Deferoxamine preconditioning to restore impaired HIF-1α-mediated angiogenic mechanisms in adipose-derived stem cells from STZ-induced type 1 diabetic rats

Objectives: Both excessive and insufficient angiogenesis are associated with progression of diabetic complications, of which poor angiogenesis is an important feature. Currently, adipose-derived stem cells (ADSCs) are considered to be a promising source to aid therapeutic neovascularization. However, functionality of these cells is impaired by diabetes which can result from a defect in hypoxia-inducible factor-1 (HIF-1), a key mediator involved in neovascularization. In the current study, we sought to explore effectiveness of pharmacological priming with deferoxamine (DFO) as a hypoxia mimetic agent, to restore the compromised angiogenic pathway, with the aid of ADSCs derived from streptozotocin (STZ)-induced type 1 diabetic rats ('diabetic ADSCs'). Materials and methods: Diabetic ADSCs were treated with DFO and compared to normal and non-treated diabetic ADSCs for expression of HIF-1α, VEGF, FGF-2 and SDF-1, at mRNA and protein levels, using qRT-PCR, western blotting and ELISA assay. Activity of matrix metalloproteinases -2 and -9 were measured using a gelatin zymography assay. Angiogenic potential of conditioned media derived from normal, DFO-treated and non-treated diabetic ADSCs were determined by in vitro (in HUVECs) and in vivo experiments including scratch assay, three-dimensional tube formation testing and surgical wound healing models. Results: DFO remarkably enhanced expression of noted genes by mRNA and protein levels and restored activity of matrix metalloproteinases -2 and -9. Compromised angiogenic potential of conditioned medium derived from diabetic ADSCs was restored by DFO both in vitro and in vivo experiments. Conclusion: DFO preconditioning restored neovascularization potential of ADSCs derived from diabetic rats by affecting the HIF-1α pathway. © 2015 John Wiley & Sons Ltd

The Effect of some Factors on the Efficacy of Pheromone Traps for Capturing Leopard Moth in Walnut Orchards

Introduction: The leopard moth, Zeuzera pyrina (L.) (Lepidoptera, Cossidae), is an important xylophagous pest on different trees such as walnut, Juglans regia (L.), which weaken the host trees by canalling in the trunks. Regarding type of damage and host range of the pest, it is so difficult to control. Catching of the male moths by traps containing sex pheromone lures is a promising method to control the pest. There are many factors influencing potential of pheromone traps. Some of these factors, such as trap shape, trap color and height of trap placement, can be manipulated by the user or grower. One of the components of the pheromone blend of leopard moth is (E, Z)-2, 13-octadecadien-1-ol acetate, which seems to be the pheromone compound of the current clearwing moth Synanthedon tipuliformis (Clerck) (Sesiidae). Materials and Methods: Separate experiments were conducted in walnut orchards (each having more than 10 years of damage history and area of 0.5-3.5 ha) of Najaf-Abad County (Isfahan, Iran). Pheromone traps were placed on the trees in late April. The traps were inspected at 5-day intervals until late July. The number of catched lthe moth males was recorded and also, during the growth season average of the capturing per trap per night was calculated. Lures were replaced monthly. In the present study the effects of four factors (pheromone type, trap shape, trap color and height of trap placement) on the number of catched leopard moth males were evaluated. The experiments for different factors were conducted as follows: 1. Pheromone type and lures of pheromones of S. tipuliformis and Z. pyrina, both of which were provided from Agriscense Company, were compared. 2. Trap shape, four shapes (cylindrical or standard, delta, wing and funnel) were compared. The experiment was conducted using white-colored traps placed at the height of 1.5m above the ground and had S. tipuliformis lures. 3. Trap color, three separate experiments were conducted as follows: delta traps with 3 colors (white, yellow and green), delta traps with 6 colors (white, yellow, green, glossy gray, normal gray and red), cylindrical traps with 4 colors (green, yellow, with and normal gray). In the experiments, traps with height of 2m above the ground had S. tipuliformis lures. 4. Height of trap placement, two separate experiments with delta and cylindrical traps were conducted for comparison of three placement heights of 1.5, 3 and more than 6 meters above the ground. The experiment was conducted using white - colored traps which had S. tipuliformis lures. All above experiments were conducted in a randomized complete block design with three or four replications. Analysis of variance was performed after data transformation, using log (x+1). In all experiments means were compared using Tukey̕s test, except for the experiment of pheromone type, which compared using t-student test. Results and Discussion: In the present study the catching was started on 4th of May and lasted till 26 July. During 105 nights of trapping, traps, which were continuing lures of S. tipuliformis and Z. pyrina, were captured in total 77 and 7 leopard moth males, respectively. Means of capturing per trap per night were 0.18 and 0.01, respectively. Results showed that catching of male moths were significantly higher on traps containing lures of S. tipuliformis than those containing lures of Z. pyrina. In comparison to shape of traps, significantly the highest catches belonged to funnel trap. The superiority of the funnel traps could be related to its specific structure. Comparison of 6 colors with delta traps showed that green and yellow colors had the highest and the lowest catching, respectively. However, in comparison of 4 colors with cylindrical trap, the highest and the lowest catching belonged to normal gray and yellow, respectively. In the present study, ranking of trap colors changed by trap shape. The phenomenon might be related to the interactions between the factors of traps̕ shape and color. In the experiments of height of trap placement, the highest catching was obtained at the height more than 6 m with both delta and wing traps. It shows that the amount of catching is increasing by increasing in the height of trap placement. Conclusion: Pheromone traps could be used as a useful method for catching leopard moths males. However efficacy of the pheromone traps was influenced by several factors, including type of pheromone, trap shape, trap color and height of trap placement

