In vivo biosensing of subcellular pyruvate pools reveals photosynthesis-dependent metabolite dynamics in Nicotiana benthamiana.

Pyruvate is central to metabolism across biology. It acts as a metabolic hub linking key pathways including glycolysis, the Krebs cycle, fermentation, and synthesis of amino acids, fatty acids, isoprenoids and nucleotides. Even though the central role of pyruvate is well established biochemically, there is a remarkable void in our understanding of how pyruvate levels behave within cells, where pyruvate is distributed across different compartments, and differential changes in pyruvate pools may occur rapidly upon changes in metabolic fluxes. Recently, this problem has been addressed by the development of a genetically-encoded pyruvate biosensor to provide first insights into the pyruvate dynamics in animal cells. Here, we establish in vivo biosensing of pyruvate in plants. We provide advanced characterisation of the biosensor properties and demonstrate the functionality of the sensor in the cytosol, the mitochondria and the chloroplasts of Nicotiana benthamiana epidermal cells. Finally, we harnessed the tool to investigate the impact of photosynthesis on pyruvate with unprecedented spatial and temporal resolution, revealing pronounced changes in cytosolic pyruvate pools. While highlighting the current limitations of the biosensor, this study provides proof-of-concept for how the dynamics and regulation of central carbon metabolites can be revealed in the context of living plant tissues.

Glutamate imbalance in key structure of the default mode network in adults with attention-deficit/hyperactivity disorder.

The default mode network (DMN) is atypically active in patients with ADHD, likely contributing to the inattention patterns observed in patients with the disorder. Nonetheless, magnetic resonance spectroscopy (MRS) studies have rarely targeted the posterior cingulate cortex, a key DMN region, and little is known about the biochemical setting within this network in patients with ADHD. We aimed to assess the differences in metabolite profiles of the posterior cingulate cortex-a key region of the DMN-between patients with ADHD and controls. Five brain metabolites-glutamate, inositol, N-acetyl aspartate, choline, and creatine-were measured through MRS in the posterior cingulate cortex of patients and controls in a 3.0 T scanner. Between-group comparison of neurometabolite concentrations in PCC was performed using multivariate analysis of covariance. A total of 88 patients and 44 controls were included in the analysis. Patients with ADHD showed lower levels of glutamate in the posterior cingulate cortex compared to controls (p = 0.003). Lower concentrations of glutamate in the posterior cingulate cortex suggest that a glutamatergic imbalance within the posterior cingulate cortex might play a role in the pathogenesis of ADHD. Further understanding of the causes and consequences of such glutamate decrease might help explain how some glutamate-related drug effects impact on ADHD symptomatology.

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.

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.

Obstructive Sleep Apnea and Wake-up Stroke - A 12 Months Prospective Longitudinal Study.

BACKGROUND: Wake-up stroke (WUS) are strokes that are noted upon awakening in patients previously going to bed in a normal state of health. The role of sleep abnormalities in WUS is uncertain. The objective was to determine clinical characteristics, respiratory abnormalities during sleep and outcomes in patients with WUS versus non-WUS. METHODS: At baseline, patients with ischemic stroke were examined clinically and with a portable sleep recorder. Apnea-Hypopnea Index greater than or equal to 20 defined a cut-off severity index. At follow-up (3 and 12-months), patients were re-evaluated clinically and with questionnaires: Epworth Sleepiness Scale, Modified Rankin (MR) and Modified Barthel Index Results: Among all (N = 102, 64% male), hypertension (73%), Type 2 diabetes (29.4), heart disease (16.7%), physical inactivity (69.6%), smoking (32.4%) and alcohol consumption (17.6) were found. Apnea-Hypopnea Index (AHI) greater than 5 (92.9%), AHI greater than 15 (44.7%), AHI greater than or equal to 20 (35.3%) and AHI greater than 30 (11.8%) were registered. Cases with and without WUS did not differ regarding polygraphic findings. Long apneas (apnea duration > 20 s) was equally found in patients with WUS (23.1%) and non-WUS (23.7%). Type 2 diabetes mellitus (T2D) was independently associated with WUS (OR = 2.76; CI: 1.10-6.05; P = .03). Prospectively, symptom severity was not different between WUS and non-WUS. Overall, patients with OSA (IAH≥20) evolved with worse functional performance (MR, P = .02). CONCLUSIONS: Wake-up stroke occurred in approximately 1 of 3 of cases. Irrespective of WUS, half of the patients had moderate to severe sleep apnea; those with OSA (AHI≥ 20) evolved with worse functional performance after 1 year. WUS was associated with TDM reinforcing a relationship with cerebral small vessel disease.

