Oxidative damage in neurodegeneration: roles in the pathogenesis and progression of Alzheimer disease.

Alzheimer disease (AD) is associated with multiple etiologies and pathological mechanisms, among which oxidative stress (OS) appears as a major determinant. Intriguingly, OS arises in various pathways regulating brain functions, and it seems to link different hypotheses and mechanisms of AD neuropathology with high fidelity. The brain is particularly vulnerable to oxidative damage, mainly because of its unique lipid composition, resulting in an amplified cascade of redox reactions that target several cellular components/functions ultimately leading to neurodegeneration. The present review highlights the "OS hypothesis of AD," including amyloid beta-peptide-associated mechanisms, the role of lipid and protein oxidation unraveled by redox proteomics, and the antioxidant strategies that have been investigated to modulate the progression of AD. Collected studies from our groups and others have contributed to unraveling the close relationships between perturbation of redox homeostasis in the brain and AD neuropathology by elucidating redox-regulated events potentially involved in both the pathogenesis and progression of AD. However, the complexity of AD pathological mechanisms requires an in-depth understanding of several major intracellular pathways affecting redox homeostasis and relevant for brain functions. This understanding is crucial to developing pharmacological strategies targeting OS-mediated toxicity that may potentially contribute to slow AD progression as well as improve the quality of life of persons with this severe dementing disorder.

Characterizing the amyloid core region of the tumor suppressor protein p16INK4a using a limited proteolysis and peptide-based approach.

The human tumor suppressor p16INK4a is a small monomeric protein that can form amyloid structures. Formation of p16INK4a amyloid fibrils is induced by oxidation which creates an intermolecular disulfide bond. The conversion into amyloid is associated with a change from an all α-helical structure into ß-sheet fibrils. Currently, structural insights into p16INK4a amyloid fibrils are lacking. Here, we investigate the amyloid-forming regions of this tumor suppressor using isotope-labeling limited-digestion mass spectrometry analysis. We discover two key regions that likely form the structured core of the amyloid. Further investigations using thioflavin-T fluorescence assays, electron microscopy, and solution nuclear magnetic resonance spectroscopy of shorter peptide regions confirm the self-assembly of the identified sequences that include methionine and leucine repeat regions. This work describes a simple approach for studying protein motifs involved in the conversion of monomeric species into aggregated fibril structures. It provides insight into the polypeptide sequence underlying the core structure of amyloid p16INK4a formed after a unique oxidation-driven structural transition.

Near UV and Visible Light-Induced Degradation of Bovine Serum Albumin and a Monoclonal Antibody Mediated by Citrate Buffer and Fe(III): Reduction vs Oxidation Pathways.

Light exposure during manufacturing, storage, and administration can lead to the photodegradation of therapeutic proteins. This photodegradation can be promoted by pharmaceutical buffers or impurities. Our laboratory has previously demonstrated that citrate-Fe(III) complexes generate the •CO2- radical anion when photoirradiated under near UV (λ = 320-400 nm) and visible light (λ = 400-800 nm) [Subelzu, N.; Schöneich, C. Mol. Pharmaceutics 2020, 17 (11), 4163-4179; Zhang, Y. Mol. Pharmaceutics 2022, 19 (11), 4026-4042]. Here, we evaluated the impact of citrate-Fe(III) on the photostability and degradation mechanisms of disulfide-containing proteins (bovine serum albumin (BSA) and NISTmAb) under pharmaceutically relevant conditions. We monitored and localized competitive disulfide reduction and protein oxidation by high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) analysis depending on the reaction conditions. These competitive pathways were affected by multiple factors, including light dose, Fe(III) concentration, protein concentration, the presence of oxygen, and light intensity.

Sensitivity of different organs and tissues as biomarkers of oxidative stress in juvenile tambaqui (Colossoma macropomum) submitted to fasting.

