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Manganese hydroxido (Mn-OH) complexes supported by a tripodal N,N,N″-[nitrilotris(ethane-2,1-diyl)]tris(P,P-diphenylphosphinic amido) ([poat]3-) ligand have been synthesized and characterized by spectroscopic techniques including UV-vis and electron paramagnetic resonance (EPR) spectroscopies. X-ray diffraction (XRD) methods were used to confirm the solid-state molecular structures of {Na2[MnIIpoat(OH)]}2 and {Na[MnIIIpoat(OH)]}2 as clusters that are linked by the electrostatic interactions between the sodium counterions and the oxygen atom of the ligated hydroxido unit and the phosphinic (P=O) amide groups of [poat]3-. Both clusters feature two independent monoanionic fragments in which each contains a trigonal bipyramidal Mn center that is comprised of three equatorial deprotonated amide nitrogen atoms, an apical tertiary amine, and an axial hydroxido ligand. XRD analyses of {Na[MnIIIpoat(OH)]}2 also showed an intramolecular hydrogen bonding interaction between the MnIII-OH unit and P=O group of [poat]3-. Crystalline {Na[MnIIIpoat(OH)]}2 remains as clusters with Na+---O interactions in solution and is unreactive toward external substrates. However, conductivity studies indicated that [MnIIIpoat(OH)]- generated in situ is monomeric and reactivity studies found that it is capable of cleaving C-H bonds, illustrating the importance of solution-phase speciation and its direct effect on chemical reactivity. Synopsis: Manganese-hydroxido complexes were synthesized to study the influence of H-bonds in the secondary coordination sphere and their effects on the oxidative cleavage of substrates containing C-H bonds.

Glaucoma leads to vision loss due to retinal ganglion cell death. Astrocyte reactivity contributes to neurodegeneration. Our recent study found that lipoxin B4 (LXB4), produced by retinal astrocytes, has direct neuroprotective actions on retinal ganglion cells. In this study, we aimed to investigate how the autacoid LXB4 influences astrocyte reactivity in the retina under inflammatory cytokine-induced activation and during ocular hypertension. The protective activity of LXB4 was investigated in vivo using the mouse silicone-oil model of chronic ocular hypertension. By employing a range of analytical techniques, including bulk RNA-seq, RNAscope in-situ hybridization, qPCR, and lipidomic analyses, we discovered the formation of lipoxins and expression of the lipoxin pathway in rodents (including the retina and optic nerve), primates (optic nerve), and human brain astrocytes, indicating the presence of this neuroprotective pathway across various species. Findings in the mouse retina identified significant dysregulation of the lipoxin pathway in response to chronic ocular hypertension, leading to an increase in 5-lipoxygenase (5-LOX) activity and a decrease in 15-LOX activity. This dysregulation was coincident with a marked upregulation of astrocyte reactivity. Reactive human brain astrocytes also showed a significant increase in 5-LOX. Treatment with LXB4 amplified the lipoxin biosynthetic pathway by restoring and amplifying the generation of another member of the lipoxin family, LXA4, and mitigated astrocyte reactivity in mouse retinas and human brain astrocytes. In conclusion, the lipoxin pathway is functionally expressed in rodents, primates, and human astrocytes, and is a resident neuroprotective pathway that is downregulated in reactive astrocytes. Novel cellular targets for LXB4s neuroprotective action are inhibition of astrocyte reactivity and restoration of lipoxin generation. Amplifying the lipoxin pathway is a potential target to disrupt or prevent astrocyte reactivity in neurodegenerative diseases, including retinal ganglion cell death in glaucoma.

Lipoxygenases (LOXs) catalyze the regioselective dioxygenation of polyunsaturated fatty acids (PUFAs), generating fatty acid hydroperoxides (FAHPs) with diverse industrial applications. Bacterial LOXs have garnered significant attention in recent years due to their broad activity towards PUFAs, yet knowledge about the structural factors influencing their substrate preferences remains limited. Here, we characterized a bacterial LOX from Burkholderia thailandensis (Bt-LOX), and identified key residues affecting its substrate preference and regioselectivity through site-directed mutagenesis. Bt-LOX preferred ω-6 PUFAs and exhibited regioselectivity at the ω-5 position. Mutations targeting the substrate binding pocket and the oxygen access channel led to the production of three active variants with distinct catalytic properties. The A431G variant bifurcated dioxygenation between the ω-5 and ω-9 positions, while F446V showed reduced regioselectivity with longer PUFAs. Interestingly, L445A displayed altered substrate specificity, favoring ω-3 over ω-6 PUFAs. Furthermore, L445A shifted the regioselectivity of dioxygenation to the ω-2 position in ω-3 PUFAs, and, for some substrates, facilitated dioxygenation closer to the carboxylic acid terminus, suggesting an altered substrate orientation. Among these variants, L445A represents a significant milestone in LOX research, as these alterations in substrate specificity, dioxygenation regioselectivity, and substrate orientation were achieved by a single mutation only. These findings illuminate key residues governing substrate preference and regioselectivity in Bt-LOX, offering opportunities for synthesizing diverse FAHPs and highlighting the potential of bacterial LOXs as biocatalysts with widespread applications. [Display omitted] • B. thailandensis LOX preferred ω-6 PUFAs and exhibited regioselectivity at the ω-5 position. • The A431G variant bifurcated dioxygenation between the ω-5 and ω-9 positions. • The L445A substitution altered substrate specificity, regioselectivity and substrate orientation. • The L445A variant enabled dioxygenation at the ω-2 position in ω-3 PUFAs, a new LOX specificity. • Changing determinant residues facilitated the synthesis of diverse fatty acid hydroperoxides. [ABSTRACT FROM AUTHOR]

Human arachidonate 15-lipoxygenase type B is a lipoxygenase that catalyzes the peroxidation of arachidonic acid at carbon-15. The corresponding murine ortholog however has 8-lipoxygenase activity. Both enzymes oxygenate polyunsaturated fatty acids in S-chirality with singular reaction specificity, although they generate a different product pattern. Furthermore, while both enzymes utilize both esterified fatty acids and fatty acid hydro(pero)xides as substrates, they differ with respect to the orientation of the fatty acid in their substrate-binding pocket. While ALOX15B accepts the fatty acid "tail-first," Alox8 oxygenates the free fatty acid with its "head-first." These differences in substrate orientation and thus in regio- and stereospecificity are thought to be determined by distinct amino acid residues. Towards their biological function, both enzymes share a commonality in regulating cholesterol homeostasis in macrophages, and Alox8 knockdown is associated with reduced atherosclerosis in mice. Additional roles have been linked to lung inflammation along with tumor suppressor activity. This review focuses on the current knowledge of the enzymatic activity of human ALOX15B and murine Alox8, along with their association with diseases. [ABSTRACT FROM AUTHOR]

Key message: Lipoxygenase activity and localization vary throughout the development of Larix kaempferi ovules, with the highest enzyme activity observed in ovules at the cellular stage and the most intense immunogold reaction noted at the mature archegonium stage of gametophyte development. Lipoxygenases are a family of oxidoreductases with a significant role in biological systems, widespread in living organisms e.g. mammals, fish, corals, plants, mosses, algae, fungi, yeasts, and bacteria. Lipoxygenase activity in plants leads to the formation of phytooxylipins, i.e. signaling molecules, which play a crucial role in many significant physiological processes such as male and female gametophyte maturation, germination and seedling growth, pathogen resistance, abiotic stress response, fruit ripening, and senescence. The activity and localization of lipoxygenase change during plant growth and development. The localization of lipoxygenase in a developing ovule of Larix kaempferi was analyzed using the immunogold labeling method, and the activity was determined spectrophotometrically with linolenic acid as a substrate. Among the investigated stages, the immunogold reaction was the most intense at the mature archegonium stage in the ovule. Lipoxygenase was found in all parts of the L. kaempferi ovule. The largest number of immunogold particles was detected in the integument cells of all the analyzed stages of ovule development. Only one isoform of lipoxygenase with an optimum at pH 8 was active in the ovules during female gametophyte maturation. The highest enzyme activity was determined at the cellular stage, whereas the mature archegonium stage was characterized by its lowest level, which means that LOX activity in developing ovules of the Japanese larch is not correlated with the number of antibody-labeled molecules of the enzyme. [ABSTRACT FROM AUTHOR]

