Partial purification of linoleic acid isomerase enzyme from Lactobacillus paracasei bacteria isolated from milk

Abstract Conjugated Linoleic Acid (CLA) has attracted the attention of many researchers, especially that of microbial origin due to its biological importance to the consumer. The current study aims to extract LA Isomerase enzyme from Lactobacillus paracasei bacteria from milk and to use the enzyme in the production of CLA. Selective media, including MRS and MRS-Dagatose, were used in isolating local strains. The selected bacterial isolates were tested for their ability to produce LA-Isomerase enzyme. The isolate with high enzymatic activity was selected. After extraction and partial purification of the enzyme, the optimal conditions for the production of conjugated fatty acid were studied, and the reaction products were diagnosed using GC-MS technology. It was found that 11 isolates have the ability to produce CLA at different concentrations, H1 isolate showed the highest production of conjugated fatty acid at a concentration of 120.45 g.ml-1, this isolate was selected as the source for enzyme extraction. The enzymatic activity of the crude extract and partially purified with ammonium sulfate was estimated using color methods at wavelength of 233 nm. The effect of the optimum conditions (pH, temperature, linoleic acid concentration and enzyme concentration) on the CLA product was studied using the partially purified LA Isomerase enzyme, the optimum conditions for production were 6.5, 45 °C, 100 g.ml-1 and 0.7 ml, respectively. The GC-MS technique showed the presence of a number of reaction products that are isomers of conjugated linoleic acid (C9T11, T9T12, T10C12) with different concentrations.

Resumo O Ácido Linoleico Conjugado (CLA) tem chamado a atenção de diversos pesquisadores, principalmente aquele de origem microbiana, devido à sua importância biológica para o consumidor. O presente estudo visa extrair a enzima LA Isomerase da bactéria Lactobacillus paracasei do leite e usar essa enzima na produção de CLA. Meios seletivos, incluindo MRS e MRS-Dagatose, foram usados no isolamento de cepas locais. Os isolados bacterianos selecionados foram testados quanto à sua capacidade de produzir a enzima LA-Isomerase. Foi selecionado o isolado com alta atividade enzimática. Após a extração e purificação parcial da enzima, as condições ideais para a produção de ácido graxo conjugado foram estudadas e os produtos da reação foram identificados usando a tecnologia GC-MS. Verificou-se que 11 isolados possuem capacidade de produzir CLA em diferentes concentrações. O isolado H1 apresentou a maior produção de ácido graxo conjugado, na concentração de 120,45 g.ml-1, e este isolado foi selecionado como fonte para extração enzimática. A atividade enzimática do extrato bruto e parcialmente purificado com sulfato de amônio foi estimada por métodos de coloração em comprimento de onda de 233 nm. O efeito das condições ótimas (pH, temperatura, concentração de ácido linoleico e concentração de enzima) no produto CLA foi estudado usando a enzima LA Isomerase parcialmente purificada e as condições ótimas para produção foram 6,5, 45 °C, 100 g.ml-1 e 0,7 mL, respectivamente. A técnica de GC-MS mostrou a presença de uma série de produtos de reação que são isômeros do ácido linoleico conjugado (C9T11, T9T12, T10C12) com diferentes concentrações.

Partial purification of linoleic acid isomerase enzyme from Lactobacillus paracasei bacteria isolated from milk / Purificação parcial da enzima isomerase do ácido linoleico da bactéria Lactobacillus paracasei isolada do leite

Conjugated Linoleic Acid (CLA) has attracted the attention of many researchers, especially that of microbial origin due to its biological importance to the consumer. The current study aims to extract LA Isomerase enzyme from Lactobacillus paracasei bacteria from milk and to use the enzyme in the production of CLA. Selective media, including MRS and MRS-Dagatose, were used in isolating local strains. The selected bacterial isolates were tested for their ability to produce LA-Isomerase enzyme. The isolate with high enzymatic activity was selected. After extraction and partial purification of the enzyme, the optimal conditions for the production of conjugated fatty acid were studied, and the reaction products were diagnosed using GC-MS technology. It was found that 11 isolates have the ability to produce CLA at different concentrations, H1 isolate showed the highest production of conjugated fatty acid at a concentration of 120.45 g.ml-1, this isolate was selected as the source for enzyme extraction. The enzymatic activity of the crude extract and partially purified with ammonium sulfate was estimated using color methods at wavelength of 233 nm. The effect of the optimum conditions (pH, temperature, linoleic acid concentration and enzyme concentration) on the CLA product was studied using the partially purified LA Isomerase enzyme, the optimum conditions for production were 6.5, 45 °C, 100 µg.ml-1 and 0.7 ml, respectively. The GC-MS technique showed the presence of a number of reaction products that are isomers of conjugated linoleic acid (C9T11, T9T12, T10C12) with different concentrations.

O Ácido Linoleico Conjugado (CLA) tem chamado a atenção de diversos pesquisadores, principalmente aquele de origem microbiana, devido à sua importância biológica para o consumidor. O presente estudo visa extrair a enzima LA Isomerase da bactéria Lactobacillus paracasei do leite e usar essa enzima na produção de CLA. Meios seletivos, incluindo MRS e MRS-Dagatose, foram usados no isolamento de cepas locais. Os isolados bacterianos selecionados foram testados quanto à sua capacidade de produzir a enzima LA-Isomerase. Foi selecionado o isolado com alta atividade enzimática. Após a extração e purificação parcial da enzima, as condições ideais para a produção de ácido graxo conjugado foram estudadas e os produtos da reação foram identificados usando a tecnologia GC-MS. Verificou-se que 11 isolados possuem capacidade de produzir CLA em diferentes concentrações. O isolado H1 apresentou a maior produção de ácido graxo conjugado, na concentração de 120,45 g.ml-1, e este isolado foi selecionado como fonte para extração enzimática. A atividade enzimática do extrato bruto e parcialmente purificado com sulfato de amônio foi estimada por métodos de coloração em comprimento de onda de 233 nm. O efeito das condições ótimas (pH, temperatura, concentração de ácido linoleico e concentração de enzima) no produto CLA foi estudado usando a enzima LA Isomerase parcialmente purificada e as condições ótimas para produção foram 6,5, 45 °C, 100 µg.ml-1 e 0,7 mL, respectivamente. A técnica de GC-MS mostrou a presença de uma série de produtos de reação que são isômeros do ácido linoleico conjugado (C9T11, T9T12, T10C12) com diferentes concentrações.

