Oxidase Activity Research Articles

Impact of ultrasonic pretreatment on the color and antioxidant capacity of vacuum freeze-dried strawberries

To enhance the color quality and antioxidant potential of vacuum-freeze-dried strawberry tablets, ultrasonic waves were used as a freezing pretreatment. Results revealed that the * values of freeze − dried strawberries following ultrasonic pretreatment improved significantly by ∼60% and the C* values increased by 34.36%–51.65%. These findings suggest that ultrasonic pretreatment results in a brighter coloration of the strawberry samples. The ultrasound − assisted freezing treatment groups exhibited a significantly higher free radical scavenging ability compared with the nonultrasonic treatment group. This enhancement was attributed to the ultrasonic pretreatment, which increased the concentrations of total phenolic compounds, flavonoids, and anthocyanins, and effectively inhibited the activities of polyphenol oxidase and peroxidase, thereby reducing the content of red pigment in the strawberry samples. Furthermore, confocal laser scanning microscopy revealed a uniform distribution of sugar within the samples following ultrasonic treatment. Cluster analysis indicated that the primary indices influencing the color of the samples were total phenolic content and peroxidase activity. Therefore, the ultrasonic pretreatment method effectively enhances product color, inhibits enzyme activity, and increases anthocyanin content. This technology holds promising development prospects.

An Unusual Application of Multifunctional Nanozyme Derived from COF: Augmenting Chemoimmunotherapy while Attenuating Cardiotoxicity

AbstractEnhancing therapeutic efficacy while reducing the adverse effects of drugs is crucial in cancer treatments. In this study, nitrogen‐doped carbon nanospheres are synthesized with variable enzymatic activities through the pyrolysis of a covalent organic framework (COF). These nanospheres show increased peroxidase (POD) and oxidase (OXD) activities in the tumor microenvironment, generating reactive oxygen species (ROS) that help eliminate tumor cells. Under normal conditions, they display catalase (CAT) and superoxide dismutase (SOD) activities, promoting antioxidative protection. Their porous structure and π–π/hydrophobic interactions allow for the loading of the chemotherapeutic agent doxorubicin (DOX). The nanospheres are then enveloped by tumor cell membranes, stabilizing the drug load and enabling targeted drug delivery with pH‐sensitive release. This targeted system induces ferroptosis in tumor cells and activates a strong anti‐tumor immune response, leading to enhanced immunotherapy for breast cancer and reduced tumor metastasis to the lungs. Importantly, the varied enzymatic activities of the nanomedicine system help mitigate the cardiotoxicity of DOX chemotherapy, improving cancer treatment while minimizing side effects. This approach introduces a nano‐enzyme drug system with distinctive activities and innovative preparation, setting a new standard for efficient, low‐toxicity cancer therapy.

Polyphenol Oxidase Activity on Guaiacyl and Syringyl Lignin Units.

The natural heterogeneity of guaiacyl (G) and syringyl (S) compounds resulting from lignin processing hampers their direct use as plant-based chemicals and materials. Herein, we explore six short polyphenol oxidases (PPOs) from lignocellulose-degrading ascomycetes for their capacity to react with G-type and S-type phenolic compounds. All six PPOs catalyze the ortho-hydroxylation of G-type compounds (guaiacol, vanillic acid, and ferulic acid), forming the corresponding methoxy-ortho-diphenols. Remarkably, a subset of these PPOs is also active towards S-compounds (syringol, syringic acid, and sinapic acid) resulting in identical methoxy-ortho-diphenols. Assays with 18O2 demonstrate that these PPOs in particular catalyze ortho-hydroxylation and ortho-demethoxylation of S-compounds and generate methanol as a co-product. Notably, oxidative (ortho-)demethoxylation of S-compounds is a novel reaction for PPOs, which we propose occurs via a distinct reaction mechanism compared to aryl-O-demethylases. We further show that addition of a reducing agent can steer the PPO reaction to form methoxy-ortho-diphenols from both G- and S-type substrates rather than reactive quinones that lead to unfavorable polymerization. Application of PPOs opens for new routes to reduce the heterogeneity and methoxylation degree of mixtures of G and S lignin-derived compounds.

