LOX Activity Research Articles

Pharmacological evaluation and molecular docking of new di-tert-butylphenol compound, LQFM-091, a new dual 5-LOX/COX inhibitor

Dual 5-LOX/COX inhibitors are potential new dual drugs to treat inflammatory conditions. This research aimed to design, synthesis and to evaluate the anti-inflammatory and antinociceptive effects of the new compound, which is derived from nimesulide and darbufelone lead compounds. The new dual inhibitor 5-LOX/COX has the possible advantage of gastrointestinal safety. A voltammetric experiment was conducted to observe the drug's antioxidative effect. A formalin test, a hot plate test and carrageenan-induced mechanical hyperalgesia were employed to evaluate the analgesic nature of LQFM-091. To evaluate anti-inflammatory activity, we measured edema, leukocyte count, myeloperoxidase activity and cytokines levels in carrageenan-induced inflammation tests. We elucidated the underlying mechanisms by assessing the interaction the with COXs and LOX enzymes by colorimetric screening assay and molecular docking. The lethal dose (LD50) was estimated using 3T3 Neutral Red Uptake assay. Our results indicate that the LQFM-091 prototype is a powerful antioxidant, as well as able to inhibit COX-1, COX-2 and LOX activities. LQFM091 was classified in GHS category 4 (300<LD50<2000mg/Kg). This prototype showed analgesic activity in the formalin test and decreased carrageenan-induced mechanical hyperalgesia. Furthermore, LQFM-091 reduced the paw edema induced by carrageenan and reduced the leukocyte count, myeloperoxidase activity, TNF-α and IL-1β levels in the pleural exudate. Another interesting finding was the absence of gastrointestinal lesions. These data indicate that LQFM-091 produced antinociceptive and anti-inflammatory effects while maintaining gastrointestinal safety. Furthermore, this compound presented a safe toxicological profile. Blocked COXs and LOX enzymes are important targets for manipulating the mechanism of this compound.

Molecular characterization of lysyl oxidase‐mediated extracellular matrix remodeling during mouse decidualization

The establishment of decidualization is a prerequisite of successful pregnancy. Lysyl oxidase (Lox) is a copper-containing amine oxidase which catalyzes cross-linking of collagen and elastin in the ECM. Lox is expressed in the subluminal stroma surrounding the implanting blastocyst on day 5 of pregnancy. From days 6 to 8, the signals for Lox mRNA and protein are strongly detected in the decidual cells. The expression of Lox is under the control of estrogen via the GSK-3β/β-catenin/c-myc pathway. Dtprp is decreased by the inhibition of Lox activity. Furthermore, the inhibition of Lox activity decreases stromal cell migration and embryo adhesion. Our findings highlight the crucial role of Lox in endometrial stromal cells and deepen our understanding of decidualization.

Yield, grain size, protein content and water use efficiency of null-LOX malt barley in a semiarid Mediterranean agroecosystem

The recent development of null-LOX barley (i.e. barley that is defective in the synthesis of two lipoxygenase enzymes, LOX-1 and LOX-2), which is oriented to produce a better quality beer, has increasingly drawn the attention of malt industry. However, the inhibition of LOX activity in such barley is raising concerns about the efficiency of its defense mechanisms to biotic and more specifically to abiotic stresses. A 2-year experiment was carried out in a typical Mediterranean environment of central southern Greece in order to: (i) explore the negative effect of drought on yield, yield components, grain size, grain protein content and water use efficiency in malt barley cultivars presenting a total loss of function of LOX-1 and LOX-2 activity, compared to a local malt barley cultivar having a full functional LOXs activity and (ii) introduce a new concept in water use efficiency, which focuses on the proportion of grains with acceptable size for malting, because according to malting industries one of the most limiting factors for the sustainable production of malt barley in the Mediterranean region is the inadequate grain size of malt barley, due to the frequent occurrence of terminal drought stress. The total loss of function of LOX-1 and LOX-2 activity did not cause any obvious advantage for the traditional malt barley cultivar over null-LOX cultivars, in terms of grain yield, yield components, grain size, grain protein content and water use efficiency. Increasing water availability with irrigation generally resulted in higher proportion of plump grains (>2.8mm) and lower proportion of thin grains (< 2.2mm), especially when water availability during the post-anthesis period was decreased and the temperature was increased. It was demonstrated that grain protein content of null-LOX cultivars was less responsive to soil water availability compared to grain size. Finally, it was shown that the new expressions of water use efficiency were able to discriminate the different adaptive responses of the studied barley cultivars to soil water availability.

