Polyfunctional sugar-free white chocolate fortified with Lacticaseibacillus rhamnosus GG co-encapsulated with beet residue extract (Beta vulgaris L.).

Chocolate is a worldwide consumed food. This study investigated the fortification of sugar-free white chocolate with Lacticaseibacillus rhamnosus GG microcapsule co-encapsulated with beet residue extract. The chocolates were evaluated for moisture, water activity, texture, color properties, melting, physicochemical, and probiotic stability during storage. Furthermore, the survival of L. rhamnosus GG and the bioaccessibility of phenolic compounds were investigated under in vitro simulated gastrointestinal conditions. Regarding the characterization of probiotic microcapsules, the encapsulation efficiency of L. rhamnosus GG was > 89 % while the encapsulation efficiency of phenolic compounds was > 62 %. Chocolates containing probiotic microcapsules were less hard and resistant to breakage. All chocolates had a similar melting behavior (endothermic peaks between 32.80 and 34.40 °C). After 120 days of storage at 4 °C, probiotic populations > 6.77 log CFU/g were detected in chocolate samples. This result demonstrates the potential of this matrix to carry L. rhamnosus GG cells. Regarding the resistance of probiotic strains during gastric simulation, the co-encapsulation of L. rhamnosus GG with beet extract contributed to high counts during gastrointestinal transit, reaching the colon (48 h) with viable cell counts equal to 11.80 log CFU/g. Finally, one of our main findings was that probiotics used phenolic compounds as a substrate source, which may be an observed prebiotic effect.

High-throughput analysis reveals disturbances throughout the cell caused by Arabidopsis UCP1 and UCP3 double knockdown.

Three genes encoding mitochondrial uncoupling proteins (UCPs) have been described in Arabidopsis thaliana (UCP1 to UCP3). In plants, UCPs may act as an uncoupler or as an aspartate/glutamate exchanger. For instance, much of the data regarding UCP functionality were obtained for the UCP1 and UCP2 isoforms compared with UCP3. Here, to get a better understanding about the concerted action of UCP1 and UCP3 in planta, we investigated the transcriptome and metabolome profiles of ucp1 ucp3 double mutant plants during the vegetative phase. For that, 21-day-old mutant plants, which displayed the most evident phenotypic alterations compared to wild type (WT) plants, were employed. The double knockdown of UCP1 and UCP3, isoforms unequivocally present inside the mitochondria, promoted important transcriptional reprogramming with alterations in the expression of genes related to mitochondrial and chloroplast function as well as those responsive to abiotic stress, suggesting disturbances throughout the cell. The observed transcriptional changes were well integrated with the metabolomic data of ucp1 ucp3 plants. Alterations in metabolites related to primary and secondary metabolism, particularly enriched in the Alanine, Aspartate and Glutamate metabolism, were detected. These findings extend our knowledge of the underlying roles played by UCP3 in concert with UCP1 at the whole plant level.

Application of nanoparticles entrapped orange essential oil to inhibit the incidence of phytopathogenic fungi during storage of agroecological maize seeds.

The demand for sustainable, healthy, and pesticide-free food has grown in recent years. Agroecological seeds cannot receive chemical treatment, as pesticides present toxicological and environmental risks, requiring the development of alternative methods for disease control, such as the use of essential oils. In this study, orange essential oil was extracted and encapsulated in zein nanoparticles by the nanoprecipitation method. The nanoparticles were tested for the antifungal activity on agroecological maize seeds and for the mycelial sensitivity of Stenocarpella macrospora. The synthesized nanoparticles presented good encapsulation efficiency (99 %) of orange essential oil rich in D-limonene, conferring high antioxidant activity to the loaded nanoparticles. The release profile indicated a pseudo-Fickian mechanism governed by diffusion, explained according to the Korsmeyer-Peppas model. The dynamic light scattering, and transmission electron microscopy showed spherical nanoparticles with particle size lower than 200 nm. The nanoparticles containing orange essential oil inhibited the incidence of Fusarium during the storage of agroecological maize seeds. The mycelial sensitivity against Stenocarpella macrospora showed that the encapsulated essential oil was more effective in inhibiting the fungus when compared to the non-encapsulated oil. Therefore, the nanoparticles containing encapsulated orange essential oil can be effectively applied as an antifungal material for the conservation of agroecological maize seeds, contributing to the development of sustainable agricultural biotechnology with pesticide-free products.

