Evaluation of the toxicity of magnetite (Fe3O4) nanoparticle aqueous suspensions on bio-indices of sea water (barnacle) and fresh water (rotifer)

Due to the inevitable achievement of nanoparticles to aquatic ecosystems, the limitation of existing reports, and the inadequate understanding of their possible biological reactions with aquatic organisms, this study in the pioneering step was aimed to toxicity assessment of aqueous suspension of chemical magnetic nanoparticles (Fe3O4) in zooplanktonic species such as barnacle larvae Amphibalanus amphitrite (sea water index) and rotifer Brachionus rotundiformis (fresh water index). For this purpose, serial concentrations (0, 10, 50, 100, 200, 500 mg / l) of magnetite nanoparticles were prepared in 24 well plates with 5 replicates. After that, the zooplanktons (50 barnacle nauplii and 20 neonate rotifer to each well) were introduced to plates and the sensitivity of the samples were evaluated for the toxicity of nanoparticles at a time interval of 12-48 hours. The results of this study showed that the toxicity effects of chemical magnetite nanoparticles on barnacle nauplii larvae and neonate rotifer were increased with increasing time and concentration of magnetite nanoparticles. Differences between control and treatment groups were significant (P1000 mg/l). Regardless of species variation in barnacles and rotifers, according to the results, magnetite nanoparticles are in the group of non-toxic contaminants for these zooplanktonic organisms

Cloning of the GH gene from the beluga sturgeon (Huso huso) into a lentivial & none viral constructs and its expression in HEK cell lines

Caviar-producing fish with their economically valuable product are important in fisheries. The cDNA growth hormone (GH) of Beluga sturgeon (Huso huso) was constructed using total RNA from pituitary glands. To construct the recombinant and active lentiviruses carring GH gene, this DNA sequence was inserted into the cloning vector pTZ57R/T and subsequently cutted from pTZ57R/T by endonuclease enzyme and incorporated into lentivirus vector pNL-EGFP/CMV-WPRE on upstream of an IRES cassette. We also insert a reporter EGFP gene downstream of IRES so transfection and transduction steps can be traced. Using this vector plus virus packaging and envelope vectors, HEK293T cells was co-transfected by DNA-Lipofectamine complexes method. Cell supernatant full of virions was collected 48 hours later and concentrated using Amicon columns to obtain a high-titer virus stock. Nearly 1/5 of this stock was applied to a new batch of cultured HEK-293T. After 72h expression of EGFP gene was detected and the cells was collected for further analysis. Total RNA of these transduced cells was extracted and GH mRNA expression was revealed by RT-PCR. Results showed that, lentiviral vectors (LV) as a gene transfer system provide efficient delivery, integration and long-term expression by establishing a stable provirus in target cells and could be important tool in aquaculture and fisheries biotechnology research to increase the growth rate of farmed fish by transferring growth hormone (GH) transgenes into fish

Cloning of the GH gene from the beluga sturgeon (Huso huso) into a lentivial & none viral constructs and it’s expression in HEK cell lines

Caviar-producing fish with their economically valuable product are important in fisheries. The cDNA growth hormone (GH) of Beluga sturgeon (Huso huso) was constructed using total RNA from pituitary glands. To construct the recombinant and active lentiviruses carring GH gene, this DNA sequence was inserted into the cloning vector pTZ57R/T and subsequently cutted from pTZ57R/T by endonuclease enzyme and incorporated into lentivirus vector pNL-EGFP/CMV-WPRE on upstream of an IRES cassette. We also insert a reporter EGFP gene downstream of IRES so transfection and transduction steps can be traced. Using this vector plus virus packaging and envelope vectors, HEK293T cells was co-transfected by DNA-Lipofectamine complexes method. Cell supernatant full of virions was collected 48 hours later and concentrated using Amicon columns to obtain a high-titer virus stock. Nearly 1/5 of this stock was applied to a new batch of cultured HEK-293T. After 72h expression of EGFP gene was detected and the cells was collected for further analysis. Total RNA of these transduced cells was extracted and GH mRNA expression was revealed by RT-PCR. Results showed that, lentiviral vectors (LV) as a gene transfer system provide efficient delivery, integration and long-term expression by establishing a stable provirus in target cells and could be important tool in aquaculture and fisheries biotechnology research to increase the growth rate of farmed fish by transferring growth hormone (GH) transgenes into fish

Early Hereditary Diffuse Gastric Cancer (eHDGC) is Characterized by Subtle Genomic Instability and Active DNA Damage Response

Diffuse gastric cancer (DGC) is one of the two primary types of stomach cancer. Carriers of germline mutations in the gene encoding E-cadherin are predisposed to DGC. The primary aim of the present study was to determine if genomic instability is an early event in DGC and how it may lead to disease progression. Chromosomal aberrations in early intramucosal hereditary diffuse gastric cancer (eHDGC) were assessed using array comparative genomic hybridization (array CGH). Notably, no aneuploidy or other large-scale chromosomal rearrangements were detected. Instead, all aberrations affected small regions (< 4.8 Mb) and were predominantly deletions. Analysis of DNA sequence patterns revealed that essentially all aberrations possessed the characteristics of common fragile sites. These results and the results of subsequent immunohistochemical examinations demonstrated that unlike advanced DGC, eHDGCs is characterized by low levels of genomic instability at fragile sites. Furthermore, they express an active DNA damage response, providing a molecular basis for the observed indolence of eHDGC. This finding is an important step to understanding the pathology underlying natural history of DGC and supports a revision of the current definition of eHDGC as a malignant disease.This research was supported by HS and JC Anderson Charitable Trust and the Health Research Council of New Zealand

In vitro cytotoxic and apoptotic activities of Allium paradoxum (M. bieb.) G. Don extract on human breast cancer cell line

Researchers from all pharmaceutical fields are trying to find new drugs from natural origin with less toxicity. In northern Hyrcanian forests Iran, Allium paradoxum (M. Bieb.) G. Don has traditionally used as food and vegetable. Previously studies reports, this plant has a medicinal potential for anti-oxidant and anti-hemolytic activities. In this regard, we evaluated the anti-tumor activity of hydroalcoholic extract of A. paradoxum (M. Bieb.) G. Don in different concentrations on human breast cancer cell line (MCF-7). MTT assay was performed with MCF-7 cancer cell line and also evaluation of apoptotic effect, Bax and Bcl-2 expression in MCF7 cells were analyzed by real time RT-PCR. The results showed that the A. paradoxum (M. Bieb.) G. Don extracts decrease the viability of MCF-7 cell line in a dose-dependent manner and the most effective concentration of this extracts after 24 h treatment was 100 µM. Apoptosis induction was confirmed by fluorescence microscopy and plant extracts display a pro-apoptotic effect by down-regulated and up-regulated the expression of Bcl-2 and BAX in tumor cells, respectively. In conclusion, the study was confirmed pro-apoptotic and cytotoxicity effect of A. paradoxum (M. Bieb.) G. Don extract against MCF-7 cell lines. Based on being natural, low cost, accessibility, and noteworthy advantages of this product, it seems that A. paradoxum (M. Bieb.) G. Don has a potential source for isolation of novel anticancer agents for a drug. © 2018, National Institute of Science Communication and Information Resources (NISCAIR). All rights reserved