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Astaxanthin (AST) is a natural marine carotenoid with a variety of biological activities. This study aimed to demonstrate the possible mechanisms by which AST improves skeletal muscle atrophy in cancer cachexia. In this study, the effects of different doses of AST (30 mg/kg b.w., 60 mg/kg b.w. and 120 mg/kg b.w.) on skeletal muscle functions were explored in mice with cancer cachexia. The results showed that AST (30, 60 and 120 mg/kg b.w.) could effectively protect cachexia mice from body weight and skeletal muscle loss. AST dose-dependently ameliorated the decrease in myofibres cross-sectional area and increased the expression of myosin heavy chain (MHC). AST treatment decreased both the serum and muscle level of IL-6 but not TNF-α in C26 tumor-bearing cachexia mice. Moreover, AST alleviated skeletal muscle atrophy by decreasing the expression of two muscle-specific E3 ligases MAFBx and MuRF-1. AST improved mitochondrial function by downregulating the levels of muscle Fis1, LC3B and Bax, upregulating the levels of muscle Mfn2 and Bcl-2. In conclusion, our study show that AST might be expected to be a nutritional supplement for cancer cachexia patients.
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Caquexia , Músculo Esquelético , Atrofia Muscular , Xantofilas , Animais , Xantofilas/farmacologia , Caquexia/tratamento farmacológico , Caquexia/etiologia , Atrofia Muscular/tratamento farmacológico , Atrofia Muscular/etiologia , Músculo Esquelético/efeitos dos fármacos , Músculo Esquelético/metabolismo , Músculo Esquelético/patologia , Camundongos , Masculino , Proteínas Musculares/metabolismo , Interleucina-6/metabolismo , Camundongos Endogâmicos BALB C , Proteínas Ligases SKP Culina F-Box/metabolismo , Proteínas Ligases SKP Culina F-Box/genética , Neoplasias/complicações , Neoplasias/tratamento farmacológico , Fator de Necrose Tumoral alfa/metabolismo , Proteínas com Motivo Tripartido/metabolismo , Proteínas com Motivo Tripartido/genética , Cadeias Pesadas de Miosina/metabolismo , Ubiquitina-Proteína Ligases/metabolismo , Ubiquitina-Proteína Ligases/genética , Linhagem Celular TumoralRESUMO
Alginate is a commercially important polysaccharide composed of mannuronic acid and its C5 differential isomer guluronic acid. Comprehensive research on alginate and alginate lyases requires efficient and precise analytical methods for alginate oligosaccharides. In this research, high-performance anion exchange chromatography (HPAEC) in parallel with pulsed amperometric detection (PAD) and mass spectrometry (MS) was applied to the analysis of oligosaccharides obtained by alginate lyase. By optimizing the chromatographic conditions including mobile phase concentration, flow rate, and elution gradient, the analysis of a single sample could be completed in 30 min. Seven unsaturated alginate oligosaccharides were separated and identified through their analysis time observed with PAD, including all structurally different unsaturated disaccharides and trisaccharides. The quantitative analysis of seven oligosaccharides was performed based on the quantitative capability of PAD. The method exhibited adequate linearity and precision parameters. All the calibration curves showed good linearity at least in the concentration range of 0.002 to 0.1 mg/mL. The HPAEC-PAD/MS method provides a general and efficient online method to analyze alginate oligosaccharides.