Effect of some fatty acids on apoptosis related genes in human breast cancer

Breast cancer, the second most common cancer after lung cancer, is the most common cancer type diagnosed in women. No definitive treatment has been established for breast cancer yet, but essential fatty acids offer a promising option. Omega fatty acids are classified in the essential fatty acids that the body cannot produce and, therefore, must be taken through the foods of animal or plant origin. Although in the literature the omega fatty acids have been shown to exhibit significant positive effects in inhibiting various tumor types, their mechanism of action, the apoptotic pathways they employ, and the genes they control have not been clarified yet. In this study, various doses and combinations of omega-3 [Eicosapentaenoic acid (EPA), Docosahexaenoic acid (DHA)] and omega-6 [Linoleic acid (LA)] fatty acids were administered to human breast cancer MCF7 cell line for 24 h, and using the enzyme-linked immunosorbent assay (ELISA) method, the protein expression levels of the following apoptosis-related genes were determined: phospho-p53 (Ser15), p53, Bad, phospho-Bad (Ser112), cleaved Caspase-3 (Asp175), and cleaved PARP (Asp214). Even though there was no significant difference observed in the expressions of phospho-p53 (Ser15) and p53 at all doses, other protein expressions were found to increase significantly, suggesting that Omega-3 and -6 can mediate apoptotic pathway to induce cell death in breast cancer cells.

Protective effect of linoleic acid on liver toxicity induced by methotrexate / Efecto protector del ácido linoleico sobre la toxicidad hepática inducida por metotrexato

SUMMARY: We aimed to investigate the protective effect of linoleic acid on liver toxicity induced by methotrexate. The study was carried out in partnership with the Department of Anatomy and Department of Medical Pharmacology of Çukurova University Faculty of Medicine, using the laboratory facilities of the Department of Medical Pharmacology. Human hepatocyte cell line (CRL- 11233) cells obtained from the American Type Culture Collection Organization (ATCC) were used. Expressions of apoptotic pathway markers, apoptosis inducing factor (AIF), BAX, BCL 2, GADD 153, 78-kDa glucose-regulated protein (GRP78), and CASPASE-3 were evaluated. All analyzes were examined in four groups (Group 1; control, Group 2; linoleic acid given, Group 3; methotrexate given and Group 4; linoleic acid and methotrexate given). The mean ± standard error values of the obtained results as nanogram / milliliter (ng / ml) are in Group I, Group II, Group III and Group IV, respectively; AIF values, 0.4150 ± 0.1208, 0.3633 ± 0.2389, 1.792 ± 0.3611 and 1.077 ± 0.1646, BAX values, 0.900 ± 0.1864, 1.002 ± 0.2098, 8.352 ± 1.467 and 4.295 ± 1.522, BCL 2 values, 13.93 ± 1.198, 13.92 ± 1.739, 2.938 ± 1.059 and 9.250 ± 1.492, GADD 153, 0.7333 ± 0.1751, 0.7067 ± 0.2115, 1.650 ± 0.2950 and 1.237 ± 0.1805, GRP78, 0.4767 ± 0.1804, 0.5233 ± 0.1590, 2.183 ± 0.2639 and 1.112 ± 0.2693, CASPASE-3 values , 1.127 ± 0.2033, 0.8317 ± 0.3392, 13.50 ± 1.871 and 8.183 ± 1.030. It was determined that linoleic acid has a protective effect on methotrexate-induced liver toxicity.

Nuestro objetivo fue investigar el efecto protector del ácido linoleico sobre la toxicidad hepática inducida por metotrexato. El estudio se llevó a cabo en colaboración con el Departamento de Anatomía y el Departamento de Farmacología Médica de la Facultad de Medicina de la Universidad de Çukurova, utilizando las instalaciones del laboratorio del Departamento de Farmacología Médica. Se usaron células de la línea celular de hepatocitos humanos (CRL-11233) obtenidas de la American Type Culture Collection Organisation (ATCC). Se evaluaron las expresiones de marcadores de vías apoptóticas, factor inductor de apoptosis (AIF), BAX, BCL 2, GADD 153, proteína regulada por glucosa de 78 kDa (GRP78) y CASPASE-3. Todos los análisis se examinaron en cuatro grupos (Grupo 1; control, Grupo 2; se administró ácido linoleico, Grupo 3; se administró metotrexato y Grupo 4; se administró ácido linoleico y metotrexato). Los valores medios ± error estándar de los resultados obtenidos como nanogramo/mililitro (ng/ml) se encuentran en el Grupo I, Grupo II, Grupo III y Grupo IV, respectivamente; Valores de AIF, 0,4150 ± 0,1208, 0,3633 ± 0,2389, 1,792 ± 0,3611 y 1,077 ± 0,1646, valores de Bax, 0,900 ± 0,1864, 1,002 ± 0,2098, 8,352 ± 1,467 y 4,295 ± 1,522, BCL 2 valores, 13,93 ± 1,199. 2,938 ± 1,059 y 9,250 ± 1,492, GADD 153, 0,7333 ± 0,1751, 0,7067 ± 0,2115, 1,650 ± 0,2950 y 1,237 ± 0,1805, Grp78, 0,4767 ± 0,1804, 0,5233 ± 0,1590, 2,183, ± 1,263. 1,127 ± 0,2033, 0,8317 ± 0,3392, 13,50 ± 1,871 y 8,183 ± 1,030. Se determinó que el ácido linoleico tiene un efecto protector sobre la toxicidad hepática inducida por metotrexato.