The present study was conducted to evaluate the effects of fasting on responses of oxidative biomarkers and antioxidant defenses using different organs and tissues of Colossoma macropomum. The fish were divided into two groups: fed (control) and fasting (7 days). After 7 days, the fish were sampled for assessment of oxidative stress biomarkers (MDA-lipid peroxidation and PCO-protein carbonyl) and antioxidant defenses (SOD-superoxide dismutase; CAT-catalase; GPX-glutathione peroxidase; and GST-glutathione-S -transferase) in the liver, intestine, gills, muscle, brain, and plasma. The results showed an increase in MDA, PCO, SOD, and GPX concentrations in the liver and intestine of fasting fish. In contrast, in the branchial tissue, there was a reduction in the activity of SOD and CAT enzymes in fasting fish. There was also a reduction in CAT activity in the muscle of fasting fish, while in the brain, there were no changes in oxidative stress biomarkers. Plasma showed a relatively low antioxidant response. In conclusion, our results confirm that a 7-day fasting period induced tissue-specific antioxidant responses, but the increase in antioxidant responses was only for the SOD and GPX enzymes of the liver and intestine. Additionally, the liver and intestine were the most responsive tissues, whereas the plasma was the least sensitive to oxidative stress.

Protein Kinase D Plays a Crucial Role in Maintaining Cardiac Homeostasis by Regulating Post-Translational Modifications of Myofilament Proteins.

Protein kinase D (PKD) enzymes play important roles in regulating myocardial contraction, hypertrophy, and remodeling. One of the proteins phosphorylated by PKD is titin, which is involved in myofilament function. In this study, we aimed to investigate the role of PKD in cardiomyocyte function under conditions of oxidative stress. To do this, we used mice with a cardiomyocyte-specific knock-out of Prkd1, which encodes PKD1 (Prkd1loxP/loxP; αMHC-Cre; PKD1 cKO), as well as wild type littermate controls (Prkd1loxP/loxP; WT). We isolated permeabilized cardiomyocytes from PKD1 cKO mice and found that they exhibited increased passive stiffness (Fpassive), which was associated with increased oxidation of titin, but showed no change in titin ubiquitination. Additionally, the PKD1 cKO mice showed increased myofilament calcium (Ca2+) sensitivity (pCa50) and reduced maximum Ca2+-activated tension. These changes were accompanied by increased oxidation and reduced phosphorylation of the small myofilament protein cardiac myosin binding protein C (cMyBPC), as well as altered phosphorylation levels at different phosphosites in troponin I (TnI). The increased Fpassive and pCa50, and the reduced maximum Ca2+-activated tension were reversed when we treated the isolated permeabilized cardiomyocytes with reduced glutathione (GSH). This indicated that myofilament protein oxidation contributes to cardiomyocyte dysfunction. Furthermore, the PKD1 cKO mice exhibited increased oxidative stress and increased expression of pro-inflammatory markers interleukin (IL)-6, IL-18, and tumor necrosis factor alpha (TNF-α). Both oxidative stress and inflammation contributed to an increase in microtubule-associated protein 1 light chain 3 (LC3)-II levels and heat shock response by inhibiting the mammalian target of rapamycin (mTOR) in the PKD1 cKO mouse myocytes. These findings revealed a previously unknown role for PKD1 in regulating diastolic passive properties, myofilament Ca2+ sensitivity, and maximum Ca2+-activated tension under conditions of oxidative stress. Finally, we emphasized the importance of PKD1 in maintaining the balance of oxidative stress and inflammation in the context of autophagy, as well as cardiomyocyte function.

Volatile Compound Markers in Beef Irradiated with Accelerated Electrons.