The human genome involves six functional arachidonic acid (AA) lipoxygenase (ALOX) genes, and the corresponding enzymes (ALOX15, ALOX15B, ALOX12, ALOX12B, ALOXE3, ALOX5) have been implicated in cell differentiations and in the pathogenesis of inflammatory, hyperproliferative, metabolic, and neurological disorders. Humans express two different AA 15-lipoxygenating ALOX isoforms, and these enzymes are called ALOX15 (15-LOX1) and ALOX15B (15-LOX2). Chromosomal localization, sequence alignments, and comparison of the enzyme properties suggest that pig and mouse ALOX15 orthologs (leukocyte-type 12-LOX) on the one hand and rabbit and human ALOX15 orthologs on the other (reticulocyte-type 15-LOX1) belong to the same enzyme family despite their different reaction specificities with AA as a substrate. In contrast, human ALOX12 (platelet-type 12-LOX), as well as pig and mouse ALOX15 (leukocyte-type 12-LOX), belong to different enzyme families, although they exhibit a similar reaction specificity with AA as a substrate. The complex multiplicity of mammalian ALOX isoforms and the controversial enzyme nomenclatures are highly confusing and prompted us to summarize the current knowledge on the biological functions, enzymatic properties, and allosteric regulation mechanisms of mammalian ALOX15, ALOX15B, and ALOX12 orthologs that belong to three different enzyme sub-families. [ABSTRACT FROM AUTHOR]

Diabetic retinopathy (DR) is a salient cause of blindness worldwide. There is still an immense need to understand the pathophysiology of DR to discover better diagnostic and therapeutic modalities. Human retinal endothelial cells (HRECs) were treated with 15-HETE or D-glucose, then miRNAs were isolated, and a microarray was performed. MirWALK 2 and Ingenuity Pathway Analysis (IPA) were used to analyze the microarray results. Exosomal miRNAs from 15-HETE-treated HRECs were isolated, microarrayed, and then imported into IPA for further analysis. The microarray results showed that 15-HETE downregulated 343 miRNAs and upregulated 297 miRNAs in HRECs. High glucose treatment induced a differential expression of HREC-miRNAs where 185 miRNAs were downregulated and 244 were upregulated. Comparing the impact of 15-HETE versus DG or diabetic mouse retina elaborated commonly changing miRNAs. Pathway and target analysis for miRNAs changed in 15-HETE-treated HRECs revealed multiple targets and pathways that may be involved in 15-HETE-induced retinal endothelial dysfunction. The HREC-exosomal miRNAs were differentially expressed after 15-HETE treatment, with 34 miRNAs downregulated and 45 miRNAs upregulated, impacting different cellular pathways. Here, we show that 15-HETE induces various changes in the cellular and exosomal miRNA profile of HRECs, highlighting the importance of targeting the 12/15 lipoxygenase pathway in DR. [ABSTRACT FROM AUTHOR]

Background: Alzheimer's disease (AD) is the most common human neurodegenerative disorder worldwide. Owing to its chronic nature, our limited understanding of its pathophysiological mechanisms, and because of the lack of effective anti-AD drugs, AD represents a significant socio-economic challenge for all industrialized countries. Neuronal cell death is a key factor in AD pathogenesis and recent studies have suggested that neuronal ferroptosis may play a major patho-physiological role. Since ferroptosis involves free radical-mediated lipid peroxidation, we hypothesized that enteral administration of the radical scavenger butylated hydroxytoluene (BHT) might slow down or even prevent the development of AD-related symptoms in an in vivo animal AD model. Material and methods: To test this hypothesis, we employed the rat model of streptozotocin-induced AD and administered butylated hydroxytoluene orally at a dose of 120 mg/kg body weight. Following BHT treatment, neuronal cell death was induced by bilateral stereotactic intraventricular injection of streptozotocin at a dose of 3.0 mg/kg body weight. Three weeks after surgery, we assessed the learning capabilities and the short-term memory of three experimental groups using the conventional y-maze test: (i) streptozotocin-treated rats (BHT pre-treatment), (ii) streptozotocin-treated rats (no BHT pre-treatment), (iii) sham-operated rats (BHT pre-treatment but no streptozotocin administration). After the y-maze test, the animals were sacrificed, hippocampal tissue was prepared and several biochemical (malonyl dialdehyde formation, glutathione homeostasis, gene expression patterns) and histochemical (Congo-red staining, Nissl staining, Perls staining) readout parameters were quantified. Results: Intraventricular streptozotocin injection induced the development of AD-related symptoms, elevated the degree of lipid peroxidation and upregulated the expression of ferroptosis-related genes. Histochemical analysis indicated neuronal cell death and neuroinflammation, which were paralleled by aberrant intraneuronal iron deposition. The streptozotocin-induced alterations were significantly reduced and sometimes even abolished by oral BHT treatment. Conclusion: Our data indicate that oral BHT treatment attenuated the development of AD-related symptoms in an in vivo rat model, most probably via inhibiting neuronal ferroptosis. These findings suggest that BHT might constitute a promising candidate as anti-AD drug. However, more work is needed to explore the potential applicability of BHT in other models of neurodegeneration and in additional ferroptosis-related disorders. [ABSTRACT FROM AUTHOR]

Background: Yellow lupine (Lupinus luteus L.) is a high-protein crop of considerable economic and ecological significance. It has the ability to fix atmospheric nitrogen in symbiosis with Rhizobium, enriching marginal soils with this essential nutrient and reducing the need for artificial fertilizers. Additionally, lupine produces seeds with a high protein content, making it valuable for animal feed production. However, drought negatively affects lupine development, its mutualistic relationship with bacteria, and overall yield. To understand how lupine responds to this stress, global transcriptome sequencing was conducted, along with in-depth biochemical, chromatography, and microscopy analyses of roots subjected to drought. The results presented here contribute to strategies aimed at mitigating the effects of water deficit on lupine growth and development. Results: Based on RNA-seq, drought-specific genes were identified and annotated to biological pathways involved in phytohormone biosynthesis/signaling, lipid metabolism, and redox homeostasis. Our findings indicate that drought-induced disruption of redox balance characterized by the upregulation of reactive oxygen species (ROS) scavenging enzymes, coincided with the accumulation of lipid-metabolizing enzymes, such as phospholipase D (PLD) and lipoxygenase (LOX). This disruption also led to modifications in lipid homeostasis, including increased levels of triacylglycerols (TAG) and free fatty acids (FFA), along with a decrease in polar lipid content. Additionally, the stress response involved alterations in the transcriptional regulation of the linolenic acid metabolism network, resulting in changes in the composition of fatty acids containing 18 carbons. Conclusion: The first comprehensive global transcriptomic profiles of lupine roots, combined with the identification of key stress-responsive molecules, represent a significant advancement in understanding lupine's responses to abiotic stress. The increased expression of the Δ12DESATURASE gene and enhanced PLD activity lead to higher level of linoleic acid (18:2), which is subsequently oxidized by LOX, resulting in membrane damage and malondialdehyde (MDA) accumulation. Oxidative stress elevates the activities of superoxide dismutase (SOD), ascorbate peroxidase (APX), and catalase (CAT), while the conversion of FFAs into TAGs provides protection against ROS. This research offers valuable molecular and biochemical candidates with significant potential to enhance drought tolerance. It enables innovative strategies in lupine breeding and crop improvement to address critical agricultural challenges. [ABSTRACT FROM AUTHOR]

Practical Application The lipoxygenase (LOX) activity of major tropical fish used for surimi production, including threadfin bream (TB), lizardfish (LZ), and goatfish (GF), was measured in the gills, skin, and muscle. The highest LOX activity was observed in the LZ samples (p < 0.05), with the gills exhibiting the greatest activity at 376.56 U/mg (p < 0.05). The highest peroxide value was detected in the TB samples, particularly in the gills (p < 0.05). Docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) were the main polyunsaturated fatty acids in all the tissues and surimi. The total lipid and DHA contents of washed mince reduced considerably after the screw press process. Although LOX activity decreased during surimi production, a residual activity of 21.33 U/g was observed in the finished surimi. LOX was partially purified and characterized from LZ gills. The purification was conducted using two successive chromatographic steps, Sephacryl S‐200 and diethylaminoethyl (DEAE)–sepharose, resulting in a 3.52% yield and a 22.43‐fold increase in purity. The optimum activity was found at 25°C and pH 7.5, with pH stability between 6.0 and 8.5. The relatively high thermal stability at 4°C–10°C suggested that LOX might contribute to fish lipid oxidation during cold storage. The enzyme was thermally inactivated at 60°C. The preferred substrate was EPA. LOX from the LZ gills was inhibited by 1 mM ethylenediaminetetraacetic acid and activated by 1 mM Fe2+, Na+, and Ca2+.Elucidating lipoxygenase activity and lipid oxidation in various tropical fish tissues, as well as understanding the characteristics of LOX, can help take appropriate postharvest actions to afford high‐quality surimi. [ABSTRACT FROM AUTHOR]

