Erucic Acid-Rich Yellow Mustard Oil Improves Insulin Resistance in KK-Ay Mice.

Obesity is a major risk factor for some metabolic disorders including type 2 diabetes. Enhancement of peroxisome proliferator-activated receptor (PPAR) γ, a master regulator of adipocyte differentiation, is known to increase insulin-sensitive small adipocytes. In contrast, decreased PPARγ activity is also reported to improve insulin resistance. We have previously identified erucic acid as a novel natural component suppressing PPARγ transcriptional activity. In this study, we investigated the effect of erucic acid-rich yellow mustard oil (YMO) on obese/diabetic KK-Ay mice. An in vitro luciferase reporter assay and mesenchymal stem cell (MSC) differentiation assay revealed that 25 µg/mL YMO significantly inhibited PPARγ transcriptional activity and differentiation of MSCs into adipocytes but promoted their differentiation into osteoblasts. In KK-Ay mice, dietary intake of 7.0% (w/w) YMO significantly decreased the surrogate indexes for insulin resistance and the infiltration of macrophages into adipose tissue. Furthermore, 7.0% YMO increased bone mineral density. These results suggest that YMO can ameliorate obesity-induced metabolic disorders.

A new pyrimidine alkaloid from the roots of Tadehagi triquetrum (L.) H.Ohashi.

Tadehagi triquetrum (L.) H.Ohashi, also known as Desmodium triquetrum (Fabaceae) is the most important plant in the herbal remedies. The present study focus on the isolation, in-silico and in-vitro studies of the two alkaloids C1 (5-(4-[(methylcarbamoyl) amino]-2-oxopyrimidin-1(2H)-yl) tetrahydrofuran-2-yl) methyl methyl carbamate is novel alkaloid and C2 13-Docosenamide is a known alkaloid. The chemical structures of compounds have been elucidated based on comprehensive techniques like GCMS, IR and NMR. In order to know the molecular mechanisms for the two compounds, in silico molecular docking study has been performed. Both compounds have shown perfect binding affinity to the enzymes TNF α, IL-4, IL-13 and 5 LOX Enzyme. The compounds also exhibited comparable G-scores and Glide energy values in comparison with the standard dexamethasone. In addition both the compounds have been tested for in vitro antioxidant assay by using ABTS and DPPH method and the results were compared with standard ascorbic acid.

Winter camelina seeds as a raw material for the production of erucic acid-free oil.

Camelina oil is increasingly popular as consumption as oil. Erucic acid is an unwanted fatty acid in oil. First studies on several genotypes have shown that this oil contains varying amounts of eriuc acid. The aim of the study was to analyses content of eriuc acid in all genotypes camelina. Hypothesis was that the content of erucic acid in winter forms is lower than in spring ones. A field experiment with 65 spring genotypes and 9 winter genotypes of camelina was conducted in Poland from 2016 to 2018. The analyses based on two chromatographic methods, i.e. UPLC-DAD and GC-MS, showed no differences in the results for the camelina samples. The average percentage content of the erucic acid in the spring genotypes was 3.432%, and in the winter genotypes was 0.1%. Our three-year research shows that some winter varieties can be used as low erucic acid forms.

The impact of reducing fatty acid desaturation on the composition and thermal stability of rapeseed oil.

Oilseed rape (Brassica napus) is the third largest source of vegetable oil globally. In addition to food uses, there are industrial applications that exploit the ability of the species to accumulate the very-long-chain fatty acid (VLCFA) erucic acid in its seed oil, controlled by orthologues of FATTY ACID ELONGASE 1 (Bna.FAE1.A8 and Bna.FAE1.C3). The proportion of polyunsaturated fatty acids (PUFAs) in rapeseed oil is predicted to affect its thermal stability and is controlled by orthologues of FATTY ACID DESATURASE 2, particularly Bna.FAD2.C5. Our aim was to develop rapeseed lines combining high erucic and low PUFA characters and to assess the impact on thermal stability of the oil they produce. The new type of rapeseed oil (high erucic low polyunsaturate; HELP) contained a substantially greater proportion of erucic acid (54%) compared with high erucic rapeseed oil (46%). Although the total VLCFA content was greater in oil from HELP lines (64%) than from high erucic rapeseed (57%), analysis of triacylglycerol composition showed negligible incorporation of VLCFAs into the sn-2 position. Rancimat analysis showed that the thermal stability of rapeseed oil was improved greatly as a consequence of reduction of PUFA content, from 3.8 and 4.2 h in conventional low erucic and high erucic rapeseed oils, respectively, to 11.3 and 16.4 h in high oleic low PUFA (HOLP) and HELP oils, respectively. Our results demonstrate that engineering of the lipid biosynthetic pathway of rapeseed, using traditional approaches, enables the production of renewable industrial oils with novel composition and properties.