Therapeutic effects of alpha lipoic acid and/or caffeine-loaded chitosan nanoparticles on memory impairment and neurochemical changes in high-fat diet-induced obese rats

The therapeutic effects of alpha lipoic acid (LA) and/or caffeine-loaded chitosan nanoparticles (CCNPs) on obesity-induced memory impairment were evaluated in the present study. Rats were divided into control rats, obese rats induced by high fat diet (HFD) and obese rats treated with LA and/or CCNPs. Obesity was confirmed by measuring the body mass index (BMI). Memory and cognitive functions were evaluated by novel object recognition test (NORT). The levels of serotonin (5-HT), dopamine (DA), norepinephrine (NE), lipid peroxidation (MDA), nitric oxide (NO), reduced glutathione (GSH), interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), leptin (LEP) and ghrelin (GHR) and the activities of monoamine oxidase (MAO), acetylcholinesterase (AchE) and Na+,K+,ATPase were determined in the cortex and hippocampus. The cerebral histopathological alterations were examined in obese rats.Obese rats showed impaired memory and exhibited significant neurochemical changes, including decreased levels of 5-HT, DA, GSH, GHR, and Na+,K+-ATPase activity, as well as an increase in AchE, MAO, MDA, NO, IL-1β, TNF-α, and LEP. LA and/or CCNPs treatment reduced BMI and improved memory. LA or CCNPs alleviated the cortical and hippocampal neurochemical changes and histopathological changes induced by obesity. Furthermore, LA and CCNPs exhibited antioxidant and anti-inflammatory properties, which likely contributed to their effects. However, no synergistic effect was observed between LA and CCNPs. These findings suggest that LA or CCNPs may be a potential therapy against obesity and its adverse effects on memory, mediated by their ability to restore monoamine levels and exhibit antioxidant and anti-inflammatory properties.

PSVIII-23 Attenuation of oxidative stress and inflammatory response improves gut integrity in piglets after infection

Abstract Enterotoxigenic Escherichia coli (ETEC) is a primary cause of post-weaning diarrhea in piglets, leading to serious health issues and economic loss in the swine industry. Increased oxidative stress and inflammation have detrimental effects on gut health. The present study aimed to investigate the regulation of oxidative stress and inflammatory response in a piglet model challenged with ETEC and to evaluate whether the chosen probiotic could improve intestinal function in piglets. A total of 24 weaned piglets susceptible to ETEC F4 infection were randomly divided into four groups: Group 1 (control), Group 2 (E. coli infection), Group 3 (E. coli infection received antibiotic avilamycin), and Group 4 (E. coli infection received B. licheniformis HG76). Group 2, Group 3 and Group 4 were infected with E. coli by an oral gavage of ETEC F4 (5 mL, 1 × 106 CFU/mL). Five days after infection, piglets in all groups were euthanized. The blood and intestine samples (a mid-jejunum segment) were collected, and serum was prepared. ETEC infection significantly increased serum and jejunal malondialdehyde (MDA) concentrations and serum diamine oxidase (DAO) activity (P < 0.05). ETEC infection also significantly decreased tight-junction proteins (ZO1, occludin), superoxide dismutase-1 (SOD), and heme oxygenase-1 (HO-1) expression (P < 0.05). Furthermore, ETEC infection significantly increased the expression of pro-inflammatory cytokines [interlueken (IL) -6, tumor necrosis factor (TNF) -α] in the intestine (P < 0.05). Antibiotic or probiotic supplementation mitigated oxidative stress and inflammation, as evidenced by decreased oxidative stress biomarkers, increased expression of antioxidant enzymes (SOD, HO-1) and downregulated the expression of proinflammatory cytokines (IL-6, TNF-α; P < 0.05). Antibiotic or probiotic supplementation also improved intestinal integrity, as demonstrated by decreased DAO activity and increased tight-junction proteins expression (P < 0.05). These results suggest that attenuation of oxidative stress and inflammatory response can improve piglet gut integrity after ETEC infection.