Antioxidant effect of blueberry flour on the digestibility and storage of Bologna-type mortadella.

The aim of the study was to add blueberry flour (BF) to Bologna-type mortadella as a natural antioxidant and to evaluate its activity during in vitro digestion and refrigerated storage. Five treatments of mortadella were prepared: without antioxidant, with sodium erythorbate and with the addition of three levels of BF: 0.05 %, 0.075 % and 0.1 %. Twenty-three phenolic compounds were quantified in blueberry fruits and twenty-eight in BF, with prevalence of chlorogenic acid. The presence of BF did not affect the proximal composition of the mortadella, but it had a small effect on pH, hardness (texture profile) and instrumental color, as well as reduced lipid oxidation during refrigerated storage (2-8 °C) for 90 days. During in vitro digestion, the addition of BF increased the content of total phenolic compounds and the antioxidant activity of mortadella (p < 0.05), among all simulated stages. At a concentration of 0.05 %, BF can be used as a synthetic antioxidant substitute in Bologna-type mortadella, enhancing the use of blueberry fruits in the form of flour and enriching the product with natural antioxidants.

Microcalorimetric growth behavior of E. coli ATCC 25922 in an MCDSC.

Isothermal microcalorimetry can provide a general analytical tool for the characterization of bacterial growth. Methodologies and equipment have been studied to expand the application and disseminate the use of the technique. The MCDSC is a microcalorimeter capable of measuring in the range of 0.2 µW that can operate at a temperature range of -20 to 140 °C or under isothermal conditions. Here, we present the first investigation of MCDSC for E. coli growth with the Baranyi and Roberts modeling application. This study presented the calorimetric E. coli fingerprint at MCDSC and compares it with the plate count technique, giving the data more biological meaning. The calorimeter was able to accurately detect growth metabolism and discriminate E. coli at different inoculum densities. Additionally, the MCDSC can offer a new point of view for evaluating microbial growth, such as the significant reduction in error due to dispersed data by the viable counting method.

Goldenberry flour as a natural antioxidant in Bologna-type mortadella during refrigerated storage and in vitro digestion.

Goldenberry is rich in phenolic compounds; however, no study has been carried out to assess its potential in meat products. The objective of the research was to characterize goldenberry (Physalis peruviana) fruits and flour and investigate its effect on the quality of Bologna-type mortadella, before and after in vitro digestion. Five treatments were performed: without antioxidant, with sodium erythorbate, and with three levels of goldenberry flour. The phenolic profile analysed by LC-MS/MS quantified 10 compounds in the fruit and 23 in the flour. Goldenberry flour did not affect the centesimal composition of the mortadella, but it had an effect on pH, hardness (texture profile) and color, as well as reduced lipid oxidation (peroxide value and TBARS) for 90 days. There was also an increase (P < 0.05) in the antioxidant activity and in the content of phenolic compounds after in vitro digestion. Thus, goldenberry flour can be used as a natural antioxidant to replace sodium erythorbate in Bologna-type mortadella.

Metabolism and Signaling of Plant Mitochondria in Adaptation to Environmental Stresses.