This study focuses on the behavior of volatile organic compounds in beef after irradiation with 1 MeV accelerated electrons with doses ranging from 0.25 kGy to 5 kGy to find reliable dose-dependent markers that could be used for establishing an effective dose range for beef irradiation. GC/MS analysis revealed that immediately after irradiation, the chemical yield and accumulation rate of lipid oxidation-derived aldehydes was higher than that of protein oxidation-derived aldehydes. The nonlinear dose-dependent relationship of the concentration of volatile organic compounds was explained using a mathematical model based on the simultaneous occurrence of two competing processes: decomposition of volatile compounds due to direct and indirect action of accelerated electrons, and accumulation of volatile compounds due to decomposition of other compounds and biomacromolecules. A four-day monitoring of the beef samples stored at 4 °C showed that lipid oxidation-derived aldehydes, protein oxidation-derived aldehydes and alkanes as well as alcohol ethanol as an indicator of bacterial activity were dose-dependent markers of biochemical processes occurring in the irradiated beef samples during storage: oxidative processes during direct and indirect action of irradiation, oxidation due to the action of reactive oxygen species, which are always present in the product during storage, and microbial-enzymatic processes. According to the mathematical model of the change in the concentrations of lipid oxidation-derived aldehydes over time in the beef samples irradiated with different doses, it was found that doses ranging from 0.25 kGy to 1 kGy proved to be most effective for beef irradiation with accelerated electrons, since this dose range decreases the bacterial content without considerable irreversible changes in chemical composition of chilled beef during storage.

Natural antioxidants (rosemary and parsley) in microwaved ground meat patties: effects of in vitro digestion.

BACKGROUND: Minimizing food oxidation remains a challenge in several environments. The addition of rosemary extract (150 mg kg-1) and lyophilized parsley (7.1 g kg-1) at equivalent antioxidant activity (5550 µg Trolox equivalents kg-1) to meat patties was assessed in terms of their effect during microwave cooking and after being subjected to an in vitro digestion process. RESULTS: Regardless of the use of antioxidants, cooking caused a decrease of the fat content as compared to raw samples, without noticing statistical differences in the fatty acid distribution between raw and cooked samples [44%, 47% and 6.8%, of saturated fatty acid (SFA), monounsaturated fatty acid (MUFA) and polyunsaturated fatty acid (PUFA), respectively]. However, the bioaccessible lipid fraction obtained after digestion was less saturated (around 34% SFA) and more unsaturated (35% MUFA +30% PUFA). Cooking caused, in all types of samples, an increased lipid [thiobarbituric acid reactive substances (TBARS)] and protein (carbonyls) oxidation values. The increase of TBARS during in vitro digestion was around 7 mg malondialdehyde (MDA) kg-1 for control and samples with parsley and 4.8 mg MDA kg-1 with rosemary. The addition of parsley, and particularly of rosemary, significantly increased the antioxidant activity (DPPH) of cooked and digested microwaved meat patties. CONCLUSION: Whereas rosemary was effective in minimizing protein oxidation during cooking and digestion as compared to control samples, parsley could only limit it during digestion. Lipid oxidation was only limited by rosemary during in vitro digestion. © 2024 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Emerging challenges and efficacy of polyphenols-proteins interaction in maintaining the meat safety during thermal processing.

Polyphenols are well documented against the inhibition of foodborne toxicants in meat, such as heterocyclic amines, Maillard's reaction products, and protein oxidation, by means of their radical scavenging ability, metal chelation, antioxidant properties, and ability to form protein-polyphenol complexes (PPCs). However, their thermal stability, low polarity, degree of dispersion and polymerization, reactivity, solubility, gel forming properties, low bioaccessibility index during digestion, and negative impact on sensory properties are all questionable at oil-in-water interface. This paper aims to review the possibility and efficacy of polyphenols against the inhibition of mutagenic and carcinogenic oxidative products in thermally processed meat. The major findings revealed that structure of polyphenols, for example, molecular size, no of substituted carbons, hydroxyl groups and their position, sufficient size to occupy reacting sites, and ability to form quinones, are the main technical points that affect their reactivity in order to form PPCs. Following a discussion of the future of polyphenols in meat-based products, this paper offers intervention strategies, such as the combined use of food additives and hydrocolloids, processing techniques, precursors, and structure-binding relationships, which can react synergistically with polyphenols to improve their effectiveness during intensive thermal processing. This comprehensive review serves as a valuable source for food scientists, providing insights and recommendations for the appropriate use of polyphenols in meat-based products.

Chemical and physical changes induced by cold plasma treatment of foods: A critical review.