The growing demand for low-cost and functional snacks in many developing nations called for interest in the use of locally grown crops as substitutes for costly imported wheat flour. The amino acid composition, antioxidant and anti-inflammatory properties of the cookies from whole wheat, sweet potato, rice bran and peanut (56.25:18.75:5:20; 37.50:37.50:5:20; 18.75:56.25:5:20% as WPRG 1, WPRG 2, WPRG 3) composite flour blends, respectively, were obtained in this study. The 100% whole wheat and 100% refined flours served as control 1 and 2, respectively. The level of hydrophobic and aromatic amino acids was significantly (p < 0.05) high in WPRG 2 (~ 30 and ~ 10%), respectively when compared to others. However, the branched chain amino acids and Fischer ratio was significantly (p < 0.05) high in WPRG 1 (11.40% and 1.29), respectively, which could have contributed to their improved bioactivities. Notably, the composite cookie samples WPRG 1, 2 and 3 had higher hydroxyl (73.86 − 84.16%), DPPH (76.52 − 84.60%) radical scavenging as well as ferric reducing antioxidant (0.64–0.87 mmolFe2+/mg) properties than the control samples WWF and CWF, respectively. On the contrary, the metal chelating activities of the cookies WPRG 1–3 were not significantly (p > 0.05) different from control samples WWF and CWF. The improved amino acid profile and enhanced antioxidant properties of the composite cookies might have effectively influenced their anti-inflammatory properties (IC50; < 26 μg/ml) when compared to the control samples (IC50; ~ 40 μg/ml), respectively. Hence, the cookies that comprised of antioxidants and anti-inflammatory potentials needed in human health, were acceptable by the consumers. [ABSTRACT FROM AUTHOR]

9-Carbon aldehydes such as (2E)-nonenal, (3Z)-nonenal, and (2E,6Z)-nonadienal are important melon and cucumber fragrance compounds. Currently, these molecules are produced either synthetically, which faces consumer aversion, or through biotransformation using plant-extracted enzymes, which is costly and inefficient. In this study, we constructed a Saccharomyces cerevisiae platform for the whole cell biotransformation of polyunsaturated fatty acids (PUFAs) to 9-carbon aldehydes. Heterologous expression of lipoxygenase (LOX) from Nicotiana benthamiana and hydroperoxide lyase (HPL) from Cucumis melo (melon) in S. cerevisiae enabled the production of (2E)-nonenal from readily available polyunsaturated fatty acid substrates. A 5.5-fold increase in (2E)-nonenal titer was then achieved utilizing genetic and reaction condition enhancement strategies. The highest titer of (2E)-nonenal was more than 0.11 mM, with about 9% yield. This platform can potentially be used to produce a variety of other aldehyde products by customizing with LOX and HPL enzymes of different regio-selectivities. Key points: • Establishment of a S. cerevisiae whole-cell biotransformation platform for cost-efficient, high-yield conversion of PUFAs into high value 9-carbon aldehyde compounds • 5.5-Fold improvement of (2E)-nonenal titer to > 0.11 mM achieved by enhancing reaction conditions and gene expression levels of LOX and HPL [ABSTRACT FROM AUTHOR]

Gastric and duodenal ulcers are increasingly becoming global health burdens. The side effects of conventional treatments such as non-steroid anti-inflammatory drugs (NSAIDs), proton pump inhibitors (PPIs), antibiotics, and cytoprotective agents have necessitated the search for new medications. Plants are a rich source of active metabolites and herbal medicines have been used in the treatment of ulcers and cancers. In this study, we used in silico methods like molecular docking and MM-GBSA calculations to evaluate the effects of some anti-ulcer and anti-inflammatory phytochemicals on some key enzymes, cyclooxygenase (COX), and lipoxygenase (LOX), which are implicated in the protection and destruction of the gastric mucosa. The phytochemicals were retrieved from the literature and docked toward the binding sites of the three enzymes (COX-1, COX-2, and 5-LOX). Five compounds, rhamnetin, kaempferol, rutin, rosmarinic acid, and chlorogenic acid were observed to putatively bind to cyclooxygenase 2 (COX-2) and 5-lipoxygenase (5-LOX) but not to cyclooxygenase 1 (COX-1). The interaction mechanisms between these phytochemicals and the target proteins are discussed. The compounds' drug metabolism, pharmacokinetics, and toxicity have been evaluated to assess their suitability as potential next-generation anti-ulcer and anti-inflammatory drugs. [ABSTRACT FROM AUTHOR]

The impact of four different harvest years and two cultivation locations (CLs) of pea seeds on their protein wet fractionation yield, volatile and non-volatile beany flavors, and functionality were investigated. Both harvest years and CLs significantly affected protein recovery, but protein purity was primarily influenced by CLs. Seed age emerged as a dominant factor causing the reduction in linolenic/linoleic acid content and lipoxygenase (LOX) activity which surpassed the effect of harvest years in the seeds but not in their proteins. CL significantly affected fatty acid composition in both seeds and proteins, whereas its effect on LOX activity was discernible only in the proteins. Volatile beany compounds in the proteins were affected by both harvest years and CLs, correlating with their polyunsaturated fatty acid (PUFA) content and LOX activity. Both factors minimally impacted the emulsification capacity of the proteins but imposed a significant effect on their rheological properties. Altogether, the results revealed that seed crop years and especially locations affect pea protein quality, calling for proper adaptation strategies. [ABSTRACT FROM AUTHOR]

Lipoxygenases (LOXs) from lower organisms have substrate flexibility and function versatility in fatty acid oxidation, but it is not clear how these LOXs acquired the ability to execute multiple functions within only one catalytic domain. This work studied a multifunctional LOX from red alga Pyropia haitanensis (PhLOX) which combined hydroperoxidelyase (HPL) and allene oxide synthase (AOS) activity in its active pocket. Molecular docking and site-directed mutagenesis revealed that Phe642 and Phe826 jointly regulated the double peroxidation of fatty acid, Gln777 and Asn575 were essential to the AOS function, and the HPL activity was improved when Asn575, Gln777, or Phe826 was replaced by leucine. Phylogenetic analysis indicated that Asn575 and Phe826 were unique amino acid sites in the separated clades clustered with PhLOX, whereas Phe642 and Gln777 were conserved in plant or animal LOXs. The N-terminal START/RHO_alpha_C/PITP/Bet_v1/CoxG/CalC (SRPBCC) domain of PhLOX was another key variable, as the absence of this domain disrupted the versatility of PhLOX. Moreover, the functions of two homologous LOXs from marine bacterium Shewanella violacea and red alga Chondrus crispus were examined. The HPL activity of PhLOX appeared to be inherited from a common ancestor, and the AOS function was likely acquired through mutations in some key residues in the active pocket. Taken together, our results suggested that some LOXs from red algae attained their versatility by amalgamating functional domains of ancestral origin and unique amino acid mutations. [ABSTRACT FROM AUTHOR]

High‐pressure processing (HPP) is considered as gentle preservation technique for especially heat‐sensitive food ingredients. So far, the focus has been on the fact that it is called a nonthermal process and high pressure can affect bioavailability, but it is questionable whether the high pressure affects the ingredients themselves. By using an o/w‐microemulsion (ME) as a model system, it was possible to investigate the influence of pressure, especially on lipophilic compounds (e.g., carotenoids and vitamin E), without the complexity of a food matrix. The ME consisted of Capryol® TM 90, Tween® 80 or 20, Transcutol® HP and distilled water. Lipophilic and hydrophilic compounds were introduced to the oil phase and to the aqueous phase, respectively. Storage experiments confirmed the applicability for β‐carotene and α‐tocopherol. HPP of MEs, performed for 10 min at room temperature (RT) and up to 600 MPa, resulted in pressure stability of β‐carotene (exceptional at 400 MPa; −11%) and α‐tocopherol. Multicomponent ME showed that both had a positive effect on the stability of chlorophyll a/b during HPP. An ME environment was used to facilitate co‐oxidation of β‐carotene via lipoxygenase (LOX) from an Edamame‐based crude enzyme extract and lyophilized LOX‐1 from soybeans during storage (RT and 4°C, dark conditions) and HPP treatment. A loss of β‐carotene occurred after addition of linoleic acid, whereas effects of added α‐tocopherol could be related to β‐carotene protection. Overall, the introduced ME for studying HPP effects on lipophilic food ingredients showed promising results as versatile model system for future investigations regarding interactions of phytochemicals. [ABSTRACT FROM AUTHOR]

Abstract Background Yellow lupine (Lupinus luteus L.) is a high-protein crop of considerable economic and ecological significance. It has the ability to fix atmospheric nitrogen in symbiosis with Rhizobium, enriching marginal soils with this essential nutrient and reducing the need for artificial fertilizers. Additionally, lupine produces seeds with a high protein content, making it valuable for animal feed production. However, drought negatively affects lupine development, its mutualistic relationship with bacteria, and overall yield. To understand how lupine responds to this stress, global transcriptome sequencing was conducted, along with in-depth biochemical, chromatography, and microscopy analyses of roots subjected to drought. The results presented here contribute to strategies aimed at mitigating the effects of water deficit on lupine growth and development. Results Based on RNA-seq, drought-specific genes were identified and annotated to biological pathways involved in phytohormone biosynthesis/signaling, lipid metabolism, and redox homeostasis. Our findings indicate that drought-induced disruption of redox balance characterized by the upregulation of reactive oxygen species (ROS) scavenging enzymes, coincided with the accumulation of lipid-metabolizing enzymes, such as phospholipase D (PLD) and lipoxygenase (LOX). This disruption also led to modifications in lipid homeostasis, including increased levels of triacylglycerols (TAG) and free fatty acids (FFA), along with a decrease in polar lipid content. Additionally, the stress response involved alterations in the transcriptional regulation of the linolenic acid metabolism network, resulting in changes in the composition of fatty acids containing 18 carbons. Conclusion The first comprehensive global transcriptomic profiles of lupine roots, combined with the identification of key stress-responsive molecules, represent a significant advancement in understanding lupine’s responses to abiotic stress. The increased expression of the Δ12DESATURASE gene and enhanced PLD activity lead to higher level of linoleic acid (18:2), which is subsequently oxidized by LOX, resulting in membrane damage and malondialdehyde (MDA) accumulation. Oxidative stress elevates the activities of superoxide dismutase (SOD), ascorbate peroxidase (APX), and catalase (CAT), while the conversion of FFAs into TAGs provides protection against ROS. This research offers valuable molecular and biochemical candidates with significant potential to enhance drought tolerance . It enables innovative strategies in lupine breeding and crop improvement to address critical agricultural challenges.

