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Metal ions have been shown to play a critical role in amyloid-β (Aβ) neurotoxicity and plaque formation which are key hallmarks of Alzheimer's disease. Amyloid-β peptides can bind both copper and hemin and this interaction modulates the redox chemistry of these metals. The characterization of the binding of hemin through UV-Vis spectroscopic titration with Aβ(4-16) shows a significantly higher affinity than that with Aβ(1-16). Also, the characterization of the hemin-catalyzed oxidation through different assays (peroxidase-like activity, ascorbate oxidation, HPLC-MS analysis of peptide oxidation) displays a greater reactivity in the presence of Aβ(4-16) when compared to that of Aβ(1-16). Since the Aβ(4-16) peptide sequence contains the typical amino-terminal copper and nickel binding motif (ATCUN), this leads to investigate the potential formation of ternary hemin/copper/Aβ(4-16) adducts. The evaluation of K 1 and K 2 (constants that regulate the formation of five-coordinated high-spin complex and of six-coordinated low-spin complex, respectively) for mixed systems indicates that the presence of copper stabilizes the 1:1 hemin-Aβ(4-16) species, partially hindering the formation of the low-spin complex. On the other hand, the formation of the ternary hemin/copper/Aβ(4-16) complex gives rise to a less efficient catalyst, resulting in a reduction of the overall oxidative reactivity. These results suggest that the reactivity of metal ions is finely modulated by the formation of complexes with amyloid peptides and this property is also regulated differently by the various in vivo relevant isoforms., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)

Brush border membrane (BBM) enzymes greatly affect the bioaccessibility and bioavailability of food nutrients. Despite their physiological importance, a step simulating the final stage of intestinal digestion has not yet been included in the harmonized protocols for in vitro digestion, primarily due to the challenges of replicating the dynamics of intestinal degradation. Herein, we propose an advancement toward a more physiologically relevant method, complementing the harmonized static gastric-duodenal digestion INFOGEST model with the missing small intestinal phase. BBM hydrolase activity, incubation time, at pH 7.2 were established to reproduce the small intestinal conditions. Skim milk powder, as a model of protein food, was subjected to the in vitro static digestion. Immediately after the duodenal phase, digesta were supplemented with BBM vesicles purified from pig jejunum. To comply with the dynamic nature of intestinal digestion and balance the spontaneous inactivation of hydrolases, BBM supplements were added every two hours throughout 6 h incubation time. Peptide degradation was monitored at each stage of digestion by amino acid analysis, free α-amino group assay, HPLC, LC-MS/MS. Hydrolysis by BBM peptidases led to a significant increase of free amino acids, reflecting the known level of amino acid adsorption (>90 %) in humans after eating milk proteins. LC-MS/MS analysis demonstrated that BBM hydrolases erode progressively the peptides released by gastro-duodenal processing up to stable sequence motifs. The approach described is particularly relevant when the endpoint is identifying the peptide sequences that cannot be further hydrolysed by digestive enzymes or to determine the amino acid bio-accessibility., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

The health of humans, animals and the environment is interconnected. Adopting a One Health approach means intervening promptly to prevent the main diseases that affect animal health to guarantee the safety of livestock production. Exposure to toxic trace elements in sheep can lead to increased accumulation in different biological substrate, developing both acute and chronic diseases in humans and livestock. The aim of this study was to evaluate the bioaccumulation of arsenic (As), cadmium (Cd), lead (Pb) and mercury (Hg) in Sicilian Barbaresca sheep using the following biological substrates: milk, blood and fleece. An inductively coupled plasma mass spectrometer (ICP-MS) was used for As, Cd and Pb, and a direct mercury analyser (DMA-80) was used for Hg determination. In addition, the role of the haematological parameters as possible indicators of different biodistribution was evaluated. A statistically significant value was observed from our analysed metals in the substrates: arsenic (p < 0.001), cadmium (p < 0.01), lead (p < 0.001) and mercury (p < 0.0001). The correlation analysis showed a relationship between milk and blood for arsenic (p < 0.0001) and lead (p < 0.0001), and no correlation for the metals was observed between milk/blood and the haematological parameters analysed for the low concentration observed in the present study comforting the final consumer., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Tritordeum is an amphiploides species resulting from the hybridization between durum wheat ( T. durum ) and wild barley ( H. chilense ). This new cereal is considered a natural crop as it is obtained by traditional breeding techniques. Given its appreciable organoleptic characteristics, agronomic features, presence of interesting components, and good technological properties, Tritordeum is of promising interest for the development of health-oriented foods. In this study, we evaluated two registered Tritordeum cultivars, Bulel and Aucan. T. durum (Provenzal) was employed as the positive control. The extracted proteins were digested by gastric/pancreatic proteases, and their biological effects on Caco-2 differentiated on transwell inserts were determined. Changes in cell viability, monolayer permeability, organization of F-actin microfilaments, and ER stress triggered by protein-digested samples (DPs) were inspected. Our results showed that exposure to Provenzal-DPs promptly disrupted the tight junction barrier. Conversely, Aucan-DPs did not enhance monolayer permeability, whereas Bulel-DPs exerted only slight effects. Provental-DPs-induced toxicity was also confirmed by changes in cell viability and by the deep reorganization of the enterocyte cytoskeleton. In contrast, Aucan-DPs and Bulel-DPs did not affect monolayer viability and cytoskeleton structure. Overall, our findings suggest that both Tritordeum cultivars could be potential candidates for mitigating the toxicity of wheat flour.