CD11b-NOX2 mutual regulation-mediated microglial exosome release contributes to rotenone-induced inflammation and neurotoxicity in BV2 microglia and primary cultures

Epidemiological studies have revealed a potent association between chronic exposure to rotenone, a commonly used pesticide, in individuals and the incidence of Parkinson's disease (PD). We previously identified the contribution of the activation of microglial NADPH oxidase (NOX2) in rotenone-induced neurotoxicity. However, the regulation of NOX2 activation remains unexplored. Integrins are known to be bidirectionally regulated in the plasma membrane through the inside-out and outside-in signaling. CD11b is the α-chain of integrin macrophage antigen complex-1. This study aimed to investigate whether CD11b mediates rotenone-induced NOX2 activation. We observed that rotenone exposure increased NOX2 activation in BV2 microglia, which was associated with elevated CD11b expression. Silencing CD11b significantly reduced rotenone-induced ROS production and p47phox phosphorylation, a key step for NOX2 activation. Furthermore, the Src-FAK-PKB and Syk-Vav1-Rac1 signaling pathways downstream of CD11b were found to be essential for CD11b-mediated NOX2 activation in rotenone-intoxicated microglia. Interestingly, we also found that inhibition of NOX2 decreased rotenone-induced CD11b expression, indicating a crosstalk between CD11b and NOX2. Subsequently, the inhibition of the CD11b-NOX2 axis suppressed rotenone-induced microglial activation and exosome release. Furthermore, inhibiting exosome synthesis in microglia blocked rotenone-induced gene expression of proinflammatory factors and related neurotoxicity. Finally, blocking the CD11b-NOX2 axis and exosome synthesis or endocytosis mitigated microglial activation and dopaminergic neurodegeneration in rotenone-intoxicated midbrain primary cultures. Our findings highlight the crucial involvement of the CD11b-NOX2 axis in rotenone-induced inflammation and neurotoxicity, offering fresh perspectives on the underlying mechanisms of pesticide-induced neuronal damage.

The elicitation effects of diode and He-Ne laser irradiations on the alleviation of nutrient-deficiency induced damage in anthocyanin-producing red-fleshed apple cell suspension

Purpose We explored the elicitation role of the laser irradiations on the alleviation of nutrient-deficiency induced damage in anthocyanin-producing red-fleshed apple cell suspension in continuous production of anthocyanin. Methods Anthocyanin-producing red-fleshed apple cells were irradiated by 4 intensity levels of red He-Ne (RHNL) and blue diode (BDL) lasers for 20 min. Results Nutrient deficiency indicated negative effect on total soluble proteins (TSP), superoxidase dismutase (SOD) activity, and total phenolics content (TPC) while it displayed a positive effect on malondialdehyde (MDA), total flavonoids content (TFC), O2 -, H2O2 - , and lipoxygenase (LOX) and polyphenol oxidase (PPO) activities in light controls, illustrating oxidative stress. The laser irradiations on suspension cells indicated variable effects on measured parameters and were time of growth-, levels of intensity-, and laser type-dependent. Likewise, the elicitation effects of lasers relied on a critical threshold among ROS generation and antioxidative system which determines the fate of cells against oxidative stress. The same trend was displayed by RHNL at 6.46 mWcm−2 intensity and BDL at 13.73 mWcm−2. These intensities resulted in a significant increase in SOD, APX, POD, and CAT activities and TSP, TPC, TFC, proline, and glycine betaine accumulation, while induced decrease in LOX, and PPO activities and MDA, and ROS generation, alleviating cellular injury from prolonged nutrient deficiency by diminishing lipid peroxidation and oxidative damages of cell membrane. Conclusion Results suggested that lasers application on mitigating nutrient deficiency stress relied on establishing a suitable balance between ROS generation and antioxidative system, which enables the nutrient-starved anthocyanin-producing cells to continuously produce anthocyanin.

Low voltage electrostatic field combined with ice-temperature to improve the quality of litchi during storage

Litchi is popular among consumers due to its delicious taste, however, the extremely short shelf life limits its commercial value. Few studies have been conducted to develop new technologies to extend the shelf life of litchi. Therefore, this study applied a novel technique (low voltage electrostatic field, LVEF) combined with ice-temperature (0 °C) treatment on litchi and evaluated its quality characteristics during storage. The results demonstrated that low voltage electrostatic field with ice-temperature storage (LVEF-IT) decreased malondialdehyde content, relative electrolyte leakage, and polyphenol oxidase and peroxidase activities, successfully delayed the start of peel browning. Additionally, it preserved the fruit pulp’s excellent quality, which included high concentrations of soluble solids, ascorbic acid, and soluble protein. These findings imply that LVEF-IT treatment may be a novel technique for extending the storage time of litchi.