Abstract The growing demand for low-cost and functional snacks in many developing nations called for interest in the use of locally grown crops as substitutes for costly imported wheat flour. The amino acid composition, antioxidant and anti-inflammatory properties of the cookies from whole wheat, sweet potato, rice bran and peanut (56.25:18.75:5:20; 37.50:37.50:5:20; 18.75:56.25:5:20% as WPRG 1, WPRG 2, WPRG 3) composite flour blends, respectively, were obtained in this study. The 100% whole wheat and 100% refined flours served as control 1 and 2, respectively. The level of hydrophobic and aromatic amino acids was significantly (p 0.05) different from control samples WWF and CWF. The improved amino acid profile and enhanced antioxidant properties of the composite cookies might have effectively influenced their anti-inflammatory properties (IC50;

Naphthalene (NA), a polycyclic aromatic hydrocarbon, is an environmental pollutant from different sources exhibiting toxicities via free radical generation. However, NA has been used in the industry as surfactants, solvents, resins, and in medicine – as an anti-viral, anti-bacterial, and antiinflammatory drug. Malondialdehyde (MDA), a by-product in lipid peroxidation and prostaglandin synthesis, is a biomarker in lipid peroxidation evaluation and cyclooxygenase (COX) and lipoxygenase (LOX) activities assessment via inhibition.

Rice bran, a byproduct of rice processing, holds great nutritional value. However, its application in the food industry is greatly restricted due to its high susceptibility to rancidity and spoilage during storage. Currently, it is primarily used as animal feed with low added value. The presence of lipase and lipoxygenase is the major cause of rapid rancidity of rice bran, and other factors such as water activity, microorganisms and phenolic substances also affect the rancidity reaction. Stabilization treatment of rice bran is a key prerequisite to achieving high-value utilization of the product. This paper provides an overview of the rancidity mechanism of rice bran, the common methods for stabilizing rice bran and their impact on its quality and nutritional characteristics, with a view to providing guidance and reference for the research and development of rice bran as a food-grade raw material.

Abstract Background Enhancing crops’ drought resilience is necessary to maintain productivity levels. Plants interact synergistically with microorganisms like Beauveria bassiana to improve drought tolerance. Therefore, the current study investigates the effects of biopriming with B. bassiana on drought tolerance in Malva parviflora plants grown under regular irrigation (90% water holding capacity (WHC)), mild (60% WHC), and severe drought stress (30% WHC). Results The results showed that drought stress reduced the growth and physiological attributes of M. parviflora. However, those bioprimed with B. bassiana showed higher drought tolerance and enhanced growth, physiological, and biochemical parameters: drought stress enriched malondialdehyde and H2O2 contents. Conversely, exposure to B. bassiana reduced stress markers and significantly increased proline and ascorbic acid content under severe drought stress; it enhanced gibberellic acid and reduced ethylene. Bioprimed M. parviflora, under drought conditions, improved antioxidant enzymatic activity and the plant’s nutritional status. Besides, ten Inter-Simple Sequence Repeat primers detected a 25% genetic variation between treatments. Genomic DNA template stability (GTS) decreased slightly and was more noticeable in response to drought stress; however, for drought-stressed plants, biopriming with B. bassiana retained the GTS. Conclusion Under drought conditions, biopriming with B. bassiana enhanced Malva’s growth and nutritional value. This could attenuate photosynthetic alterations, up-regulate secondary metabolites, activate the antioxidant system, and maintain genome integrity.

Abstract Background Oil bodies or lipid droplets (LDs) in the cytosol are the subcellular storage compartments of seeds and the sites of lipid metabolism providing energy to the germinating seeds. Major LD-associated proteins are lipoxygenases, phospholipaseD, oleosins, TAG-lipases, steroleosins, caleosins and SEIPINs; involved in facilitating germination and enhancing peroxidation resulting in off-flavours. However, how natural selection is balancing contradictory processes in lipid-rich seeds remains evasive. The present study was aimed at the prediction of selection signatures among orthologous clades in major oilseeds and the correlation of selection effect with gene expression. Results The LD-associated genes from the major oil-bearing crops were analyzed to predict natural selection signatures in phylogenetically close-knit ortholog clusters to understand adaptive evolution. Positive selection was the major force driving the evolution and diversification of orthologs in a lineage-specific manner. Significant positive selection effects were found in 94 genes particularly in oleosin and TAG-lipases, purifying with excess of non-synonymous substitution in 44 genes while 35 genes were neutral to selection effects. No significant selection impact was noticed in Brassicaceae as against LOX genes of oil palm. A heavy load of deleterious mutations affecting selection signatures was detected in T-lineage oleosins and LOX genes of Arachis hypogaea. The T-lineage oleosin genes were involved in mainly anther, tapetum and anther wall morphogenesis. In Ricinus communis and Sesamum indicum > 85% of PLD genes were under selection whereas selection pressures were low in Brassica juncea and Helianthus annuus. Steroleosin, caleosin and SEIPINs with large roles in lipid droplet organization expressed mostly in seeds and were under considerable positive selection pressures. Expression divergence was evident among paralogs and homeologs with one gene attaining functional superiority compared to the other. The LOX gene Glyma.13g347500 associated with off-flavor was not expressed during germination, rather its paralog Glyma.13g347600 showed expression in Glycine max. PLD-α genes were expressed on all the tissues except the seed,δ genes in seed and meristem while β and γ genes expressed in the leaf. Conclusions The genes involved in seed germination and lipid metabolism were under strong positive selection, although species differences were discernable. The present study identifies suitable candidate genes enhancing seed oil content and germination wherein directional selection can become more fruitful.

12‐hydroxyeicosatetraenoic acid (12‐HETE), a major metabolite of arachidonic acid, is converted by 12/15‐lipoxygenase and implicated in diabetic retinopathy (DR). Our previous study demonstrated a positive correlation between 12‐HETE and the prevalence of DR. However, reasons for the increased production of 12‐HETE are unclear, and the underlying mechanisms through which 12‐HETE promotes DR are unknown. This study aimed to elucidate the correlation between 12‐HETE and DR onset, investigate potential mechanisms through which 12‐HETE promotes DR, and seek explanations for the increased production of 12‐HETE in diabetes. We conducted a prospective cohort study, which revealed that higher serum 12‐HETE levels could induce DR. Additionally, G protein‐coupled receptor 31 (GPR31), a high‐affinity receptor for 12‐HETE, was expressed in human retinal microvascular endothelial cells (HRMECs). 12‐HETE/GPR31‐mediated HRMEC inflammation occurred via the p38 MAPK pathway. 12‐HETE levels were significantly higher in the retina of mice with high‐fat diet (HFD)‐ and streptozotocin (STZ)‐induced diabetes than in those with only STZ‐induced diabetes and healthy controls. They were positively correlated with the levels of inflammatory cytokines in the retina, indicating that HFD could induce increased 12‐HETE synthesis in patients with diabetes in addition to hyperglycemia. Conclusively, 12‐HETE is a potential risk factor for DR. The 12‐HETE/GPR31 axis plays a crucial role in HRMEC dysfunction and could be a novel target for DR prevention and control. Nevertheless, further research is warranted to provide comprehensive insights into the complex underlying mechanisms of 12‐HETE in DR. [ABSTRACT FROM AUTHOR]