The interaction of mitochondria with cellular components evolved differently in plants and mammals; in plants, the organelle contains proteins such as ALTERNATIVE OXIDASES (AOXs), which, in conjunction with internal and external ALTERNATIVE NAD(P)H DEHYDROGENASES, allow canonical oxidative phosphorylation (OXPHOS) to be bypassed. Plant mitochondria also contain UNCOUPLING PROTEINS (UCPs) that bypass OXPHOS. Recent work revealed that OXPHOS bypass performed by AOXs and UCPs is linked with new mechanisms of mitochondrial retrograde signaling. AOX is functionally associated with the NO APICAL MERISTEM transcription factors, which mediate mitochondrial retrograde signaling, while UCP1 can regulate the plant oxygen-sensing mechanism via the PRT6 N-Degron. Here, we discuss the crosstalk or the independent action of AOXs and UCPs on mitochondrial retrograde signaling associated with abiotic stress responses. We also discuss how mitochondrial function and retrograde signaling mechanisms affect chloroplast function. Additionally, we discuss how mitochondrial inner membrane transporters can mediate mitochondrial communication with other organelles. Lastly, we review how mitochondrial metabolism can be used to improve crop resilience to environmental stresses. In this respect, we particularly focus on the contribution of Brazilian research groups to advances in the topic of mitochondrial metabolism and signaling.

Production of methylcellulose films functionalized with poly-ε-caprolactone nanocapsules entrapped ß-carotene for food packaging application.

Nowadays, there is a worldwide demand in the production of innovative packaging that release active compounds to increase the shelf life of perishable food products. Therefore, this study produced methylcellulose films functionalized with poly-ε-caprolactone nanocapsules entrapped ß-carotene. The nanoparticles were produced by the nanoprecipitation method, and 10, 30, and 50 % of nanoparticles colloidal solution was added in the methylcellulose filmogenic solution. The films were characterized by the mechanical, physicochemical properties, antioxidant activity, and release of ß-carotene from the polymeric matrix to a food simulant. The results demonstrated satisfactory mechanical properties; however, the addition of nanoparticles decreased the Young's Modulus and increased the elongation at break. Regarding light transmission, the incorporation of ß-carotene nanoparticles promoted a decrease in the percentage of ultraviolet ray's transmittance through the film matrix, as well as visible light. The incorporation of nanoparticles improved the antioxidant activity of the films, which was proportional to the concentration of ß-carotene used in the formulation. The release of ß-carotene reached a maximum value of 10.93 µg g-1 film containing 70 % nanoparticles, which was a desired profile for food application. Finally, the methylcellulose films functionalized with poly-ε-caprolactone nanocapsules can release ß-carotene, and therefore, can be considered as a novel nanomaterial for food conservation, with a potential to increase the shelf life of perishable food products.

The double knockdown of the mitochondrial uncoupling protein isoforms reveals partial redundant roles during Arabidopsis thaliana vegetative and reproductive development.

Mitochondrial uncoupling proteins (UCPs) are specialized proteins capable of dissipating the proton electrochemical gradient generated in respiration independent of ATP synthesis. Three UCP coding genes with distinct expression patterns have been identified in Arabidopsis thaliana (namely UCP1, UCP2 and UCP3). Here, we generated T-DNA double-insertion mutants (ucp1 ucp2, ucp1 ucp3 and ucp2 ucp3) to investigate the functionality of the Arabidopsis UCP isoforms. A strong compensatory effect of the wild-type UCP gene was found in the double-knockdown lines. Higher levels of reactive oxygen species (ROS) were observed in vegetative and reproductive organs of double mutant plants. This exacerbated oxidative stress in plants also increased lipid peroxidation but was not compensated by the activation of the antioxidant system. Alterations in O2 consumption and ADP/ATP ratio were also observed, suggesting a change in mitochondrial energy-generating processes. Deficiencies in double-mutants were not limited to mitochondria and also changed photosynthetic efficiency and redox state. Our results indicate that UCP2 and UCP3 have complementary function with UCP1 in plant reproductive and vegetative organ/tissues, as well as in stress adaptation. The partial redundancy between the UCP isoforms suggests that they could act separately or jointly on mitochondrial homeostasis during A. thaliana development.

Disasters with oil spills in the oceans: Impacts on food safety and analytical control methods.