Effects and mechanisms of Polygonati Rhizoma polysaccharide on potassium oxonate and hypoxanthine-induced hyperuricemia in mice

Hyperuricemia, a prevalent metabolic disturbance intricately linked to gout and chronic kidney disease (CKD), may be relieved by traditional Chinese medicine Polygonati Rhizoma. It is derived from the rhizomes of Polygonatum sibiricum, Polygonatum kingianum, and Polygonatum cyrtonema, which are rich in polysaccharides and are effective hyperuricemia alleviators. This study investigated the potential of Polygonatum sibiricum polysaccharide (PSP) in managing hyperuricemia. PSP (125, 250, and 500 mg/kg, i.g.) or allopurinol was administered to hyperuricemia mice treated with potassium oxonate and hypoxanthine for two weeks. PSP effectively decreased serum uric acid levels by inhibiting xanthine oxidase and adenosine deaminase activity and expression in the liver and modulating uric acid-related transporters (URAT1, OAT1, and OAT3) in the kidney. PSP lowered serum creatinine and blood urea nitrogen levels, alleviating hyperuricemia-induced renal tubular epithelial-mesenchymal fibrosis. In vitro, PSP promoted mitochondrial biogenesis via the PGC-1α/NRF1/TFAM pathway, suppressed reactive oxygen species production, and prevented cytochrome C and dynamin-related protein 1 dysregulation in HK-2 cells. Furthermore, PSPA (Mw 4.0 kDa) and PSPB (Mw 112.2 kDa) isolated from PSP exhibit different uric acid-lowering mechanisms. In conclusion, our findings highlight the therapeutic potential of PSP and its nephroprotective effects in hyperuricemia, thereby supporting its development as a therapeutic agent for hyperuricemia.

Thymol, a Dietary Monoterpene, Abrogates Hexachlorobenzene-Induced Hepatic Dysfunction Via Different Mechanisms

Thymol (2-isopropyl-5-methyl-phenol) is a natural monoterpene phenolic antioxidant that has anti-inflammatory and anti-apoptotic potentials in experimental studies. Literatures are scarce on effects of hexachlorobenzene-induced hepatic damage. Therefore, this study investigated the ameliorating potential of thymol (THY) on hexachlorobenzene (HCB)-induced hepatic damage as well as the effects on the antioxidant status, polyamine catabolism, inflammatory and apoptotic processes. Thirty-two adult male rats were daily treated orally by gavage for 25 days and allocated into four groups; control group received corn oil, HCB alone group (15 mg/kg b.wt), THY-treated group (100 mg/kg b.wt) and HCB + THY-treated group. The results showed that HCB significantly reduced the body weight with concurrent increase in relative liver weight accompanied by widespread histological aberrations. Furthermore, HCB-treated rats revealed increases in hepatic putrescine oxidase, spermine oxidase, and myeloperoxidase activities, tumor necrosis-α and interleukin-1β levels, caspase-3 activity, induced oxidative damage as evidenced by elevated malondialdehyde (MDA) levels and significant reduction in antioxidant enzyme activities and reduced glutathione (GSH). However, co-administration of THY with HBC abated the hepatic damage by preventing the generation and release of reactive oxygen species, improving the antioxidant system, down-regulated polyamine catabolism, inflammatory and apoptotic responses. The findings of this study revealed that HCB acts as a hepatotoxicant and thymol might be a possible future therapeutic agent for HCB-induced hepatic damage.

Sensitive colorimetric detection of heparin using reverse modulation of peroxidase- and oxidase-mimetic activities in Fe3O4@PDA@MnO2 nanocomposites

Heparin, a widely studied glycosaminoglycan, plays crucial roles in the regulation of various physiological and pathological processes. Therefore, it's important to develop highly selective and sensitive methods for convenient monitoring of heparin levels in biological systems. We report the design and synthesis of Fe3O4@PDA@MnO2 nanoparticles (FPM-NPs), which exhibit dual enzymatic activities, enabling quantitative detection of heparin. The FPM-NPs feature a unique tri-layer spherical shell structure, possessing both peroxidase-like and oxidase-like activities, and catalyze the oxidation of 3,3′,5,5′-tetramethylbenzidine (TMB) in the presence or absence of H2O2. Remarkably, upon co-incubated with heparin, the oxidase activity of FPM-NPs decreases, while the peroxidase activity increases. By leveraging these dual enzymatic properties of FPM-NPs, a highly sensitive and specific colorimetric detection of heparin is achieved, with a detection limit reaching 6.51 nM and a good linear response to quantify heparin ranging 10–800 nM. Additionally, the developed FPM-NPs are successfully applied to measure heparin in fetal bovine serum samples. We also extend this detection method to a paper-based chip, enabling portable detection of heparin through grayscale analysis of mobile phone photographs. The multi-nanozyme-based heparin detection approach provides a new perspective for future research on expanding the application of nanocomposite materials in biomedical detection and analysis.