Cold plasma treatment is an innovative technology in the food processing and preservation sectors. It is primarily employed to deactivate microorganisms and enzymes without heat and chemical additives; hence, it is often termed a "clean and green" technology. However, food quality and safety challenges may arise during cold plasma processing due to potential chemical interactions between the plasma reactive species and food components. This review aims to consolidate and discuss data on the impact of cold plasma on the chemical constituents and physical and functional properties of major food products, including dairy, meat, nuts, fruits, vegetables, and grains. We emphasize how cold plasma induces chemical modification of key food components, such as water, proteins, lipids, carbohydrates, vitamins, polyphenols, and volatile organic compounds. Additionally, we discuss changes in color, pH, and organoleptic properties induced by cold plasma treatment and their correlation with chemical modification. Current studies demonstrate that reactive oxygen and nitrogen species in cold plasma oxidize proteins, lipids, and bioactive compounds upon direct contact with the food matrix. Reductions in nutrients and bioactive compounds, including polyunsaturated fatty acids, sugars, polyphenols, and vitamins, have been observed in dairy products, vegetables, fruits, and beverages following cold plasma treatment. Furthermore, structural alterations and the generation of volatile and non-volatile oxidation products were observed, impacting the color, flavor, and texture of food products. However, the effects on dry foods, such as seeds and nuts, are comparatively less pronounced. Overall, this review highlights the drawbacks, challenges, and opportunities associated with cold plasma treatment in food processing.

Endogenous reactive oxygen species (ROS)-driven protein oxidation regulates emulsifying and foaming properties of liquid egg white.

Highly oxidative reactive oxygen species (ROS) attack protein structure and regulate its functional properties. The molecular structures and functional characteristics of egg white (EW) protein (EWP) during 28 d of aerobic or anaerobic storage were explored to investigate the "self-driven" oxidation mechanism of liquid EW mediated by endogenous ROS signaling. Results revealed a significant increase in turbidity during the storage process, accompanied by protein crosslinking aggregation. The ROS yield initially increased and then decreased, leading to a substantial increase in carbonyl groups and tyrosine content. The free sulfhydryl groups and molecular flexibility in EWP exhibited synchronicity with ROS production, reflecting the self-repairing ability of cysteine residues in EWP. Fourier-transform infrared spectroscopy indicated stable crosslinking between EWP molecules in the early oxidation stage. However, continuous ROS attacks accelerated EWP degradation. Compared with the control group, the aerobic-stimulated EWP showed a significant decrease in foaming capacity from 30.5 % to 9.6 %, whereas the anaerobic-stimulated EWP maintained normal levels. The emulsification performance exhibited an increasing-then-decreasing trend. In conclusion, ROS acted as the predominant factor causing deterioration of liquid EW, triggering moderate oxidation that enhanced the superior foaming and emulsifying properties of EWP, and excessive oxidation diminished the functional characteristics by affecting the molecular structure.

Methionine oxidation of clusterin in Alzheimer's disease and its effect on clusterin's binding to beta-amyloid.

Clusterin is a secreted glycoprotein that participates in multiple physiological processes through its chaperon function. In Alzheimer's disease, the brain functions under an increased oxidative stress condition that causes an elevation of protein oxidation, resulting in enhanced pathology. Accordingly, it is important to determine the type of human brain cells that are mostly prone to methionine oxidation in Alzheimer's disease and specifically monitoring the methionine-oxidation levels of clusterin in human and mice brains and its effect on clusterin's function. We analyzed the level of methionine sulfoxide (MetO)-clusterin in these brains, using a combination of immunoprecipitation and Western-blott analyses. Also, we determine the effect of methionine oxidation on clusterin ability to bind beta-amyloid, in vitro, using calorimetric assay. Our results show that human neurons and astrocytes of Alzheimer's disease brains are mostly affected by methionine oxidation. Moreover, MetO-clusterin levels are elevated in postmortem Alzheimer's disease human and mouse brains in comparison to controls. Finally, oxidation of methionine residues of purified clusterin reduced its binding efficiency to beta-amyloid. In conclusion, we suggest that methionine oxidation of brain-clusterin is enhanced in Alzheimer's disease and that this oxidation compromises its chaperon function, leading to exacerbation of beta-amyloid's toxicity in Alzheimer's disease.