Flat enzyme-based lactate biofuel cell integrated with power management system: Towards long term in situ power supply for wearable sensors

Flat enzyme-based lactate biofuel cell (ELBC) integrated with power management system (PMS) was developed as a potential power supply for wearable sensors. Kinetic models of power output and chemical reagent concentration (lactate oxidase LOx and lactate) were developed to determine the limiting factor of the ELBC performance. Given the lactate concentration (0–40mM) in human sweat, the optimum LOx amount coated on the anode ranged from 6U to 54U based on the experimental maximum power output. In the ELBC-PMS entity simulation test, power discharge/recharge frequency was calculated for self-sustained sensing and data transmission, indicating that the ELBC could support wearable sensors (power requirement: 1–1350mW; signal transfer frequency: 1–1320times/h). The electrochemical activity of LOx coated anode was validated using the cyclic voltammetry (CV). The coefficient of variance and regression statistical analysis revealed the high stability and long lifespan (2weeks) of ELBC without the need of lactate refill.

Lysyl Oxidase Is a Strong Determinant of Tumor Cell Colonization in Bone.

Lysyl oxidase (LOX) is a secreted copper-dependent amine oxidase whose primary function is to drive collagen crosslinking and extracellular matrix stiffness. LOX in colorectal cancer synergizes with hypoxia-inducible factor-1 (HIF-1) to promote tumor progression. Here we investigated whether LOX/HIF1 endows colorectal cancer cells with full competence for aggressive colonization in bone. We show that a high LOX expression in primary tumors from patients with colorectal cancer was associated with poor clinical outcome, irrespective of HIF-1 In addition, LOX was expressed by tumor cells in the bone marrow from colorectal cancer patients with bone metastases. In vivo experimental studies show that LOX overexpression in colorectal cancer cells or systemic delivery of the conditioned medium from LOX-overexpressing colorectal cancer cells promoted tumor cell dissemination in the bone marrow and enhanced osteolytic lesion formation, irrespective of HIF-1 Conversely, silencing or pharmacologic inhibition of LOX activity blocked dissemination of colorectal cancer cells in the bone marrow and tumor-driven osteolytic lesion formation. In vitro, tumor-secreted LOX supported the attachment and survival of colorectal cancer cells to and in the bone matrix, and inhibited osteoblast differentiation. LOX overexpression in colorectal cancer cells also induced a robust production of IL6. In turn, both LOX and IL6 were acting in concert to promote RANKL-dependent osteoclast differentiation, thereby creating an imbalance between bone resorption and bone formation. Collectively, our findings show that LOX supports colorectal cancer cell dissemination in the bone marrow and they reveal a novel mechanism through which LOX-driven IL6 production by colorectal cancer cells impairs bone homeostasis. Cancer Res; 77(2); 268-78. ©2016 AACR.

The two-row malting barley cultivar 'New Sachiho Golden' with null lipoxygenase-1 improves flavor stability in beer and was developed by marker-assisted selection.

Lipoxygenase-1 (LOX-1) null ‘New Sachiho Golden’ is a two-row malting barley (Hordeum vulgare L.) cultivar released in 2015 that was developed at the Tochigi Prefectural Agricultural Experimental Station by backcross breeding using the high-yield leading cultivar ‘Sachiho Golden’ as a recurrent parent and the LOX-1 null mutant ‘Daikei LM1’ as a non-recurrent parent. To develop ‘New Sachiho Golden’ we used a simple LOX activity assay and marker-assisted selection. This is the first LOX-1 null malting barley cultivar in Japan that is resistant to barley yellow mosaic virus (types I–III). Agronomic characteristics and malting qualities of ‘New Sachiho Golden’ were similar to those of ‘Sachiho Golden’, except that ‘New Sachiho Golden’ had no LOX activity in ungerminated grains and had clearly lower LOX activity during malting than ‘Sachiho Golden’. The concentrations of a trans-2-nonenal (T2N) precursor in wort and beer made from ‘New Sachiho Golden’ were significantly lower than in those made from ‘Sachiho Golden’, both before and after storage.