Biochemical and structural insights of a recombinant AA16 LPMO from the marine and sponge-symbiont Peniophora sp

Lytic polysaccharide monooxygenases (LPMOs) are copper-dependent enzymes that oxidize polysaccharides, leading to their cleavage. LPMOs are classified into eight CAZy families (AA9-11, AA13-17), with the functionality of AA16 being poorly characterized. This study presents biochemical and structural data for an AA16 LPMO (PnAA16) from the marine sponge symbiont Peniophora sp. Phylogenetic analysis revealed that PnAA16 clusters separately from previously characterized AA16s. However, the structural modelling of PnAA16 showed the characteristic immunoglobulin-like fold of LPMOs, with a conserved his-brace motif coordinating a copper ion. The copper-bound PnAA16 showed greater thermal stability than its apo-form, highlighting copper's role in enzyme stability. Functionally, PnAA16 demonstrated oxidase activity, producing 5 μM H₂O₂ after 30 min, but showed 20 times lower peroxidase activity (0.27 U/g) compared to a fungal AA9. Specific activity assays indicated that PnAA16 acts only on cellohexaose, generating native celloligosaccharides (C3 to C5) and oxidized products with regioselective oxidation at C1 and C4 positions. Finally, PnAA16 boosted the activity of a cellulolytic cocktail for cellulose saccharification in the presence of ascorbic acid, hydrogen peroxide, or both. In conclusion, the present work provides insights into the AA16 family, expanding the understanding of their structural and functional relationships and biotechnological potential.

Para-coumaric acid and cinnamic acid enhance resistance of Agaricus bisporus mushrooms to Brown blotch disease caused by Pseudomonas tolaasii

The effects of postharvest pre-treatments with para-coumaric acid (PCA) and cinnamic acid (CA) against brown blotch disease in Agaricus bisporus mushrooms caused by Pseudomonas tolaasii were investigated. PCA and CA at the concentration of 300 mg L−1 provided the highest inhibition ratio of 28.9% and 20.9%, respectively, on the in vitro growth of P. tolaasii. In addition, PCA and CA pre-treatments dramatically reduced the P. tolaasii-caused brown blotch disease in postharvest A. bisporus. Biochemical analysis revealed that PCA and CA inhibited P. tolaasii-induced activation of polyphenol oxidase and melanin production, increased activities of β-N-acetylglucosaminidase, β-1,3-glucanase and chitinase, promoted phenylpropanoid biosynthetic enzyme activities and metabolite production, enhanced antioxidant enzyme activities, and decreased malondialdehyde content in A. bisporus mushrooms. Collectively, these results demonstrated that PCA and CA can induce resistance to postharvest brown blotch disease in A. bisporus mushrooms, and have good prospects for disease control in edible mushrooms.

Salicylic Acid Treatment Ameliorates Postharvest Quality Deterioration in 'France' Prune (Prunus domestica L. 'Ximei') Fruit by Modulating the Antioxidant System.

The potential of salicylic acid (SA) in delaying postharvest fruit senescence has been extensively documented; nevertheless, its effect on antioxidant activity and quality of 'France' prune fruit is largely unknown. The study investigated the effects of SA (0.5 mM) on postharvest quality deterioration of 'France' prune fruit. Results indicated that SA impeded the increase in respiration rate and weight loss, and mitigated the decrease of soluble solids content (SSC), titratable acidity (TA) content, firmness, and hue angle. SA sustained the ascorbate-glutathione cycle by inducing the production of ascorbic acid (AsA) and glutathione (GSH) and attenuates flavonoids, total phenols, and anthocyanins degradation by inhibiting polyphenol oxidase (PPO) activity and PdPPO. Moreover, SA significantly improved superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), peroxidase (POD), and glutathione reductase (GR) activities and gene expression levels, sustained higher 2,2'-Azinobis-(3-ethylbenzthiazoline-6-sulphonate) (ABTS) and 1,1-diphenyl-2-picryl-hydrazyl (DPPH) free radical scavenging capacity, ferric reducing antioxidant power (FRAP), and hydroxyl radical (·OH) inhibition capacity, and impeded the production of hydrogen peroxide (H2O2) and superoxide anion (O2•-). Overall, SA improved the antioxidant capacity by inducing the synthesis of defense response-related substances and promoting antioxidant enzyme activities to sustain the storage quality of 'France' prune fruit.