The Effects of Four Different Thawing Methods on Quality Indicators of Amphioctopus neglectus.

Amphioctopus neglectus is a species of octopus that is favored by consumers due to its rich nutrient profile. To investigate the influence of different thawing methods on the quality of octopus meat, we employed four distinct thawing methods: air thawing (AT), hydrostatic thawing (HT), flowing water thawing (FWT), and microwave thawing (MT). We then explored the differences in texture, color, water retention, pH, total volatile basic nitrogen (TVB-N), total sulfhydryl content, Ca2+-ATPase activity, and myofibrillar protein, among other quality indicators in response to these methods, and used a low-field nuclear magnetic resonance analyzer to assess the water migration that occurred during the thawing process. The results revealed that AT had the longest thawing time, leading to oxidation-induced protein denaturation, myofibrillar protein damage, and a significant decrease in water retention. Additionally, when this method was utilized, the content of TVB-N was significantly higher than in the other three groups. HT, to a certain extent, isolated the oxygen in the meat and thus alleviated protein oxidation, allowing higher levels of Ca2+-ATPase activity, sulfhydryl content, and springiness to be maintained. However, HT had a longer duration: 2.95 times that of FWT, resulting in a 9.84% higher cooking loss and a 28.21% higher TVB-N content compared to FWT. MT had the shortest thawing time, yielding the lowest content of TVB-N. However, uneven heating and in some cases overcooking occurred, severely damaging the protein structure, with a concurrent increase in thawing loss, W value, hardness, and shear force. Meanwhile, FWT improved the L*, W* and b* values of octopus meat, enhancing its color and water retention. The myofibrillar protein (MP) concentration was also the highest after FWT, with clearer subunit bands in SDS-PAGE electrophoresis, indicating that less degradation occurred and allowing greater springiness, increased Ca2+-ATPase activity, and a higher sulfhydryl content to be maintained. This suggests that FWT has an inhibitory effect on oxidation, alleviating protein oxidation degradation and preserving the quality of the meat. In conclusion, FWT outperformed the other three thawing methods, effectively minimizing adverse changes during thawing and successfully maintaining the quality of octopus meat.

Preservation effects of photodynamic inactivation-mediated antibacterial film on storage quality of salmon fillets: Insights into protein quality.

The preservation effects of a photodynamic inactivation (PDI)-mediated polylactic acid/5-aminolevulinic acid (PLA/ALA) film on the storage quality of salmon fillets were investigated. Results showed that the PDI-mediated PLA/ALA film could continuously generate reactive oxygen species by consuming oxygen to inactivate native pathogens and spoilage bacteria on salmon fillets. Meanwhile, the film maintained the content of muscle proteins and their secondary and tertiary structures, as well as the integrity of myosin by keeping the activity of Ca2+-ATPase, all of which protected the muscle proteins from degradation. Furthermore, the film retained the activity of total superoxide dismutase (T-SOD), suppressed the accumulation of lipid peroxides (e.g., MDA), which greatly inhibited four main types of protein oxidations. As a result, the content of flavor amino acids and essential amino acids in salmon fillets was preserved. Therefore, the PDI-mediated antimicrobial packaging film greatly preserves the storage quality of aquatic products by preserving the protein quality.

Effect of constant-current pulsed electric field thawing on proteins and water-holding capacity of frozen porcine longissimus muscle.

This study explored the effect of constant-current pulsed electric field thawing (CC-T) on the proteins and water-holding capacity of pork. Fresh meat (FM), and frozen meat after constant-voltage thawing (CV-T), air thawing (AT) and water immersion thawing (WT) were considered as controls. The results indicated that CC-T had a higher thawing rate than conventional thawing during ice-crystal melting stage (-5 to -1 °C). It also showed a lower water migration and thawing loss, maintaining pH and shear force closer to FM. Meanwhile, CC-T decreased myoglobin oxidation, resulting in a favorable surface color. The results of protein solubility, differential scanning calorimetry, total sulfhydryl, carbonyl and surface hydrophobicity demonstrated that CC-T reduced myofibrillar protein oxidative denaturation by suppressing the formation of disulfide and carbonyl bonds, thus enhancing solubility and thermal stability. Additionally, microstructural observation found that CC-T maintained a relatively intact muscle fiber structure by reducing muscle damage and myosin filament denaturation.

