Structural and antigenic preservation of plant samples by microwave-enhanced fixation, using dedicated hardware, minimizing heat-related effects

We explored the use of microwave technology in fixation with the objective of achieving quicker fixation regimes, lower concentrations of toxic and volatile reagents, and enhanced antigen detection. We used a modified domestic microwave oven (900 W) and a low-power (5 W) microwave bench. The work was done on plant materials. The oven was supplemented with a cooling device, a stirring system, and a record of the sample temperature and the time of effective irradiation. The sample, immersed in a fixative solution of 1% paraformaldehyde (PFA) in PBS, was irradiated for only 10 minutes. The sample temperature did not exceed 37 degrees C. In these mild conditions, the quality of the (ultra)structural preservation of the samples, morphometrically assessed, was at the same level as obtained with the same fixative, using conventional methods. On the contrary, samples fixed in the same conditions without irradiation showed a poor structural preservation. The antigenic preservation of the irradiated samples was excellent, since the labeling levels of two nucleolar proteins, detected by immunogold, were three times higher than in conventionally fixed samples. In the so-called microwave bench, the pathway of microwaves is guided, so that low-power microwaves directly hit the sample and there is no dispersion of energy. Temperature of fixative did not increase after microwave irradiation. Fixation in the bench with either 4% PFA, or 1% PFA, for 20 minutes resulted in structural preservation of samples similar in quality as obtained with conventional fixation and in a similar or better level of antigen preservation. Therefore, controlling temperature and effective irradiation is crucial in order to obtain optimal structural and antigen preservation with microwave-enhanced fixation. The dramatic differences observed between microwave-irradiated samples and samples fixed in the same conditions without irradiation, strongly support the existence of specific effects of microwaves on fixation, independent from the mere heating of the samples.     

Relationship between flavour descriptors and overall grading of analytical panels for virgin olive oil

An analytical panel is used to evaluate the quality of virgin olive oil. Two regression procedures—step-wise multiple regression and regression on principal components—have been used to construct equations that relate profile to overall grading of 57 samples of virgin olive oil collected from all over Spain. Every equation has an adjusted R² > 0.94. Forty new samples evaluated by another panel were used to verify the models. Regression on the principal components correctly classified 87.5% of samples, against 95% classified by step-wise multiple regression. Kurtosis, outliers, multicollinearity and singularity were also studied before building the regression models.     

Degradation of polycyclic aromatic hydrocarbons by the Chilean white-rot fungus Anthracophyllum discolor

The degradation of three- and four-ring polycyclic aromatic hydrocarbons (PAHs) in Kirk medium by Anthracophyllum discolor, a white-rot fungus isolated from the forest of southern Chile, was evaluated. In addition, the removal efficiency of three-, four- and five-ring PAHs in contaminated soil bioaugmented with A. discolor in the absence and presence of indigenous soil microorganisms was investigated. Production of lignin-degrading enzymes and PAH mineralization in the soil were also determined. A. discolor was able to degrade PAHs in Kirk medium with the highest removal occurring in a PAH mixture, suggesting synergistic effects between PAHs or possible cometabolism. A high removal capability for phenanthrene (62%), anthracene (73%), fluoranthene (54%), pyrene (60%) and benzo(a)pyrene (75%) was observed in autoclaved soil inoculated with A. discolor in the absence of indigenous microorganisms, associated with the production of manganese peroxidase (MnP). The metabolites found in the PAH degradation were anthraquinone, phthalic acid, 4-hydroxy-9-fluorenone, 9-fluorenone and 4,5-dihydropyrene. A. discolor was able to mineralize 9% of the phenanthrene. In non-autoclaved soil, the inoculation with A. discolor did not improve the removal efficiency of PAHs. Suitable conditions must be found to promote a successful fungal bioaugmentation in non-autoclaved soils.     

Density and level set-XFEM schemes for topology optimization of 3-D structures

As the capabilities of additive manufacturing techniques increase, topology optimization provides a promising approach to design geometrically sophisticated structures. Traditional topology optimization methods aim at finding conceptual designs, but they often do not resolve sufficiently the geometry and the structural response such that the optimized designs can be directly used for manufacturing. To overcome these limitations, this paper studies the viability of the extended finite element method (XFEM) in combination with the level-set method (LSM) for topology optimization of three dimensional structures. The LSM describes the geometry by defining the nodal level set values via explicit functions of the optimization variables. The structural response is predicted by a generalized version of the XFEM. The LSM–XFEM approach is compared against results from a traditional Solid Isotropic Material with Penalization method for two-phase “solid–void” and “solid–solid” problems. The numerical results demonstrate that the LSM–XFEM approach describes crisply the geometry and predicts the structural response with acceptable accuracy even on coarse meshes.     