Both β-amyloid (Aβ) peptides and oxidative stress conditions play key roles in Alzheimer's disease. Hemin contributes to the development of the disease as it possesses redox properties and its level increases in pathological conditions or traumatic brain injuries. The aim of this work was to deepen the investigation of the reactivity of the hemin-Aβ 16 complex, considering its ability to catalyze oxidation and nitration reactions. We performed kinetic studies in the presence of hydrogen peroxide and nitrite with phenolic and catechol substrates, as well as mass spectrometry studies to investigate the modifications occurring on the peptide itself. The kinetic constants were similar for oxidation and nitration reactions, and their values suggest that the hemin-Aβ 16 complex binds negatively charged substrates with higher affinity. Mass spectrometry studies showed that tyrosine residue is the endogenous target of nitration. Hemin degradation analysis showed that hemin bleaching is only partly prevented by the coordinated peptide. In conclusion, hemin has rich reactivity, both in oxidation and nitration reactions on aromatic substrates, that could contribute to redox equilibrium in neurons. This reactivity is modulated by the coordination of the Aβ 16 peptide and is only partly quenched when oxidative and nitrative conditions lead to hemin degradation.

The aim of our study was to analyze circadian rhythm of the hematological profile of horses housed in a loose box and paddock during the different seasons (spring, summer, autumn, and winter). Blood samples were performed every 4 h for 48 consecutive hours. Red blood cells (RBCs), hemoglobin (HGB), hematocrit (HCT), white blood cells (WBCs), platelets (PLTs), and leukocyte subpopulations (neutrophils, basophils, eosinophils, lymphocytes, and monocytes) were analyzed, and, at the same time, environmental conditions were recorded. A statistically significant effect of housing conditions ( p < 0.0001) was observed on all hematological values except for WBC during winter and for neutrophils ( p < 0.0001) during spring and autumn. A statistically significant effect of season ( p < 0.0001) was found for RBC, HCT, and PLT and for all leukocyte cells ( p < 0.0001) except for basophils. The single Cosinor method revealed a daily rhythm of hematological parameters during spring in both groups, and a daily rhythm for lymphocytes and neutrophils was observed during spring and summer in horses kept in a loose box and during winter in horses housed in a paddock. Our results revealed that the response of the immune system is regulated by circadian physiology. Knowledge of the periodic temporal structure of mammals should be considered when evaluating animals' adaptation to temporizations imposed by the environment.