Characterization of expression and enzyme activity of lipoxygenases during fruit softening and superficial scald development in ‘Wujiuxiang’ pear

Lipoxygenases (LOXs; EC 1.13.11.12) have been implicated in fruit ripening and senescence; however, little is known regarding the specific LOX genes involved in fruit softening and scald develop- ment in pear. In this study, two 9-lipoxygenase genes, PcLOX1 and PcLOX7 , were characterized in silico in the Pyrus communis draft genome. The expression pattern of PcLOX1 and PcLOX7 , LOX ac-tivity, respiration rate and ethylene production were analyzed during fruit softening and superficial scald development in ‘Wujiuxiang’ pear with or without 1-MCP treatment. The results demonstrated that PcLOX1 and PcLOX7 belong to the type-1 LOX subfamily, and their expression levels, enzyme activity, respiration rate and ethy-lene production were increased during fruit softening and scald de­velopment. In contrast, 1-MCP treatment could effectively suppress respiration rate and ethylene production and inhibit expression of PcLOX1 and PcLOX7 and LOX enzyme activity, resulting in de- layed fruit softening and scald development. These findings indicate that PcLOX1 and PcLOX7 are involved in fruit ripening and scald development in ‘Wujiuxiang’ pear.

Comparative anti-psoriatic efficacy studies of clobetasol loaded chitin nanogel and marketed cream

In the present study chitin nanogel loaded with anti-psoriatic drug clobetasol was developed (CLCNG) for its topical delivery in psoriasis. CLCNG had the particle size of 132±14nm, with gel like consistency, stability in refrigerator, having higher drug release properties at acidic pH. CLCNG exhibited significant toxicity towards HaCaT and THP-1cell lines by MTT assay. The uptake of nanogel by HaCaT cell lines was confirmed by fluorescent microscopy. CLCNG at 0.35mg/ml exhibited significant anti-inflammatory activity with an average of 65% and 70% inhibition in COX and LOX activities expressed in THP-1 cells. In vitro skin permeation studies revealed the increased transdermal flux with fragmented stratum corneum and loosened epidermal layers in CLCNG treated samples, compared with control drug solution. The in vivo anti-psoriatic studies done on imiquimod model confirmed the potential benefits of the nanogel for the topical delivery of clobetasol in psoriasis.

VOLATILE COMPOUNDS IN GOLDEN DELICIOUS APPLE FRUIT (Malus domestica) DURING COLD STORAGE

Biosynthesis of volatile compounds (VC), as well as activity of related enzymes (lipoxygenase LOX, alcohol acyltransferase AAT, and alcohol dehydrogenase ADH), and fatty acids (palmitic, stearic, oleic, linoleic and linolenic acids) were assessed in Golden Delicious fruit apples (Malus domestica Borkh.) during 1 ºC storage at different atmosphere conditions. Three atmosphere conditions were used: 21 % O2 and &gt; 1 % CO2 (Regular Atmosphere, RA), 3 % CO2 and 2 % O2 (Controlled Atmosphere, CA), and CA, with 7 d under RA conditions (CA + RA), to evaluate the effect of shorts periods under air storage. CA conditions inhibited the production of butyl acetate and hexyl acetate esters, and increased hexanol concentration. Production of the branched ester 2-methyl butyl acetate did not decrease under CA conditions. As a result of 7 d under RA, butyl acetate and hexyl acetate in CA + RA increased, mainly after one month of storage. Storage under CA conditions inhibited LOX and AAT activity at some stages whereas ADH activity increased during CA storage. LOX activity showed high correlation with production of aldehydes (r2 = 0.85) and cis-2-hexenal (r2 = 0.94), during storage of apples under CA conditions. Good correlation was found between AAT activity and total esters and butyl acetate content under CA storage of apples (r2 = 0.92 and r2 = 0.93, respectively). While most fatty acids increased in concentration during RA and CA storage, linolenic acid content decreased. No correlation between volatile compounds content and fatty acid production was found.