Colorimetric detection of benzoate in the beverage using the peroxidase-like properties of single-atom Fe-N-C nanozymes

Overuse of benzoate as a preservative is bad for human health. Therefore, we need to achieve quantitative detection of benzoate. Colorimetric nanozyme sensors have garnered significant interest owing to their exceptional efficacy and affordability. In this article, Fe-N-C dodecahedral nanozymes with superior peroxidase-like catalytic activity are prepared using ZIF-8 as a precursor. Therefore, we construct a colorimetric sensor for benzoate detection with a detection limit of 0.33 μM through integrating the inhibitory impact of benzoate on D-amino acid oxidase activity with the Fe-N-C-catalyzed 3,3′,5,5′-tetramethylbenzidine (TMB)–H2O2 system, and this reaction is completed within 10 min. It is noteworthy that a recovery rate of 96.8–109 % has been attained for the detection of benzoate in beverages.

CoNiCoNC tumor therapy by two-ways producing H2O2 to aggravate energy metabolism, chemokinetics, and ferroptosis

The effectiveness of chemokinetic therapy nanozymes is severely constrained because of the low H2O2 levels in the tumor microenvironment. Unlike other self-produced H2O2 nanozymes, the N-CNTs-encapsulated CoNi alloy (CoNiCoNC) with glucose oxidase and lactate oxidase activities has two ways to produce H2O2. It can facilitate the transformation of glucose and lactic acid into H2O2 simultaneously. First, the H2O2 generation pathway is favorable for aggravating energy metabolism. Second, some produced H2O2 can be decomposed by CoNiCoNC to H2O and O2 with the 4e− pathway to alleviate the TME hypoxia. Third, H2O2 can be catalyzed to form OH to enhance reactive oxygen species (ROS) content. Through proteomic analysis, nanozymes substantially impact the metabolic pathways of cancer cells because of their aggravating energy metabolism. The high levels of ROS can cause mitochondrial lipid peroxidation and cellular ferroptosis. Consequently, the two-way H2O2-selective nanoenzymatic platform realizes the synergistic effect of starvation therapy and chemokinetics.

Effect of ultrasound pretreatment combined with different drying methods on the quality and bioactive compounds of broccoli stems

SummaryThe effects of ultrasound (US) pretreatment combined with drying process on storage quality maintenance and bioactive compound accumulation of broccoli stems were investigated. Results indicated that US pretreatment (λ = 8.5 mm) delayed the decline of L* value and ascorbic acid content but inhibited the increase of a* and b* values, browning index, and microbial counts of fresh‐cut broccoli stems. Furthermore, the US application enhanced total phenolics content by activating phenylalanine ammonia‐lyase, cinnamate‐4‐hydroxylase, and 4‐coumarate‐CoA ligase in phenylpropane metabolism. Meanwhile, it also promoted γ‐aminobutyric acid (GABA) accumulation by upregulating activities of diamine oxidase, polyamine oxidase, and aminoaldehyde dehydrogenase in GABA shunt and polyamine degradation pathway, respectively, which in turn, accelerated the biosynthesis of GABA from polyamines and glutamate. In addition, compared with other three drying methods, the combined vacuum freeze drying (VFD) could further retain the sensory attributes and bioactive compounds to a maximum extent in dehydrated broccoli stems. Hence, current findings illustrated that US pretreatment combined with VFD process is beneficial for commercial quality maintenance and nutritional value enhancement, thus providing new perspectives on the comprehensive utilisation of broccoli stems.

Copper Single-Atom Nanozyme Mimicking Galactose Oxidase with Superior Catalytic Activity and Selectivity.