More than 5.8 million tonnes of oil have been spilled into the oceans. Some oil disasters marked history, causing multiple social and economic consequences in addition to catastrophic environmental impacts. Recently, Brazil and Mauritius faced oil disasters that have severely impacted seafood sanitary credibility. One of the components of the oil composition are the polycyclic aromatic hydrocarbons (PAH), which are the main contamination markers of petrogenic origin. There is enough evidence to correlate the intake of food contaminated with PAH with increased risks of developing cancer. The set PAH4, composed of benzo[a]anthracene, benzo[a]pyrene, benzo[b]fluoranthene, and chrysene, and the set PAH8, composed of benzo[a]anthracene, benzo[a]pyrene, benzo[b]fluoranthene, benzo[k]fluoranthene, benzo[ghi]perylene, chrysene, dibenzo[a,h]anthracene, and indeno[1,2,3-cd]pyrene are recognized as markers of food chemical contamination. After oil disasters in the oceans, the risk to the health of seafood consumers tends to be of special concern, Countries like the European Union set maximum levels for benzo[a]pyrene (5 µg kg-1) and PAH4 (30 µg kg-1) in bivalve mollusks. Levels of concern established by countries that have faced oil disasters are given special attention in this review. Laboratory analysis of PAH in food samples is very challenging because it deals with quite different kinds of matrices. Furthermore, analytical results are usually related to the closure or reopening of cultivated areas and fishing points. Therefore, the progress of the analytical methods for PAH in seafood is covered in detail. Chemical laboratory measurements provide essential data to assess the potential risks to human health due to consumption of seafood contaminated with PAH. The main human health risk assessment approaches in a seafood contamination scenario with PAH are reviewed and discussed, providing an insightful and guiding tool to each step of the risk assessment framework.

Mitochondrial retrograde signaling through UCP1-mediated inhibition of the plant oxygen-sensing pathway.

Mitochondrial retrograde signaling is an important component of intracellular stress signaling in eukaryotes. UNCOUPLING PROTEIN (UCP)1 is an abundant plant inner-mitochondrial membrane protein with multiple functions including uncoupled respiration and amino-acid transport1,2 that influences broad abiotic stress responses. Although the mechanism(s) through which this retrograde function acts is unknown, overexpression of UCP1 activates expression of hypoxia (low oxygen)-associated nuclear genes.3,4 Here we show in Arabidopsis thaliana that UCP1 influences nuclear gene expression and physiological response by inhibiting the cytoplasmic PLANT CYSTEINE OXIDASE (PCO) branch of the PROTEOLYSIS (PRT)6 N-degron pathway, a major mechanism of oxygen and nitric oxide (NO) sensing.5 Overexpression of UCP1 (UCP1ox) resulted in the stabilization of an artificial PCO N-degron pathway substrate, and stability of this reporter protein was influenced by pharmacological interventions that control UCP1 activity. Hypoxia and salt-tolerant phenotypes observed in UCP1ox lines resembled those observed for the PRT6 N-recognin E3 ligase mutant prt6-1. Genetic analysis showed that UCP1 regulation of hypoxia responses required the activity of PCO N-degron pathway ETHYLENE RESPONSE FACTOR (ERF)VII substrates. Transcript expression analysis indicated that UCP1 regulation of hypoxia-related gene expression is a normal component of seedling development. Our results show that mitochondrial retrograde signaling represses the PCO N-degron pathway, enhancing substrate function, thus facilitating downstream stress responses. This work reveals a novel mechanism through which mitochondrial retrograde signaling influences nuclear response to hypoxia by inhibition of an ancient cytoplasmic pathway of eukaryotic oxygen sensing.

Comprehensive In Silico Analysis and Transcriptional Profiles Highlight the Importance of Mitochondrial Dicarboxylate Carriers (DICs) on Hypoxia Response in Both Arabidopsis thaliana and Eucalyptus grandis.