Thermoregulation is an important mechanism to ensure thermal homeostasis. In horses, different housing conditions could influence this mechanism by exposing animals to different microclimates. The circadian rhythm of body temperature is well known. For this reason, we wanted to investigate the daily rhythm of cutaneous (jugular, shoulder, inner thigh and croup) temperature in comparison to rectal temperature in athletic horses kept under two different housing conditions. Ten clinically healthy Italian Saddle horses were divided into two equal groups: the first group was housed in individual boxes, and the second group was kept in paddocks. In all horses, rectal and cutaneous temperature measurements were performed every 4 h for 48 consecutive hours during different seasons (spring, summer, autumn and winter), respectively, by means of digital and infrared thermometers. Ambient temperature, relative humidity and ventilation were recorded simultaneously. A general linear model (GLM) was applied to the recorded temperature values displaying a significant effect of season, time of day, site of recording and housing condition (p < 0.001). No statistical differences were found between the two days of monitoring (p = 0.49). A trigonometric statistical model (single cosinor method) was applied to investigate the circadian rhythm of rectal and cutaneous temperatures in the two different groups. Our results showed circadian rhythmicity of rectal temperature during all seasons and in both groups. Cutaneous temperature shows daily rhythmicity that was different in the various regions and was influenced by housing conditions and seasons. Application of GLM also showed a statistically significant effect of season and site of recording (p < 0.0001) on all circadian parameters and of management condition on amplitude and robustness (p < 0.0001). No statistical differences between the two days of monitoring were observed (p = 0.68). These findings are probably due to the animals' thermoregulatory mechanisms ensuring heat exchange between the body and the environment, and that was influenced by the microclimate. In particular, the microclimate influenced the thermodispersion disrupting the daily rhythmicity of some body regions, in horses kept in boxes. Only the temperate climatic conditions observed in summer guarantee the expression of the daily rhythmicity of all body surfaces in both management conditions. These results contribute to the knowledge of the mechanisms of homeostasis and control of body temperature in the athletic horse kept in different housing conditions, with a chronophysiological interpretation that completes the proper management of thermal well-being in horses., (© 2022. The Author(s) under exclusive licence to International Society of Biometeorology.)

Piezoelectric energy harvesters (PEHs) are a reduced, but fundamental, source of power for embedded, remote, and no-grid connected electrical systems. Some key limits, such as low power density, poor conversion efficiency, high internal impedance, and AC output, can be partially overcome by matching their internal electrical impedance to that of the applied resistance load. However, the applied resistance load can vary significantly in time, since it depends on the vibration frequency and the working temperature. Hence, a real-time tracking of the applied impedance load should be done to always harvest the maximum energy from the PEH. This paper faces the above problem by presenting an active control able to track and follow in time the optimal working point of a PEH. It exploits a non-conventional AC-DC converter, which integrates a single-stage DC-DC Zeta converter and a full-bridge active rectifier, controlled by a dedicated algorithm based on pulse-width modulation (PWM) with maximum power point tracking (MPPT). A prototype of the proposed converter, based on discrete components, was created and experimentally tested by applying a sudden variation of the resistance load, aimed to emulate a change in the excitation frequency from 30 to 70 Hz and a change in the operating temperature from 25 to 50 °C. Results showed the effectiveness of the proposed approach, which allowed to match the optimal load after 0.38 s for a ΔR of 47 kΩ and after 0.15 s for a ΔR of 18 kΩ.

By virtue of exclusive nutrient composition and nutritional properties, seed germ flours from both European carob (Ceratonia siliqua L.) and South American algarrobo (Prosopis alba and Prosopis nigra) or vinal (Prosopis ruscifolia) have potential as a high nutritional value and health-promoting ingredient for food formulations. In order to define their compositional and functional properties, we investigated the germ protein content of carob compared to the P. alba, P. nigra and P. ruscifolia counterparts, applying proteomics and complementary methods. The mono- and two-dimensional electrophoretic profiles of Prosopis spp. were very similar among one another, while C. siliqua exhibited significant differences. Attenuated total reflectance-Fourier transform infrared (ATR-FTIR) spectroscopy highlighted a dominant β-sheet structural conformation for C. siliqua, suggesting that carob germ flour might more suited than Prosopis germ flour for baking and food technological applications. In contrast, Prosopis spp. contained a more adequate nutritional value than C. siliqua, in terms of essential amino acid complement. Both carob and algarrobo germ flour samples were highly digestible, as demonstrated by in vitro simulated gastrointestinal digestion, releasing high amounts of free amino acids and only minor proportions of low molecular weight peptides., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