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Alginatos , Espectrometria de Massas , Oligossacarídeos , Alginatos/química , Oligossacarídeos/análise , Oligossacarídeos/química , Cromatografia por Troca Iônica/métodos , Espectrometria de Massas/métodos , Cromatografia Líquida de Alta Pressão/métodos , Polissacarídeo-Liases/química , Polissacarídeo-Liases/metabolismo , Ácidos Hexurônicos/química , Ácidos Hexurônicos/análise , Limite de DetecçãoRESUMO
Gangliosides play an imperative role in cell signaling, neuronal recovery, apoptosis, and other physiological processes. For example, GM3 can regulate hypothalamic leptin resistance and control energy homeostasis, GD3 can mediate cell proliferation and differentiation and induce apoptosis, and GQ1b can stimulate neurogenesis. Therefore, the present study sought to establish and optimize the targeted analysis method for ganglioside subclasses and their molecular species using hydrophilic interaction liquid chromatography-triple quadrupole-MS/MS (HILIC-QQQ-MS/MS). Additionally, the fragmentation pattern of different ganglioside subclasses and their retention time patterns were analyzed, providing more accurate qualitative results. The limit of quantitation (LOQ) was as low as 10-4 ng. Moreover, the molecular species of gangliosides in the liver, cortex, and hypothalamus of C57BL/6 mice were analyzed using the established method. A total of 23 ganglioside subclasses with 164 molecular species, including 40 O-acetylated ganglioside molecular species and 28 NeuGc ganglioside molecular species, were identified using the semi-quantitative analysis method of an external standard curve corrected by an internal standard. In addition to NeuGc gangliosides, the contents of ganglioside subclasses were more abundant in the mouse brain than those in the mouse liver; especially, the contents of unsaturated gangliosides in the hypothalamus were much higher than those in the liver. Among them, O-acetylated gangliosides were detected only in the cortex and hypothalamus at a concentration of up to 100 µg/mg protein (40 molecular species). Overall, the proposed method expanded the detectable number of ganglioside subclasses and molecular species in biological samples and provided more opportunities for further study of the biological functions of gangliosides.
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Patients with ulcerative colitis (UC) have higher rates of depression. However, the mechanism of depression development remains unclear. The improvements of EPA and DHA on dextran sulfate sodium (DSS)-induced UC have been verified. Therefore, the present study mainly focused on the effects of EPA and DHA on UC-induced depression in C57BL/6 mice and the possible mechanisms involved. A forced swimming test and tail suspension experiment showed that EPA and DHA significantly improved DSS-induced depressive-like behavior. Further analysis demonstrated that EPA and DHA could significantly suppress the inflammation response of the gut and brain by regulating the NLRP3/ASC signal pathway. Moreover, intestine and brain barriers were maintained by enhancing ZO-1 and occludin expression. In addition, EPA and DHA also increased the serotonin (5-HT) concentration and synaptic proteins. Interestingly, EPA and DHA treatments increased the proportion of dominant bacteria, alpha diversity, and beta diversity. In conclusion, oral administration of EPA and DHA alleviated UC-induced depressive-like behavior in mice by modulating the inflammation, maintaining the mucosal and brain barriers, suppressing neuronal damage and reverting microbiota changes.
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Colite Ulcerativa , Humanos , Camundongos , Animais , Sulfato de Dextrana/toxicidade , Camundongos Endogâmicos C57BL , Colite Ulcerativa/metabolismo , Transdução de Sinais , Inflamação/metabolismo , Modelos Animais de Doenças , Colo/metabolismoRESUMO
BACKGROUND: α-l-Fucose confers unique functions for fucose-containing biomolecules such as human milk oligosaccharides. α-l-Fucosidases can serve as desirable tools in the application of fucosylated saccharides. Discovering novel α-l-fucosidases and elucidating their enzyme properties are always worthy tasks. RESULTS: A GH95 family α-l-fucosidase named Afc95A_Wf was cloned from the genome of the marine bacterium Wenyingzhuangia fucanilytica and expressed in Escherichia coli. It exhibited maximum activity at 40 °C and pH 7.5. Afc95A_Wf defined a different substrate specificity among reported α-l-fucosidases, which was capable of hydrolyzing α-fucoside in CNP-fucose, Fucα1-2Galß1-4Glc and Galß1-4(Fucα1-3)Glc, and showed a preference for α1,2-fucosidic linkage. It adopted Asp residue in the amino acid sequence at position 391, which was distinct from the previously acknowledged residue of Asn. The predicted tertiary structure and site-directed mutagenesis revealed that Asp391 participates in the catalysis of Afc95A_Wf. The differences in the substrate specificity and catalytic site shed light on that Afc95A_Wf adopted a novel mechanism in catalysis. CONCLUSION: A GH95 family α-l-fucosidase (Afc95A_Wf) was cloned and expressed. It showed a cleavage preference for α1,2-fucosidic linkage to α1,3-fucosidic linkage. Afc95A_Wf demonstrated a different substrate specificity and a residue at an important catalytic site compared with known GH95 family proteins, which revealed the occurrence of diversity on catalytic mechanisms in the GH95 family. © 2024 Society of Chemical Industry.