Chemical Characterization and Release of Polyphenols from Pecan Nut Shell [Carya illinoinensis (Wangenh) C. Koch] in Zein Microparticles for Bioactive Applications.

The pecan nut [Carya illinoinensis (Wangenh) C. Koch] is a natural source of polyphenols with antioxidant properties. In this study, the encapsulation of aqueous and hydroalcoholic extracts of pecan nut shell were evaluated for the release of bioactive compounds and antioxidant potential in order to explore food applications using zein as encapsulating agent. The extracts showed high contents of total phenolics, condensed tannins and high antioxidant activity. Concentrations of proanthocyanidins were 9-fold higher in hydroalcoholic extracts. The LC-DAD analysis showed that catechins were the major phenolic compounds in samples, with epigallocatechin levels up to 138.62 mg mL-1. Zein microparticles loaded with aqueous extract released 2.3 times more phenolic compounds than the hydroalcoholic extracts and the DSC thermograms showed that extracts of pecan nut shell remained thermally stable up to 240 °C. The zein microcapsules obtained in this study were efficiently encapsulated and represent an interesting additive due its high antioxidant capacity, physicochemical characteristics and morphology. The use of zein microparticles combined with natural extracts constitute a step forward in the improvement of current technology for delivering phenolic compounds with applications in functional foods and nutraceuticals.

Overexpression of mitochondrial uncoupling protein 1 (UCP1) induces a hypoxic response in Nicotiana tabacum leaves.

Mitochondrial uncoupling protein 1 (UCP1) decreases reactive oxygen species production under stress conditions by uncoupling the electrochemical gradient from ATP synthesis. This study combined transcriptome profiling with experimentally induced hypoxia to mechanistically dissect the impact of Arabidopsis thaliana UCP1 (AtUCP1) overexpression in tobacco. Transcriptomic analysis of AtUCP1-overexpressing (P07) and wild-type (WT) plants was carried out using RNA sequencing. Metabolite and carbohydrate profiling of hypoxia-treated plants was performed using (1)H-nuclear magnetic resonance spectroscopy and high-performance anion-exchange chromatography with pulsed amperometric detection. The transcriptome of P07 plants revealed a broad induction of stress-responsive genes that were not strictly related to the mitochondrial antioxidant machinery, suggesting that overexpression of AtUCP1 imposes a strong stress response within the cell. In addition, transcripts that mapped into carbon fixation and energy expenditure pathways were broadly altered. It was found that metabolite markers of hypoxic adaptation, such as alanine and tricarboxylic acid intermediates, accumulated in P07 plants under control conditions at similar rates to WT plants under hypoxia. These findings indicate that constitutive overexpression of AtUCP1 induces a hypoxic response. The metabolites that accumulated in P07 plants are believed to be important in signalling for an improvement in carbon assimilation and induction of a hypoxic response. Under these conditions, mitochondrial ATP production is less necessary and fermentative glycolysis becomes critical to meet cell energy demands. In this scenario, the more flexible energy metabolism along with an intrinsically activated hypoxic response make these plants better adapted to face several biotic and abiotic stresses.

Overexpression of UCP1 in tobacco induces mitochondrial biogenesis and amplifies a broad stress response.