Effect of three unsaturated fatty acids on the protein oxidation and structure of myofibrillar proteins from rainbow trout (Oncorhynchus mykiss).

In this study, the impact of three unsaturated fatty acids (Oleic acid: OA, Eicosapentaenoic acid: EPA, Docosahexaenoic acid: DHA) on the oxidation and structure of rainbow trout myofibrillar protein (MP) was explored. The findings revealed a notable increase in carbonyl content (P < 0.05) and a significant decrease in total sulfhydryl content (P < 0.05) of MP with the concentration increase of the three unsaturated fatty acids. Endogenous fluorescence spectroscopy and surface hydrophobicity analyses showed that unsaturated fatty acids can cause unfolding and exposure of hydrophobic groups in MP. In addition, SDS-PAGE showed that disulfide bonds were associated with MP cross-linking and aggregate size induced by unsaturated fatty acids. Overall, three unsaturated fatty acid treatments facilitated the oxidation of myofibrillar proteins, and the extent of protein oxidation was closely associated with the concentration of unsaturated fatty acids.

Metabolomic profile of muscles from tilapia cultured in recirculating aquaculture systems and traditional aquaculture in ponds and protein stability during freeze-thaw cycles.

Muscle protein stability during freeze-thaw (F-T) cycles was investigated with tilapia cultured in recirculating aquaculture systems (RAS) and traditional aquaculture in ponds (TAP). This study found that fatty acids (eg., palmitic acid) were enriched in TAP, while antioxidants (eg., glutathione) were enriched in RAS. Generally, proteins in the RAS group exhibited greater stability against denaturation during the F-T cycle, suggested by a less decrease in haem protein content (77% in RAS and 86% in TAP) and a less increase in surface hydrophobicity of sarcoplasmic protein (63% in RAS and 101% in TAP). There was no significant difference in oxidative stability of myofibrillar protein between the two groups. This study provides a theoretical guide for the quality control of tilapia cultured in RAS during frozen storage.

A solution for fillet quality: Slaughter age's effect on protein mechanism and oxidation.

Physico-chemical properties of fish flesh are reliable predictors of fillet quality and nutritional value. In our study, the age-related variations of the chemical composition, pH, water activity (aw), water holding capacity (WHC), color and texture analysis, protein thermal stability, myofibrillar fragmentation index (MFI), glycogen content, protein oxidation and protein profiles were investigated in Oncorhynchus mykiss (rainbow trout) fillet. The results revealed that protein denaturation temperatures (Tmax1 and Tmax2) decreased by 2 % and 11.6 % depending on fish age. Tmax1 and Tmax2 values in the same groups were raised 71 % at 11 months' fish and this increase was 58 % at 23 months' fish. An age-related reduction by 66.6 % and 31.25 % was noticed for protein oxidation markers sulfhydryl groups and disulfide bonds. MFI value increased by 86.6 % connected with age. The characteristics of fish meat quality are complex and are influenced by various factors that affect the degree of freshness of the product and its acceptance in the market. Taking into account the different demands of the consumer, this study has shown that age at slaughter has an impact on final product quality and that the recommended age at slaughter, taking into account market weight, positively affects meat quality.

Effects of Different Heat Treatments on Yak Milk Proteins on Intestinal Microbiota and Metabolism.