Targeted Lipidomics Reveals Lipid Modulation of Kaixuan Bushen Method on Psoriasis Vulgaris / 中国实验方剂学杂志

ObjectiveThrough the targeted lipidomics， we explored the intervention mechanism of Kaixuan Bushen method on psoriasis vulgaris （PV） from the perspective of lipid metabolism， providing reference for the diagnosis and treatment of PV. MethodTwenty-six patients with PV admitting the outpatient clinic of the Department of Dermatology， Guang'anmen Hospital， China Academy of Chinese Medical Sciences from September 2019 to November 2020 were selected as the research object （observation group）， and 26 sex- and age-matched healthy volunteers in the same period were recruited as control group. Venous blood was collected for lipid index and targeted lipidomics detection in the control and observation groups at inclusion. After 12 weeks of continuous treatment of Kaixuan Bushen method， the psoriasis area and severity index （PASI） was measured and compared before and after treatment to assess the clinical efficacy， while venous blood was collected again in the observation group to compare the blood lipid level and lipid metabolism of patients before and after treatment. Targeted lipidomics analysis was performed by ultra performance liquid chromatography tandem mass spectrometry （UPLC-MS/MS） on an ACQUITY UPLC BEH C8 column （2.1 mm×100 mm， 1.7 μm） with mobile phase of 5 mmol∙L-1 ammonium formate in acetonitrile-water （6∶4， A）-5 mmol∙L-1 ammonium formate in acetonitrile-isopropanol （1∶9， B） for gradient elution and flow rate of 0.26 mL∙min-1. Conditions of MS were electrospray ionization （ESI）， positive and negative ion modes， and scanning range of m/z 50-1 200. Then principal component analysis （PCA） and partial least squares-discriminant analysis （PLS-DA） models were developed to screen differential metabolites， and the differential metabolites were identified and the pathways were enriched. ResultAfter 12 weeks of treatment with Kaixuan Bushen method， PASI score decreased by more than 50% in a total of 22 out of 26 patients with PV， suggesting the total effective rate was 84.62%. The serum triglyceride level of patients with PV was significantly higher than that of healthy individuals （P<0.05）， and the triglyceride level was significantly reduced after treatment （P<0.05）. Targeted lipidomics analysis screened a total of 43 potential biomarkers for PV， of which 42 were up-regulated and 1 was down-regulated， involving 7 signaling pathways such as linoleic acid metabolism， glycerophospholipid metabolism and unsaturated fatty acid biosynthesis. Moreover， there were 14 response makers for clinical efficacy of Kaixuan Bushen method on PV， of which 6 were up-regulated and 8 were down-regulated， involving five signaling pathways such as linoleic acid metabolism， glycerophospholipid metabolism and sphingolipids metabolism. In a comparison between healthy individuals and patients with PV and PV before and after treatment， the common differential metabolites were screened as phosphatidylcholine （PC） 38∶0 and ceramide （Cer） 42∶1， and the common pathways were linoleic acid and glycerophospholipid metabolic pathways. ConclusionThe disorder of lipid metabolism in PV are largely due to abnormal sphingolipid， glycerophospholipid and linoleic acid metabolic pathways， of which Kaixuan Bushen method can regulate the glycerophospholipid and linoleic acid metabolism， thereby improving psoriatic lesions.

Production of hispidin polyphenols from medicinal mushroom Sanghuangporus vaninii in submerged cultures / 中草药·英文版

OBJECTIVE@#The medicinal mushroom Sanghuangporus vaninii produces pharmaceutically valuable hispidin polyphenols in natural habitats. However, due to the slow growth in nature, S. vaninii grown in the field (sclerotia) is not reliable for pharmaceutical purposes. Although higher biomass of fungal mycelia can be obtained in submerged cultures, the accumulation of hispidin polyphenols is rare.@*METHODS@#In this study, the polyunsaturated fatty acids (PUFAs), linoleic acid (LA), linolenic acid (ALA), and methyl jasmonate (MeJa) were employed as the stimulant agents to coordinate the accumulation of biomass and hispidin polyphenols in its submerged cultures.@*RESULTS@#The addition of LA and ALA promoted the mycelial accumulation, while the addition of MeJa inhibited the growth of S. vaninii concomitant with reduced total polyphenols. UPLC-Triple-TOF-MS analysis revealed an increased production of hispidin, phellinstatin, pinnilidine, and its derivatives upon the addition of LA and ALA, and hypholomine B and its isomer, 3,14'-bihispidinyl, and phelligridin E upon the addition of MeJa on day 13. Intriguingly, total polyphenols from the MeJa-supplementing cultures harbored a high capacity in scavenging free radicals. Chemical structural analysis showed that hispidin polyphenols had higher antioxidant activity due to more hispidin moieties induced by MeJa.@*CONCLUSION@#The supplement of PUFAs affects the synthesis and composition of hispidin polyphenols in S. vaninii. Our results provide a possibility to coordinate the production of hispidin polyphenols via submerged cultures of S. vaninii.