Due to the low stability and high cost of some natural enzymes, nanozymes have been developed as enzyme-imitating nanomaterials. Single-atom nanozymes are a class of nanozymes with metal centers that mimic the structure of metal-based natural enzymes. Herein, Cu-N-C single-atom nanozyme (SAN) is synthesized with excellent peroxidase- and enhanced oxidase-like activities to mimic the action of natural galactose oxidase. Cu-SAN demonstrates stereospecific activity akin to that of natural galactose oxidase by oxidizing D-galactose and primary alcohol but not L-Galactose or other carbohydrates. The SAN can catalyze the oxidation of galactose in the presence of oxygen, producing hydrogen peroxide as a sub-product. The produced hydrogen peroxide then oxidizes 3,3',5,5'-tetramethylbenzidine catalyzed by the SAN, yielding the typical blue product. The relationship between absorbance and galactose concentration is linear in the 1-60µm range with a detection limit as low as 0.23µm. This strategy can be utilized in the diagnosis of galactosemia disorder and detection of galactose in some dairy and other commercial products. DFT calculations clarify the high activity of the Cu sites in the POD-like reaction and explain the selectivity of the Cu-SAN oxidase-like reaction toward D-galactose.

Glucose-Dependent Insulinotropic Polypeptide Inhibits AGE-Induced NADPH Oxidase-Derived Oxidative Stress Generation and Foam Cell Formation in Macrophages Partly via AMPK Activation.

Glucose-dependent insulinotropic polypeptide (GIP) of the incretin group has been shown to exert pleiotropic actions. There is growing evidence that advanced glycation end products (AGEs), senescent macromolecules formed at an accelerated rate under chronic hyperglycemic conditions, play a role in the pathogenesis of atherosclerotic cardiovascular disease in diabetes. However, whether and how GIP could inhibit the AGE-induced foam cell formation of macrophages, an initial step of atherosclerosis remains to be elucidated. In this study, we address these issues. We found that AGEs increased oxidized low-density-lipoprotein uptake into reactive oxygen species (ROS) generation and Cdk5 and CD36 gene expressions in human U937 macrophages, all of which were significantly blocked by [D-Ala2]GIP(1-42) or an inhibitor of NADPH oxidase activity. An inhibitor of AMP-activated protein kinase (AMPK) attenuated all of the beneficial effects of [D-Ala2]GIP(1-42) on AGE-exposed U937 macrophages, whereas an activator of AMPK mimicked the effects of [D-Ala2]GIP(1-42) on foam cell formation, ROS generation, and Cdk5 and CD36 gene expressions in macrophages. The present study suggests that [D-Ala2]GIP(1-42) could inhibit the AGE-RAGE-induced, NADPH oxidase-derived oxidative stress generation in U937 macrophages via AMPK activation and subsequently suppress macrophage foam cell formation by reducing the Cdk5-CD36 pathway.

Pyrroloquinoline Quinone Alleviates Intestinal Inflammation and Cell Apoptosis via the MKK3/6-P38 Pathway in a Piglet Model.

This study investigates the underlying mechanism through which dietary supplementation of pyrroloquinoline quinone disodium (PQQ) alleviates intestinal inflammation and cell apoptosis in piglets challenged with lipopolysaccharide (LPS). Seventy-two barrows were divided into three groups: control (CTRL), LPS challenged (LPS), and LPS challenged with PQQ supplementation (PQQ + LPS). On d 7, 11, and 14, piglets received intraperitoneal injections of LPS or 0.9% of NaCl (80 μg/kg). After a 4 h interval following the final LPS injection on d 14, blood samples were obtained, and all piglets were euthanized for harvesting jejunal samples. The results showed that dietary supplementation of PQQ improved the damage of intestinal morphology, increased the down-regulated tight junction proteins, and reduced the increase of serum diamine oxidase activity, the intestinal fatty acid binding protein, and TNF-α levels in piglets challenged with LPS (p < 0.05). The proteomics analysis revealed a total of 141 differentially expressed proteins (DEPs), consisting of 64 up-regulated DEPs and 77 down-regulated DEPs in the PQQ + LPS group compared to the LPS group. The KEGG pathway analysis indicated enrichment of the tight junction pathway and the apoptosis pathway (p < 0.05). Compared to the LPS group, the piglets in the PQQ + LPS group had increased levels of Bcl-2 protein, reduced positive apoptosis signals, and a decrease in the abundance of MKK 3/6 and p-p38 proteins (p < 0.05). In conclusion, dietary supplementation of PQQ could alleviate jejunal inflammatory damage and cell apoptosis in piglets challenged with LPS through the MKK3/6-p38 signaling pathway.