Engineering Rhodosporidium toruloides for the production of very long-chain monounsaturated fatty acid-rich oils.

Erucic acid (cis-docosa-13-enoic acid, C22:1∆13) and nervonic acid (cis-tetracosa-15-enoic acid, C24:1 ∆15) are important renewable feedstocks in plastic, cosmetic, nylon, and lubricant industries. Furthermore, nervonic acid is also applied to the treatment of some neurological diseases. However, the production of these two very long-chain fatty acids (VLCFA) is very limited as both are not present in the main vegetable oils (e.g., soybean, rapeseed, sunflower, and palm). Ectopic integration and heterologous expression of fatty acid elongases (3-ketoacyl-CoA synthases, KCS) from different plants in Rhodosporidium toruloides resulted in the de novo synthesis of erucic acid and nervonic acid in this oleaginous yeast. Increasing KCS gene copy number or the use of a push/pull strategy based on the expression of elongases with complementary substrate preferences increased significantly the amount of these two fatty acids in the microbial oils. Oil titers in 7-L bioreactors were above 50 g/L, and these two VLCFA represented 20-30% of the total fatty acids. This is the first time that microbial production of these types of oils is reported.

Novel function of N,N-bis(2-chloroethyl)docos-13-enamide for reversal of multidrug resistance in tongue cancer.

Multidrug resistance (MDR) is a key element in the failure of chemotherapies, and development of agents to overcome MDR is crucial to improving cancer treatments. The overexpression of glutathione-S-transferases (GSTs) is one of the major mechanisms of MDR. Because some agents used in traditional Chinese medicine have strong antitumor effects coupled with low toxicity; we investigated the ability of N,N-bis(2-chloroethyl)docos-13-enamide (compound J), the synthesized analog of a highly unsaturated fatty acid from Isatis tinctoria L., to reverse the MDR induced by adriamycin (ADM) in TCA8113/ADM cells. We found that compound J significantly increased the cytotoxicity of ADM in TCA8113/ADM cells, with a reversal fold of 2.461. Analysis of the mechanisms through which compound J reversed MDR indicated that compound J significantly decreased the activity of GSTs and enhanced the depletion of GSH in TCA8113/ADM cells, but did not affect the P-glycoprotein (P-gp) efflux. Taken together, our data suggested that compound J was an excellent candidate for reversing MDR in cancer therapy.

[Study on the chemical constituents from Patrinia scabra].

OBJECTIVE: To study the chemical constituents from Patrinia scabra Bunge. METHODS: Chemical constituents were isolated and purified by silica gel and Sephadex LH-20 column chromatography. Their structures were elucidated on the basis of spectroscopic analysis and chemical evidence. RESULTS: Seven compounds were isolated and purified. Their structures were identified as villosol (I), patriscabrol (II), protocatechuic acid (III), beta-daucosterol (IV), oleic acid (V), beta-sitosterol (VI), erucic acid (VII). CONCLUSION: Compounds I, III, V, VII are isolated from this plant for the first time.

Eliminating expression of erucic acid-encoding loci allows the identification of "hidden" QTL contributing to oil quality fractions and oil content in Brassica juncea (Indian mustard).

Oil content and oil quality fractions (viz., oleic, linoleic and linolenic acid) are strongly influenced by the erucic acid pathway in oilseed Brassicas. Low levels of erucic acid in seed oil increases oleic acid content to nutritionally desirable levels, but also increases the linoleic and linolenic acid fractions and reduces oil content in Indian mustard (Brassica juncea). Analysis of phenotypic variability for oil quality fractions among a high-erucic Indian variety (Varuna), a low-erucic east-European variety (Heera) and a zero-erucic Indian variety (ZE-Varuna) developed by backcross breeding in this study indicated that lower levels of linoleic and linolenic acid in Varuna are due to substrate limitation caused by an active erucic acid pathway and not due to weaker alleles or enzyme limitation. To identify compensatory loci that could be used to increase oil content and maintain desirable levels of oil quality fractions under zero-erucic conditions, we performed Quantitative Trait Loci (QTL) mapping for the above traits on two independent F1 doubled haploid (F1DH) mapping populations developed from a cross between Varuna and Heera. One of the populations comprised plants segregating for erucic acid content (SE) and was used earlier for construction of a linkage map and QTL mapping of several yield-influencing traits in B. juncea. The second population consisted of zero-erucic acid individuals (ZE) for which, an Amplified Fragment Length Polymorphism (AFLP)-based framework linkage map was constructed in the present study. By QTL mapping for oil quality fractions and oil content in the ZE population, we detected novel loci contributing to the above traits. These loci did not co-localize with mapped locations of the fatty acid desaturase 2 (FAD2), fatty acid desaturase 3 (FAD3) or fatty acid elongase (FAE) genes unlike those of the SE population wherein major QTL were found to coincide with mapped locations of the FAE genes. Some of the new loci identified in the ZE population could be detected as 'weak' contributors (with LOD < 2.5) in the SE population in which their contribution to the traits was "masked" due to pleiotropic effects of erucic acid genes. The novel loci identified in this study could now be used to improve oil quality parameters and oil content in B. juncea under zero-erucic conditions.

