p53 can potentially be targeted with novel drugs designed to bind to a mutation and restore its stability or wild-type conformation. For the current study, Hartree–Fock calculations were used to investigate the solvent-induced effects of five different solvent media (acetone, ethanol, methanol, dimethyl sulfoxide, and water) on the thermochemical parameters and relative energies, and on the multinuclear nuclear magnetic resonance shielding tensors of oxygen, nitrogen, and phosphorus nuclei, of GAT. To understand how the solvent affects the mutation region (the “hot spot”) of p53, the relative energies of GAT in selected solvent media were determined. Some biological evidence suggested the structural stabilities of hot spots of GAT have the optimum temperature and solvent type for mutation. All the authors’ findings are in accordance with common biological phenomena. Another important objective of this study was to compare the hydration Gibbs free energies of CUA and GAT in water using two different approaches where the solvent was treated as a continuum of the constant at different levels of Hartree–Fock theory. The Gibbs hydration energy values obtained in water with the polarized continuum model directly applied on the isolated CUA and GAT sequences were compared with those determined from the hydrated models with four, six, and eight water molecule clusters around the hot spots uracil and adenine. The clustered structures of water molecules around the hot spots of GAT (in DNA level) and CUA (in transcriptional level) were found to be energetically favored. The results of this study provide a reliable insight into the nature of mutation processes, which is of utmost importance for the study of biochemical structures, and provide a basis for drug design

Induction Effects Of Bacteroides Fragilis Derived Outer Membrane Vesicles On Toll Like Receptor 2, Toll Like Receptor 4 Genes Expression And Cytokines Concentration In Human Intestinal Epithelial Cells

Objective Gastrointestinal (GI) tract, like other mucosal surface, is colonized with a microbial population known as gut microbiota. Outer membrane vesicles (OMVs) which are produced by gram negative bacteria could be sensed by Toll like receptors (TLRs). The interaction between gut microbiota and TLRs affects homeostasis and immune responses. In this study, we evaluated TLR2, TLR4 genes expression and cytokines concentration in Caco-2 cell line treated with Bacteroides fragilis (B. fragilis) and its OMVs. Materials And Methods In this experimental study, OMVs were extracted using sequential centrifugation and their physicochemical properties were evaluated as part of quality control assessment. Caco-2 cells were treated with B. fragilis and its OMVs (180 and 350 µg/ml). Quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) was performed to assess TLR2 and TLR4 mRNA expression levels. Pro-inflammatory (IFNᵧ) and anti-inflammatory (IL- 4 and IL-10) cytokines were evaluated by ELISA. Results B. fragilis significantly decreased TLR2 and slightly increased TLR4 mRNA levels in Caco-2 cell line. The TLR2 mRNA level was slightly increased at 180 and 350 µg/ml of OMVs. Conversely, the TLR4 mRNA level was decreased at 180 µg/ml of OMVs, while it was significantly increased at 350 µg/ml of OMVs. Furthermore, B. fragilis and its OMVs significantly increased and decreased IFNᵧ concentration, respectively. Anti-inflammatory cytokines were increased by B. fragilis and its OMVs. Conclusion B. fragilis and its OMVs have pivotal role in the cross talk between gut microbiota and the host especially in the modulation of the immune system. Based on the last studies on immunomodulatory effect of B. fragilis derived OMVs on immune cells and our results, we postulate that B. fragilis derived OMVs could be possible candidates for the reduction of immune responses

The differential DNA hypermethylation patterns of 2 microRNA-137 and microRNA-342 locus in early 3 colorectal lesions and tumours

Colorectal cancer (CRC) is the third most commonly diagnosed cancer worldwide, representing 13% of all cancers. The role of epigenetics in cancer diagnosis and prognosis is well established. MicroRNAs in particular influence numerous cancer associated processes including apoptosis, proliferation, differentiation, cell-cycle controls, migration/invasion and metabolism. MiRNAs-137 and 342 are exon- and intron-embedded, respectively, acting as tumour-suppressive microRNA via hypermethylation events. Levels of miRNAs 137 and 342 have been investigated here as potential prognostic markers for colorectal cancer patients. The methylation status of miRNA-137 and miRNA-342 was evaluated using methylation-specific (MSP) polymerase chain reaction (PCR) on freshly frozen tissue derived from 51 polyps, 8 tumours and 14 normal colon mucosa specimens. Methylation status of miRNA-137 and miRNA-342 was significantly higher in tumour lesions compared to normal adjacent mucosa. Surprisingly, the methylation frequency of miR-342 (76.3%) among colorectal cancer patients was significantly higher compared to miR-137 (18.6%). Furthermore, normal tissues, adjacent to the lesions (N-Cs), displayed no observable methylation for miRNA-137, whereas 27.2% of these N-Cs showed miRNA-342 hypermethylation. MiRNA-137 hypermethylation was significantly higher in male patients and miR-342 hypermethylation correlated with patient age. Methylation status of miRNA-137 and miRNA-342 has both diagnostic and prognostic value in CRC prediction and prevention

Synergistic effect of microRNA and albumin-bound nanoparticles for inhibition of glioblastoma cancer cell proliferation

The functional significance of upregulation miR-34a in combination with albumin-bound paclitaxel nanoparticles in U251 glioblastoma cell line has been evaluated. The MTT assay determined that miR34a and albumin-bound paclitaxel nanoparticles can reduce cell viability, but the combination of both factors has a stronger effect on cell viability. The application of qRT-PCR has demonstrated that the transduction of miR-34a could lead to exogenous upregulation of miR-34a level and downregulation of SURVIVIN. Moreover, treatment of U251 cells with miR-34a and nanoparticles together considerably inhibit SURVIVIN expression compared to miR-34a and nanoparticles alone. Flow cytometry showed that upon miR-34a overexpression cell cycle arrested in G1 phase, while treatment with nanoparticles increased the cell population in G2 phase. Upregulation of miR-34a along with treatment with nanoparticles elevated the number of cells arrested in G1/ G2 phases of the cell cycle. Expression of miR-34a with albumin-bound paclitaxel nanoparticles reduced cell viability, downregulated SURVIVIN and enhanced cell cycle arrest in G1/G2 phases. Thus, the upregulation of miR-34a with these nanoparticles are potential candidates therapeutic for glioblastoma cancer