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Protein oxidation is a topic of indisputable scientific interest given the impact of oxidized proteins on food quality and safety. Carbonylation is regarded as one of the most notable post-translational modifications in proteins and yet, this reaction and its consequences are poorly understood. From a mechanistic perspective, primary protein carbonyls (i.e. α-aminoadipic and γ-glutamic semialdehydes) have been linked to radical-mediated oxidative stress, but recent studies emphasize the role alternative carbonylation pathways linked to the Maillard reaction. Secondary protein carbonyls are introduced in proteins via covalent linkage of lipid carbonyls (i.e. protein-bound malondialdehyde). The high reactivity of protein carbonyls in foods and other biological systems indicates the intricate chemistry of these species and urges further research to provide insight into these molecular mechanisms and pathways. In particular, protein carbonyls are involved in the formation of aberrant and dysfunctional protein aggregates, undergo further oxidation to yield carboxylic acids of biological relevance and establish interactions with other biomolecules such as oxidizing lipids and phytochemicals. From a methodological perspective, the routine dinitrophenylhydrazine (DNPH) method is criticized not only for the lack of accuracy and consistency but also authors typically perform a poor interpretation of DNPH results, which leads to misleading conclusions. From a practical perspective, the biological relevance of protein carbonyls in the field of food science and nutrition is still a topic of debate. Though the implication of carbonylation on impaired protein functionality and poor protein digestibility is generally recognized, the underlying mechanism of such connections requires further clarification. From a medical perspective, protein carbonyls are highlighted as markers of protein oxidation, oxidative stress and disease. Yet, the specific role of specific protein carbonyls in the onset of particular biological impairments needs further investigations. Recent studies indicates that regardless of the origin (in vivo or dietary) protein carbonyls may act as signalling molecules which activate not only the endogenous antioxidant defences but also implicate the immune system. The present paper concisely reviews the most recent advances in this topic to identify, when applicable, potential fields of interest for future studies.

The accumulation in the leaves and young stems of phenolic compounds, such as hydrolyzable and condensed tannins, constitutes a defense mechanism of plants against herbivores. Among other stressing factors, chronic herbivory endangers “Quercus ilex”, a tree playing a central role in Mediterranean forests. This work addressed the connections between the chemical defenses of “Q. ilex” leaves and their susceptibility to herbivory, quantitative traits whose relationships are modulated by environmental and genetic factors that could be useful as molecular markers for the selection of plants with improved fitness. A search for natural variants detected the polymorphism D165H in the effector domain of QiMYB-like-1, a TT2-like transcription factor whose family includes members that control the late steps of condensed tannins biosynthesis in different plant species. QiMYB-like-1 D165H polymorphism was screened by PCR-RFLP in trees from six national parks in Spain where “Q. ilex” has a relevant presence, revealing that, unlike most regions that match the Hardy-Weinberg equilibrium, homozygous plants are over-represented in “Monfragüe” and “Cabañeros”, among the best examples to represent the continental Mediterranean (cM) ecosystem. Accordingly, the averages of two stress-related quantitative traits measured in leaves, herbivory index and accumulation of condensed tannins, showed asymmetric distributions depending on the clustering of trees based on ecological and genetic factors. Thus, the impact of herbivory was greater in managed forests with a low density of trees from the cM region, among which QiMYB-like-1 D165 homozygotes stand out, whereas condensed tannins accumulation was higher in leaves of QiMYB-like-1 H165 homozygotes from low-density forests, mainly in the Pyrenean (Py) region. Besides, the correlation between the contents of condensed tannins and total tannins vanished after clustering by the same factors: the cM region singularity, forest tree density, and QiMYB-like-1 genotype, among which homozygous shared the lowest link. The biogeographical and genetic constraints that modulate the contribution of condensed tannins to chemical defenses also mediated their interactions with the herbivory index, which was found positively correlated with total phenolics or tannins, suggesting an induction signal by this biotic stress. In contrast, a negative correlation was observed with condensed tannins after tree clustering by genetics factors where associations between tannins were lost. Therefore, condensed tannins might protect “Q. ilex” from defoliation in parks belonging to the cM ecosystem and carrying genetic factor(s) linked to the QiMYB-like-1 D165H polymorphism.

