Impacts of the unsaturation degree of long-chain fatty acids on the volatile fatty acid profiles of rumen microbial fermentation in goats in vitro

This study investigated the impacts of the degree of unsaturation （unsaturity） of long-chain fatty acids on volatile fatty acid （VFA） profiles of rumen fermentation in vitro. Six types of long-chain fatty acids, including stearic acid （C18：0, control group）, oleic acid （C18：1, n-9）, linoleic acid （C18：2, n-6）, a-linolenic acid （C18：3, n-3）, arachidonic acid （C20：4, n-6） and eicosapentaenoic acid （C20：5, n-3）, were tested. Rumen fluid from three goats fitted with ruminal fistulae was used as inoculum and the inclusion rate of long-chain fatty acid was at 3% （w/w） of substrate. Samples were taken for VFA analysis at 0, 3, 6, 9, 12, 18 and 24 h of incubation, respectively. The analysis showed that there were significant differences in the total VFA among treatments, sampling time points, and treatment×time point interactions （P〈0.01）. a-Linolenic acid had the highest total VFA （P〈0.01） among different long-chain fatty acids tested. The molar proportion of acetate in total VFA significantly differed among treatments （P〈0.01） and sampling time points （P〈0.01）, but not treatment×time point interactions （P〉0.05）. In contrast, the molar proportion of propionate did not differ among treatments during the whole incubation （P〉0.05）. However, for butyrate molar proportions, significant differences were found not only among sampling time points but also among treatments and treatment×time point interactions （P〈0.01）, with eicosapentaenoic acid having the highest value （P〈0.01）. Additionally, no statistically significant differences were found in the acetate to propionate ratios among treatments groups （P〉0.05）, even the treatments stearic acid and a-linolenic acid were numerically higher than the others. The inclusion of 3% long-chain unsaturated fatty acids differing in the degree of unsaturation brought out a significant quadratic regression relation between the total VFA concentration and the double bond number of fatty acid. In conclusion, the a-linolenic acid with 3 double bonds appeared better for improving rumen microbial fermentation and the total VFA concentration.

Identification of a Long-Chain Fatty Acid Elongase from Nannochloropsis sp. Involved in the Biosynthesis of Fatty Acids by Heterologous Expression in Saccharomyces cerevisiae

The marine microalga Nannochloropsis sp.contains various elongases and desaturases that are critical for biosynthesis of polyunsaturated fatty acids.A full-length cDNA encoding a long-chain fatty acid elongase,named Ns FAE,was cloned from Nannochloropsis sp..The open reading frame of Ns FAE(GenBank accession no.MF680548)consisted of 1068 bp and encoded a predicted protein of 355 amino acids with molecular mass 38.8 k Da.The deduced polypeptide showed 43%–44%identity to fatty acyl elongases from other algae.RT-PCR experiments indicated that the Ns FAE gene exhibited the highest expression in Nannochloropsis sp.at 72 h(i.e.,during the third growth stage)and the expression was significantly lower in the other four growth stages.Plasmid p Ns FAE-CRISPR and a recombinant DNA fragment(ADH1p-Ns FAE-CYCt)were transformed into Saccharomyces cerevisiae strain BY4742 using the CRISPR-Cas system.Yeast transformants containing Ns FAE produced three fatty acids not normally present in wild-type BY4742-linoleic acid,linolenic acid and eicosadienoic acid-indicating that Ns FAE encodes a functional elongase enzyme.

The effects of the unsaturated degree of long-chain fatty acids on the rumen microbial protein content and the activities of transaminases and dehydrogenase in vitro

This study investigated the effects of the degree of unsaturation（unsaturity） of long-chain fatty acids on microbial protein content and the activities of transaminases and dehydrogenase in vitro using goat rumen fluid as the cultural medium.Six types of fatty acids,stearic acid（C18：0,group A,control group）,oleic acid（C18：1,n-9,group B）,linoleic acid（C18：2,n-6,group C）,α-linolenic acid（C18：3,n-3,group D）,arachidonic acid（C20：4,n-6,group E）,and eicosapentaenoic acid（C20：5,n-3,group F）,were tested,and the inclusion ratio of each fatty acid was 3%（w/w） in total of culture substrate.Samples were taken at 0,3,6,9,12,18 and 24 h,respectively,during culture for analyses.Compared with stearic acid,linoleic acid,a-linolenic acid,and arachidonic acid increased the bacterial protein content,while oleic acid and eicosapentaenoic acid had no significant effects;the protozoal protein content was reduced for all the unsaturated fatty acids,and the magnitude of the reduction appeared to be associated with the degree of unsaturity of fatty acids.The total microbial protein content was dominantly accounted by the protozoal protein content（about 4-9 folds of the bacterial protein）,and only increased by linoleic acid,but reduced by oleic acid,arachidonic acid and eicosapentaenoic acid.There were no significant effects in the activities of both glutamic oxaloacetic transaminase（GOT） and glutamic-pyruvic transaminase（GPT） for all the other fatty acids,except for arachidonic acid which enhanced GOT activity and oleic acid which enhanced GPT activity.The total dehydrogenase activity was positively correlated with the degree of unsaturation of fatty acids.In conclusion,the inclusion of 3%of long-chain unsaturated fatty acids increased bacterial protein content,whereas reduced protozoal protein content and enhanced dehydrogenase activity.The fatty acids with more than three double bonds had detrimental effects on the microbial protein content.This work demonstrates for the first time the effect rule of the unsaturation degree of long-chain fatty acids on the rumen microbial protein in vitro.