Improving the production of conjugated linoleic acid from sunflower oil by lactic acid bacteria spp: Effect of calcium carbonate supplementation in fermentation medium

Conjugated linoleic acid (CLA) is a polyunsaturated fatty acid with various positional and geometric isomers of linoleic acid (LA) present mainly in food items and produced endogenously in non-ruminants and humans or through fermentation process. It is associated with health-beneficial effects and subject to more research on its natural sources (ruminant-derived foods) and strategies to increase the content in various foodstuffs. Although several studies have reported the most common intake value of 0.8 g/day (0.6 to 3.0 g/day), research for raising in situ concentration should focus on strategies such as in vitro bioconversion of its precursors by bacteria and supplementation of LA rich-oils in foods fermentation process. In this study, the ability of some lactic acid bacteria (LAB) from diverse samples to produce CLA from sunflower oil and the effect on production yield of fermentation medium supplementation with carbonate calcium were investigated. Results showed that ten Limosilactobacillus fermentum and Enterococcus faecium produced trans-10, cis-12-CLA isomer accounted for at least 85% of total CLA ranging from 4.64 to 5.22 µg/mL. Despite the fermentation medium supplementation with CaCO3 enhanced the production yield, the residual LA inhibitory effect on bacteria growth governing CLA biosynthesis process was not mitigated. So, although our LAB strains can produce CLA, the more the LA concentration goes up, the more the conversion rate downgrades. Further studies on strains behavior in a wide range of LA concentrations will help establish a stable relationship between bacteria and LA in the presence of CaCO3.

RNA-seq Was Used to Obtain the Key Genes in the Regulation of Conjugated Linoleic Acid on Intramuscular Fat Metabolism in Royal Chicken / 中国生物化学与分子生物学报

Transcriptome sequencing was used to obtain the differentially expressed genes of conjugated linoleic acid (CLA) on the basal diet of Royal chickens on intramuscular fat metabolism in related signal pathways and candidate genes that may play important roles by bioinformatics analysis, which could provide a further understanding of the molecular mechanism of CLA on intramuscular fat deposition. In this study, 55-day-old healthy Royal chickens were selected as experimental animals, and 0%, 1% and 2% of CLA were added to the basic diet. The pre-feeding period was 1 week and the normal feeding period was 6 weeks. The breast muscle tissues were collected for transcriptome sequencing, and the sequencing data were subjected for differential expression analysis, such as GO function and KEGG pathway enrichment analysis of differentially expressed genes. Differentially expressed genes related to breast muscle lipid metabolism were screened out, and qRT-PCR was used to verify differentially expressed genes. The results showed that a total of 1 065 differentially expressed genes were obtained, of which 703 were up-regulated genes and 362 were down-regulated genes. GO enrichment results show that differentially expressed genes are concentrated in cellular processes, single-biological processes, biological regulation and metabolic processes in biological processes. Enrichment analysis of KEGG signaling pathway shows that differentially expressed genes were significantly enriched in focal adhesion, unsaturated fatty acid biosynthesis, fatty acid biosynthesis, and steroid biosynthesis. Totally 11 candidate genes were found mainly related to intramuscular fat metabolism, including FADS1, FADS2, ELOVL5, ACOX2, SLC27A1, FABP5, LPL, LOC107050163, ENSGALG00000030996, ENSGALG00000005043 and ENSGALG00000048882. Six genes were randomly selected for qRT-PCR verification, and their relative expression changes were consistent with the sequencing results. This study screened the differentially expressed genes related to CLA affecting breast muscle lipid metabolism in Royal chickens, and analyzed 11 genes related to fat metabolism, laying the foundation for revealing the molecular mechanism of CLA regulating intramuscular fat deposition.

Walnut ointment promotes full-thickness burning wound healing: role of linoleic acid

Purpose: To investigate the active ingredients of walnut ointment (WO) and its mechanism in repairing wounds. Methods: The ingredients of WO were detected by gas chromatography­mass spectrometry. The effect of linoleic acid (LA) was tested by in vitro Alamar Blue (AB) reagent. Image J software, histological and immunohistochemical analysis were used to confirm the healing effect of LA in the porcine skin model. The animals were euthanized after the experiment by injection of pentobarbital sodium. Results: LA, 24% in WO, promotes keratinocytes and fibroblasts proliferation, which were 50.09% and 15.07% respectively higher than control (p < 0.05). The healing rate of the LA group (96.02% ± 2%, 98.58% ± 0.78%) was higher than the saline group (82.11% ± 3.37%, 88.72% ± 1.73%) at week 3 and week 4 (p < 0.05). The epidermal thickness of the LA was 0.16 ± 0.04 mm greater and the expression of the P63 and CK10 proteins was stronger in the LA group than the control (p < 0.05). Conclusions: LA, which is the main components in WO can promote full-thickness burning wounds (FBWs) by stimulating cell proliferation and differentiation.

Functional and chemical qualities of Vitis labrusca grape seed oil extracted by supercritical CO 2 / Calidad funcional y química del aceite de semilla de Vitis labrusca extraído mediante CO 2 supercrítico / Qualidade funcional e química do óleo de semente de Vitis labrusca extraído por CO 2 supercrítico

Abstract Grape seed oil, which is usually extracted with highly toxic organic solvents that are harmful to human health, is produced from tons of grape pomace waste, generated during winemaking. Sometimes, this waste is used to make compost or is burnt, which causes environmental contamination. The functional qualities, antioxidant capacity (AC), α-tocopherol and total phenolic compounds content (TPC) of Black Borgoña (Vitis labrusca) grape seed oil, extracted by supercritical CO2, were evaluated. The high content of linoleic acid (ω-6) and monounsaturated fatty acids contributed to the beneficial effect on the functional quality indices, which were 0.20, 0.23, 11.80 for IA, IT and H:H, respectively. In addition, a POV of 6.23 ± 0.08 milliequivalents of peroxide/kg oil and an anisidine index of 2.70 ± 0.05 indicated a good quality oil. Also, a high concentration of a-tocopherol (9.82 ± 0.02 mg/100 g oil) and a high TPC ("4.14 ± 3.24 mg GAE/kg oil) were obtained. This study demonstrated that supercritical CO2 extraction is a suitable method for the delivery of a high-quality grape seed oil.