Plant dicarboxylate carriers (DICs) transport a wide range of dicarboxylates across the mitochondrial inner membrane. The Arabidopsis thalianaDIC family is composed of three genes (AtDIC1, 2 and 3), whereas two genes (EgDIC1 and EgDIC2) have been retrieved in Eucalyptus grandis. Here, by combining in silico and in planta analyses, we provide evidence that DICs are partially redundant, important in plant adaptation to environmental stresses and part of a low-oxygen response in both species. AtDIC1 and AtDIC2 are present in most plant species and have very similar gene structure, developmental expression patterns and absolute expression across natural Arabidopsis accessions. In contrast, AtDIC3 seems to be an early genome acquisition found in Brassicaceae and shows relatively low (or no) expression across these accessions. In silico analysis revealed that both AtDICs and EgDICs are highly responsive to stresses, especially to cold and submergence, while their promoters are enriched for stress-responsive transcription factors binding sites. The expression of AtDIC1 and AtDIC2 is highly correlated across natural accessions and in response to stresses, while no correlation was found for AtDIC3. Gene ontology enrichment analysis suggests a role for AtDIC1 and AtDIC2 in response to hypoxia, and for AtDIC3 in phosphate starvation. Accordingly, the investigated genes are induced by submergence stress in A. thaliana and E. grandis while AtDIC2 overexpression improved seedling survival to submergence. Interestingly, the induction of AtDIC1 and AtDIC2 is abrogated in the erfVII mutant that is devoid of plant oxygen sensing, suggesting that these genes are part of a conserved hypoxia response in Arabidopsis.

Knockdown of Mitochondrial Uncoupling Proteins 1 and 2 (AtUCP1 and 2) in Arabidopsis thaliana Impacts Vegetative Development and Fertility.

Mitochondrial uncoupling proteins (UCPs) are mitochondrial inner membrane proteins that dissipate the proton electrochemical gradient generated by the respiratory chain complexes. In plants, these proteins are crucial for maintaining mitochondrial reactive oxygen species (ROS) homeostasis. In this study, single T-DNA insertion mutants for two (AtUCP1 and AtUCP2) out of the three UCP genes present in Arabidopsis thaliana were employed to elucidate their potential roles in planta. Our data revealed a significant increase in the Adenosine triphosphate (ATP)/Adenosine diphosphate (ADP) ratios of both mutants, indicating clear alterations in energy metabolism, and a reduced respiratory rate in atucp2. Phenotypic characterization revealed that atucp1 and atucp2 plants displayed reduced primary root growth under normal and stressed conditions. Moreover, a reduced fertility phenotype was observed in both mutants, which exhibited an increased number of sterile siliques and a lower seed yield compared with wild-type plants. Reciprocal crosses demonstrated that both male fertility and female fertility were compromised in atucp1, while such effect was exclusively observed in the male counterpart in atucp2. Most strikingly, a pronounced accumulation of hydrogen peroxide in the reproductive organs was observed in all mutant lines, indicating a disturbance in ROS homeostasis of mutant flowers. Accordingly, the atucp1 and atucp2 mutants exhibited higher levels of ROS in pollen grains. Further, alternative oxidase 1a was highly induced in mutant flowers, while the expression profiles of transcription factors implicated in gene regulation during female and male reproductive organ/tissue development were perturbed. Overall, these data support the important role for AtUCP1 and AtUCP2 in flower oxidative homeostasis and overall plant fertility.

Correcting optic capture with 2 flanged 6-0 sutures after intrascleral haptic fixation with ViscoNeedling.

A 6-0 polypropylene suture was introduced in the eye through an ophthalmic viscosurgical device syringe in a completely closed system. The procedure was used to correct optic capture of the intraocular lens by the pupil after 2 cases of Yamane technique.

Chitosan packaging functionalized with Cinnamodendron dinisii essential oil loaded zein: A proposal for meat conservation.

The nanoencapsulation of essential oils for biodegradable films functionalization is a viable alternative for the production of active food packaging. In this study, the Cinnamodendron dinisii Schwanke essential oil was nanoencapsulated using zein as wall material, and applied in chitosan matrix to produce an active nanocomposite film packaging for food conservation. The chemical composition of the Cinnamodendron dinisii Schwanke essential oil showed a variety of unexplored bioactive compounds, and 1,8-cineole was the major compound. The oil nanoencapsulation produced stable and homogeneous nanoparticles with zeta potential close to 30 mV and polydispersity index lower than 0.2. The nanoparticles size showed a size variation between 70 and 110 nm. The chitosan films obtained functionalized with nanoparticles demonstrated antioxidant activity and antimicrobial activity. The active packaging containing zein nanoparticles was efficient in the conservation of ground beef, stabilizing the deterioration reactions and preserving the color.

