Receptor priming of major group human rhinoviruses for uncoating and entry at mild low-pH environments

Receptor priming of low-pH-triggered virus entry has been described for an enveloped virus (15). Here we show with major group human rhinoviruses (HRV) and its intercellular adhesion molecule-1 receptor that nonenveloped viruses follow this novel cell entry principle. In vitro the receptor primed HRV for efficient uncoating at mild low pH (5.5 to 6.0). Agents preventing endosomal acidification reduced or blocked rhinovirus cell infection, while nocodazole had no effect on infection of any serotype tested. The entry inhibitory effect of lysosomotropic agents was overcome by exposing cell-internalized HRV to mild low pH (5.5 to 6.0). We therefore conclude that receptor priming of major group HRV must occur in vivo as well. Cooperation of a cellular receptor and low pH in virus uncoating will polarize the exit of the genome to the receptor-bound, membrane-proximal region of the virus particle during acidification of endosomes. This process must be required for efficient penetration of the cellular membrane by viruses.     

Subcutaneous administration CpG-ODNs acts as a potent adjuvant for an HIV-1-<Emphasis Type="Italic">tat</Emphasis>-based vaccine candidate to elicit cellular immunity in BALB/c mice

Objective

To evaluate the combined effects of CpG oligodeoxynucleotides (CpG-ODNs) adjuvant and subcutaneous injection route on efficacy of a HIV-1-tat DNA vaccine candidate using BALB/c mice as an animal model.

Results

Evaluation of cellular and humoral immunity of mice injected subcutaneously with HIV-1-tat gene cloned into a pcDNA3.1 vector indicated that significant levels of IFN-γ cytokine secretion (900 pg/ml), lymphocyte proliferation (2.5 stimulation index) and IgG_2a (1.45 absorbance 450 nm) production could be achieved. These indicators of stimulated cellular immunity were elicited 2 weeks after the last injection (P < 0.05).

Conclusions

Formulation of HIV-1-tat DNA vaccine candidate with CpG-ODNs as an adjuvant while administrated subcutaneously are a promising approach to induce effective cellular immunity responses against HIV-1 infection.

     

Induction of tetraploidy in garden cress: morphological and cytological changes

Plant Cell, Tissue and Organ Culture (PCTOC) - Garden cress (Lepidium sativum L., Brassicaceae) is one of the most popular leafy vegetables which is widely used, and has also various medicinal...     

Epigenetic modulation of BRCA-1 and MGMT genes,and histones H4 and H3 are associated with breast tumors

Aberrant patterns in promoter methylation of tumor-suppressor genes and posttranslational modifications of histone proteins are considered as major features of malignancy. In this study, we aimed to investigate promoter methylation of three tumor-suppressor genes (BRCA-1, MGMT, and P16) and three histone marks (H3K9ac, H3K18ac, and H4K20me3) in patients with breast tumors. This case-control study included 27 patients with malignant breast tumors (MBT) and 31 patients with benign breast tumors (BBT). The methylation-specific PCR was used for determining promoter methylation of BRCA-1, MGMT, and P16 genes. Western blot analysis was performed to detect histone lysine acetylation (H3K9ac and H3K18ac) and lysine methylation (H4K20me3). BRCA-1 promoter methylation was detected in 44.4% of the MBT whereas this alteration was found in 9.7% of BBT (P = 0.005). The Kaplan-Meier analysis indicated that hypermethylation in BRCA-1 promoter was significantly associated with poor overall survival of patients with breast cancer (P = 0.039). MGMT promoter methylation was identified in 18.5% of MBT and 0.0% of the BBT (P = 0.01). The frequency of P16 promoter methylation was 25.8% in BBT and 11.1% in MBT (P = 0.12). As compared with BBT, MBT samples displayed the aberrant patterns of histones marks with hypomethylation of H4K20 and hypoacetylation of H3K18 (P = 0.03 and P = 0.04, respectively). There was a negative significant correlation between H3K9ac levels and tumor size in MBT group (r = −0.672; P = 0.008). The present findings suggest that promoter hypermethylation of MGMT and BRCA-1 genes along with alterations in H3K18ac and H4K20me3 levels may have prognostic values in patients with breast cancer. Moreover, the detection of these epigenetic modifications in breast tumors could be helpful in finding new methods for breast cancer therapy.     

Effect of colchicine-induced polyploidy on morphological characteristics and essential oil composition of ajowan (<Emphasis Type="Italic">Trachyspermum ammi</Emphasis> L.)