Lipoxygenase Inhibitory Activities of Some Plant Extracts and Chemical Compounds

Lipoxygenases (LOXs, linoleate: oxygen reductase, E.C. 1.13.11.12) are a family of non-heme ironcontaining dioxygenases. LOXs are associated with several inflammation-related diseases such as arthritis, asthma, cardiovascular, kidney, skin and allergic diseases, neurodegenerative disorders, cancer and metabolic syndrome. In this study, the inhibitory effects of ethanolic extracts prepared from 12 different plants and 12 different chemical compounds were investigated on the activity of LOX which has an important value in the health area. All the plant extracts and chemical substances used in our study showed LOX inhibitory effect. The enzyme inhibitory activities of the extracts and chemical compounds were increasing in a dose-dependent manner. The results obtained in the present study indicate that plant extracts and chemical compounds examined can be a potential source of novel antiinflammatory therapeutics.

Pb accumulation, Pb tolerance, antioxidants, thiols, and organic acids in metallicolous and non-metallicolous Peganum harmala L. under Pb exposure

The aim of the present study was to compare the effect of lead (Pb) exposure (0–50mgL−1 Pb) on Pb accumulation, growth, and the concentrations of thiol compounds, organic acids, antioxidant enzyme activities and other biochemical parameters between a non-metallicolous and a metallicolous harmal (Peganum harmala L.) population. Seedlings were grown in hydroponics, and exposed to 0, 5, 10, 25 and 50mgL−1 Pb for 14 days. Lead decreased dry weight to the same extent in both populations, in a concentration-dependent way. Pb accumulation was consistently higher in the metallicolous population than in the non-metallicolous one, both in roots and shoots, showing that the metallicolous plants were more tolerant to plant-internal Pb. Under Pb exposure the concentrations of malondialdehyde (MDA) and H2O2, as well as the LOX and GPX activities, were higher in the non-metallicolous population, albeit inconsiderably. The activities of antioxidant enzymes such as superoxide dismutase (SOD), ascorbate peroxidase (APX), catalase (CAT), as well as the concentrations of the non-enzymatic antioxidants GSH, ascorbic acid, and proline, were enhanced under Pb exposure, and overall higher in the metallicolous population, though only significantly so under Pb exposure. The concentrations of total thiols, non-protein thiols and protein bound thiols were also increased under Pb exposure. In particular, Pb strongly enhanced the average protein cysteine content in both populations, but much more so in the metallicolous plants than in the non-metallicolous ones. Furthermore, Pb-exposed plants showed significantly increased concentrations of organic acids. Citrate accumulation was strongly induced under Pb exposure, particularly in the metallicolous plants, suggesting that citrate is the most obvious candidate ligand for Pb in the vacuole. Overall, the quantitatively most remarkable differences in Pb response between the populations lie in the degrees of Pb-induced accumulation of proline and protein thiol.

Effects of abiotic stress and hormones on the expressions of five 13-CmLOXs and enzyme activity in oriental melon (Cucumis melo var. makuwa Makino)

AbstractLipoxygenases (LOXs) are a group of non-heme, iron-containing enzymes and extensively involved in plant growth and development, ripening and senescence, stress responses, biosynthesis of regulatory molecules and defense reaction. In our previous study, 18 LOXs in melon genome were screened and identified, and five 13-LOX genes (CmLOX08, CmLOX10, CmLOX12, CmLOX13 and CmLOX18) were predicted to locate in chloroplast. Phylogenetic analysis result showed that the five genes have high homology with jasmonic acid (JA) biosynthesis-related LOXs from other plants. In addition, promoter analysis revealed that motifs of the five genes participate in gene expression regulated by hormones and stresses. Therefore, we analyzed the expressions of the five genes and LOX activity in leaves of four-leaf stage seedlings of oriental melon cultivar Yumeiren under abiotic stress: wounding, cold, high temperature and hydrogen peroxide (H2O2), and signal molecule treatments: methyl jasmonate (MeJA), abscisic acid (ABA) and salicylic acid (SA). Real time qPCR revealed that wounding and H2O2 induced the expressions of all the five genes. Only CmLOX08 was induced by cold while only CmLOX13 was suppressed by high temperature. ABA induced the expressions of CmLOX10 and CmLOX12 while inhibited CmLOX13 and CmLOX18. MeJA increased the 3 genes expressions except CmLOX08 and CmLOX13, whereas SA decreased the effect, apart from CmLOX12. All the abiotic stresses and signal molecules treatments increased the LOX activity in leaves of oriental melon. In summary, the results suggest that the five genes have diverse functions in abiotic stress and hormone responses, and might participate in defense response. The data generated in this study will be helpful in subcellular localization and transgenic experiment to understand their precise roles in plant defense response.