Spouted bed drying of skimmed milk: Multivariable optimization of the conditions to improve physicochemical properties of the dried milk

In the present research, the influence of the inlet parameters of a laboratory spouted bed (inlet air temperature and milk flow rate) on the physical properties (moisture content, wettability, dispersibility, particle size and density), chemical properties (protein, cholesterol, and triglyceride content) and morphology of the milk powder was evaluated and compared with properties of the commercial spray dried powder. The effect of these two inlet parameters was significant on all of the physical and chemical properties. Particle morphology of the spouted bed dried samples (flat, solid, and irregular) was entirely different from commercial spray dried milk powder (spherical, and hollow). The optimum values of the inlet air temperature and milk flow rate, calculated by RSM, were obtained 80 °C and 2.8 ml/min, respectively. The small error between properties predicted by the models and the experimental data indicated that the performed optimization was satisfactory. © 2021 Elsevier Lt

Comprehensive evaluation of gene expression in negative and positive trigger-based targeting niosomes in HEK-293 cell line

An efficient gene delivery system has some critical factors that enhance the efficiency of nanocarrier. These factors are low production cost, high bioavailability, high encapsulation efficiency, controllable release, and targeting ability. Niosome (the nonionic surfactant vesicles) was considered as a promising gene delivery system. Niosome can increase stability and uptake of active agents. We used all mentioned factors in one optimized formulation entitled plasmidloaded magnetic niosomes (PMN). To increase the bioavailability of niosomes, we used ergosterol (a natural lipid) instead of cholesterol in structure of niosome. Also, cetyl trimethyl ammonium bromide (CTAB) in different concentrations was used to improve encapsulation of plasmid and compared to niosomes that did not have CTAB (negative niosome). Afterward, magnetic nanoparticle (Fe3O4@SiO2) was synthesized and loaded into niosome to obtain targeting ability. Prepared formulations were evaluated regarding size, zeta potential, morphology, encapsulation of magnetic nanoparticles and plasmid (Pm-cherry-N1), release rate, and transfection efficiency. Results demonstrated that optimum formulation (Nio/CTAB3/Fe/P) has a nanometric size (118 ± 2.31 nm, positive zeta potential (+25 ± 0.67 mV), high loading of plasmid (72), and good gene expression (35). Interestingly, after applying a magnetic field below the cell plate, we obtained ac increased gene expression from 35 to 42. These results showed that this new formulation would have a promising future and also can be used for delivering the other drugs and active agents. © 2020, Iranian Journal of Pharmaceutical Research. All rights reserved

Niosomal virosome derived by vesicular stomatitis virus glycoprotein as a new gene carrier

Virosomes as membranous vesicles with viral fusion protein in their membrane are versatile vehicles for cargo delivery. The vesicular stomatitis virus glycoprotein (VSV-G) is a common fusogenic protein used in virosome preparation. This glycoprotein has been used in liposomal systems so far, but in this study, we have tried to use the niosomal form instead of liposome for. Niosomes are vesicular systems composed of non-ionic surfactants. Niosomes were constructed by the thin-film hydration method. VSV-G gene in pMD2.G plasmid was expressed in the HEK293T cell line and then was reconstituted in the niosome bilayer. The formation of niosomal virosomes was confirmed with different methods such as SDS-PAGE gel, western blotting, and transmission electron microscopy (TEM). The efficiency of niosomal virosome was investigated with the pmCherry reporter gene. SDS-PAGE and western blotting proved the expression and successful insertion of protein into the bilayer. The TEM images showed the spike projection of VSV-G on the surface of niosomes. The transfection results showed high efficiency of niosomal virosomes as a novel carrier. This report has verified that niosome could be used as an efficient bilayer instead of liposome to construct virosomes. © 2020 The Italian Diabetes Society, the Italian Society for the Study of Atherosclerosis, the Italian Society of Human Nutrition and the Department of Clinical Medicine and Surgery, Federico II Universit

Evaluation the effect of Myrtus communis L. extract on several underlying mechanisms involved in wound healing: An in vitro study

Objective: Myrtus communis L. (Myrtle) has been widely used in traditional remedies for curing several medical conditions including wounds. However, studies addressing its detailed molecular mechanisms especially those involved in the angiogenic process are still limited. Hence, the current study was aimed to investigate the effect of ethanol extract of myrtle leaves ME on some molecular mechanisms involved in the wound healing process. Methods: Firstly, the effect of ME on the viability of HUVECs, 3T3 fibroblast cell line and J774A.1 macrophage was evaluated using MTT assay. Moreover, the effect of ME on the migration of HUVECs was evaluated by the scratch assay. The protein expression of HIF-1α and VEGF in HUVECs was analyzed using western blotting. To evaluate the anti-inflammatory effect of ME on lipopolysaccharide (LPS)-stimulated J774A.1 macrophage, the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) at mRNA level was assessed using qRT-PCR. Furthermore, in vitro antimicrobial and antioxidant activities of ME were accomplished. Results: At concentration lower than 6 μg/mL, ME showed no significant toxic effect on all three cell lines. ME significantly reduced the expression of iNOS and COX-2 at mRNA level in LPS-stimulated J774A.1 macrophage as well as increased expression of HIF-1α and VEGF at the protein level in HUVECs. The migratory potential of HUVECs was enhanced by ME. The ME showed potent antioxidant activity at concentrations � 15 μg/mL. Staphylococcus aureus and Bacillus subtilis were more sensitive to ME than other spices. Conclusion: The present study may provide more detailed information about the underlying mechanisms involved in wound healing effect of ME. © 2018 South African Association of Botanist