BACKGROUND: Uncoupling protein one (UCP1) is a mitochondrial inner membrane protein capable of uncoupling the electrochemical gradient from adenosine-5'-triphosphate (ATP) synthesis, dissipating energy as heat. UCP1 plays a central role in nonshivering thermogenesis in the brown adipose tissue (BAT) of hibernating animals and small rodents. A UCP1 ortholog also occurs in plants, and aside from its role in uncoupling respiration from ATP synthesis, thereby wasting energy, it plays a beneficial role in the plant response to several abiotic stresses, possibly by decreasing the production of reactive oxygen species (ROS) and regulating cellular redox homeostasis. However, the molecular mechanisms by which UCP1 is associated with stress tolerance remain unknown. RESULTS: Here, we report that the overexpression of UCP1 increases mitochondrial biogenesis, increases the uncoupled respiration of isolated mitochondria, and decreases cellular ATP concentration. We observed that the overexpression of UCP1 alters mitochondrial bioenergetics and modulates mitochondrial-nuclear communication, inducing the upregulation of hundreds of nuclear- and mitochondrial-encoded mitochondrial proteins. Electron microscopy analysis showed that these metabolic changes were associated with alterations in mitochondrial number, area and morphology. Surprisingly, UCP1 overexpression also induces the upregulation of hundreds of stress-responsive genes, including some involved in the antioxidant defense system, such as superoxide dismutase (SOD), glutathione peroxidase (GPX) and glutathione-S-transferase (GST). As a consequence of the increased UCP1 activity and increased expression of oxidative stress-responsive genes, the UCP1-overexpressing plants showed reduced ROS accumulation. These beneficial metabolic effects may be responsible for the better performance of UCP1-overexpressing lines in low pH, high salt, high osmolarity, low temperature, and oxidative stress conditions. CONCLUSIONS: Overexpression of UCP1 in the mitochondrial inner membrane induced increased uncoupling respiration, decreased ROS accumulation under abiotic stresses, and diminished cellular ATP content. These events may have triggered the expression of mitochondrial and stress-responsive genes in a coordinated manner. Because these metabolic alterations did not impair plant growth and development, UCP1 overexpression can potentially be used to create crops better adapted to abiotic stress conditions.

Investigating the etiology of Haff disease: Optimization and validation of a sensitive LC-MS/MS method for palytoxins analysis in directly associated freshwater and marine food samples from Brazil.

Haff disease typically develops after eating contaminated marine or freshwater species, especially fish. Despite still having an unknown etiology, recent reports have suggested its possible correlation with palytoxins. Therefore, the present work aimed to optimize and perform a validation of a sensitive method using liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) for the analysis of palytoxin and some of its analogs, with the main purpose of investigating their presence in marine and freshwater food samples associated with Haff disease in Brazil. The method optimization was performed using a central composite rotatable design and fish samples fortified with the palytoxin standard. Then, the optimized method was validated for different food matrices, including freshwater and marine fish, mollusks, and crustaceans. The sample preparation involved a solid-liquid extraction using methanol and water, solid-phase extraction using Strata-X cartridges, and on-column palytoxin oxidation. The detection of the main oxidized fragments (amino and amide aldehydes) was achieved by LC-MS/MS with electrospray ionization in positive mode, using a C18 column, as well as acetonitrile and water as mobile phases, both acidified with 0.1 % of formic acid. After optimization and validation, the etiological investigation involved the analysis of 16 Brazilian Haff disease-related food samples (in natura and leftover meals) from 2022. The method was demonstrated to be appropriate for quantitative analysis of freshwater and marine species. So far, it has proven to be one of the most sensitive methods related to palytoxin detection (LOD 10 µg/kg), being able to work in a range that includes the provisional ingestion limit (30 µg/kg). Regarding the Haff disease-related samples analysis, there is a strong indication of palytoxin contamination since the amino aldehyde (common fragment for all palytoxins) was detected in 15 of the 16 samples. Selected results were confirmed using liquid chromatography coupled with high-resolution mass spectrometry (LC-HRMS).

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.

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.

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.

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.

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.

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.

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.