Effect of N-Feruloylserotonin and Methotrexate on Severity of Experimental Arthritis and on Messenger RNA Expression of Key Proinflammatory Markers in Liver

Rheumatoid arthritis (RA) is a chronic inflammatory disease, leading to progressive destruction of joints and extra-articular tissues, including organs such as liver and spleen. The purpose of this study was to compare the effects of a potential immunomodulator, natural polyphenol N-feruloylserotonin (N-f-5HT), with methotrexate (MTX), the standard in RA therapy, in the chronic phase of adjuvant-induced arthritis (AA) in male Lewis rats. The experiment included healthy controls (CO), arthritic animals (AA), AA given N-f-5HT (AA-N-f-5HT), and AA given MTX (AA-MTX). N-f-5HT did not affect the body weight change and clinical parameters until the 14th experimental day. Its positive effect was rising during the 28-day experiment, indicating a delayed onset of N-f-5HT action. Administration of either N-f-5HT or MTX caused reduction of inflammation measured as the level of CRP in plasma and the activity of LOX in the liver. mRNA transcription of TNF-α and iNOS in the liver was significantly attenuated in both MTX and N-f-5HT treated groups of arthritic rats. Interestingly, in contrast to MTX, N-f-5HT significantly lowered the level of IL-1β in plasma and IL-1β mRNA expression in the liver and spleen of arthritic rats. This speaks for future investigations of N-f-5HT as an agent in the treatment of RA in combination therapy with MTX.

Purification and properties of lipoxygenase from opium poppy seedlings(Papaver somniferum L.)

Lipoxygenase (linoleate:oxygen oxidoreductase; EC 1.13.11.12; LOX) is a key enzyme in the signaling pathway leading to the production of biologically active compounds involved in plant growth development and stress responses. In this study the novel LOX enzyme from opium poppy (Papaver somniferum L.) seedlings was purified to electrophoretic homogeneity and characterized. LOX activity during the germination of opium poppy seeds was analyzed, and the highest LOX activity was determined on the fourth day of germination. Opium poppy LOX was purified from 4-day-old poppy seedlings after ammonium sulfate precipitation followed by hydrophobic chromatography (Phenyl-Sepharose CL-4B), ion exchange chromatography (Q-Sepharose), and affinity chromatography (linoleyl-aminopropyl agarose) for the first time. The relative molecular weight of purified LOX was estimated to be 78 kDa by immunoblotting. The highest enzyme activity occurred at pH 6.5 and 0.75 mM calcium ion concentration. LOX showed preferential activity towards linoleic acid followed by linolenic acid as a substrate. To investigate the positional specificity of the LOX reaction, purified LOX was incubated with linoleic acid, and the products were analyzed by high-performance liquid chromatography. Opium poppy LOX converted linoleic acid to 13-hydroperoxy-(9Z,11E)-octadecadienoic acid (63.5%) and, to a lesser extent, 9-hydroperoxy-(10E,12Z)-octadecadienoic acid (36.5%).