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BACKGROUND: Alginate lyases are important tools for alginate biodegradation and oligosaccharide production, which have great potential in food and biofuel fields. The alginate polysaccharide utilization loci (PUL) typically encode a series of alginate lyases with a synergistic action pattern. Exploring valuable alginate lyases and revealing the synergistic effect of enzymes in the PUL is of great significance. RESULTS: An alginate PUL was discovered from the marine bacterium Wenyingzhuangia fucanilytica CZ1127T , and a repertoire of alginate lyases within it was cloned, expressed and characterized. The four alginate lyases in PUL demonstrated similar optimal reaction conditions: maximum enzyme activity at 35-50 °C and pH 8.0-9.0. The results of action pattern indicated that they were two PL7 endolytic bifunctional enzymes (Aly7A and Aly7B), a PL6 exolytic bifunctional enzyme (Aly6A) and a PL17 exolytic M-specific enzyme (Aly17A). Ultra-performance liquid chromatography-mass spectrometry was employed to reveal the synergistic effect of the four enzymes. The end products of Aly7A were further degraded by Aly7B and eventually generated oligosaccharides, from disaccharide to heptasaccharide. The oligosaccharide products were completely degraded to monosaccharides by Aly6A, but it was unable to directly degrade alginate. Aly17A could also produce monosaccharides by cleaving the M-blocks of oligosaccharide products, as well as the M-blocks of polysaccharides. The combination of these enzymes resulted in the complete degradation of alginate to monosaccharides. CONCLUSION: A new alginate PUL was mined and four novel alginate lyases in the PUL were expressed and characterized. The four cooperative alginate lyases provide novel tools for alginate degradation and biological fermentation. © 2023 Society of Chemical Industry.
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Alginatos , Flavobacteriaceae , Alginatos/metabolismo , Flavobacteriaceae/metabolismo , Monossacarídeos , Oligossacarídeos/metabolismo , Especificidade por Substrato , Concentração de Íons de HidrogênioRESUMO
BACKGROUND: Lack of n-3 polyunsaturated fatty acids during the period of maternity drastically lowers the docosahexaenoic acid (DHA) level in the brain of offspring and studies have demonstrated that different molecular forms of DHA are beneficial to brain development. The aim of this study was to investigate the effect of short-term supplementation with DHA-enriched phosphatidylserine (PS) and phosphatidylcholine (PC) on DHA levels in the liver and brain of congenital n-3-deficient mice. RESULTS: Dietary supplementation with DHA significantly changed the fatty acid composition of various phospholipid molecules in the cerebral cortex and liver while DHA-enriched phospholipid was more effective than DHA triglyceride (TG) in increasing brain and liver DHA. Both DHA-PS and DHA-PC could effectively increase the DHA levels, but DHA in the PS form was superior to PC in the contribution of DHA content in the brain ether-linked PC (ePC) and liver lyso-phosphatidylcholine molecular species. DHA-PC showed more significant effects on the increase of DHA in liver TG, PC, ePC, phosphatidylethanolamine (PE) and PE plasmalogen (pPE) molecular species and decreasing the arachidonic acid level in liver PC plasmalogen, ePC, PE and pPE molecular species compared with DHA-PS. CONCLUSION: The effect of dietary interventions with different molecular forms of DHA for brain and liver lipid profiles is different, which may provide theoretical guidance for dietary supplementation of DHA for people. © 2024 Society of Chemical Industry.
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To enhance the bioavailability of bioactives with varying efficacy in the gastrointestinal tract (GIT), a co-delivery system of solid-in-oil-in-water (S/O/W) emulsion was designed for the co-encapsulation of two bioactives in this paper. S/O/W emulsions were fabricated utilizing fucoxanthin (FUC)-loaded nanoparticles (NPs) as the solid phase, coconut oil containing curcumin (Cur) as the oil phase, and carboxymethyl starch (CMS)/propylene glycol alginate (PGA) complex as the aqueous phase. The high entrapment efficiency of Cur (82.3-91.3%) and FUC (96.0-96.1%) was found in the CMS/PGA complex-stabilized S/O/W emulsions. Encapsulation of Cur and FUC within S/O/W emulsions enhanced their UV and thermal stabilities. In addition, S/O/W emulsions prepared with CMS/PGA complexes displayed good stability. More importantly, the formed S/O/W emulsion possessed programmed sequential release characteristics, delivering Cur and FUC to the small intestine and colon, respectively. These results contributed to designing co-delivery systems for the programmed sequential release of two hydrophobic nutrients in the GIT.