In this work, we explored the "deep" seed peanut proteome by using both two dimensional electrophoresis (2-DE)-based analysis run under reducing and non-reducing condition (protein-centric) and LC-MS/MS gel-free proteomic (peptide-centric). The former approach allowed to identify high molecular weight disulfide-linked Ara h 1 and Ara h 3 heteroligomers and Ara h 1 homoligomers linked through covalent bonds other than disulfides. The occurrence of these protein complexes revealed natural interactions between Ara(s) subunits with a possible involvement in the allergenic potential of peanut. The second approach, also referred to as shot-gun proteomics, allowed the identification of 149 gene products, including low-abundance proteins escaped the 2-DE detection. Interestingly, we identified 60 proteins never catalogued previously. The complementary exploitation of two proteomic approaches enabled the access to new relevant information about the complexity of the peanut proteome, with special emphasis to the complement of allergens (allergome)., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Wheat (Triticum spp. L.), rye (Secale cereal L.), and barley (Hordeum vulgare L.) seeds contain peptides toxic to celiac patients. Maize (Zea mays L.) and rice (Oryza sativa L.) are distant relatives of wheat as well as sorghum (Sorghum bicolor (L.) Moench) and are known to be safe for celiacs. Both immunochemical studies and in vitro and in vivo challenge of wheat-free sorghum food products support this conclusion, although molecular evidence is missing. The goal of the present study was to provide biochemical and genetic evidence that sorghum is safe for celiac patients. In silico analysis of the recently published sorghum genome predicts that sorghum does not contain peptides that are toxic for celiac patients. Aqueous/alcohol-soluble prolamins (kafirins) from different sorghum varieties, including pure lines and hybrids, were evaluated by SDS-PAGE and HPLC analyses as well as an established enzyme-linked immunosorbent assay (ELISA) based on the R5 antibody. These analyses provide molecular evidence for the absence of toxic gliadin-like peptides in sorghum, confirming that sorghum can be definitively considered safe for consumption by people with celiac disease.

Traumatic brain injury has received significant attention in recent years. Advances in diagnosis and management have resulted in opportunities to improve patient outcomes; however, controversies in diagnosis and management have resulted in increased interactions between the medical and legal communities. This article highlights some of the areas of controversy in traumatic brain injury litigation with the hope that synchronous resolutions of both legal and medical issues will ultimately benefit patient care. It is imperative that the neuroscience community engage the legal community to facilitate an understanding of the issues and their ramifications. Proactive communication and understanding between medical and legal specialties offer the potential to maximize efficiencies in our health care and legal systems., (Copyright © 2010 Elsevier Inc. All rights reserved.)

We have established primary cell culture of the marine demosponge Dysidea avara and Suberites domuncula. Microbial contamination was controlled by the use of a pool of antibiotics confirming the goodness of this procedure. Effect of pH, temperature and light was studied to establish the better growth conditions. The comparison of lipid composition of sponge and cells suggested a series of experiments to optimise the medium. A glucose dose-dependent experiment showed that the ideal glucose concentration is 1 g l(-1). Supplementing the medium with unsaturated fatty acid and retinol, no promotion of growth was observed, but the compounds were totally metabolised by cells. Increments from 70 to 160% in the number of cells were observed, supplementing the medium with different concentration of cholesterol. These results suggest that the analysis of the chemical composition of sponge and cells give indication on the composition of the nutrient media.

The effects of picture-plane rotations on times taken to name familiar objects (RTs) may reflect a process of mental rotation to stored viewpoint-specific representations: the rotate-to-recognize hypothesis. Alternatively, mental rotation might be used after stored object representations are activated by a misoriented stimulus in order to verify a weak or distorted shape percept: the double-checking hypothesis. We tested these two accounts of rotation effects in object recognition by having subjects verify the orientations (to within 90 degrees) and basic-level names of 14-msec, backward-masked depictions of common objects. The stimulus-mask interval (SOA) varied from 14 to 41 msec, permitting interpolation of the SOA required for 75% accuracy (SOAc). Whereas the SOAc to verify orientation increased with rotation up to 180 degrees, the SOAc to verify identity was briefer and asymptoted at approximately 60 degrees. We therefore reject the rotate-to-recognize hypothesis, which implies that SOAc should increase steadily with rotation in both tasks. Instead, we suggest that upright and near-upright stimuli are matched by a fast direct process and that misoriented stimuli are matched at a featural level by a slightly slower view-independent process. We also suggest that rotation effects on RTs reflect a postrecognition stage of orientation verification: the rotate-to-orient hypothesis, a version of double-checking that also explains the well-known reduction in orientation effects on RTs when naming repeated objects.

A novel paradigm, description-depiction classification (DDC), was used to test whether knowledge of object orientation in the picture plane precedes or follows object identification. Undergraduate students were asked to verify the identities and orientations of depicted objects against preceding descriptions, e.g. 'UPRIGHT CAR'. The results showed that identity mismatches were verified faster and more accurately than orientation mismatches, regardless of whether subjects had to discriminate small (90 deg) or large (180 deg) differences in orientation. These findings suggest there is primacy for identity information in that (a) subjects determine object identity before they determine object orientation, and (b) even when orientation is determined, identity information tends to dominate the response. This is the signature of a perceptual-cognitive system that has evolved to rapidly identify objects, not their orientations, contra to theories of recognition that assume orientation is determined before an object has been classified at the basic level.