Biological activities of polyphenols-enriched propolis from Argentina arid regions

BackgroundPropolis is a bioactive natural product collected by honeybees (Apis mellifera) from plant sources. PurposeThis study was undertaken to determine the effect of propolis extracts from arid region of Argentina, on the activity/expression of pro-inflammatory enzymes, and as potential free radical scavenger, antifungal and anthelmintic agent as well as to get a first insight into the polyphenolic profile of the active fractions. Study design/methodsTwo propolis samples were collected in different time from hives located in Tucumán, Argentina. They are representative of the collection time of the raw material for phytotherapeutical purposes. Ethanolic extracts from both propolis were obtained. The PEEs were analyzed for total polyphenol (TP), non-flavonoid phenols (NFP) and flavonoid (FP) content followed by HPLC-DAD analysis and identification of components by HPLC-MS/MSn. The potentiality as anti-inflammatory (LOX, COX, iNOS enzymes), antioxidant, antifungal and nematicidal was determined. ResultsPEEs contain high levels of TP, NFP and FP, including cinnamic acid, caffeic acid prenyl ester, caffeoyl dihydrocaffeate and caffeic acid 3,4-dihydroxyphenethyl ester, liquiritigenin, 2′,4′-dihydroxychalcone and 2′,4′-dihydroxy-3′-methoxychalcone. The PEEs in vitro reduced the activity of LOX and COX-2. Pretreatment of RAW 264.7 cells with PEEs before the induction of inflammatory state, inhibited NO overproduction and the iNOS protein expression was significantly decreased. The PEEs exhibited antioxidant, antifungal (Candida sp.) and nematicidal effect (C. elegans). ConclusionThese findings show the potential use of characterized PEEs from arid regions of Argentina as phytomedicine.

Modulation of oxidized-LDL receptor-1 (LOX1) contributes to the antiatherosclerosis effect of oleanolic acid

Oleanolic acid (OA) is a bioactive pentacyclic triterpenoid. The current work studied the effects and possible mechanisms of OA in atherosclerosis. Quails (Coturnix coturnix) were treated with high fat diet with or without OA. Atherosclerosis was assessed by examining lipid profile, antioxidant status and histology in serum and aorta. Human umbilical vein endothelial cells (HUVECs) were exposed to 200μg/mL ox-LDL for 24h, then cell viability was assessed with MTT assay; reactive oxygen species (ROS) was assessed with DCFDA staining. Expression levels of LOX-1, NADPH oxidase subunits, nrf2 and ho-1 were measured with real time PCR and western blotting. Furthermore, LOX-1 was silenced with lentivirus and the expression levels assessment was repeated. OA treatment improved the lipid profile and antioxidant status in quails fed with high fat diet. Histology showed decreased atherosclerosis in OA treated animals. Ox-LDL exposure decreased viability and induced ROS generation in HUVECs, and this progression was alleviated by OA pretreatment. Moreover, elevated expression of LOX-1, NADPH oxidase subunits, nrf2 and ho-1 were observed in ox-LDL exposed HUVECs. OA pretreatment prevented ox-LDL induced increase of LOX-1 and NADPH oxidase subunits expression, while further increased nrf2 and ho-1 expression. Silencing of LOX-1 abolished ox-LDL induced effects in cell viability, ROS generation and gene expression. OA could alleviate high fat diet induced atherosclerosis in quail and ox-LDL induced cytotoxicity in HUVECs; the potential mechanism involves modulation of LOX-1 activity, including inhibition of expression of NADPH oxidase subunits and increase of the expression of nrf2 and ho-1.

Matrix Remodeling Promotes Pulmonary Hypertension through Feedback Mechanoactivation of the YAP/TAZ-miR-130/301 Circuit.

Pulmonary hypertension (PH) is a deadly vascular disease with enigmatic molecular origins. We found that vascular extracellular matrix (ECM) remodeling and stiffening are early and pervasive processes that promote PH. In multiple pulmonary vascular cell types, such ECM stiffening induced the microRNA-130/301 family via activation of the co-transcription factors YAP and TAZ. MicroRNA-130/301 controlled a PPAR?-APOE-LRP8 axis, promoting collagen deposition and LOX-dependent remodeling and further upregulating YAP/TAZ via a mechanoactive feedback loop. In turn, ECM remodeling controlled pulmonary vascular cell crosstalk via such mechanotransduction, modulation of secreted vasoactive effectors, and regulation of associated microRNA pathways. In vivo, pharmacologic inhibition of microRNA-130/301, APOE, or LOX activity ameliorated ECM remodeling and PH. Thus, ECM remodeling, as controlled by the YAP/TAZ-miR-130/301 feedback circuit, is an early PH trigger and offers combinatorial therapeutic targets for this devastating disease.

