Adenovirus-mediated siRNA targeting TNF-α and overexpression of bone morphogenetic protein-2 promotes early osteoblast differentiation on a cell model of Ti particle-induced inflammatory response in vitro

Wear particles are phagocytosed by macrophages and other inflammatory cells, resulting in cellular activation and release of proinflammatory factors, which cause periprosthetic osteolysis and subsequent aseptic loosening, the most common causes of total joint arthroplasty failure. During this pathological process, tumor necrosis factor-alpha (TNF-α) plays an important role in wear-particle-induced osteolysis. In this study, recombination adenovirus (Ad) vectors carrying both target genes [TNF-α small interfering RNA (TNF-α-siRNA) and bone morphogenetic protein 2 (BMP-2)] were synthesized and transfected into RAW264.7 macrophages and pro-osteoblastic MC3T3-E1 cells, respectively. The target gene BMP-2, expressed on pro-osteoblastic MC3T3-E1 cells and silenced by the TNF-α gene on cells, was treated with titanium (Ti) particles that were assessed by real-time PCR and Western blot. We showed that recombinant adenovirus (Ad-siTNFα-BMP-2) can induce osteoblast differentiation when treated with conditioned medium (CM) containing RAW264.7 macrophages challenged with a combination of Ti particles and Ad-siTNFα-BMP-2 (Ti-ad CM) assessed by alkaline phosphatase activity. The receptor activator of nuclear factor-κB ligand was downregulated in pro-osteoblastic MC3T3-E1 cells treated with Ti-ad CM in comparison with conditioned medium of RAW264.7 macrophages challenged with Ti particles (Ti CM). We suggest that Ad-siTNFα-BMP-2 induced osteoblast differentiation and inhibited osteoclastogenesis on a cell model of a Ti particle-induced inflammatory response, which may provide a novel approach for the treatment of periprosthetic osteolysis.

Lipoic acid, but not tempol, preserves vascular compliance and decreases medial calcification in a model of elastocalcinosis

Vascular calcification decreases compliance and increases morbidity. Mechanisms of this process are unclear. The role of oxidative stress and effects of antioxidants have been poorly explored. We investigated effects of the antioxidants lipoic acid (LA) and tempol in a model of atherosclerosis associated with elastocalcinosis. Male New Zealand white rabbits (2.5-3.0 kg) were fed regular chow (controls) or a 0.5% cholesterol (chol) diet+104 IU/day vitamin D2 (vitD) for 12 weeks, and assigned to treatment with water (vehicle, n=20), 0.12 mmol·kg-1·day-1 LA (n=11) or 0.1 mmol·kg-1·day-1 tempol (n=15). Chol+vitD-fed rabbits developed atherosclerotic plaques associated with expansive remodeling, elastic fiber disruption, medial calcification, and increased aortic stiffness. Histologically, LA prevented medial calcification by ∼60% and aortic stiffening by ∼60%. LA also preserved responsiveness to constrictor agents, while intima-media thickening was increased. In contrast to LA, tempol was associated with increased plaque collagen content, medial calcification and aortic stiffness, and produced differential changes in vasoactive responses in the chol+vitD group. Both LA and tempol prevented superoxide signals with chol+vitD. However, only LA prevented hydrogen peroxide-related signals with chol+vitD, while tempol enhanced them. These data suggest that LA, opposite to tempol, can minimize calcification and compliance loss in elastocalcionosis by inhibition of hydrogen peroxide generation.

CTLA4 enhances the osteogenic differentiation of allogeneic human mesenchymal stem cells in a model of immune activation

Allogeneic mesenchymal stem cells (allo-MSCs) have recently garnered increasing interest for their broad clinical therapy applications. Despite this, many studies have shown that allo-MSCs are associated with a high rate of graft rejection unless immunosuppressive therapy is administered to control allo-immune responses. Cytotoxic T-lymphocyte-associated protein 4 (CTLA4) is a co-inhibitory molecule expressed on T cells that mediates the inhibition of T-cell function. Here, we investigated the osteogenic differentiation potency of allo-MSCs in an activated immune system that mimics the in vivo allo-MSC grafting microenvironment and explored the immunomodulatory role of the helper T cell receptorCTLA4 in this process. We found that MSC osteogenic differentiation was inhibited in the presence of the activated immune response and that overexpression of CTLA4 in allo-MSCs suppressed the immune response and promoted osteogenic differentiation. Our results support the application of CTLA4-overexpressing allo-MSCs in bone tissue engineering.

Application of the Hsu model to soybean grain hydration

A comparative analysis of the theoretical-experimental study, developed by Hsu on the hydration of Amsoy 71 soybean grain, was performed through several soaking experiments using CD 202 soybean at 10, 20, 30, 40, and 50 °C, measuring moisture content over time. The results showed that CD 202 soybean equilibrium moisture content, Xeq, does not depend on temperature and is 21% higher than that found by Hsu, suggesting that soybean cultivar exerts great influence on Xeq. The Hsu model was numerically solved and its parameters were adjusted by the least squares method, with maximum deviations of +/- 10% relative to the experimental values. The limiting step in the mass transfer process during hydration corresponds to water diffusion inside the grain, leading to radial moisture gradients that decrease over time and with an increase in temperature. Regardless of the soybean cultivar, diffusivity increases as temperature or moisture content increases. However, the values of this transport property for Amsoy 71 were superior to those of CD 202, very close at the beginning of hydration at 20 °C and almost three times higher at the end of hydration at 50 °C.