Introduction: Green tea possesses gastroprotective effects via different mechanisms including lipoxygenase inhibition. We compared its protective effects with licofelone (as a potent lipoxygenase inhibitor) on reducing the incidence of gastric ulcers caused by indomethacin. Methods: 48 rats received an aqueous extract of green tea (GTAE; 50, 100, and 200 mg/ kg), licofelone (30 mg/kg), zileuton (100 mg/kg), or 0.18% Tween 80 in the presence of indomethacin (100 mg/kg). Two groups received only GTAE (200 mg/kg) or indomethacin (100 mg/kg). The gastric ulcer index and Malondialdehyde (MDA) in gastric tissues were evaluated. To investigate the analgesic effect in acute and chronic phases, 24 rats received GTAE (200 mg/kg), licofelone (30 mg/kg), indomethacin (30 mg/kg), or 0.18% Tween 80 in the presence of formalin (2.5%). The behavior of rats was monitored for 30 minutes (minutes 0 to 5 and 25 to 30) for licking and biting feet and tails. Results: Indomethacin (100 mg/kg) produced clear macroscopic lesions compared to the control group. GTAE (100 and 200 mg/kg), licofelone, and Zileuton showed a significant decrease in wound score compared to indomethacin. GTAE (100 mg/kg and 200), licofelone, and zileuton displayed a significant decrease in the MDA content of gastric tissue compared to the indomethacin group. Notably, GTAE exerted greater benefits than licofelone. Besides, GTAE (200 mg/kg) showed a significant decrease observed in both acute and chronic stages of pain compared to licofelone (30 mg/kg). Conclusion: GTAE (200 mg/kg) possesses anti-ulcer and analgesic effects similar to licofelone. The exact mechanism is probably via inhibition of lipoxygenase (LOX) and antioxidant effects. [ABSTRACT FROM AUTHOR]

Argemone mexicana belonging to family Papaveraceae is a traditional medicinal plant widely utilized by tribal people in India for treating various ailments like skin infections, wounds and inflammation. This plant is very rich in alkaloidal content, which has a great potential in the treatment of anti-inflammatory disorders. Therapeutically promising bioactive molecules are often produced by endophytic fungi associated with medicinal plants. In this investigation, endophytic fungi were isolated from various parts of A. mexicana and screened for alkaloidal content. Among these, one of the fungal isolate, Acremonium alternatum AMEF-5 producing maximum alkaloids showed significant anti-inflammatory activity. Fractionation of this crude fungal extract through column chromatography yielded eight fractions, which were further screened for anti-inflammatory activities. Fraction 3 exhibited significant anti-inflammatory activity by the inhibition of lipoxygenase enzyme (IC50 15.2 ± 0.09 µg/ml), scavenging of the nitric oxide radicals (IC50 11.38 ± 0.35 µg/ml), protein denaturation (IC50 14.93 ± 0.4 µg/ml), trypsin inhibition (IC50 12.06 ± 0.64 µg/ml) and HRBC stabilization (IC50 11.9 ± 0.22 µg/ml). The bioactive alkaloid in fraction 3 was identified as aconitine which was confirmed by UV, FTIR, HPLC, HRMS, 1H NMR, and 13C NMR analysis. This study demonstrates that endophytic fungi serve a potential source for sustainable production of therapeutically important alkaloids. [ABSTRACT FROM AUTHOR]

Copyright of Food & Fermentation Industries is the property of Food & Fermentation Industries and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Black soybeans with green cotyledon have long been widely consumed in the East due to their high content of health-promoting anthocyanins and lutein. However, major anti-nutritional and allergenic components such as lectin, Kunitz trypsin inhibitor (KTI), P34, lipoxygenase, and stachyose are contained in mature seed. The objective of this research is to breed a soybean line with a black seed coat, a green cotyledon, and the penta null genotype (lele-titi-p34p34-lox1lox1lox2lox2lox3lox3-rs2rs2) for all five components. The F2 plant strain with penta null genotype for lectin, KTI, P34, lipoxygenase, and stachyose components was developed. The breeding line has purple flower, determinate growth habit, brown pod, black seed coat and green cotyledon. The stem height of the breeding line was 57.0 cm and 100 seed weight was 31.5 g. This is the first soybean breeding line with black seed coat, green cotyledon and penta null genotypes for lectin, KTI, P34, lipoxygenase, and stachyose factors. This line will be used as parent to improve a black soybean cultivar with green cotyledon that have significantly reduced anti-nutritional and allergenic traits. [ABSTRACT FROM AUTHOR]

In the current study, two commercial industrial hemp (IH) fiber varieties (V1: CFX-2 and V2: Henola) were assessed for their ability to regulate salt-induced oxidative stress metabolism. For 30 days, plants were cultivated in greenhouse environments with five different salinity treatments (0, 50, 80, 100, 150, and 200 mM NaCl). Hydrogen peroxide (H2O2), malondialdehyde (MDA), and lipoxygenase (LOX) and antioxidant enzymes (superoxide dismutase (SOD), catalase, guaiacol peroxidase (GPOD), ascorbate peroxidase (APX), glutathione reductase (GR), and glutathione-S-transferase (GST)) were assessed in fully expanded leaves. At 200 and 100 mM NaCl concentrations, respectively, 30 days after saline treatment, plants in V1 and V2 did not survive. At 80 mM NaCl, the leaves of V2 showed higher concentrations of H2O2, MDA, and LOX than those of V1. Higher SOD, CAT, GPOD, APX, GR, and GST activity in the leaves of V1 up to 100 mM NaCl resulted in lower levels of H2O2 and MDA. At 80 mM NaCl, V2 demonstrated the total failure of the antioxidant defense mechanism. These results reveal that V1 demonstrated stronger salt tolerance than V2, in part due to better antioxidant metabolism. [ABSTRACT FROM AUTHOR]

Inflammation is a physiological process that involves a variety of inflammatory mediators such as PGE2, Nitric oxides, PLA2 etc., blocking these mediators could be helpful in designing and development of promising anti‐inflammatory compounds. Non‐steroidal anti‐inflammatory drugs (NSAIDs) have been used for the treatment of inflammatory‐related disorders via variety of pathways that include inhibition of COX‐1, COX‐2, LOX, NO etc. Chalcone derivatives, characterized by their unique chemical structure, have emerged as potential candidates for modulating inflammatory pathways. Literature suggested that modifications in chalcone derivatives by targeting different molecular targets could leads to the development of potential anti‐inflammatory agents. In this review, we have summarised representative chalcone derivatives against different anti‐inflammatory targets and discussed their structure‐activity relationship along with relevant data. [ABSTRACT FROM AUTHOR]

Background: Oil bodies or lipid droplets (LDs) in the cytosol are the subcellular storage compartments of seeds and the sites of lipid metabolism providing energy to the germinating seeds. Major LD-associated proteins are lipoxygenases, phospholipaseD, oleosins, TAG-lipases, steroleosins, caleosins and SEIPINs; involved in facilitating germination and enhancing peroxidation resulting in off-flavours. However, how natural selection is balancing contradictory processes in lipid-rich seeds remains evasive. The present study was aimed at the prediction of selection signatures among orthologous clades in major oilseeds and the correlation of selection effect with gene expression. Results: The LD-associated genes from the major oil-bearing crops were analyzed to predict natural selection signatures in phylogenetically close-knit ortholog clusters to understand adaptive evolution. Positive selection was the major force driving the evolution and diversification of orthologs in a lineage-specific manner. Significant positive selection effects were found in 94 genes particularly in oleosin and TAG-lipases, purifying with excess of non-synonymous substitution in 44 genes while 35 genes were neutral to selection effects. No significant selection impact was noticed in Brassicaceae as against LOX genes of oil palm. A heavy load of deleterious mutations affecting selection signatures was detected in T-lineage oleosins and LOX genes of Arachis hypogaea. The T-lineage oleosin genes were involved in mainly anther, tapetum and anther wall morphogenesis. In Ricinus communis and Sesamum indicum > 85% of PLD genes were under selection whereas selection pressures were low in Brassica juncea and Helianthus annuus. Steroleosin, caleosin and SEIPINs with large roles in lipid droplet organization expressed mostly in seeds and were under considerable positive selection pressures. Expression divergence was evident among paralogs and homeologs with one gene attaining functional superiority compared to the other. The LOX gene Glyma.13g347500 associated with off-flavor was not expressed during germination, rather its paralog Glyma.13g347600 showed expression in Glycine max. PLD-α genes were expressed on all the tissues except the seed,δ genes in seed and meristem while β and γ genes expressed in the leaf. Conclusions: The genes involved in seed germination and lipid metabolism were under strong positive selection, although species differences were discernable. The present study identifies suitable candidate genes enhancing seed oil content and germination wherein directional selection can become more fruitful. [ABSTRACT FROM AUTHOR]