Improvement of Mass Transfer by Freezing Pre-treatment and Ultrasound Application on the Convective Drying of Beetroot (<Emphasis Type="Italic">Beta vulgaris</Emphasis> L.)

The effects of freezing pre-treatment and ultrasound application during drying on microstructure, drying curves, and bioactive compounds of beetroot have been evaluated. Raw and previously frozen (at ? 20 °C) beetroots were convectively dried (40 °C and 1 m/s) with and without ultrasound application using two acoustic densities (16.4 and 26.7 kW/m³), and a diffusional model was proposed to simulate the drying curves. Freezing pre-treatment and ultrasound application caused significant disruptions in the beetroot microstructure and reduced the drying time, enhancing the mass transfer. The external mass transfer coefficient significantly (p < 0.05) increased by 28–49% when ultrasound was applied; moreover, the effective diffusion coefficient significantly (p < 0.05) increased by 60–73% and 204–211%, respectively, due to the ultrasound application on the drying of raw and pre-frozen samples. Freezing caused significant (p < 0.05) increases in betalain and total polyphenol contents and antioxidant activity compared with the raw sample (16–57%), probably due to the release of free forms from the food matrix; meanwhile, drying had the opposite effect (8–54% decrease). Significantly (p < 0.05) higher decreases (32–81%) in bioactive compounds and antioxidant activity were observed when drying was assisted by ultrasound compared with dying without ultrasound. Therefore, freezing pre-treatment and ultrasound application enhanced mass transfer during drying. However, significant changes in quality parameters of the final product were observed.     

The Effects of Residual Oxygen on the Storage Life of Retail-Ready Fresh Beef Steaks Masterpackaged Under a CO2 Atmosphere

ABSTRACT:  Retail-packed rump ( Gluteus medius , GM) and striploins ( Longissiumus dorsi, LD) steaks were masterpackaged under carbon dioxide (CO₂) and stored at 1 °C ± 1 °C for 14, 28, 35, and 42 d. A commercial oxygen (O₂) scavenger (ATCO HV1000®, Standa Industrie, Caen, France) was used in the masterpacks to achieve an O₂-free atmosphere. Similar packages without the O₂ scavengers were also prepared. At each storage time, 2 masterpacks of each treatment were opened and the retail trays were placed in a display case at 4 °C ± 2 °C for 1 and 48 h for microbiological and sensorial evaluations. The low growth rate of aerobic psychrotrophic flora on the stored beefsteaks demonstrated the bacteriostatic effect of CO₂ during storage. The maximum level of psychrotrophic aerobic bacteria reached during storage was approximately 10⁶ CFU/g. The steaks stored in masterpacks with scavengers bloomed to the desirable red color associated with freshly cut meat in the display case for all of the storage periods, except in the case of GM steaks, which showed a cycle of transient discoloration. GM and LD steaks were well accepted (65% and 82%, respectively) after 42 d under CO₂ at 1 °C ± 1 °C. The GM and LD steaks stored without the O₂ scavenger showed variable fractions of discoloration that significantly detracted from the appearance of the samples.     

Targeting and ultrasound imaging of microbubble-based contrast agents

Preparation and characterization of targeted microbubbles (ultrasound contrast agents) is described. Specific ligands were attached to the microbubble shell, and ligand-coated microbubbles were selectively attached to various targets, using either an avidin-biotin model system or an antigen-antibody system for targeting to live activated endothelial cells. Firm attachment of microbubbles to the target was achieved. Forces necessary to detach microbubbles from the target were estimated to exceed dozens of pN. Microbubbles were bound to the target even in the rapidly moving stream of the aqueous medium. Down to 20 ng of the ultrasound contrast material on the target surface could be detected by the ultrasound imaging with a commercial medical imaging system. At high bubble density on the target surface, strong ultrasound image attenuation was observed.     

