Glucose effects on metformin-inhibited cell invasion.

<p><b>(</b>A & B) AGS cells were treated with metformin for 24 hours and the cell invasion was measured in the Culturex 96 well BME invasion assay kit by counting the number of cells invading underside of BME (*<i>p</i> ≤ 0.01, **<i>p</i> ≤ 0.001 and ***<i>p</i> ≤ 0.0001). (C) Effect of high- and low-glucose media on cell invasion analyzed by JMP and levels not connected by same letter are significantly different (<i>p</i> ≤ 0.05).</p

Metformin effects on EMT markers in two glucose levels.

<p>(A) The cells were treated with metformin for 72 hours in two glucose concentrations and the desired genes were quantified by real-time qPCR. For normalization, GAPDH was amplified in each sample. Each column shows the mean and SD of three independent experiments, performed in triplicate. (B) Western blot analysis of cells treated with 10 mM metformin for 24 to 72 hours in two different glucose concentrations. The GAPDH band, which is considered as control, confirms the integrity and equal loading of protein.</p

Toward Chemical Perfection of Graphene-Based Gene Carrier via Ugi Multicomponent Assembly Process

The graphene-based materials with unique, versatile, and tunable properties have brought new opportunities for the leading edge of advanced nanobiotechnology. In this regard, the use of graphene in gene delivery applications is still at early stages. In this study, we successfully designed a new complex of carboxylated-graphene (G-COOH) with ethidium bromide (EtBr) and used it as a nanovector for efficient gene delivery into the AGS cells. G-COOH, with carboxyl functions on its surface, in the presence of EtBr, formaldehyde, and cyclohexylisocyanide were participated in Ugi four component reaction to fabricate a stable amphiphilic graphene-EtBr (AG-EtBr) composite. The coupling reaction was confirmed by further analyses with FT-IR, AFM, UV–vis, Raman, photoluminescence, EDS, and XPS. The AG-EtBr nanocomposite was able to interact with a plasmid DNA (pDNA). This nanocomposite has been applied for transfection of cultured mammalian cells successfully. Moreover, the AG-EtBr composites showed a remarkable decreased cytotoxicity in compared to EtBr. Interestingly, the advantages of AG-EtBr in cell transfection are more dramatic (3-fold higher) than Lipofectamine2000 as a commercial nonviral vector. To the best of our knowledge, this is the first report in which EtBr is used as an intercalating agent along with graphene to serve as a new vehicle for gene delivery application

MicroRNA expression in serum samples of sulfur mustard veterans as a diagnostic gateway to improve care

<div><p>Sulfur mustard is a vesicant chemical warfare agent, which has been used during Iraq-Iran-war. Many veterans and civilians still suffer from long-term complications of sulfur mustard exposure, especially in their lung. Although the lung lesions of these patients are similar to Chronic Obstructive Pulmonary Disease (COPD), there are some differences due to different etiology and clinical care. Less is known on the molecular mechanism of sulfur mustard patients and specific treatment options. microRNAs are master regulators of many biological pathways and proofed to be stable surrogate markers in body fluids. Based on that microRNA expression for serum samples of sulfur mustard patients were examined, to establish specific microRNA patterns as a basis for diagnostic use and insight into affected molecular pathways. Patients were categorized based on their long-term complications into three groups and microRNA serum levels were measured. The differentially regulated microRNAs and their corresponding gene targets were identified. Cell cycle arrest, ageing and TGF-beta signaling pathways showed up to be the most deregulated pathways. The candidate microRNA miR-143-3p could be validated on all individual patients. In a ROC analysis miR-143-3p turned out to be a suitable diagnostic biomarker in the mild and severe categories of patients. Further microRNAs which might own a link to the biology of the sulfur mustard patients are miR-365a-3p, miR-200a-3p, miR-663a. miR-148a-3p, which showed up only in a validation study, might be linked to the airway complications of the sulfur mustard patients. All the other candidate microRNAs do not directly link to COPD phenotype or lung complications. In summary the microRNA screening study characterizes several molecular differences in-between the clinical categories of the sulfur mustard exposure groups and established some useful microRNA biomarkers. qPCR raw data is available via the Gene Expression Omnibus <a href="https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE110797" target="_blank">https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE110797</a>.</p></div

The 9 top affected cellular pathways influenced from the 29 microRNAs of the normal-severe comparison.

<p>The 9 top affected cellular pathways influenced from the 29 microRNAs of the normal-severe comparison.</p

The 9 top affected cellular pathways influenced from the 15 microRNAs of the normal-mild comparison.

<p>The 9 top affected cellular pathways influenced from the 15 microRNAs of the normal-mild comparison.</p

Box plots of raw data microRNA expression values and controls.

<p>On the x axis the different measurement groups for each experiment are shown: 'control' stands for reference genes including miR-103a-3p, miR-423-5p and miR-191-5p. 'control2' denotes non-miRNA coding reference genes. 'IPC' lists the inter plate calibrators. 'targets' comprises all the measured individual microRNAs. As can be seen in this figure the distribution of the target genes is nearly consistent in all tested samples even on the raw data level. The y axis denotes the Ct values.</p

Graphical abstract.

<p>Graphical abstract.</p

Differential microRNAs (29) of the normal-severe group.

<p>The values shown here are from the right part of the workflow in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0194530#pone.0194530.g002" target="_blank">Fig 2</a>. dCt: delta Ct, FC: fold change and sampling p: sampling p value which was finally considered.</p

The 9 top affected cellular pathways influenced from the 9 microRNAs of the mild-severe intersection.

<p>The 9 top affected cellular pathways influenced from the 9 microRNAs of the mild-severe intersection.</p