Copyright of Shipin Kexue/ Food Science is the property of Food Science Editorial Department and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Lipoxygenase (LOX) is associated with responses to plant hormones, environmental stresses, and signaling substances. Methyl jasmonate (MeJA) treatment triggers the production of LOX, polyphenol oxidase, and protease inhibitors in various plants, producing resistance to herbivory. To examine the response of MtLOX24 to MeJA, the phenotypic and physiological changes in Medicago truncatula MtLOX24 overexpression and lox mutant plants were investigated. Additionally, wild-type R108, the MtLOX24-overexpressing line L4, and the mutant lox-1 were utilized as experimental materials to characterize the differentially expressed genes (DEGs) and metabolic pathways in response to MeJA. The results indicate that after treatment with 200 µM of MeJA, the damage in the mutants lox-1 and lox-2 was more serious than in the overexpressing lines L4 and L6, with more significant leaf wilting, yellowing, and oxidative damage in lox-1 and lox-2. Exogenous application of MeJA induced H2O2 production and POD activity but reduced CAT activity in the lox mutants. Transcriptome analysis revealed 10,238 DEGs in six libraries of normal-growing groups (cR108, cL4, and clox1) and MeJA-treated groups (R108, L4, and lox1). GO and KEGG functional enrichment analysis demonstrated that under normal growth conditions, the DEGs between the cL4 vs. cR108 and the clox-1 vs. cR108 groups were primarily enriched in signaling pathways such as plant–pathogen interactions, flavonoid biosynthesis, plant hormone signal transduction, the MAPK signaling pathway, and glutathione metabolism. The DEGs of the R108 vs. cR108 and L4 vs. cL4 groups after MeJA treatment were mainly enriched in glutathione metabolism, phenylpropanoid biosynthesis, the MAPK signaling pathway, circadian rhythm, and α-linolenic acid metabolism. Among them, under normal growth conditions, genes like PTI5, PR1, HSPs, PALs, CAD, CCoAOMT, and CYPs showed significant differences between L4 and the wild type, suggesting that the expression of these genes is impacted by MtLOX24 overexpression. CDPKs, CaMCMLs, IFS, JAZ, and other genes were also significantly different between L4 and the wild type upon MeJA treatment, suggesting that they might be important genes involved in JA signaling. This study provides a reference for the study of the response mechanism of MtLOX24 under MeJA signaling. [ABSTRACT FROM AUTHOR]

Lipid metabolism is a complex process crucial for energy production resulting in high levels of acyl-coenzyme A (acyl-CoA) molecules in the cell. Acyl-CoAs have also been implicated in inflammation, which could be possibly linked to lipoxygenase (LOX) biochemistry by the observation that an acyl-CoA was bound to human platelet 12-lipoxygenase via cryo-EM. Given that LOX isozymes play a pivotal role in inflammation, a more thorough investigation of the inhibitory effects of acyl-CoAs on lipoxygenase isozymes was judged to be warranted. Subsequently, it was determined that C18 acyl-CoA derivatives were the most potent against h12-LOX, human reticulocyte 15-LOX-1 (h15-LOX-1), and human endothelial 15-LOX-2 (h15-LOX-2), while C16 acyl-CoAs were more potent against human 5-LOX. Specifically, oleoyl-CoA (18:1) was most potent against h12-LOX (IC50 = 32 μM) and h15-LOX-2 (IC50 = 0.62 μM), stearoyl-CoA against h15-LOX-1 (IC50 = 4.2 μM), and palmitoleoyl-CoA against h5-LOX (IC50 = 2.0 μM). The inhibition of h15-LOX-2 by oleoyl-CoA was further determined to be allosteric inhibition with a Ki of 82 +/- 70 nM, an α of 3.2 +/- 1, a β of 0.30 +/- 0.07, and a β/α = 0.09. Interestingly, linoleoyl-CoA (18:2) was a weak inhibitor against h5-LOX, h12-LOX, and h15-LOX-1 but a rapid substrate for h15-LOX-1, with comparable kinetic rates to free linoleic acid (kcat = 7.5 +/- 0.4 s-1, kcat/KM = 0.62 +/- 0.1 µM-1s-1). Additionally, it was determined that methylated fatty acids were not substrates but rather weak inhibitors. These findings imply a greater role for acyl-CoAs in the regulation of LOX activity in the cell, either through inhibition of novel oxylipin species or as a novel source of oxylipin-CoAs.

Abstract Glaucoma leads to vision loss due to retinal ganglion cell death. Astrocyte reactivity contributes to neurodegeneration. Our recent study found that lipoxin B4 (LXB4), produced by retinal astrocytes, has direct neuroprotective actions on retinal ganglion cells. In this study, we aimed to investigate how the autacoid LXB4 influences astrocyte reactivity in the retina under inflammatory cytokine-induced activation and during ocular hypertension. The protective activity of LXB4 was investigated in vivo using the mouse silicone-oil model of chronic ocular hypertension. By employing a range of analytical techniques, including bulk RNA-seq, RNAscope in-situ hybridization, qPCR, and lipidomic analyses, we discovered the formation of lipoxins and expression of the lipoxin pathway in rodents (including the retina and optic nerve), primates (optic nerve), and human brain astrocytes, indicating the presence of this neuroprotective pathway across various species. Findings in the mouse retina identified significant dysregulation of the lipoxin pathway in response to chronic ocular hypertension, leading to an increase in 5-lipoxygenase (5-LOX) activity and a decrease in 15-LOX activity. This dysregulation was coincident with a marked upregulation of astrocyte reactivity. Reactive human brain astrocytes also showed a significant increase in 5-LOX. Treatment with LXB4 amplified the lipoxin biosynthetic pathway by restoring and amplifying the generation of another member of the lipoxin family, LXA4, and mitigated astrocyte reactivity in mouse retinas and human brain astrocytes. In conclusion, the lipoxin pathway is functionally expressed in rodents, primates, and human astrocytes, and is a resident neuroprotective pathway that is downregulated in reactive astrocytes. Novel cellular targets for LXB4’s neuroprotective action are inhibition of astrocyte reactivity and restoration of lipoxin generation. Amplifying the lipoxin pathway is a potential target to disrupt or prevent astrocyte reactivity in neurodegenerative diseases, including retinal ganglion cell death in glaucoma.

The enzyme soybean lipoxygenase (SLO) provides a prototype for deep tunneling mechanisms in hydrogen transfer catalysis. This work combines room temperature X-ray studies with extended hydrogen-deuterium exchange experiments to define a catalytically-linked, radiating cone of aliphatic side chains that connects an active site iron center of SLO to the protein-solvent interface. Employing eight variants of SLO that have been appended with a fluorescent probe at the identified surface loop, nanosecond fluorescence Stokes shifts have been measured. We report a remarkable identity of the energies of activation (Ea) for the Stokes shifts decay rates and the millisecond C-H bond cleavage step that is restricted to side chain mutants within an identified thermal network. These findings implicate a direct coupling of distal protein motions surrounding the exposed fluorescent probe to active site motions controlling catalysis. While the role of dynamics in enzyme function has been predominantly attributed to a distributed protein conformational landscape, the presented data implicate a thermally initiated, cooperative protein reorganization that occurs on a timescale faster than nanosecond and represents the enthalpic barrier to the reaction of SLO.

The antiplatelet effect of polyunsaturated fatty acids is primarily attributed to its metabolism to bioactive metabolites by oxygenases, such as lipoxygenases (LOX). Platelets have demonstrated the ability to generate 15-LOX-derived metabolites (15-oxylipins); however, whether 15-LOX is in the platelet or is required for the formation of 15-oxylipins remains unclear. This study seeks to elucidate whether 15-LOX is required for the formation of 15-oxylipins in the platelet and determine their mechanistic effects on platelet reactivity. In this study, 15-HETrE, 15-HETE, and 15-HEPE attenuated collagen-induced platelet aggregation, and 15-HETrE inhibited platelet aggregation induced by different agonists. The observed anti-aggregatory effect was due to the inhibition of intracellular signaling including αIIbβ3 and protein kinase C activities, calcium mobilization, and granule secretion. While 15-HETrE inhibited platelets partially through activation of peroxisome proliferator-activated receptor β (PPARβ), 15-HETE also inhibited platelets partially through activation of PPARα. 15-HETrE, 15-HETE, or 15-HEPE inhibited 12-LOX in vitro, with arachidonic acid as the substrate. Additionally, a 15-oxylipin-dependent attenuation of 12-HETE level was observed in platelets following ex vivo treatment with 15-HETrE, 15-HETE, or 15-HEPE. Platelets treated with DGLA formed 15-HETrE and collagen-induced platelet aggregation was attenuated only in the presence of ML355 or aspirin, but not in the presence of 15-LOX-1 or 15-LOX-2 inhibitors. Expression of 15-LOX-1, but not 15-LOX-2, was decreased in leukocyte-depleted platelets compared to non-depleted platelets. Taken together, these findings suggest that 15-oxylipins regulate platelet reactivity; however, platelet expression of 15-LOX-1 is low, suggesting that 15-oxylipins may be formed in the platelet through a 15-LOX-independent pathway.