Resumen El aceite de semilla de uva que generalmente se extrae con disolventes orgánicos altamente tóxicos y perjudiciales para la salud humana, se produce a partir de toneladas de residuos de orujo de uva, generados durante la elaboración del vino. A veces, estos residuos se utilizan para hacer compost o se queman, lo que provoca la contaminación del medio ambiente. Se evaluaron las cualidades funcionales, la capacidad antioxidante (AC), el contenido de a-tocoferol y los compuestos fenólicos totales (TPC) del aceite de semilla de uva Borgoña Negra (Vitis labrusca), extraído mediante CO2 supercrítico. El alto contenido de ácido linoleico (ω-6) y de ácidos grasos monoinsaturados contribuyó al efecto beneficioso sobre los índices de calidad funcional que fueron de 0.20, 0.23, ''.80 para IA, IT y H:H, respectivamente. Además, un POV de 6.23 ± 0.08 miliequivalentes de peróxido/ kg de aceite y un índice de anisidina de 2.70 ± 0.05 indicaban una buena calidad del aceite. También se obtuvo una alta concentración de α-tocoferol (9.82 ± 0.02 mg/100 g de aceite) y un alto TPC ("4.14 ± 3.24 mg de GAE/ kg de aceite). Este estudio demostró que la extracción con CO2 supercrítico es un método adecuado para obtener un aceite de semilla de uva de alta calidad.

Resumo O óleo de semente de uva é geralmente extraído com solventes orgânicos altamente tóxicos que são prejudiciais à saúde humana, é produzido a partir de toneladas de resíduos de bagaço de uva, gerados durante a vinificação. Às vezes, esses resíduos são usados para fazer adubo ou são queimados, o que causa contaminação ambiental. Foram avaliadas as qualidades funcionais, capacidade antioxidante (AC), a-tocoferol e o teor total de compostos fenólicos (TPC) do óleo de semente de uva Borgoña Negra (Vitis labrusca), extraído por CO2 supercrítico. O alto teor de ácido linoleico (ω-6) e ácidos graxos monoinsaturados contribuiu para o efeito benéfico sobre os índices de qualidade funcional que foram 0.20, 0.23, 11.80 para IA, IT e H:H, respectivamente. Além disso, um POV de 6.23 ± 0.08 miliequivalentes de peróxido/ kg de óleo e um índice de anisidina de 2.70 ± 0.05 indicava uma boa qualidade de óleo. Também foi obtida uma alta concentração de α-tocoferol (9.82 ± 0.02 mg/100 g de óleo) e um alto TPC ("4.14 ± 3.24 mg de óleo GAE/ kg). Este estudo mostrou que a extração de CO2 supercrítico é um método adequado para a entrega de um óleo de semente de uva de alta qualidade.

Linoleic acid ameliorates inflammation in rheumatoid arthritis rats via regulation of TLR4/NF-κB signaling pathway / 中国药理学通报

Aim To explore the effects of linoleic acid on the joint swelling in rats caused by rheumatoid arthritis (RA), and to reveal possible mechanism underlying the inhibitory effect of linoleic acid on TLR4/NF-κB signaling pathway in RA. Methods The RA rat model was constructed and divided into control group, model group, linoleic acid (0. 1 mL) group, linoleic acid (0.2 mL) group, linoleic acid (0.4 mL) group and methotrexate (MTX) group. After the model was successfully established, the corresponding drugs were given by gavage for seven days. The control group and the model group were given normal saline. The changes of rat joint swelling were measured; joint pathological damage was assessed by HE staining; the protein expression levels and the mRNA expression levels of TNF-α, IL-1β, IL-6 and IL-10 were determined by ELISA and qRT -PCR; the protein expression levels of TLR4 and p-p65 were determined by immunohisto-chemistry and Western blot. Results Compared with model group, linoleic acid significantly alleviated the joint swelling of RA rats; linoleic acid significantly inhibited the joint pathological damage, and markedly reduced the protein expression levels and mRNA expression levels of TNF-α, IL-1β, IL-6 and IL-10; linoleic acid inhibited the protein expression levels of TLR4 and p-p65 in the TLR4/NF-κB signaling pathway. Conclusions Linoleic acid inhibits the protein expression levels of TLR4 and p-p65 in the TLR4/NF-κB signaling pathway, thereby inhibiting the expression of inflammatory factors TNF-α, IL-1β, IL-6 and IL-10.

Grain production fatty acid and oil profile from sunflower cultivars receiving different boron doses / Produção de grãos, perfil de óleo e ácido graxo em cultivares de girassol submetidos a diferentes doses de boro

Among the main factors that affect the productivity of crops is deficiency of nutrients. Boron (B) is an essential micronutrient for plants, and sunflower is one of the most sensitive plants to deficiency of the element. Its inadequate supply can impair sunflower plants' metabolism and grain and oil yield. The objective of this study was to assess the effect of different boron doses on the production of sunflower grains and the content and quality of the oil obtained from them. The experimental design was randomized blocks in a factorial scheme with three cultivars (Helio251, BRS323, BRS324) and four B doses (0, 2.5, 5.0, 8.0 kg ha-1). Two harvests were performed, the first in the R5 reproductive stage and the second at the end of the R9 cycle. In both cases, the levels of B in the capitulum were measured. At the end of the cycle, the grain yield, crude protein and oil content in the grains and fatty acid profile were analyzed. The cultivars responded differently to the treatments with B. The boron fertilization influenced the grain yield and oil content, but was not correlated with the profile of the majority unsaturated fatty acids and crude protein in the grains. Variations were observed in the fatty acid profile between the cultivars, an important aspect that needs to be evaluated according to the purpose of the production. In soil with lower availability of B, cultivar BRS323 was most efficient in B uptake, grain yield and oil content and quality.