Cloning and characterization of lipoxygenase gene from germinating seedlings of green gram (Vigna radiata L.)

Plant LOXs play a significant role in plant physiological processes such as growth, senescence, defence and stress-related responses. In the present study a full length lipoxygenase cDNA (VrLOX) has been cloned from green gram germinating seedlings using 5′- and 3′-RACE PCR method and nomenclatured VrLOX. The ORF (2601) encodes 867 amino acid residues with an approximate molecular weight of 97.43 kDa and an isoelectric point of 5.93. The VrLOX comprises of a putative PLAT domain and an iron-binding catalytic domain, and the sequence displayed a significant identity to known plant LOXs deposited in Protein Data Bank. Bioinformatics analysis demonstrated that this gene is most probably a LOX putative protein of type 13-LOX, and shared highest identity (91 %) with the Glycine max LOX-10. Enzymatic assay showed LOX activity was more on day 3 at pH 6.5 and higher MDA levels were observed on day 5 after germination. Semi-quantitative RT-PCR analysis indicated highest VrLOX mRNA expression on day 3 after germination, and could be induced by methyl jasmonate (MeJA). The 3-dimensional structure of the VrLOX was elucidated using in silico approach and molecular dynamics simulations.

RETRACTED: Effect of intermittent oven drying on lipid oxidation, fatty acids composition and antioxidant activities of walnut

Compared with sun drying and direct oven drying methods, intermittent oven drying for walnut (Juglans regia L.) drying was employed, and the consequent changes in lipid oxidation attributes, such as acid value (AV), peroxide value (POV), saponification value (SV), fatty acid composition, antioxidant activity (reducing power, superoxide anion scavenging activity, and DPPH free radical scavenging activity), lipoxygenase activity, and total phenols content (TPC) and total flavonoids content (TFC) were investigated in this study. Sun drying resulted in the highest AV, POV and SV, at the end of drying, that was 0.8 g/kg, 2.4 meq/kg, 230.9 g/kg, respectively, followed by direct oven drying and intermittent oven drying. Antioxidant activities and related compounds, TPC and TFC, were initially increased then declined during drying, and a significant correlation was found between antioxidant activity and TPC and TFC (P < 0.05). The LOX activities of all samples showed an increasing trend, although those of the sun-dried samples were the lowest (0.43 μkat/mg). The walnuts dried with the intermittent oven drying method had much more linoleic acid than those dried by direct oven drying.

Healthy effect of different proportions of marine ω-3 PUFAs EPA and DHA supplementation in Wistar rats: Lipidomic biomarkers of oxidative stress and inflammation

Dietary intervention with ω-3 marine fatty acids may potentially modulate inflammation and oxidative stress markers related with CVD, metabolic syndrome and cancer. The aim of this study was to evaluate whether different proportions of ω-3 EPA and DHA intake provoke a modulation of the production of lipid mediators and then, an influence on different indexes of inflammation and oxidative stress in a controlled dietary animal experiment using Wistar rats. For such scope, a lipidomic SPE-LC-ESI-MS/MS approach previously developed was applied to determine lipid mediators profile in plasma samples. The effect of ω-3 fatty acids associated to different ratios EPA:DHA was compared with the effect exerted by ω-3 ALA supplementation from linseed oil and ω-6 LA from soybean oil. CRP showed a tendency to greater inflammatory status in all ω-3-fed animals. Interestingly, ratios 1:1 and 2:1 EPA:DHA evidenced a noteworthy healthy effect generating a less oxidative environment and modulating LOX and COX activities toward a decrease in the production of proinflammatory ARA eicosanoids and oxidative stress biomarkers from EPA and DHA. In addition, the ability of 1:1 and 2:1 fish oil diets to reduce lipid mediator levels was in concurrence with the protective effect exerted by decreasing inflammatory markers as ω-6/ω-3 ratio in plasma and membranes. It was also highlighted the effect of a higher DHA amount in the diet reducing the healthy benefits described in terms of inflammation and oxidative stress. Results support the antiinflammatory and antioxidative role of fish oils and, particularly, the effect of adequate proportions EPA:DHA.