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Curcumina , Emulsões , Xantofilas , Emulsões/química , Curcumina/química , Xantofilas/química , Composição de Medicamentos , Nanopartículas/química , Tamanho da Partícula , Estabilidade de Medicamentos , Água/química , Sistemas de Liberação de Medicamentos , Portadores de Fármacos/químicaRESUMO
The purpose of this study was to compare the influences of gamma-poly glutamic acid (γ-PGA) (1, 2, 3, and 4 %) to see which could outperform conventional cryoprotectant mixture (4 % sorbitol + 4 % sucrose) on cooked crayfish properties, such as physicochemical, textural qualities, oxidation reaction, water distributions, and microstructure integrity, during different freeze-thaw cycles. Crayfish quality characteristics improved significantly as γ-PGA concentration increased compared to control samples.Adding γ-PGA 4 % reduced the carbonyl content from 4.20 to 3.00 nmol/ mg protein during fluctuation-1 (F1), and from 4.15 to 2.80 nmol/ mg protein during fluctuation-2 (F2) compared to control samples. Furthermore, it increased the total sulfhydryl content from 4.15 and 4.76 to 6.19 and 6.47 mol/105 g protein during F1 and F2 and after five freeze-thaw cycles (FTC). This suggests that this concentration was more effective at controlling protein changes than other concentrations. γ-PGA generally enhanced the water-holding capacity by preventing protein denaturation and limiting ice crystal recrystallization. As a result, microstructure stability was evident, texture degradation was avoided, and the crayfish's color was preserved.
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Astacoidea , Ácido Poliglutâmico/análogos & derivados , Água , Animais , Temperatura , Congelamento , Água/químicaRESUMO
In order to overcome the problem that traditional W1/O/W2 double emulsions do not have targeted release performance, thereby better meeting the health needs of consumers, ovalbumin fibrils/pectin-based bilayer-stabilized double emulsion (OP-BDE) co-encapsulated with Lactobacillus plantarum and curcumin was constructed with pectin as the outer protective shell, which was expected to be used in the development of novel functional foods. The effects of pectin coating on the viability of Lactobacillus plantarum under conditions including storage, pasteurization, freeze-thaw cycles and in vitro simulated digestion were investigated. Results showed that pectin as protective shell could significantly enhance the tolerance of Lactobacillus plantarum to various environmental factors. Besides, the adsorption of pectin endowed OP-BDE with higher lipolysis and stronger protective effect on curcumin, remarkably improving the photostability and bioaccessibility of curcumin. In addition, in vitro simulated gastrointestinal release study indicated that OP-BDE possessed programmed sequential release property, allowing curcumin and Lactobacillus plantarum to be released in small intestine and colon, respectively. OP-BDE is the first reported co-delivery emulsion system with programmed release characteristic. This study provides new insights into OP-BDE in constructing co-delivery systems and programmed sequential release of active substances, and has potential reference and application value in actual food production.