Dentre os principais fatores que afetam a produtividade das culturas está a deficiência de nutrientes. O Boro (B) é um micronutriente essencial para plantas, e o girassol é uma das mais sensíveis à deficiência desse elemento. Seu fornecimento inadequado pode prejudicar o metabolismo, a produção de óleo e grãos das plantas de girassol. O objetivo deste estudo foi avaliar o efeito de diferentes doses de boro na produção de grãos, conteúdo a qualidade do óleo de girassol. O experimento foi conduzido em delineamento de blocos ao acaso em esquema fatorial com três cultivares (Helio251, BRS323, BRS324) e quatro doses de B (0; 2,5; 5,0; 8,0 kg ha-1). Foram realizadas duas coletas, a primeira na fase reprodutiva R5 e a segunda no final do ciclo R9. Em ambos os casos, os níveis de B no capítulo foram medidos. No final do ciclo, foram analisados a produção de grãos, teor de proteína bruta e óleo nos grãos e perfil de ácidos graxos. Os cultivares responderam diferentemente aos tratamentos com B. A adubação com boro influenciou o rendimento de grãos e teor de óleo, mas não se correlacionou com o perfil da maioria dos ácidos graxos insaturados e da proteína bruta nos grãos. Foram observadas variações no perfil de ácidos graxos entre cultivares, um aspecto importante que precisa ser avaliado de acordo com a finalidade da produção. No solo com menor disponibilidade de B, o cultivar BRS323 foi mais eficiente na absorção de B, rendimento de grãos, teor e qualidade de óleo.

Enrichment diets of pigs with oil blends and its effects on performance, carcass characteristics and fatty acid profile / Dietas enriquecidas com blends de óleos para suínos em terminação e seus efeitos sobre o desempenho, as características de carcaça e o perfil de ácidos graxos

The addition of different oil blends in the feed of finishing pigs was evaluated. Twenty-four castrated male finishing pigs were used in a randomized block design containing four treatments and six replicates. The treatments consisted of: Reference ration (RR) - 100% soybean oil feed; and the combination of the different oils: Blend1 - 50.0% soybean oil (SO), 25.0% flaxseed oil (FO), 12.5% olive oil (OO) and 12.5% canola oil (CO); Blend2 - 25.0% SO, 50.0% FO, 12.5% OO and 12.5% CO; and Blend3 - 25.0% SO, 12.5% FO, 12.5% OO and 50.0% CO. The performance, quantitative and qualitative carcass parameters, fatty acids profile and economic feasibility of the diets were evaluated. The use of blends in the diets did not influence the performance or carcass quality, but increased marbling and carcass yield. The fatty acid profile of the loin presented greater amounts of stearic acid in Blend3 and higher percentage of unsaturated fatty acids in animals fed with Blend1. The fatty tissue presented greater amounts of myristic acid in Blend1 and oleic acid in Blend3. The reference ration was the most economic. The Blends did not affect performance or carcass characteristics and improved the fatty acid profile.(AU)

Foi avaliada a utilização de diferentes blends de óleo em dietas de suínos em terminação. Foram utilizados 24 suínos, machos, castrados, distribuídos em delineamento de blocos ao acaso, com quatro tratamentos e seis repetições. Os tratamentos consistiram de: ração referência (RR) - 100% de ração com utilização de óleo de soja; e a combinação de diferentes óleos: Blend1 - 50,0% de óleo de soja (OS), 25,0% de óleo de linhaça (OL), 12,5% de óleo de oliva (OO) e 12,5% de óleo de canola (OC); Blend2 - 25,0% OS; 50,0% OL; 12,5% OO e 12,5% OC; e Blend3 - 25,0% OS; 12,5% OL; 12,5% OO e 50,0% OC. Foram avaliados os parâmetros de desempenho, a qualidade de carcaça, o perfil de ácidos graxos e a viabilidade econômica. O uso de blends nas dietas não influenciou o desempenho ou a qualidade da carcaça, mas aumentou o marmoreio e o rendimento de carcaça. O perfil de ácidos graxos do lombo apresentou maiores quantidades de ácido esteárico com a utilização do Blend3 e maior porcentagem de ácidos graxos insaturados nos animais alimentados com o Blend1. O tecido adiposo apresentou maiores quantidades de ácido mirístico quando se forneceu o Blend1 e de ácido oleico com o Blend3. A ração testemunha foi a mais econômica. As misturas não afetaram o desempenho e as características de carcaça e melhoraram o perfil de ácidos graxos da carne.(AU)

Screening of Plasma Biomarkers for Cerebral Ischemia-Reperfusion Based on Free Fatty Acid Targeted Lipidomics / 中国药学杂志

OBJECTIVE: To find disease biomarkers of cerebral ischemia by establishing cerebral ischemia-reperfusion injury model of rats and comparing with healthy rats. METHODS: Cerebral ischemia-reperfusion model of rats was established by modified Longa′s intraluminal suture method. After 24 h of modeling, plasma and brain tissue from the model group and the control group were collected for plasma biochemical indicators and metabolites analysis. Brain slices and plasma biochemical indicators were used to demonstrate the successful modeling. Gas chromatography-mass spectrometry was adopted to conduct targeted metabolomics studies on free fatty acids in rat plasma, tridecanoic acid (C13∶0) and 2,6-di-tert-butyl-4-methylphenol (BHT) were added as internal standard and antioxidant during sample preparation. Methylation was carried out using concentrated sulfuric acid/CH3OH as a derivatization reagent, and a non-polar stationary phase column was selected for chromatographic separation. RESULTS: Comparing the model group with the blank group, four differential metabolites were obtained, namely palmitic acid, stearic acid, oleic acid and linoleic acid, which were up-regulated throughout the metabolic pathway. The RESULTS of the relevant metabolic pathway analysis showed that the linoleic acid metabolic pathway had significant changes in the disease process. CONCLUSION: In the fatty acid anabolic pathway, the linoleic acid metabolic pathway changes significantly during the onset and treatment of cerebral ischemia-reperfusion. The effect of preventing and treating cerebral ischemia-reperfusion injury can be achieved by controlling the level of linoleic acid. This study evaluates the prognosis of cerebral ischemia reperfusion injury by measuring the plasma of model rats, explores its cause and pathogenesis, and laid a foundation for the study of disease pathology and the mechanism of drug action.