GABAergic Regulation of Astroglial Gliotransmission through Cx43 Hemichannels

Gamma-Aminobutyric acid (GABA) is the primary inhibitory neurotransmitter in the brain. It is produced by interneurons and recycled by astrocytes. In neurons, GABA activates the influx of Cl^- via the GABA_A receptor or efflux or K⁺ via the GABA_B receptor, inducing hyperpolarization and synaptic inhibition. In astrocytes, the activation of both GABA_A and GABA_B receptors induces an increase in intracellular Ca²⁺ and the release of glutamate and ATP. Connexin 43 (Cx43) hemichannels are among the main Ca²⁺-dependent cellular mechanisms for the astroglial release of glutamate and ATP. However, no study has evaluated the effect of GABA on astroglial Cx43 hemichannel activity and Cx43 hemichannel-mediated gliotransmission. Here we assessed the effects of GABA on Cx43 hemichannel activity in DI NCT1 rat astrocytes and hippocampal brain slices. We found that GABA induces a Ca²⁺-dependent increase in Cx43 hemichannel activity in astrocytes mediated by the GABA_A receptor, as it was blunted by the GABA_A receptor antagonist bicuculline but unaffected by GABA_B receptor antagonist {"type":"entrez-protein","attrs":{"text":"CGP55845","term_id":"875097176","term_text":"CGP55845"}}CGP55845. Moreover, GABA induced the Cx43 hemichannel-dependent release of glutamate and ATP, which was also prevented by bicuculline, but unaffected by CGP. Gliotransmission in response to GABA was also unaffected by pannexin 1 channel blockade. These results are discussed in terms of the possible role of astroglial Cx43 hemichannel-mediated glutamate and ATP release in regulating the excitatory/inhibitory balance in the brain and their possible contribution to psychiatric disorders.     

Vitis vinifera L. Bioactive Components Modulate Adipose Tissue Metabolic Markers of Healthy Rats in a Photoperiod-Dependent Manner

Scope

The beneficial health effects of (poly)phenol-rich foods such as red grapes mainly depend on both the type and concentration of (poly)phenols. Since fruit (poly)phenol content is influenced by growing conditions, the study examines the seasonal effects of red grapes (Vitis vinifera L.), grown under various cultivation conditions, on metabolic markers of adipose tissue in healthy rats.

Methods and results

For this purpose, Fischer 344 rats are exposed into three different light-dark cycles and daily supplemented with 100 mg kg⁻¹ of either conventionally or organically grown red grapes for 10 weeks (n = 6). Seasonal consumption of organic grapes (OGs), which are richer in anthocyanins, increases energy expenditure (EE) of animals exposed to long photoperiod and enhances uncoupling protein 1 (UCP1) protein expression in brown adipose tissue of animals under standard photoperiod. Additionally, red grape consumption affects the gene expression profile of white adipose tissue (WAT), upregulating browning markers of subcutaneous WAT in 12 h light (L12) and 18 h light (L18) photoperiods, and downregulating adipogenic and lipolytic markers of visceral WAT in 6 h light (L6) and L12 photoperiods.

Conclusions

These results clearly show that bioactive compounds of grapes can modulate the metabolic markers of white and brown adipose tissues in a photoperiod and depot-dependent manner, partly affecting EE when consumed out of season.     

Dry surface biofilms in the food processing industry: An overview on surface characteristics,adhesion and biofilm formation,detection of biofilms,and dry sanitization methods

Bacterial biofilm formation in low moisture food processing (LMF) plants is related to matters of food safety, production efficiency, economic loss, and reduced consumer trust. Dry surfaces may appear dry to the naked eye, however, it is common to find a coverage of thin liquid films and microdroplets, known as microscopic surface wetness (MSW). The MSW may favor dry surface biofilm (DSB) formation. DSB formation is similar in other industries, it occurs through the processes of adhesion, production of extracellular polymeric substances, development of microcolonies and maturation, it is mediated by a quorum sensing (QS) system and is followed by dispersal, leading to disaggregation. Species that survive on dry surfaces develop tolerance to different stresses. DSB are recalcitrant and contribute to higher resistance to sanitation, becoming potential sources of contamination, related to the spoilage of processed products and foodborne disease outbreaks. In LMF industries, sanitization is performed using physical methods without the presence of water. Although alternative dry sanitizing methods can be efficiently used, additional studies are still required to develop and assess the effect of emerging technologies, and to propose possible combinations with traditional methods to enhance their effects on the sanitization process. Overall, more information about the different technologies can help to find the most appropriate method/s, contributing to the development of new sanitization protocols. Thus, this review aimed to identify the main characteristics and challenges of biofilm management in low moisture food industries, and summarizes the mechanisms of action of different dry sanitizing methods (alcohol, hot air, UV-C light, pulsed light, gaseous ozone, and cold plasma) and their effects on microbial metabolism.