A Human Ex Vivo Atherosclerotic Plaque Model to Study Lesion Biology

Atherosclerosis is a chronic inflammatory disease of the vasculature. There are various methods to study the inflammatory compound in atherosclerotic lesions. Mouse models are an important tool to investigate inflammatory processes in atherogenesis, but these models suffer from the phenotypic and functional differences between the murine and human immune system. In vitro cell experiments are used to specifically evaluate cell type-dependent changes caused by a substance of interest, but culture-dependent variations and the inability to analyze the influence of specific molecules in the context of the inflammatory compound in atherosclerotic lesions limit the impact of the results. In addition, measuring levels of a molecule of interest in human blood helps to further investigate its clinical relevance, but this represents systemic and not local inflammation. Therefore, we here describe a plaque culture model to study human atherosclerotic lesion biology ex vivo. In short, fresh plaques are obtained from patients undergoing endarterectomy or coronary artery bypass grafting and stored in RPMI medium on ice until usage. The specimens are cut into small pieces followed by random distribution into a 48-well plate, containing RPMI medium in addition to a substance of interest such as cytokines or chemokines alone or in combination for defined periods of time. After incubation, the plaque pieces can be shock frozen for mRNA isolation, embedded in Paraffin or OCT for immunohistochemistry staining or smashed and lysed for western blotting. Furthermore, cells may be isolated from the plaque for flow cytometry analysis. In addition, supernatants can be collected for protein measurement by ELISA. In conclusion, the presented ex vivo model opens the possibility to further study inflammatory lesional biology, which may result in identification of novel disease mechanisms and therapeutic targets.     

The effect of oleuropein on unilateral ureteral obstruction induced-kidney injury in rats: the role of oxidative stress,inflammation and apoptosis

Molecular Biology Reports - Unilateral ureteral obstruction (UUO) induces kidney injury. Oleuropein as a major compound of olive leaves modulates the inflammatory parameters and decreases oxidative...     

Does Magnesium Supplementation Improve Body Composition and Muscle Strength in Middle-Aged Overweight Women? A Double-Blind, Placebo-Controlled, Randomized Clinical Trial

Muscle strength, an independent predictor of metabolic disorders, disability, and mortality, reduces gradually with advancing age. Little is known about the influence of nutritional intervention on muscle strength in middle-aged. The aim of the present study is to examine whether magnesium could improve body composition and muscle strength in middle-aged overweight women. In this double-blinded, placebo-controlled, randomized trial, a total of 74 healthy middle-aged overweight women (25?≤?BMI?≤?30 kg/m²) received either 250 mg magnesium in the form of magnesium oxide or placebo daily for 8 weeks. Body composition was assessed using Bioelectrical Impedance Analysis (BIA). Handgrip strength and knee extension strength were measured with isometric dynamometry. Functional mobility was assessed using Time Get Up and Go Test (TGUG). A significant increase in mean lean body mass was observed (P?=?0.05) accompanied with a significant decrease in fat mass (P?=?0.02) solely in the magnesium group at the end of 8 weeks compared to baseline values but the changes did not reach significant as compared to placebo group. Handgrip strength and TGUG improved in the magnesium group compared to baseline but they were not significant compared to placebo. There were no significant differences in increasing knee extension strength in the magnesium group as compared with placebo. Baseline values of serum magnesium and muscle strength of participants did not indicate any influences on response to magnesium supplementation. Our findings indicate that magnesium as magnesium oxide, 250 mg/day, for 8 weeks do not lead to a significant greater gain in muscle strength and function compared to placebo.     

Effect of Tricalcium Magnesium Silicate Coating on the Electrochemical and Biological Behavior of Ti-6Al-4V Alloys

In the current study, a sol-gel-synthesized tricalcium magnesium silicate powder was coated on Ti-6Al-4V alloys using plasma spray method. Composition of feed powder was evaluated by X-ray diffraction technique before and after the coating process. Scanning electron microscopy and atomic force microscopy were used to study the morphology of coated substrates. The corrosion behaviors of bare and coated Ti-6Al-4V alloys were examined using potentiodynamic polarization test and electrochemical impedance spectroscopy in stimulated body fluids. Moreover, bare and coated Ti-6Al-4V alloys were characterized in vitro by culturing osteoblast and mesenchymal stem cells for several days. Results demonstrated a meaningful improvement in the corrosion resistance of Ti-6Al-4V alloys coated with tricalcium magnesium silicate compared with the bare counterparts, by showing a decrease in corrosion current density from 1.84 μA/cm² to 0.31 μA/cm². Furthermore, the coating substantially improved the bioactivity of Ti-6Al-4Valloys. Our study on corrosion behavior and biological response of Ti-6Al-4V alloy coated by tricalcium magnesium silicate proved that the coating has considerably enhanced safety and applicability of Ti-6Al-4V alloys, suggesting its potential use in permanent implants and artificial joints.     