Drying of sicilian lemon residue: influence of process variables on the evaluation of the dietary fiber produced

In this study, the effect of the process variables of the air-drying of Sicilian lemon residues on some technological properties of the fibers produced was studied. The determination and modeling of desorption isotherms were used to establish the equilibrium moisture content at 60, 75, and 90 °C using the static method with 8 saturated salt solutions. The best fit was obtained with BET and GAB models. The drying process was conducted in a vertical tray dryer and delineated according to a central composite experimental design (2²) using the following as factors: air velocity (0.5, 0.75 and 1 m/s) and temperature (60, 75, and 90 °C), and it presented a good fit to the exponential model (R² > 99.9%). The experimental design responses evaluated were the technological properties of the fibers: water-holding, oil-holding, and swelling capacity. Since these properties were present in high levels, the lemon residues could be used to increase content of fibers in foods resulting in the addition of nutritional benefits for the consumers.

Influence of thermal treatment of wood on the aroma of a sugar cane spirit (cachaça) model-solution

The aging process of alcoholic beverages is generally conducted in wood barrels made with species from Quercus sp. Due to the high cost and the lack of viability of commercial production of these trees in Brazil, there is demand for new alternatives to using other native species and the incorporation of new technologies that enable greater competitiveness of sugar cane spirit aged in Brazilian wood. The drying of wood, the thermal treatment applied to it, and manufacturing techniques are important tools in defining the sensory quality of alcoholic beverages after being placed in contact with the barrels. In the thermal treatment, several compounds are changed by the application of heat to the wood and various studies show the compounds are modified, different aromas are developed, there is change in color, and beverages achieve even more pleasant taste, when compared to non-treated woods. This study evaluated the existence of significant differences between hydro-alcoholic solutions of sugar cane spirits elaborated from different species of thermo-treated and non-treated wood in terms of aroma. An acceptance test was applied to evaluate the solutions preferred by tasters under specific test conditions.

Effect of air-temperature and diet composition on the drying process of pellets for japanese abalone (Haliotis discus hannai) feeding

The aim of this research was to study the effect of air-temperature and diet composition on the mass transfer kinetics during the drying process of pellets used for Japanese Abalone (Haliotis discus hannai) feeding. In the experimental design, three temperatures were used for convective drying, as well as three different diet compositions (Diets A, B and C), in which the amount of fishmeal, spirulin, algae, fish oil and cornstarch varied. The water diffusion coefficient of the pellets was determined using the equation of Fick's second law, which resulted in values between 0.84-1.94×10-10 m²/s. The drying kinetics was modeled using Page, Modified Page, Root of time, Exponential, Logarithmic, Two-Terms, Modified Henderson-Pabis and Weibull models. In addition, two new models, referred to as 'Proposed' models 1 and 2, were used to simulate this process. According to the statistical tests applied, the models that best fitted the experimental data were Modified Henderson-Pabis, Weibull and Proposed model 2, respectively. Bifactorial analysis of variance ANOVA showed that Diet A (fishmeal 44%, spirulin 9%, fish oil 1% and cornstarch 36%) presented the highest diffusion coefficient values, which were favored by the temperature increase in the drying process.

Effects of the mechanical damage on the water absorption process by corn kernel

The purpose of this study was to investigate and model the water absorption process by corn kernels with different levels of mechanical damage Corn kernels of AG 1510 variety with moisture content of 14.2 (% d.b.) were used. Different mechanical damage levels were indirectly evaluated by electrical conductivity measurements. The absorption process was based on the industrial corn wet milling process, in which the product was soaked with a 0.2% sulfur dioxide (SO2) solution and 0.55% lactic acid (C3H6O3) in distilled water, under controlled temperatures of 40, 50, 60, and 70 ºC and different mechanical damage levels. The Peleg model was used for the analysis and modeling of water absorption process. The conclusion is that the structural changes caused by the mechanical damage to the corn kernels influenced the initial rates of water absorption, which were higher for the most damaged kernels, and they also changed the equilibrium moisture contents of the kernels. The Peleg model was well adjusted to the experimental data presenting satisfactory values for the analyzed statistic parameters for all temperatures regardless of the damage level of the corn kernels.

An artificial neural network model for prediction of quality characteristics of apples during convective dehydration

In this study, the effects of hot-air drying conditions on color, water holding capacity, and total phenolic content of dried apple were investigated using artificial neural network as an intelligent modeling system. After that, a genetic algorithm was used to optimize the drying conditions. Apples were dried at different temperatures (40, 60, and 80 °C) and at three air flow-rates (0.5, 1, and 1.5 m/s). Applying the leave-one-out cross validation methodology, simulated and experimental data were in good agreement presenting an error < 2.4 %. Quality index optimal values were found at 62.9 °C and 1.0 m/s using genetic algorithm.