Long-chain acyl-CoA synthetase in fatty acid metabolism involved in liver and other diseases:An update

Long-chain acyl-Co A synthetase(ACSL) family members include five different ACSL isoforms, each encoded by a separate gene and have multiple spliced variants. ACSLs on endoplasmic reticulum and mitochondrial outer membrance catalyze fatty acids with chain lengths from 12 to 20 carbon atoms to form acyl-Co As, which are lipid metabolic intermediates and involved in fatty acid metabolism, membrane modifications and various physiological processes. Gain- or lossof-function studies have shown that the expression of individual ACSL isoforms can alter the distribution and amount of intracellular fatty acids. Changes in the types and amounts of fatty acids, in turn, can alter the expression of intracellular ACSLs. ACSL family members affect not only the proliferation of normal cells, but the proliferation of malignant tumor cells. They also regulate cell apoptosis through different signaling pathways and molecular mechanisms. ACSL members have individual functions in fatty acid metabolism in different types of cells depending on substrate preferences, subcellular location and tissue specificity, thus contributing to liver diseases and metabolic diseases, such as fatty liver disease, obesity, atherosclerosis and diabetes. They are also linked to neurological disorders and other diseases. However, the mechanisms are unclear. This review addresses new findings in the classification and properties of ACSLs and the fatty acid metabolismassociated effects of ACSLs in diseases.

Effects of long-chain fatty acid supplementation on the growth performance of grower and finisher pigs: a meta-analysis

Background: Supplementation of feed with long-chain fatty acids(LCFAs) during the grower and finisher phases has long been discussed as a growth promotion strategy in pigs, but its effects are inconsistent. The purpose of our study was to comprehensively evaluate its effects on the growth performance based on the average daily gain(ADG), average daily feed intake(ADFI) and gain: feed(G:F) ratio and to unveil the roles of the basal diet, LCFA concentration and LCFA saturation.Results: We searched the Pub Med and Web of Science databases(articles published from Jan 1 st, 2000, to Sep 30 th,2018;restricted to English) and compared LCFA-supplemented diets with control diets. We retrieved 2346 studies, 18 of which(1314 pigs, 26 records) were eligible for our analysis. We used a random-effects model to calculate the weighted mean differences(WMDs) and 95% confidence intervals(CIs). LCFA supplementation in the grower-finisher phase improved the ADG(WMD = 41.74 g/d, 95% CI: 8.81 to 74.66, P = 0.013) and G:F ratio(WMD = 0.019, 95% CI: 0.006 to 0.032, P = 0.003). For supplementation solely in the finisher phase, we found a similar performance in the ADG(WMD = 39.93 g/d, 95% CI: 26.48 to 53.38, P < 0.001) and G:F ratio(WMD = 0.019, 95% CI: 0.006 to 0.032, P < 0.001) but a reduction in the ADFI(WMD =-83.863 g/d, 95% CI:-156.157 to-11.569, P = 0.023). Specifically, approximately 5%LCFA supplementation in the finisher phase had significant effects on the ADG(WMD = 51.385 g/d, 95% CI: 35.816 to66.954, P < 0.001), ADFI(WMD =-102.869 g/d, 95% CI:-189.236 to-16.502, P = 0.02) and G:F ratio(WMD = 0.028, 95%CI: 0.018 to 0.039, P < 0.001), whereas a concentration of approximately 1% exhibited no effects.Conclusions: Overall, regardless of the basal diet and saturation, LCFA supplementation greatly improves the growth performance of grower and finisher pigs, primarily by increasing the energy density.