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Curcumina , Lactobacillus plantarum , Emulsões , Curcumina/farmacologia , Pectinas , Trato GastrointestinalRESUMO
Porphyran is a favorable functional polysaccharide widely distributed in Porphyra. It displays a linear structure majorly constituted by alternating 1,4-linked α-l-galactopyranose-6-sulfate (L6S) and 1,3-linked ß-d-galactopyranose (G) units. Carbohydrate-binding modules (CBMs) are desired tools for the investigation and application of polysaccharides, including in situ visualization, on site and specific assay, and functionalization of biomaterials. However, only one porphyran-binding CBM has been hitherto reported, and its structural knowledge is lacking. Herein, a novel CBM16 family domain from a marine bacterium Aquimarina sp. BL5 was discovered and expressed. The recombinant protein AmCBM16 exhibited the desired specificity for porphyran. Bio-layer interferometry assay revealed that the protein binds to porphyran tetrasaccharide (L6S-G)2 with an association constant of 1.3 × 103 M-1. The structure of AmCBM16 was resolved by the X-ray crystallography, which displays a ß-sandwich fold with two antiparallel ß-sheets constituted by 10 ß-strands. Site-directed mutagenesis analysis demonstrated that the residues Gly-30, Trp-31, Lys-88, Lys-123, Phe-125, and Phe-127 play dominant roles in AmCBM16 binding. This study provides the first structural insights into porphyran-binding CBM.
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Flavobacteriaceae , Galactose , Sefarose/análogos & derivados , Sítios de Ligação , Proteínas de Bactérias/química , Polissacarídeos/química , Flavobacteriaceae/metabolismo , Cristalografia por Raios XRESUMO
Fibrillin is an important structural protein in connective tissues. The presence of fibrillin in sea cucumber Apostichopus japonicus is still poorly understood, which limits our understanding of the role of fibrillin in the A. japonicus microstructure. The aim of this study was to clarify the presence of fibrillin in the sea cucumber A. japonicus body wall. Herein, the presence of fibrillin in sea cucumber A. japonicus was investigated by utilizing targeted proteomics and visualization strategies. The contents of three different isoforms of fibrillin with high abundance in A. japonicus were determined to be 0.96, 2.54, and 0.15 µg/g (wet base), respectively. The amino acid sequence of fibrillin (GeneBank number: PIK56741.1) that started at position 631 and ended at position 921 was selected for cloning and expressing antigen. An anti-A. japonicus fibrillin antibody with a titer greater than 1:64â¯000 was successfully obtained. It was observed that the distribution of fibrillin in the A. japonicus body wall was scattered and dispersed in the form of fibril bundles at the microscale. It further observed that fibrillin was present near collagen fibrils and some entangled outside the collagen fibrils at the nanoscale. Moreover, the stoichiometry of the most dominant collagen and fibrillin molecules in A. japonicus was determined to be approximately 250:1. These results contribute to an understanding of the role of fibrillin in the sea cucumber microstructure.
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Pepinos-do-Mar , Stichopus , Animais , Stichopus/genética , Stichopus/química , Pepinos-do-Mar/metabolismo , Proteômica , Fibrilinas , Colágeno/químicaRESUMO
Microplastics have been an emerging threat to filtering species and the ingestion and impacts of microplastics on oysters are a cause for concern. However, much remains unknown about the effects of microplastics on flavor-related biomarkers in oysters. Herein, a laboratory microplastic exposure with concentrations of 1, 10, and 100 mg/L for 15 days was performed to investigate the impacts of microplastics on the flavor parameters of oysters. Exposure to microplastics changed the odor characteristics of oysters. Microplastic exposure had minor effects on the fatty acid composition; however, significant alterations in free amino acids and nucleotides were observed under the 1 and 10 mg/L exposure groups, respectively. The overall results indicated 10 mg/L of microplastic exposure significantly increased the equivalent umami value of oysters. These findings stressed the effects of microplastics on oysters and would be an important reference for the assessment of the potential risks associated with microplastics in marine edible species.
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The process of producing cell-cultured meat involves utilizing a significant amount of culture medium, including fetal bovine serum (FBS), which represents a considerable portion of production expense while also raising environmental and safety concerns. This study demonstrated that supplementation with Auxenochlorella pyrenoidosa protein extract (APE) under low-serum conditions substantially increased Carassius auratus muscle (CAM) cell proliferation and heightened the expression of Myf5 compared to the absence of APE. An integrated intracellular metabolomics and proteomics analysis revealed a total of 13 and 67 differentially expressed metabolites and proteins, respectively, after supplementation with APE in the medium containing 5%FBS, modulating specific metabolism and signaling pathways, which explained the application of APE for passage cell culture under low-serum conditions. Further analysis revealed that the bioactive factors in the APE were protein components. Moreover, CAM cells cultured in reconstructed serum-free media containing APE, l-ascorbic acid, insulin, transferrin, selenium, and ethanolamine exhibited significantly accelerated growth in a scale-up culture. These findings suggest a promising alternative to FBS for fish muscle cell culture that can help reduce production costs and environmental impact in the production of cultured meat.