Intramuscular fatty acid composition of steers finished on oat pastures mixture with vetch or cornmeal supplementation / Composição de ácidos graxos da gordura intramuscular de novilhos terminados em pastagem de aveia consorciada com ervilhaca ou suplementados com grão de milho moído

ABSTRACT: The effects of energetic supplementation and mixture vetch (Vica sativa L.) were studied to evaluate the intramuscular fatty acid profile of steers finished on oat (Avena sativa L.) pastures. Eighteen 21-month-old crossbred steers were evaluated. The experiment was carried out in three treatments and six repetitions: oat pasture (OA), oat pasture + vetch (OA + VET), and oat pasture + supplementation (OA + SUP). Supplementation comprised the addition of corn meal at a daily dose of 1% of the animals' body weight. Steers receiving supplementation had higher lipid content in Longissimus lumborum than did those fed with OA + VET (1.25 vs 1.02%), whereas those fed with OA (1.15%) showed intermediate values. Conjugated linoleic acid levels were higher in steers fed OA (0.57%) and OA + SUP (0.59%), whereas the highest amount of omega-3 fatty acids was observed in animals fed OA + VET (3.32%). Pasture-finishing without supplementation resulted in a lower ratio of n-6:n-3 (3.14). Steers finished in oat mixture with vetch showed healthier intramuscular fat than did those finished with cornmeal supplementation; importantly, the higher the amount of PUFAs, the better n-6:n-3 ratio.

RESUMO: Os efeitos da suplementação energética e da ervilhaca (Vica sativa L.) foram estudados para avaliar o perfil de ácidos graxos intramusculares de novilhos terminados em pastagens de aveia (Avena sativa L.). Foram avaliados 18 novilhos mestiços com 21 meses de idade. O experimento foi realizado em três tratamentos e seis repetições: pastagem de aveia (OA), pastagem de aveia + ervilhaca (OA + VET) e pastagem de aveia + suplementação (OA + SUP). A suplementação compreendeu a adição de fubá na dose diária de 1% do peso corporal dos animais. Os novilhos que receberam suplementação apresentaram maior teor lipídico no Longissimus lombar do que aqueles alimentados com OA + VET (1,25 contra 1,02%), enquanto aqueles alimentados com AO (1,15%) apresentaram valores intermediários. Os níveis de ácido linoléico conjugado foram maiores em novilhos alimentados com AO (0,57%) e OA + SUP (0,59%), enquanto a maior quantidade de ácidos graxos ômega-3 foi observada em animais alimentados com OA + VET (3,32%). Animais terminados em pastagens sem suplementação apresentaram menor proporção de n-6: n-3 (3,14). Novilhos terminados em pastagem de aveia com ervilhaca apresentaram gordura intramuscular mais saudável do que aqueles terminados com suplementação de milho moído, uma vez que apresentaram maior quantidade de PUFAs e melhor a relação n-6: n-3.

Analysis of chemical constituents of fatty acids and their esters of Coicis Semen by UPLC-Triple-TOF-MS / 中草药

Objective: To establish a rapid qualitative analysis method for fatty acids and esters in Coicis Semen by ultra-performance liquid chromatography triple quadrupole time-of-flight mass spectrometry (UPLC-Triple-TOF-MS). Methods: Agilent ZORBAX-SB C18 (250 mm × 4.6 mm, 5.0 μm) column was used. The mobile phase was acetonitrile-isopropanol (1:1) elution, the flow rate was 1 mL/min, the detection wavelength was 210 nm, the column temperature was 30 ℃, and the injection quantity was 10 μL. Electrospray ion source positive ion mode was adopted, and the scanning range was m/z 100-1 500. The sample data were collected by full scanning mode, and the fatty acid chemical composition of Coicis Semen was quickly identified according to the information obtained by high-resolution mass spectrometry combined with secondary mass spectrometry. Results: A total of 29 kinds of fatty acids and esters in Coicis Semen were detected, and the cracking rules of the compounds were analyzed. Through the mass-to-charge ratio of molecular ion peaks and fragment ions, the Scifinder and Reaxy network databases, and the literature, these compounds were different under the action of ion source by losing the structure of oleic acid, linoleic acid, palmitic acid, oxidizing oleic acid and the like. The mass-to-charge ratio of the fragment ions, and the name and structural formula of the 29 fatty acids and their ester compounds were inferred. Conclusion: The method of qualitative analysis of the fatty acids and esters of Coicis Semen established in this study is accurate, rapid and sensitive, which provides experimental basis for improving the quality control level of Coicis Semen and further elucidating the pharmacodynamic substance basis of Coicis Semen.

Structure of the Conjugated Linoleic Acid-Paclitaxel Conjugate Self-Assembled Nanoparticles / 中国药学杂志

OBJECTIVE: To prepare, the conjugated linoleic acid-paclitaxel conjugate self-assembled nanoparticles (CLA-PTX NPs) by nanoprecipitation. METHODS: The Dynamic light scattering, nuclear magnetic resonance spectroscopy, raman spectroscopy, fourier transform infrared spectroscopy and nitrogen element distribution of CLA-PTX NPs were studied. RESULTS: The hydroxyl groups (C-4 and C-10 of PTX) and the acetyl groups (C-1 and C-7 of PTX) were on the surface of CLA-PTX NPs, CLA carbon chain, the benzene ring (C-2 and C-3' of PTX) and the amide bond (C-3' of PTX) were inside the CLA-PTX NPs. CONCLUSION: It is speculated that the self-assembly of CLA-PTX is that the non-polar CLA carbon chain spontaneously aggregates inward due to hydrophobic interaction, and the hydrophilic oxygen-containing groups of PTX (hydroxyl group and carbonyl group) are on the surface of the nanoparticle to form nanoparticles.