Lysine Catabolism Through the Saccharopine Pathway: Enzymes and Intermediates Involved in Plant Responses to Abiotic and Biotic Stress.

The saccharopine pathway (SACPATH) involves the conversion of lysine into α-aminoadipate by three enzymatic reactions catalyzed by the bifunctional enzyme lysine-ketoglutarate reductase/saccharopine dehydrogenase (LKR/SDH) and the enzyme α-aminoadipate semialdehyde dehydrogenase (AASADH). The LKR domain condenses lysine and α-ketoglutarate into saccharopine, and the SDH domain hydrolyzes saccharopine to form glutamate and α-aminoadipate semialdehyde, the latter of which is oxidized to α-aminoadipate by AASADH. Glutamate can give rise to proline by the action of the enzymes Δ1-pyrroline-5-carboxylate synthetase (P5CS) and Δ1-pyrroline-5-carboxylate reductase (P5CR), while Δ1-piperideine-6-carboxylate the cyclic form of α-aminoadipate semialdehyde can be used by P5CR to produce pipecolate. The production of proline and pipecolate by the SACPATH can help plants face the damage caused by osmotic, drought, and salt stress. AASADH is a versatile enzyme that converts an array of aldehydes into carboxylates, and thus, its induction within the SACPATH would help alleviate the toxic effects of these compounds produced under stressful conditions. Pipecolate is the priming agent of N-hydroxypipecolate (NHP), the effector of systemic acquired resistance (SAR). In this review, lysine catabolism through the SACPATH is discussed in the context of abiotic stress and its potential role in the induction of the biotic stress response.

Mitochondrial uncoupling protein-dependent signaling in plant bioenergetics and stress response.

The biological function of plant mitochondrial uncoupling proteins (pUCPs) has been a matter of considerable controversy. For example, the pUCP capacity to uncouple respiration from ATP synthesis in vivo has never been fully acknowledged, in contrast to the mammalian UCP1 (mUCP1) role in uncoupling respiration-mediated thermogenesis. Interestingly, both pUCPs and mUCPs have been associated with stress response and metabolic perturbations. Some central questions that remain are how pUCPs and mUCPs compare in biochemical properties, molecular structure and cell biology under physiological and metabolically perturbed conditions. This review takes advantage of the large amount of data available for mUCPs to review the biochemical properties, 3D structure models and potential physiological roles of pUCPs during plant development and response to stress. The biochemical properties and structure of pUCPs are revisited in light of the recent findings that pUCPs catalyse the transport of metabolites across the mitochondrial inner membrane and the resolved mUCP2 protein structure. Additionally, transcriptional regulation and co-expression networks of UCP orthologues across species are analysed, taking advantage of publicly available curated experimental datasets. Taking these together, the biological roles of pUCPs are analysed in the context of their potential roles in thermogenesis, ROS production, cell signalling and the regulation of plant cellular bioenergetics. Finally, pUCPs biological function is discussed in the context of their potential role in protecting against environmental stresses.

Bioactive food packaging based on starch, citric pectin and functionalized with Acca sellowiana waste by-product: Characterization and application in the postharvest conservation of apple.

Innovative biodegradable packaging with antimicrobial and antioxidant properties was developed, and functionalized with Acca sellowiana waste by-product (feijoa peel flour, FPF). Physicochemical, morphological, antioxidant, antimicrobial properties, and in situ application in the postharvest conservation of apple were conducted with the packaging produced. The results obtained demonstrate that FPF addition had a positive influence on the packaging characteristics, for all the parameters tested. The high concentration of antioxidant compounds in the films with FPF promoted antimicrobial activity against Escherichia coli, Salmonella typhimurium, and Pseudomonas aeruginosa. The packaging produced maintained the quality of apples during storage, with constant weight after 5 days of storage. Based on our results, the bioactive, antioxidant and antimicrobial packaging functionalized with Acca sellowiana waste by-product may be considered as a new alternative to packaging in food systems.