Extraction temperature, extraction time and liquid (water) to solid ratio were optimized in order to extract antioxidant phenolic compounds from brewers’ spent grain (BSG). The extracts were analysed for their total phenol content (TPC) and antioxidant activity was measured using three different methods: 2,2-diphenyl-2-picrylhydrazyl (DPPH) free radical, 2,2´-azino-bis (3-ethylbenothiazoline-6-sulphonic acid) (ABTS), and reducing power (RP) assays. All the parameters except extraction time promoted different efficiencies for the extraction of antioxidant phenolic compounds. TPC extraction was higher at lower temperatures and lower liquid/solid ratios up to a certain point. In this sense, a decrease in TPC with increasing liquid/solid ratios took place until a 16:1 ratio and a plateau was reached beyond that ratio. The highest DPPH activity was reported for 30–35 °C and 60–90 min extraction and 60–90 min extraction with a 25 mL/g ratio. ABTS values increased as the liquid to solid ratio decreased, being positively correlated with TPC (R = 0.788; p < 0.01). The highest RP was achieved at 30–33 °C extraction temperature and 10–14 mg/mL v/w ratio and at 116–120 min extraction and 16–17 mg/mL ratio. Gallic acid accounted for the majority of the phenolic compounds found, followed by hydroxyphenylacetic acid, epicatechin, and protocatechuic acid. Sinapic, 4-hydroxy benzoic, and syringic acids were also found in lower quantities. Coumaric, vanillic, ferulic, and caffeic acids were present in very small amounts. All the extracts contained phenolics and showed in vitro antioxidant activity, but the extracts obtained by using 30 °C, 121.9 min, and 10 mL/g liquid/solid ratio exhibited the highest content in TPC and antioxidant potential. The aqueous extraction of a potentially bioactive extract from BSG was demonstrated to be an efficient and simple method to recover these value-added compounds.

Oxidative stress is considered a major mechanism causing sperm damage during cryopreservation and storage, and underlies male factor infertility. Currently, oxidative stress is no longer believed to be caused only by the overproduction of reactive oxygen species, but rather by the deregulation of redox signaling and control mechanisms. With this concept in mind, here, we describe for the first time the presence of the soluble carrier family 7 member 11 (SLC7A11) antiporter, which exchanges extracellular cystine (Cyss) for intracellular glutamate, in stallion spermatozoa, as well as its impact on sperm function using the specific inhibitor sulfasalazine. Spermatozoa incubated with Cyss exhibited an increased intracellular GSH content compared with controls (P < 0.01): 50% in fresh extended stallion spermatozoa and 30% in frozen-thawed spermatozoa. This effect was prevented by the addition of sulfasalazine to the media. Cystine supplementation also reduced the oxidation–reduction potential of spermatozoa, with sulfasalazine only preventing this effect on fresh spermatozoa that were incubated for 3 h at 37°C, but not in frozen-thawed spermatozoa. While sulfasalazine reduced the motility of frozen-thawed spermatozoa, it increased motility in fresh samples. The present findings provide new and relevant data on the mechanism regulating the redox status of spermatozoa and suggest that a different redox regulatory mechanism exists in cryopreserved spermatozoa, thus providing new clues to improve current cryopreservation technologies and treat male factor infertility. Summary Sentence The SLC7A11 antiporter that exchanges cystine by intracellular glutamate is present and functional in stallion spermatozoa, but cryopreserved spermatozoa may present altered functionality.

There is a necessity to protect the quality and authenticity of herbs and spices because of the increase in the fraud and adulteration incidence during the last 30 years. There are several aspects that make herbs and spices quite vulnerable to fraud and adulteration, including their positive and desirable sensorial and health-related properties, the form in which they are sold, which is mostly powdered, and their economic relevance around the world, even in developing countries. For these reasons, sensitive, rapid, and reliable techniques are needed to verify the authenticity of these agri-food products and implement effective adulteration prevention measures. This review highlights why spices and herbs are highly valued ingredients, their economic importance, and the official quality schemes to protect their quality and authenticity. In addition to this, the type of frauds that can take place with spices and herbs have been disclosed, and the fraud incidence and an overview of scientific articles related to fraud and adulteration based on the Rapid Alert System Feed and Food (RASFF) and the Web of Science databases, respectively, during the last 30 years, is carried out here. Next, the methods used to detect adulterants in spices and herbs are reviewed, with DNA-based techniques and mainly spectroscopy and image analysis methods being the most recommended. Finally, the available adulteration prevention measurements for spices and herbs are presented, and future perspectives are also discussed.

Background Currently, many micromammals are important targets for study. The endangered Galemys pyrenaicus is an outstanding example. Globally, their populations have suffered a substantial decline in last 20 years. In the surveyed area, the capture of desman is legally forbidden due to the high conservation concerns. Reason by non-invasive sampling through faeces is proposed for its monitoring. Furthermore, the confusion between faeces from desman and Mediterranean water shrews must be considered. Thus, the aim of this study was focused on developing RT-PCR assays to determine the presence of Galemys pyrenaicus and N. a. anomalus from non-invasive samples. Methods and results The study was conducted in the mountains of the System Central of Extremadura (Spain). A total of 186 samples were collected from 2018 to 2021 by experts where historically reported and/or our previous studies confirmed their presence. RT-PCR assays using hydrolysis probes were designed to detect genetic material from both desman and Mediterranean water shrews and its specificity was confirmed. The reliability of the method was further assessed by PCR sequencing of mitochondrial Cyb and d-loop, resulting fully compatible with the RT-PCR approach. Intraspecific phylogenetic relationship was reported to improve knowledge about mtDNA variability in the desman from the Central System. Conclusions We demonstrated that RT-PCR gives a gold opportunity to further map the species using faeces which minimizes disturbance and reports both population status and individual presence. Cost-effective RT-PCR combined with field-collected faeces allows us to better investigate the full range of occurrence of the species.