Interactions between Cannabis Fructus and human gut microflora / 中草药

Objective To explore the characteristics of the interactions between traditional laxative medicine Cannabis Fructus and human gut microflora. Methods HPLC method was used to determine the content of the main unsaturated fatty acids linoleic acid and linolenic acid in Cannabis Fructus Decoction. At the same time, solid culture and liquid culture in vitro anaerobic culture method were combined with 16 S rRNA technology to analyze the interactions between Cannabis Fructus Decotion and human gut microflora. Moreover, the metabolits of linoleic acid and linolenic acid in Cannabis Fructus Decoction by human intestinal microflora were determined using HPLC method. At the same time, the possible conjugated linolenic acid and linoleic acid were determined. Results Cannabis Fructus Decoction promoted the growth of Proteobacteria significantly, which showed that Escherichia-shigella was significantly increased (P < 0.01), but the growth of Bacteroidetes was decreased (P < 0.01), and the content of unsaturated fatty acids linoleic acid and linolenic acid in Cannabis Fructus Decoction were reduced after being incubated with human intestinal bacteria, and the metabolites were conjugated linoleic acid and conjugated linolenic acid. Conclusion The interactions between Cannabis Fructus Decoction and human intestinal microflora are obvious. The Chinese medicine can change the structure of the gut microflora, and the gut microflora can metabolize the drug components. This analysis method partially restores the pharmacokinetics process of the oral administration drug in the human intestinal tract. It could provide a new insight of the mechanism research of Cannabis Fructus.

Mechanism of Ermiao power in the treatment of rheumatoid arthritis based on urine metabolomics technology / 国际中医中药杂志

Objective To study the anti-rheumatoid arthritis mechanism of Ermiao powder by high-throughput urine metabolomics.Methods The rats were randomly divided into control group,model group and administration group with 8 rats in each group.The rat model of rheumatoid arthritis was established by intradermal injection of complete Freund's adjuvant.Rats in control group were given Ermiao power solution 0.108 g/ml by gavage.The degree of joint swelling in rats was observed and scored.On this basis,metabolic data of rat urine samples were collected for metabolomic analysis.Unsupervised and supervised pattern recognition technology was used to analyze the high-throughput biological information data and reduce the dimension.Metabolic information related to rheumatoid arthritis was screened and focused to clarify the pathogenesis of rheumatoid arthritis and the therapeutic mechanism of Ermiao power.Results Compared with the model group,the swelling degree of the foot (1.93 ± 0.11 vs.2.36 ± 0.19) in Ermiao power group significantly decreased (P＜0.01).Metabolic profiles showed that the metabolic distribution of healthy rats was significantly separated from that of model rats,and the treatment group was in the middle of the two groups.From the macro-metabolic point of view,the metabolism of model rats changed dramatically.The Ermiao power had a good intervention effect on rheumatoid arthritis.Thirteen biomarkers related to rheumatoid arthritis were identified by database matching,including linolenic acid,arachidonic acid,5,6-EET,alpha-lactose,sucrose,trehalose,prostaglandins and leukotriene C4.It involved linoleic acid metabolism,arachidonic acid metabolism,starch and galactose metabolism and sphingolipid metabolism.Conclusions The Ermiao power has significant therapeutic effect on rheumatoid arthritis rats.Regulation of the linoleic acid metabolism,arachidonic acid metabolism,starch and galactose metabolism may be the mechanism of its treatment of rheumatoid arthritis.

Conjugated linoleic acid improves glucose and lipid metabolism in diabetic mice / 南方医科大学学报

OBJECTIVE@#To analyze the effect of conjugated linoleic acid (CLA) on glucose and lipid metabolism in obese diabetic (db/db) mice.@*METHODS@#db/db mice were randomized for treatment with saline or CLA mixture administered intragastrically. The changes in body weight, dietary intake, water intake, oral glucose tolerance, triglyceride and total cholesterol were recorded after the treatments. HE staining and oil red O staining were used to assess liver pathologies and fatty acid content. The expression levels of PPARα, PPARγ, CD36, CHREBP and SREBP-1c were detected using real-time PCR and Western blotting. HepG2 cells were treated with CLA and linoleic acid and the expressions of PPARα, ACC, P-ACC, and CD36 were detected; the level of acetyl-CoA in the cell supernatant was detected using ELISA.@*RESULTS@#CLA treatment obviously reduced the dietary and water intake of db/db mice, effectively reduced the body weight and decreased serum triglyceride and cholesterol levels ( < 0.05). CLA significantly reduced fasting blood glucose, increased glucose tolerance, reduced the accumulation of lipid droplets in the liver and improved lipid metabolism in db/db mice. The mice showed significantly increased expression of PPARα ( < 0.05) and lowered CD36 expression ( < 0.001) in the liver after CLA treatment. Cellular experiments showed that CLA significantly up-regulated PPARα ( < 0.001) and P-ACC and decreased the expression of CD36 ( < 0.01). ELISA showed that acetyl-CoA was significantly up-regulated in the cells after CLA treatment ( < 0.01).@*CONCLUSIONS@#The mixture of two conjugated linoleic acid isomers can reduce fasting blood glucose, increase glucose tolerance and improve glycolipid metabolism in db/db mice by enhancing the expression of PPARα, increasing P-ACC and inhibiting CD36 expression.