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Hominidae , Soroalbumina Bovina , Animais , Células Cultivadas , Meios de Cultura , Técnicas de Cultura de Células , MúsculosRESUMO
Astaxanthin is a carotenoid that is taken orally and has antitumor and anti-inflammatory properties. Our previous research demonstrated that astaxanthin alleviated skeletal muscle atrophy during sorafenib treatment in H22 tumor-bearing mice and altered the intestinal flora composition. However, the relationship between astaxanthin's amelioration of skeletal muscle atrophy in tumor-bearing mice and its ability to regulate intestinal flora is not clear. We used broad-spectrum antibiotics to create pseudo-sterile tumor-bearing mice, which we then used in fecal bacteria transplantation experiments. Our results indicate that the role of astaxanthin in ameliorating skeletal muscle atrophy during molecularly targeted therapy in mice with tumors is dependent on the intestinal flora. Astaxanthin substantially promoted the proliferation of Blautia, Parabacteroides, and Roseburia, altered the levels of metabolites in mouse serum, and primarily affected the amino acid metabolism of mice. Astaxanthin ameliorated skeletal muscle atrophy by promoting the activation of AKT/FOXO3a, which inhibited the expression of ubiquitination-degrading Fbx32 and MuRF1 and promoted myogenesis in skeletal muscle. Our study confirms that the intestinal flora is an important target for astaxanthin to combat skeletal muscle atrophy. Our research supports the use of astaxanthin as a nutritional supplement and intestinal microecological regulator for cancer patients.
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Microbioma Gastrointestinal , Neoplasias , Camundongos , Humanos , Animais , Sorafenibe , Atrofia Muscular/metabolismo , Músculo Esquelético/metabolismo , Neoplasias/metabolismo , XantofilasRESUMO
This study aimed to investigate the function of gut microbiota in astaxanthin's adjuvant anticancer effects. Our prior research demonstrated that astaxanthin enhanced the antitumor effects of sorafenib by enhancing the body's antitumor immune response; astaxanthin also regulated the intestinal flora composition of tumor-bearing mice. However, it is presently unknown whether this beneficial effect is dependent on the gut microbiota. We first used broad-spectrum antibiotics to eradicate gut microbiota of tumor-bearing mice, followed by the transplantation of fecal microbiota. The results of this study indicate that the beneficial effects of astaxanthin when combined with molecular targeting are dependent on the presence of intestinal microbiota. Astaxanthin facilitates the infiltration of CD8+ T lymphocytes into the tumor microenvironment and increases Granzyme B production by modulating the intestinal flora. Therefore, it strengthens the body's anti-tumor immune response and synergistically boosts the therapeutic efficacy of drugs. Astaxanthin stimulates the production of cuprocytes and mucus in the intestines by promoting the proliferation of Akkermansia. In addition, astaxanthin enhances the intestinal mucosal immunological function. Our research supports the unique ability of astaxanthin to sustain intestinal flora homeostasis and its function as a dietary immune booster for individuals with tumors.
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Microbioma Gastrointestinal , Animais , Camundongos , Imunidade nas Mucosas , Intestinos/patologia , Mucosa Intestinal , XantofilasRESUMO
Trimethylamine N-oxide (TMAO), a characteristic nonprotein nitrogen compound, is widely present in seafood, which exhibits osmoregulatory effects for marine organisms in vivo and plays an important role in aquaculture and aquatic product preservation. However, much attention has been focused on the negative effect of TMAO since it has recently emerged as a putative promoter of chronic diseases. To get full knowledge and maximize our ability to balance the positive and negative aspects of TMAO, in this review, we comprehensively discuss the TMAO in aquatic products from the aspects of physiological functions for marine organisms, flavor, quality, the conversion of precursors, the influences on human health, and the seafood ingredients interaction consideration. Though the circulating TMAO level is inevitably enhanced after seafood consumption, dietary seafood still exhibits beneficial health effects and may provide nutraceuticals to balance the possible adverse effects of TMAO.