Livestock farming is currently highly questioned and is considered by society to be one of the main precursors of climate change and innumerable environmental impacts. This social concern has marked a trend in public policies in Europe, promoting strategies to reduce greenhouse gas (GHG) emissions by controlling the carbon footprint of agri-food products. However, in certain regions, the perception of the main actors in the sector about the role that livestock farming plays in this fight against climate change and how new political trends point the way toward the sustainability of agrarian systems is still uncertain. In this study, the opinions of stakeholders of the agro-livestock sector on the role that extensive livestock farming plays in the current context of the fight against climate change and the demands for public policies to facilitate the adoption of mitigation practices were examined. A participatory research process through focus groups was used in this qualitative study. Specifically, five sessions were held at the beginning of 2022; the sessions were recorded, transcribed, and anonymized for further analysis. In these sessions, projective techniques were used, such as word association and sentence completion to understand stakeholders' perceptions of the role of extensive livestock farming in climate change. Brand mapping was conducted to determine the opinion on the profitability and GHG emissions of 10 livestock systems typical of the region and of eight quality labelling systems related to sustainability. Brainstorming was carried out to assess available practices for the adaptation of livestock farms and mitigation of climate change. Finally, there was an open debate regarding the demands for public aid for the implementation of mitigation practices. The word association technique identified concepts such as ‘Equilibrium’ in extensive livestock farming and concepts such as ‘Effects’, ‘Action’ and ‘Concern’ in climate change. For the term carbon footprint, the most mentioned concept was ‘ignorance’, and for common agricultural policy, the most mentioned term was ‘injustices’. The results of the brand mapping allowed us to determine the perception of the stakeholders regarding the profitability of the different extensive farm systems and on their GHG emissions, with the most extensive and traditional ones being perceived as the lowest emitters of gases but also the least profitable. For sustainable labels, stakeholders believed that labels contribute to profitability and lower GHG emissions. Strategies to adapt to climate change and reduce the impact of farms were focused on reforestation, grazing, and soil management, adjusting the livestock stocking rate and self-production of food on farms. The best mitigating practices proposed were the maintenance of the extensive livestock farming (4.69), improvement of accesses, livestock routes and roads (4.63), making and applying compost (4.50) and the simplified administrative procedures (5.00). In the prioritization of public aids, three categories were established based on the field of action: social/organizational measures (38 votes), economic measures (44 votes) and environmental measures (22 votes). The aid related to maintaining profitability and improving marketing, followed by aid to reduce bureaucracy and direct aid to extensive livestock farming, were identified as priorities. This study offers a detailed picture of how stakeholders in the agro-livestock sector see the role that extensive livestock farming plays in the fight against climate change. The best farm management practices and priority lines of public support that policy-makers can apply have been identified in this study and emanate directly from those who receive subsidies and make the decisions in their livestock farming to ensure their implementation more successful.

Sporadic Parkinson’s disease, characterised by a decline in dopamine, usually manifests in people over 65 years of age. Although 10% of cases have a genetic (familial) basis, most PD is sporadic. Genome sequencing studies have associated several genetic variants with sporadic PD. Our aim was to analyse the promoter region of the ATG16L1 and ATG5 genes in sporadic PD patients and ethnically matched controls. Genotypes were obtained by using the Sanger method with primers designed by us. The number of haplotypes was estimated with DnaSP software, phylogeny was reconstructed in Network, and genetic divergence was explored with Fst. Seven and two haplotypes were obtained for ATG16L1 and ATG5, respectively. However, only ATG16L1 showed a significant contribution to PD and a significant excess of accumulated mutations that could influence sporadic PD disease. Of a total of seven haplotypes found, only four were unique to patients sharing the T allele (rs77820970). Recent studies using MAPT genes support the notion that the architecture of haplotypes is worthy of being considered genetically risky, as shown in our study, confirming that large-scale assessment in different populations could be relevant to understanding the role of population-specific heterogeneity. Finally, our data suggest that the architecture of certain haplotypes and ethnicity determine the risk of PD, linking haplotype variation and neurodegenerative processes.

The authors would like to acknowledge the support and funding provided by the Junta de Extremadura and FEDER Funds through the Research Project IB18114 and Grant GR21125, which made this research and its publication possible.

Carbon footprint has become a reference indicator of the environmental impact of food production. Governments are increasingly demanding a trend towards low-carbon-footprint production in order to reduce the impact on climate change. In this sense, the study of consumers’ preferences and assessment of products from the perspective of their carbon footprint is crucial to achieve a green and circular economy. This paper specifically attempted to assess consumer preference and willingness to pay for beef that has been carbon-footprint-labelled as an attribute. In order to attain this objective, a choice experiment was designed and applied to a total of 362 Spanish consumers. The results revealed a positive preference towards beef produced in low-carbon-footprint systems. In addition, the segmentation of these consumers revealed the existence of a group of consumers who prioritise environmental impact over product price.