Dairy products are susceptible to modifications in protein oxidation during heat processing, which can lead to changes in protein function, subsequently affecting intestinal health. Despite being a unique nutritional source, yak milk has not been thoroughly examined for the effects of its oxidized proteins on intestinal microbiota and metabolism. Hence, this study employed different heat treatment methods (low-temperature pasteurization, high-temperature pasteurization, and high-temperature sterilization) to induce oxidation in yak milk proteins. The study then assessed the degree of oxidation in these proteins and utilized mice as research subjects. Using metagenomics and metabolomics methods, this study examined the structure of intestinal microbial communities and metabolic products in mice consuming oxidized yak milk. The results showed a decrease in carbonyl and total thiol contents of yak milk proteins after different heat treatments, indicating that heat treatment causes oxidation in yak milk proteins. Metagenomic analysis of mouse intestinal microbiota revealed significant changes in 66 genera. In the high-temperature sterilization group (H), key differential genera included Verrucomicrobiales, Verrucomicrobiae, Akkermansiaceae, and 28 others. The high-temperature pasteurization group (M) mainly consisted of Latilactobacillus, Bacillus, and Romboutsia. The low-temperature pasteurization group (L) primarily comprised of Faecalibacterium, Chaetomium, Paenibacillaceae, Eggerthella, Sordariales, and 33 others. Functionally, compared to the control group (C), the H group upregulated translation and energy metabolism functions, the L group the M group significantly upregulated metabolism of other amino acids, translation, and cell replication and repair functions. Based on metabolomic analysis, differential changes in mouse metabolites could affect multiple metabolic pathways in the body. The most significantly affected metabolic pathways were phenylalanine metabolism, vitamin B6 metabolism, steroid hormone biosynthesis, and pantothenate and CoA biosynthesis. The changes were similar to the functional pathway analysis of mouse metagenomics, affecting amino acid and energy metabolism in mice. In summary, moderate oxidation of yak milk proteins exhibits a positive effect on mouse intestinal microbiota and metabolism. In conclusion, yak milk has a positive effect on mouse intestinal microflora and metabolism, and this study provides a scientific basis for optimizing dairy processing technology and further developing and applying yak milk.

Effects of deep, air and vacuum frying on oyster quality and protein-mediated mechanism analysis via TMT quantitative proteomics.

Fried oyster is a popular aquatic food product in East Asia, but nutrient loss during thermal processing become a significant concern. The goal of this research was to examine the impact of distinct frying techniques, including deep frying (DF), air frying (AF), and vacuum frying (VF), on the nutritional, textural and flavor characteristics of oysters. The VF method demonstrated superior retention of beneficial properties and flavor, and reduced protein and lipid oxidation compared to the DF and AF methods. Furthermore, proteomic analysis of oysters was attempted to explain the molecular mechanisms governing the influence of key differential proteins. 20 major differential proteins, including actin-2 protein, tryptophan 2,3-dioxygenase and 1-alph, involved in oyster protein oxidation were identified, annotated and analyzed to elucidate their influence mechanisms. This research provides a deeper understanding of intricate interactions between frying techniques and oyster biochemistry, which offers valuable implications for enhancing food quality in seafood industry.

Cysteine Is the Only Universally Affected and Disfavored Proteomic Amino Acid under Oxidative Conditions in Animals.

Oxidative modifications of amino acid side chains in proteins are a hallmark of oxidative stress, and they are usually regarded as structural damage. However, amino acid oxidation may also have a protective effect and may serve regulatory or structural purposes. Here, we have attempted to characterize the global redox role of the 20 proteinogenic amino acids in animals by analyzing their usage frequency in 5 plausible evolutionary paradigms of increased oxidative burden: (i) peroxisomal proteins versus all proteins, (ii) mitochondrial proteins versus all proteins, (iii) mitochondrially encoded respiratory chain proteins versus all mitochondrial proteins, (iv) proteins from long-lived animals versus those from short-lived animals, and (v) proteins from aerobic, free-living animals versus those from facultatively anaerobic animals. We have found that avoidance of cysteine in the oxidative condition was the most pronounced and significant variation in the majority of comparisons. Beyond this preeminent pattern, only local signals were observed, primarily increases in methionine and glutamine as well as decreases in serine and proline. Hence, certain types of cysteine oxidation appear to enforce its proteome-wide evolutionary avoidance despite its essential role in disulfide bond formation and metal ligation. The susceptibility to oxidation of all other amino acids appears to be generally unproblematic, and sometimes advantageous.