Cyclophilins (CYPs) belong to the immunophilin superfamily, having the peptidyl prolyl cis/trans isomerase activity that can catalyze the cis/trans isomerisation process of proline residues. Previous studies have shown their importance in plants, but no comprehensive analysis of maize CYP family has been reported. In the present study, a whole-genome-wide analysis of maize CYP family was performed and 39 ZmCYP genes (ZmCYP1 to ZmCYP39) were identified from maize genome, which were unequally distributed on maize ten chromosomes. Phylogenetic analysis revealed a weak relationship among these ZmCYP genes. Furthermore, their gene structure and motif patterns also displayed variant within the gene family. Four segmental and one tandem duplicated gene pairs were found from 39 ZmCYP genes, respectively, indicating their roles in the expansion of maize CYP family. Expression analysis of 39 ZmCYP genes in maize tissues showed their differential tissue specific expression patterns. Quantitative real-time PCR analysis of 19 selected ZmCYP genes under salinity stress indicated their stress-inducible expression profile. Heterologous expression of ZmCYP15 in E. coli enhanced tolerance against abiotic stress. Subcellular localization analysis indicated ZmCYP15 was located in nucleus and cytoplasm. Our study describes the importance of the maize CYP gene family in stress response, and provides a reference for future study and application for maize genetic improvement.     

Impacts of process parameters optimization on the performance of the annular single chamber microbial fuel cell in wastewater treatment

Energy harvest from optimized annular single chamber microbial fuel cell (ASCMFC) with novel configuration, which treats chocolate industry wastewater, was investigated. In this study, optimization of operational parameters of the ASCMFC in terms of efficiency water‐soluble organic matter reduction and capability of electricity generation was evaluated. During the experiment, effluent from the anode compartment was examined through current and power density curves for variation in temperature and pH, chemical oxygen demand (COD), and turbidity removal, and substrate concentration. The performance analyzed at different temperature ranges such as 25, 30, 35, and 40°C, which showed 88% increase by uprising temperature from 25 to 35°C. The ASCMFC was used to produce electricity by adjusting pH between 5 and 9 at resistance of 100 Ω. Under the condition of pH 7 power density (16.75 W/m³) was highest, which means natural pH is preferred to maximize microbial activities. Wastewater concentration with COD of 700 and 1400 mg/L were investigated to determine its affection on current production. Reduction of current density was observed due to decrease in wastewater concentration. Significant reduction in COD and turbidity of effluent were 91 and 78%, respectively. The coulombic efficiency of 45.1% was achieved.     

Mesenchymal stem cells as carrier of the therapeutic agent in the gene therapy of blood disorders

Nonhematopoietic stem cells as a delivery platform of therapeutic useful genes have attracted widespread attention in recent years, owing to gained a long lifespan, easy separation, high proliferation, and high transfection capacity. Mesenchymal stem/stromal cells (MSCs) are the choice of the cells for gene and cell therapy due to high self-renewal capacity, high migration rate to the site of the tumor, and with immune suppressive and anti-inflammatory properties. Hence, it has a high potential of safety genetic modification of MSCs for antitumor gene expression and has paved the way for the clinical application of these cells to target the therapy of cancers and other diseases. The aim of gene therapy is targeted treatment of cancers and diseases through recovery, change, or enhancement cell performance to the sustained secretion of useful therapeutic proteins and induction expression of the functional gene in intended tissue. Recent developments in the vectors designing leading to the increase and durability of expression and improvement of the safety of the vectors that overcome a lot of problems, such as durability of expression and the host immune response. Nowadays, gene therapy approach is used by MSCs as a delivery vehicle in the preclinical and the clinical trials for the secretion of erythropoietin, recombinant antibodies, coagulation factors, cytokines, as well as angiogenic inhibitors in many blood disorders like anemia, hemophilia, and malignancies. In this study, we critically discuss the status of gene therapy by MSCs as a delivery vehicle for the treatment of blood disorders. Finally, the results of clinical trial studies are assessed, highlighting promising advantages of this emerging technology in the clinical setting.     

The association between fat mass and obesity-associated (FTO) genotype and serum vitamin D level in breast cancer patients

The preventive effect of vitamin D against breast cancer can be influenced by gene polymorphisms. This study aimed to investigate the association between serum level of 25(OH) vitamin D and FTO genotype in breast cancer patients. A cross-sectional study was carried out on 180 newly diagnosed patients with breast cancer in Tehran, Iran. The blood samples were collected from the participants in order to assess the FTO gene rs9939609 polymorphism by the tetra-primer amplification refractory mutation system (Tetra-ARMS) PCR method. The serum level of 25(OH) vitamin D was measured using the direct competitive enzyme-linked immunosorbent assay (ELISA) method. The association between vitamin D and the FTO genotype in patients with breast cancer was assessed after adjustment for cofounders. The frequency of TT, AT and AA genotypes in the breast cancer patients were 43% (n = 77), 49% (n = 89) and 8% (n = 14), respectively. All patients with higher than 40 ng/dl of serum 25(OH) vitamin D had one or two copies of FTO rs9939609 risk allele (p = 0.019). No linear association was found between the number of FTO risk allele and the level of serum vitamin D. All patients with high serum level of 25(OH) vitamin D had one or two copies of FTO rs9939609 risk allele. FTO gene polymorphisms may counteract the beneficial effects of vitamin D in breast cancer prevention. Further studies can help to better understand the genetic factors predisposing to breast cancer and their effect on the association between vitamin D and breast cancer.