Metal complexes have pivotal applications in catalysis, photochemistry, and biological systems. Due to their unique chemical properties, metal complexes occupy a coveted place in the field of drug design. The present study includes the synthesis of symmetrical oxamides (O1–O11) and their copper complexes (C1–C11) that have potential to inhibit α‐glucosidase and lipoxygenase enzymes. To elucidate the structures of these compounds, spectroscopic investigations including mass spectrometry, 1H‐NMR spectroscopy, UV‐Visible spectroscopy and IR spectroscopy were adapted along with CHN elemental analysis. The spectroscopic investigations suggested that the synthesized complexes to be square planar with metal to ligand ratio 1 : 2. All compounds were scanned for potential inhibitory studies against α‐glucosidase and lipoxygenase enzyme. Their structure activity relationships (SAR) were also established. It was found that all synthesized compounds are strong inhibitors of α‐glucosidase when compared to standard 1‐deoxynojirimycin. It was noteworthy that presence of substituent played a significant role for the enhancing potential inhibitory skills of the compounds. The results obtained were also compared with molecular docking studies. A similar inhibition trend was observed between experimental inhibitory activity and docking energies. The current work presents a special class of Cu(II) coordination complexes from the perspective of their biological as well as structural features. [ABSTRACT FROM AUTHOR]

Traditional nutraceutical therapy is a promising approach in the management of inflammatory disorders. Fenugreek, bottle gourd, and betel leaf are rich sources of polyphenolic secondary metabolites. They are abundantly found in the Indian subcontinent for the preparation of healthy traditional functional food. Traditional evidence recommended that these herbs could control inflammation. The study was conducted with those herbs to validate their protective role against inflammatory mediators of arthritis and asthma through in‐silico and in‐vitro models. Phytochemical tests revealed the presence of flavonoids, amino acids, polyphenols, and saponins. Molecular docking was performed against six potential inflammatory bio‐marker proteins, like, LOX‐5, 15‐LOX, PLA2, IL‐6, TNFR1 & TNF‐α with the phyto‐marker compounds of test herbs and found satisfactory binding scores against inflammatory bio‐markers. Extended studies with Fenugreek and betel leaf aqueous fractions showcased 87 % and 91 % inhibition of protein denaturation at 200 μg/ml. Bottle‐gourd and betel‐leaf demonstrated 96 % and 98 % inhibition against 15‐Lipoxygenase at 100 μg/ml. TNF‐α and IL‐6 were dropped significantly by 49 % and 36 % upon treatment of betel leaf at 200 μg/ml from the LPS‐stimulated PBMC cultured cells. Findings may recommend the development of commercial anti‐inflammatory polyherbal phytopharmaceuticals for controlling polyarthritis and inflammatory complications subject to preclinical in‐vivo and clinical investigation. [ABSTRACT FROM AUTHOR]

Introduction: There is a growing interest in studying natural products for the identification of novel lead compounds for drug development for treating inflammatory diseases. Although some studies have focused anti-inflammatory activity of benzophenones and xanthones, exploring additional targets such as enzymes and cytokines, involved in their inflammatory response could provide more comprehensive understanding of the compounds' anti-inflammatory effects. In this study, four xanthones ananixanthone (1), smeathxanthone A (2), smeathxanthone B (3), and 1,3,5,8-tetrahydroxy-2-(3-methybut-2-enyl)-4-(3,7- dimethyloct-2,6-dienyl) xanthone (4); and three benzophenones guttiferone O (5), guttiferone M (6), and aristophenone A (7) from Garcinia smeathmannii (Planch. & Triana) Oliv. were investigated for their effect on nitric oxide production, cyclooxygenase, lipoxygenase inhibition, and Th1/Th2 cytokines production in activated RAW 264.7 macrophages. Methods: The Griess reagent method and the ferrous oxidation-xylenol orange assay were used to evaluate the inhibition of NO production and the 15-lipoxygenase activity respectively. Cyclooxygenase activity was assessed using the fluorometric COX activity assay kit and measurement of Th1/Th2 cytokines was performed using a flow cytometer. Results: All the tested compounds exhibited a dose-dependent inhibition of NO production with varying degrees of inhibitory effects on 15-LOX activity. Compound (6), displays the best inhibitory effect on COX-1/COX-2 activity. A general trend of the tested compounds on cytokines profiles revealed that compound (5) showed a pronounced enhancement of anti-inflammatory cytokines (IL-4 and IL-10). Conclusion: This observation supports future exploration of ananixanthone (1), guttiferone O (5), and guttiferone (6) as potential candidates for the development of anti-inflammatory drugs. [ABSTRACT FROM AUTHOR]

Hydroalcoholic extracts from Malbec and Torrontés wine pomaces (Vitis vinifera L.) originating from the high-altitude vineyards of Argentina's Calchaquí Valleys were characterized. Total phenolics, hydroxycinnamic acids, orthodiphenols, anthocyanins, non-flavonoid phenolics, total flavonoids, flavones/flavonols, flavanones/dihydroflavonols, and tannins were quantified through spectrophotometric methods, with the Malbec extract exhibiting higher concentrations in most of phytochemical groups when compared to Torrontés. HPLC-DAD identified more than 30 phenolic compounds in both extracts. Malbec displayed superior antiradical activity (ABTS cation, nitric oxide, and superoxide anion radicals), reduction power (iron, copper, and phosphomolybdenum), hypochlorite scavenging, and iron chelating ability compared to Torrontés. The cytotoxicity assessments revealed that Torrontés affected the viability of HT29-MTX and Caco-2 colon cancer cells by 70% and 50%, respectively, at the highest tested concentration (1 mg/mL). At the same time, both extracts did not demonstrate acute toxicity in Artemia salina or in red blood cell assays at 500 µg/mL. Both extracts inhibited the lipoxygenase enzyme (IC50: 154.7 and 784.7 µg/mL for Malbec and Torrontés), with Malbec also reducing the tyrosinase activity (IC50: 89.9 µg/mL), and neither inhibited the xanthine oxidase. The substantial phenolic content and diverse biological activities in the Calchaquí Valleys' pomaces underline their potentialities to be valorized for pharmaceutical, cosmetic, and food industries. [ABSTRACT FROM AUTHOR]

High-frequency pulse continuous heating (HFP) is a newly developed electric heating technique that can increase the temperature of slurry foods to over 100 °C in less than 1 s. In this study, we investigated the effect of heating soybean slurry by HFP on lipoxygenase (LOX) activity, the extraction ratio of soy protein, and aroma components of soymilk. Despite the short heating time, HFP inactivated 99 % and 100 % of LOX at 80 °C and 90 °C, respectively. However, conventional heating at 80 °C took 5 min to inactivate LOX. Furthermore, HFP increased the extraction ratio of protein, and decreased the unpleasant beany flavor. These results show that HFP can significantly reduce heating time, thereby enhancing the quality of soymilk. [ABSTRACT FROM AUTHOR]

A novel series of dihydropyrimidine/sulphonamide hybrids 3a-j with antiinflammatory properties have been developed and tested as dual mPGES-1/5- LOX inhibitors. In vitro assay, results showed that compounds 3c, 3e, 3h, and 3j were the most effective dual inhibitors of mPGES-1 and 5-LOX activities. Compound 3j was the most potent dual inhibitor with IC50 values of 0.92 ^M and 1.98 ^M, respectively. In vivo, anti-inflammatory studies demonstrated that compounds 3c, 3e, 3h, and 3e had considerable anti-inflammatory activity, with EI% ranging from 29% to 71%. Compounds 3e and 3j were equivalent to celecoxib after the first hour but exhibited stronger anti-inflammatory effects than celecoxib after the third and fifth hours. Moreover, compounds 3e and 3j significantly reduced the levels of pro-inflammatory cytokines (PGE2, TNF-a, and IL-6) with gastrointestinal safety profiles. Molecular docking simulations explored the most potent derivatives' binding affinities and interaction patterns within mPGES-1 and 5-LOX active sites. This study disclosed that compound 3j is a promising anti-inflammatory lead with dual mPGES-1/5-LOX inhibition that deserves further preclinical investigation. [ABSTRACT FROM AUTHOR]

Fucoidans are sulphated fucose rich heteropolysaccharides found in marine source seaweed. Fucoidan has gained interest due to its diverse biological activities. Dual inhibition of cyclooxygenase and lipoxygenase prevents shunting of the arachidonic acid metabolism sparing gastric toxicity observed with selective cyclooxygenase-2 inhibitors. The present study investigated dual inhibition potential of fucoidan extracted from Sargassum wightii on cyclooxygenase, lipoxygenase and its anti-inflammatory effect on carrageenan induced paw oedema. Fucoidan and diclofenac demonstrated dual inhibition towards cyclooxygenase and lipoxygenase with half-maximal inhibitory concentration of 44.03±3.74 µg/ml, 34.92±5.14 µg/ml (cyclooxygenase) and 28.26±2.06 µg/ml and 26.54±1.37 µg/ml (lipoxygenase) in lipopolysaccharide stimulated RAW 264.7 macrophage cells respectively. Fucoidan is showing concentration dependant reduction in nitric oxide level of lipopolysaccharide stimulated RAW 264.7 cells with a half-maximal inhibitory concentration of 17.88±2.43 µg/ml and 17.99±3.15 µg/ml for diclofenac. Fucoidan showed suppression of lipopolysaccharide stimulated gene expression of cyclooxygenase-2 and 5-lipoxygenase in RAW 264.7 macrophage cells. Fucoidan showed 99.62±1.69 % cell viability in RAW 264.7 macrophage cells. The acute oral toxicity study of fucoidan showed no abnormal clinical signs. The fucoidan attenuated carrageenan induced increase in paw volume in Wistar rats at the tested doses of 100 mg/kg and 200 mg/kg at 60, 120 and 240 min (p<0.05). Results suggest that the anti-inflammatory activities of fucoidan involve attenuation of eicosanoid production. [ABSTRACT FROM AUTHOR]