Synthesis and characterization of poly(3-hydroxyalkanoates) from Brassica carinata oil with high content of erucic acid and from very long chain fatty acids.

Pseudomonas aeruginosa produced medium chain length poly(3-hydroxyalkanoates) (mcl-PHAs) when grown on substrates containing very long chain fatty acids (VLCFA, C>20). Looking for low cost carbon sources, we tested Brassica carinata oil (erucic acid content 35-48%) as an intact triglyceride containing VLCFA. Oleic (C18:1), erucic (C22:1), and nervonic (C24:1) acids were also employed for mcl-PHA production as model substrates. The polymers obtained were analyzed by GC of methanolyzed samples, GPC, 1H and 13C NMR, ESI MS of partially pyrolyzed samples, and DSC. The repeating units of such polymers were saturated and unsaturated, with a higher content of the latter in the case of the PHA obtained from B. carinata oil. Statistical analysis of the ion intensity in the ESI mass spectra showed that the PHAs from pure fatty acids are random copolymers, while the PHA from B. carinata oil is either a pure polymer or a mixture of polymers. Weight-average molecular weight varied from ca. 56,000 g/mol for the PHA from B. carinata oil and oleic acid, to about 120,000 g/mol for those from erucic and nervonic acids. The PHAs from erucic and nervonic acids were partially crystalline, with rubbery characteristics and a melting point (Tm) of 50°C, while the PHAs from oleic acid and from B. carinata oil afforded totally amorphous materials, with glass transition temperatures (Tg) of -52°C and -47°C, respectively.

[Studies on the chemical constituents of Urtica dioica L. grown in Tibet Autonomous Region].

Studies on the whole herb of Urtica dioica L. grown in Nyingchi area, China's Tibet Autonomous Region, resulted in the isolated of nine compounds: beta-sitosterol, trans-ferulic acid, dotriacotane, erucic acid, ursolic acid, scopoletin, rutin, quercetin and p-hydroxylbenzalcohol. Dotriacotane, erucic acid, scopoletin, rutin and p-hydroxylbenzalcohol were obtained from Urtica L. for the first time. Their structures were confirmed by modem spectral analysis (NMR, MS, etc).

Development of high-oleic, low-linolenic acid Ethiopian-mustard (Brassica carinata) germplasm.

Seed oil of current zero erucic-acid germplasm of Ethiopian mustard ( Brassica carinata A. Braun) is characterized by a low concentration of oleic acid and high concentrations of linoleic and linolenic acids. Sources of increased oleic-acid (HO) and reduced linolenic-acid (LL) concentration have been developed separately in high erucic-acid germplasm. The objectives of the present research were to study the inheritance of the HO and LL traits in crosses HO x LL, and to develop HOLL recombinants, both in high erucic-acid and zero erucic-acid backgrounds. The HO mutant N2-3591 (about 20% oleic acid compared to 9% in conventional high erucic-acid materials), was reciprocally crossed with the LL lines N2-4961 and HF-186 (both with about 5% linolenic acid compared to 12% in standard high erucic-acid materials). Increased oleic acid concentration of N2-3591 was found to be controlled by alleles at one locus (Ol), whereas three different loci for reduced linolenic-acid concentration (Ln, Ln1 and Ln2) were identified in N2-4961 and HF-186. Crosses between N2-3591 and N2-4961 generated HOLL recombinants where levels of increased oleic-acid and reduced linolenic-acid were similar to those of the parents. However, a transgressive segregation for oleic acid was observed in crosses between N2-3591 and HF-186, where F(2) seeds with up to 29.7% oleic acid were obtained, in comparison to an upper limit of 25.1% in the N2-3591 parent grown in the same environment. The transgressive increased oleic-acid was expressed in the F(3) generation and was attributed to the presence of a second locus, designated Ol2. The transgressive trait was transferred to the zero erucic-acid line 25X-1, resulting in a zero erucic-acid germplasm with very high oleic-acid concentration (83.9% compared to 32.9% in 25X-1) and low linolenic-acid concentration (5.0% compared to 16% in 25X-1). Additionally, two other lines exhibiting different stable levels of increased oleic-acid (70.7% and 79.5%, respectively) and reduced levels of linolenic-acid (7.5% and 8.7%, respectively) were isolated.