Natural long-chain saturated fatty acids doped LNPs enabling spleen selective mRNA translation and potent cancer immunotherapy

Rationally tailored lipid nanoparticles(LNPs)with efficient and tunable delivery of mRNA in vivo are crucial for mRNA vaccines.Selective expression of antigenic protein in lymphoid tissues/organs could improve the immunostimulatory efficacy and safety of LNPs-based mRNA vaccines.Inspired by the metabolic behavior that long-chain saturated fatty acids tending to enter lymphoid tissue rather than the liver,we developed fatty acid-doped LNPs capable of mediating differential protein expressions in the liver and spleen when administered intravenously.When the molar ratio of saturated fatty acid located 60%–70%,the doped LNPs achieved the spleen selective mRNA translation.The mechanism could be attributed to the different cellular uptake behaviors of saturated fatty acids in hepatocytes.Immunization with a model antigen(ovalbumin)mRNA-loaded spleen selective LNPs,we observed enhanced antigen-specific T cell immune responses,and potent immunotherapeutic and immunoprophylactic efficacy in the mouse lymphoma model.Our natural long-chain saturated fatty acids metabolic characteristics-inspired design of LNPs for spleen-selective mRNA vaccines delivery will provide references for designing mRNA vaccines with high efficacy and safety for tumor immunotherapy.

Study on Extracting Rare Earth from Sulfate System by Long-Chain Fatty Acid

The extraction of rare earths by long-chain fatty acid in kerosene from sulphate system was described.It was demonstrated from the experimental results that the ratio of kerosene: fatty acid: isooctanol = 55 : 30: 15 ( V/V),By the saturation capability method and the slope method, the extracted reaction mechanism of the extraction of rare earth was studied.It is shown that the extraction reaction conform to the cation exchange reaction mechanism.The extracted sequence of rare earth was determined in this system and it is shown that there is no tetrad effect and the position of yttrium is between lanthanum and cerium.

Impact of Chain Length of Saturated Fatty Acids during Their Heterogeneously Catalyzed Deoxygenation

Fatty acids with different chain length were deoxygenated in the absence of hydrogen (caprylic acid (CA), lauric acid (LA) and stearic acid (SA)). The catalytic tests were carried over Pd-containing catalysts out in a batch reactor under inert gas for 6 h at 250&degC to 350&degC and pressures from 18 to 75 bar in the absence of additionally fed hydrogen. Pd-containing catalysts were tested;the best performing catalyst was 10% Pd/C with 63% undecane yield at 327&degC. These catalysts were used for a comparative decarboxylation of CA, LA and SA. At equal reaction conditions (300&degC, 6 h), the chain length of the fatty acid had a strong impact on the conversion, which was steadily increasing, whereas the alkane selectivity ran through a maximum. This work demonstrated the usability of Pd-containing catalysts for the decarboxylation of various fatty acids in the absence of additionally fed hydrogen with respect to the manufacture of hydrocarbons that can be used as blending components for fuels.

Protective effect of long-chain polyunsaturated fatty acids on hepatorenal syndrome in rats

BACKGROUND Hepatorenal syndrome(HRS)is the most prevalent form of acute kidney injury in cirrhotic patients.It is characterized by reduced renal blood flow and represents the most severe complication in cirrhotic patients with advanced disease.Previous research has indicated that antioxidants can delay the onset of a hyperdynamic circulatory state in cirrhosis and improve renal function in HRS patients.Regular omega-3 supplementation has significantly reduced the risk of liver disease.This supplementation could represent an additional therapy for individuals with HRS.AIM To evaluated the antioxidant effect of omega-3 polyunsaturated fatty acid supplementation on the kidneys of cirrhotic rats.METHODS Secondary biliary cirrhosis was induced in rats by biliary duct ligation(BDL)for 28 d.We used 24 male Wistar rats divided into the following groups:I(control);II(treated with omega-3,1 g/kg of body weight);III(BDL treated with omega-3,1 g/kg of body weight);and IV(BDL without treatment).The animals were killed by overdose of anesthetic;the kidneys were dissected,removed,frozen in liquid nitrogen,and stored in a freezer at-80℃for later analysis.We evaluated oxidative stress,nitric oxide(NO)metabolites,DNA damage by the comet assay,cell viability test,and apoptosis in the kidneys.Data were analyzed by one-way analysis of variance,and means were compared using the Tukey test,with P≤0.05.RESULTS Omega-3 significantly decreased the production of reactive oxygen species(P<0.001)and lipoperoxidation in the kidneys of cirrhotic rats treated with omega-3(P<0.001).The activity of the antioxidant enzymes superoxide dismutase and catalase increased in the BDL+omega-3 group compared to the BDL group(P<0.01).NO production,DNA damage,and caspase-9 cleavage decreased significantly in the omega-3-treated BDL group.There was an increase in mitochondrial electrochemical potential(P<0.001)in BDL treated with omega-3 compared to BDL.No changes in the cell survival index in HRS with omega-3 compared to the control group(P>0.05)were observed.CONCLUSION The study demonstrates that omega-3 can protect cellular integrity and function by increasing antioxidant enzymes,inhibiting the formation of free radicals,and reducing apoptosis.