Novel plasma phospholipid biomarkers of autism: Mitochondrial dysfunction as a putative causative mechanism

Autism is a neurological disorder that manifests as noticeable behavioral and developmental abnormalities predominantly in males between the ages of 2 and 10. Although the genetics, biochemistry and neuropathology of this disease have been extensively studied, underlying causal factors to this disease have remained elusive. Using a longitudinal trial design in which three plasma samples were collected from 15 autistic and 12 non-autistic age-matched controls over the course of 1 year, universal and unambiguous alterations in lipid metabolism were observed. Biomarkers of fatty acid elongation and desaturation (poly-unsaturated long chain fatty acids (PUFA) and/or saturated very long chain fatty acids (VLCFA)-containing ethanolamine phospholipids) were statistically elevated in all autistic subjects. In all 8 of the affected/non-affected sibling pairs, the affected sibling had higher levels of these biomarkers than the unaffected sibling. Exposure of neurons, astrocytes and hepatocytes in vitro to elevated extracellular glutamate levels resulted in lipid biomarker changes indistinguishable from those observed in autistic subjects. Glutamate stress also resulted in in vitro decreased levels of reduced glutathione (GSH), methionine and cysteine, in a similar way to the decreases we observed in autism plasma. Impaired mitochondrial fatty acid oxidation, elevated plasma VLCFAs, and glutamate toxicity as putative causal factors in the biochemistry, neuropathology, and gender bias in autism are discussed.     

Aging: A cell source limiting factor in tissue engineering

Tissue engineering has yet to reach its ideal goal, i.e. creating profitable off-the-shelf tissues and organs, designing scaffolds and three-dimensional tissue architectures that can maintain the blood supply, proper biomaterial selection, and identifying the most efficient cell source for use in cell therapy and tissue engineering. These are still the major challenges in this field. Regarding the identification of the most appropriate cell source, aging as a factor that affects both somatic and stem cells and limits their function and applications is a preventable and, at least to some extents, a reversible phenomenon. Here, we reviewed different stem cell types, namely embryonic stem cells, adult stem cells, induced pluripotent stem cells, and genetically modified stem cells, as well as their sources, i.e. autologous, allogeneic, and xenogeneic sources. Afterward, we approached aging by discussing the functional decline of aged stem cells and different intrinsic and extrinsic factors that are involved in stem cell aging including replicative senescence and Hayflick limit, autophagy, epigenetic changes, miRNAs, mTOR and AMPK pathways, and the role of mitochondria in stem cell senescence. Finally, various interventions for rejuvenation and geroprotection of stem cells are discussed. These interventions can be applied in cell therapy and tissue engineering methods to conquer aging as a limiting factor, both in original cell source and in the in vitro proliferated cells.     

Functional response of Nesidiocoris tenuis (Hemiptera: Miridae) to Trialeurodes vaporariorum (Hemiptera: Aleyrodidae): effect of different host plants

The influence of three host plants, namely cucumber, tomato and eggplant, on functional response of male, virgin and mated female predatory bug Nesidiocoris tenuis was investigated on different densities of Trialeurodes vaporariorum nymphs. The 24-h experiment conducted at laboratory conditions revealed that N. tenuis exhibited a type II functional response to T. vaporariorum on host plants. There were no significant differences between attack rates, as well as handling times estimated for each adult stage of the predator between host plants. However, on each host plant, the handling time estimated for the mated female in comparison with two other adult stages had lower values (0.7952, 0.6827 and 0.8884?h^?1 on cucumber, tomato and eggplant, respectively). Handling time estimated for the mated female on cucumber was significantly lower than that estimated for the male predator. The highest maximum handling rate (T/T_h) was estimated for the mated female followed by the virgin female and male on all host plants. For three adult stages of the predator, the highest value of this parameter was determined on tomato followed by cucumber and eggplant. Unlike virgin and mated females, the host plant significantly affected prey consumption by the male. Prey consumed by mated females was higher than those obtained for two other adult stages of the predator on each host plant. The difference in trichome density between three host plants may be responsible for the obtained results. These results revealed that N. tenuis is more effective in the biological control of T. vaporariorum on tomato in comparison with cucumber and eggplant.     

Responses of pomegranate cultivars to severe water stress and recovery: changes on antioxidant enzyme activities,gene expression patterns and water stress responsive metabolites

The biochemical and molecular responses of five commercially well-known pomegranate cultivars to severe water stress were studied. The cultivars were subjected to 14-day water stress by withholding irrigation, followed by re-watering for 7 days. Results showed clear differences in metabolites contents and activities of antioxidant enzymes among various pomegranate cultivars during severe water stress and recovery. According to our results, increased accumulation of proline in pomegranate was not related to osmotic adjustment during severe water stress. Except for ‘Ghojagh’, leaves grown under severe water stress conditions showed symptoms of oxidative stress such as reduced chlorophyll concentration. The improved performance of ‘Ghojagh’ under drought stress may be associated with an efficient osmotic adjustment. The up- or down regulated expression of cytosolic glutathione reductase (cytosolic GR) and glutathione peroxidase were observed under drought conditions. Moreover, the suppressed expression of cytosolic GR was also noted. Comparatively, ‘Rabab’ exhibited higher antioxidant capacity and an efficient ROS-scavenging mechanism under drought stress. Lower levels of membrane lipid peroxidation in ‘Ghojagh’ and ‘Rabab’ under drought stress and the marked reduction of malondialdehyde concentration after re-watering represents that these cultivars have a good tolerance to drought stress. As a first step towards the study of the biochemical and molecular responses of pomegranate plants to water stress, this research provides new information into the mechanisms of drought tolerance in the plants.