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Metilaminas , Alimentos Marinhos , Metilaminas/análise , Metilaminas/metabolismo , Humanos , Animais , Alimentos Marinhos/análise , Organismos Aquáticos/química , PeixesRESUMO
A novel alginate lyase Aly7Aq was cloned and heterologous expressed by a combination of bioinformatics and molecular biology. Aly7Aq was an M-specific alginate lyase, exhibiting optimum reaction conditions at 50 °C and pH 10.0. Aly7Aq was determined to degrade polysaccharides in a random endo-acting manner. The minimum reaction substrate was tetrasaccharide, and Aly7Aq mainly attacked the third glycosidic linkage from the reducing end of oligosaccharide substrates. The disaccharide product of Aly7Aq was ΔM and the trisaccharide products were ΔMM and ΔMG, which differed from all previously characterized M-specific alginate lyases. The degradation products demonstrated that the ±2 subsites of Aly7Aq strictly recognized M units, while the -1 subsite accommodated both M and G units. Therefore, the substrate specificity of Aly7Aq was derived from the specificity of ±2 subsites. This is the first report on the specificity at subsite ±2 of M-specific alginate lyase. The novel M-specific Aly7Aq could serve as a potential tool in the specific degradation of alginate and targeted preparation of oligosaccharide.
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Sea cucumber intestines are considered a valuable resource in the sea cucumber processing industry due to their balanced amino acid composition. Studies have reported that peptides rich in glutamate and branched-chain amino acids have anti-fatigue properties. However, the function of the sea cucumber intestine in reducing exercise-induced fatigue remains unclear. In this study, we enzymatically hydrolyzed low molecular weight peptides from sea cucumber intestines (SCIP) and administered SCIP orally to mice to examine its effects on exercise-induced fatigue using swimming and pole-climbing exhaustion experiments. The results revealed that supplementation with SCIP significantly prolonged the exhaustion time of swimming in mice, decreased blood lactate and urea nitrogen levels, and increased liver and muscle glycogen levels following a weight-loaded swimming test. Immunofluorescence analysis indicated a notable increase the proportion of slow-twitch muscle fiber and a significant decrease the proportion of fast-twitch muscle fiber following SCIP supplementation. Furthermore, SCIP upregulated mRNA expression levels of Ca2+ /Calcineurin upstream and downstream regulators, thereby contributing to the promotion of skeletal muscle fiber type conversion. This study presents the initial evidence establishing SCIP as a potential enhancer of skeletal muscle fatigue resistance, consequently providing a theoretical foundation for the valuable utilization of sea cucumber intestines.
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Calcineurina , Pepinos-do-Mar , Camundongos , Animais , Calcineurina/metabolismo , Calcineurina/farmacologia , Pepinos-do-Mar/metabolismo , Músculo Esquelético/metabolismo , Peptídeos/farmacologia , Natação/fisiologia , Transdução de Sinais , Intestinos , Peptídeo Hidrolases/metabolismoRESUMO
Customers are increasingly opting for ready-to-eat and easy-to-prepare food products, such as cooked crayfish. It is highly valued for its unique taste, tender meat, and nutritional properties. Therefore, we conducted an investigation into its quality parameters over an 8-week period at -20 °C. Parameters such as water distribution, oxidation reactions, color, microstructure, texture properties, and physicochemical parameters were examined. The physicochemical results indicated that as the storage time increased, the levels of pH and TVB-N (total volatile basic nitrogen) showed a significant increase, while the water holding capacity decreased significantly (P < 0.05). After two months of frozen storage, the carbonyl content and TBARS (thiobarbituric acid reactive substances) increased to 4.15 ± 0.16 nmol/mg protein and 1.6 ± 0.00 mg/kg, respectively. Additionally, the total sulfhydryl content decreased to 4.91 ± 0.10 mol/105 g protein, which had an impact on the quality of the crayfish. Electron microscopy revealed that with increasing storage time, the fiber structure gradually deteriorated due to water crystallization, leading to severe damage and breakage of muscle fibers. Interestingly, these changes related to storage affected color and texture parameters, thereby influencing the overall quality of the crayfish.