The understanding of the role of LXR in the regulation of macrophages during inflammation is emerging. Here, we show that LXR agonist T09 specifically increases 15-LOX abundance in primary human M2 macrophages. In time- and dose-dependent incubations with T09, an increase of 3-fold for ALOX15 and up to 15-fold for 15-LOX-derived oxylipins was observed. In addition, LXR activation has no or moderate effects on the abundance of macrophage marker proteins such as TLR2, TLR4, PPARγ, and IL-1RII, as well as surface markers (CD14, CD86, and CD163). Stimulation of M2-like macrophages with FXR and RXR agonists leads to moderate ALOX15 induction, probably due to side activity on LXR. Finally, desmosterol, 24(S),25-Ep cholesterol and 22(R)-OH cholesterol were identified as potent endogenous LXR ligands leading to an ALOX15 induction. LXR-mediated ALOX15 regulation is a new link between the two lipid mediator classes sterols, and oxylipins, possibly being an important tool in inflammatory regulation through anti-inflammatory oxylipins. [ABSTRACT FROM AUTHOR]

The persisting presence of opportunistic pathogens like Pseudomonas aeruginosa poses a significant threat to many immunocompromised cancer patients with pulmonary infections. This review highlights the complexity of interactions in the host's defensive eicosanoid signaling network and its hijacking by pathogenic bacteria to their own advantage. Human lipoxygenases (ALOXs) and their mouse counterparts are integral elements of the innate immune system, mostly operating in the pro-inflammatory mode. Taking into account the indispensable role of inflammation in carcinogenesis, lipoxygenases have counteracting roles in this process. In addition to describing the structure-function of lipoxygenases in this review, we discuss their roles in such critical processes as cancer cell signaling, metastases, death of cancer and immune cells through ferroptosis, as well as the roles of ALOXs in carcinogenesis promoted by pathogenic infections. Finally, we discuss perspectives of novel oncotherapeutic approaches to harness lipoxygenase signaling in tumors. [ABSTRACT FROM AUTHOR]

Changes in the lipid oxidation and volatile compounds of a variety of tilapia tissues (Oreochromis niloticus) including the muscle, gills, and skin during ice storage were investigated by evaluating peroxide values (PVs), lipoxygenase (LOX) activity, fatty acid (FA) composition, and volatile substances. LOX activity and PV were determined in the gills, skin, and muscles throughout 9 days of storage in ascending order to the extended storage time. The highest level of LOX activity was found in the gills, whereas the highest PV was determined in the skin. FA content of all tissues decreased during the storage period. Oleic acid was the predominant monounsaturated fatty acid, whereas linoleic acid and docosahexaenoic acid were the main polyunsaturated fatty acids and omega‐3 in all tissues. The fish gills were shown to have the highest level of volatile compounds followed by the skin and muscle, based on headspace solid‐phase microextraction coupled with gas chromatography and mass spectrometry. Principal component analysis indicated gradual changes in the volatile compound composition with increasing storage time. 2‐Butanone and nonanal in the muscle, 6‐methyl‐2‐heptanone and 2‐nonenal in the gills, and 1‐heptanol, and 1‐nonanol in the skin were found to be the potential freshness indicators. In addition, hexanal could be a general potential marker for measuring the degree of lipid oxidation in all tissues. Practical Application: Understanding the volatile compound formation related to lipid oxidation within storage time at various tissues of tilapia could be critical to the side‐stream processing to yield the desired quality. [ABSTRACT FROM AUTHOR]

Research background. Among legumes, peas are characterised by their high protein content, low glycaemic index and exceptional versatility. However, their potential as a food is often compromised by their undesirable off-flavour and taste. Hence, this study focuses on minimising off-flavours through simple pretreatments with the aim of improving the potential for the production of pea milk analogues. Pea milk analogues are a burgeoning type of plant-based milk alternatives in the growing plant-based market. Experimental approach. Pea seeds were subjected to different pretreatments: (i) dry milling, (ii) blanching followed by soaking in alkaline solution and subsequent dehulling and (iii) vacuum. Typical physicochemical properties such as pH, viscosity, colour, titratable acidity and yield were measured to obtain a brief overview of the products. Consumer acceptance test, descriptive sensory analysis, gas chromatography-mass spectrometry and gas chromatography-olfactometry were used to map the complete sensory profile and appeal of the pea milk substitutes. Results and conclusions. The L* values of the pea milk analogues were significantly lower than those of cow’s milk, while a*, b*, viscosity and pH were similar. In the descriptive sensory analysis, sweet, astringent, pea-like, cooked, hay-like, boiled corn and green notes received relatively higher scores. The vacuum-treated pea milk analogues received higher scores for flavour and overall acceptability in the consumer acceptance test. The pretreatments resulted in significant changes in the volatile profiles of the pea milk analogues. Some volatiles typically associated with off-flavour, such as hexanal, were found in higher concentrations in blanched pea milk analogues. Among the applied pretreatments, vacuum proved to be the most effective method to reduce the content of volatile off-flavour compounds. Novelty and scientific contribution. This study stands out as a rare investigation to characterise pea milk analogues and to evaluate the impact of simple pretreatments on the improvement of their sensory properties. The results of this study could contribute to the development of milk alternatives that offer both high nutritional value and strong appeal to consumers. [ABSTRACT FROM AUTHOR]

Ischemic stroke remains a leading cause of mortality and long-term disability worldwide, necessitating efforts to identify biomarkers for diagnosis, prognosis, and treatment monitoring. The present study aimed to identify novel plasma biomarkers of neurodegeneration and inflammation in a mouse model of stroke induced by distal middle cerebral artery occlusion. Using targeted lipidomic and global untargeted metabolomic profiling of plasma collected from aged male mice 24 h after stroke and weekly thereafter for 7 weeks, we discovered distinct acute and chronic signatures. In the acute phase, we observed elevations in myelin-associated lipids, including sphingomyelin (SM) and hexosylceramide (HCER) lipid species, indicating brain lipid catabolism. In the chronic phase, we identified 12-hydroxyeicosatetraenoic acid (12-HETE) as a putative biomarker of prolonged inflammation, consistent with our previous observation of a biphasic pro-inflammatory response to ischemia in the mouse brain. These results provide insight into the metabolic alterations detectable in the plasma after stroke and highlight the potential of myelin degradation products and arachidonic acid derivatives as biomarkers of neurodegeneration and inflammation, respectively. These discoveries lay the groundwork for further validation in human studies and may improve stroke management strategies.

Medicinal plants have been utilized for centuries in traditional medicine systems worldwide, providing a rich source of bioactive compounds with diverse biological activities. Lavandula officinalis, a member of the Lamiaceae family, has been recognized for its multifaceted pharmacological activities. In this current investigation, our primary objective was to scrutinize the in vitro inhibitory potential of L. officinalis essential oil (LOEO) against alpha-amylase and alpha-glucosidase, with the aim of understanding its antidiabetic effects. Additionally, the assay encompassed tyrosinase and lipoxygenase (LOX) to assess its anti-inflammatory attributes. Unraveling the underlying molecular mechanisms of these activities prompted an in-silico study. The purpose was to establish correlations between in-vitro observations and computational insights derived from molecular docking, which forecasts the interaction of LOEO molecules with their respective targets, alongside ADMET prediction. The Gas Chromatography-Mass Spectrometry (GC-MS) analysis allow to identify eighteen compounds, with the dominance of L−camphor (43.12 %), 1,8−cineole (34.27 %) and borneol (8.60 %) in LOEO. The antidiabetic evaluation revealed that LOEO exhibited noteworthy inhibitory activity against both α-amylase and α-glucosidase, displaying IC50 values of 3.14 ± 0.05 mg/mL and 2.07 ± 0.03 mg/mL, respectively. The subsequent in-silico study highlighted the particularly strong binding affinity of (E)-Farnesene, with a binding score of −7.4 kcal/mol for alpha-glucosidase, while Germacrene D displayed the highest affinity among the ligands (−7.9 kcal/mol) for the alpha-amylase target. Furthermore, the investigation into in vitro anti-inflammatory activity unveiled LOEO efficacy against tyrosinase (IC50 = 42.74 μg/mL) and LOX (IC50 = 11.58 ± 0.07 μg/mL). The in-silico analysis echoed these findings, indicating α-Cadinene's notable binding affinity of 6 kcal/mol with tyrosinase and α-Cedrene's binding score of −6.5 kcal/mol for LOX. Impressively, for both COX-1 and COX-2, α-Cedrene exhibited significant binding affinities of −7.6 and −7.3 kcal/mol, respectively. The convergence between the in vitro and in silico outcomes underscores the potential of LOEO and its constituent compounds as potent inhibitors targeting both diabetes and the inflammatory processes.