Fatty acid metabolism and acyl-CoA synthetases in the liver-gut axis

Fatty acids are energy substrates and cell components which participate in regulating signal transduction,transcription factor activity and secretion of bioactive lipid mediators.The acyl-CoA synthetases(ACSs)family containing 26 family members exhibits tissue-specific distribution,distinct fatty acid substrate preferences and diverse biological functions.Increasing evidence indicates that dysregulation of fatty acid metabolism in the liver-gut axis,designated as the bidirectional relationship between the gut,microbiome and liver,is closely associated with a range of human diseases including metabolic disorders,inflammatory disease and carcinoma in the gastrointestinal tract and liver.In this review,we depict the role of ACSs in fatty acid metabolism,possible molecular mechanisms through which they exert functions,and their involvement in hepatocellular and colorectal carcinoma,with particular attention paid to long-chain fatty acids and small-chain fatty acids.Additionally,the liver-gut communication and the liver and gut intersection with the microbiome as well as diseases related to microbiota imbalance in the liver-gut axis are addressed.Moreover,the development of potentially therapeutic small molecules,proteins and compounds targeting ACSs in cancer treatment is summarized.

Acyl-CoA synthase ACSL4:an essential target in ferroptosis and fatty acid metabolism

Long-chain acyl-coenzyme A(CoA)synthase 4(ACSL4)is an enzyme that esterifies CoA into specific polyunsaturated fatty acids,such as arachidonic acid and adrenic acid.Based on accumulated evidence,the ACSL4-catalyzed biosynthesis of arachidonoyl-CoA contributes to the execution of ferroptosis by triggering phospholipid peroxidation.Ferroptosis is a type of programmed cell death caused by iron-dependent peroxidation of lipids;ACSL4 and glutathione peroxidase 4 positively and negatively regulate ferroptosis,respectively.In addition,ACSL4 is an essential regulator of fatty acid(FA)metabolism.ACSL4 remodels the phospholipid composition of cell membranes,regulates steroidogenesis,and balances eicosanoid biosynthesis.In addition,ACSL4-mediated metabolic reprogramming and antitumor immunity have attracted much attention in cancer biology.Because it facilitates the cross-talk between ferroptosis and FA metabolism,ACSL4 is also a research hotspot in metabolic diseases and ischemia/reperfusion injuries.In this review,we focus on the structure,biological function,and unique role of ASCL4 in various human diseases.Finally,we propose that ACSL4 might be a potential therapeutic target.

植物超长链脂肪酸及角质层蜡质生物合成相关酶基因研究现状

超长链脂肪酸(very long chain fatty acids,VLCFAs)在生物体中具有广泛的生理功能,它们参与种子甘油酯、生物膜膜脂及鞘脂的合成,并为角质层蜡质的生物合成提供前体物质。角质层是覆盖在植物地上部分最表层的保护层,由角质和蜡质组成,其中蜡质又分为角质层表皮蜡和内部蜡,在植物生长发育、适应外界环境方面起重要作用。VLCFAs的合成由脂肪酰-CoA延长酶催化,该酶是由β-酮脂酰-CoA合酶、β-酮脂酰-CoA还原酶、β-羟脂酰-CoA脱水酶和反式烯脂酰-CoA还原酶组成的多酶体系。合成后的VLCFAs通过脱羰基与酰基还原作用进入角质层蜡质合成途径,形成各种蜡质组分。文章就VLCFAs及角质层蜡质合成代谢途径中相关酶基因研究进展方面做了综述,并对植物蜡质基因研究中存在的问题提出一些看法。

肾上腺脑白质营养不良临床分析(附6例报道)

目的分析肾上腺脑白质营养不良（adrenoleukodystrophy，ALD）的临床和影像学特征。方法对6例ALD患者的临床资料进行回顾性分析。结果6例患者均为男性儿童，年龄为5—12岁，初始症状表现为注意力不集中、情绪不稳定、多动、学习成绩差等，逐渐进展加重，出现肢体瘫痪、共济失调、言语障碍、视听力下降、癫痫发作、意识障碍等。MRI均可见双侧脑室后角周围、顶、枕叶白质区呈对称性蝶翼状长T1及T2信号；可累及胼胝体压部、脑桥及双侧小脑中脚。3例血浆极长链脂肪酸水平升高。脑脊液和脑电图改变无特异性。目前尚无特效治疗方法。结论ALD以儿童脑型常见，依据临床症状和典型的MRI表现可以进行临床诊断。

植物芥酸的研究

芥酸等是重要的VLCFA ,主要存在于少数植物的种子油中。植物中芥酸等VLCFA生物合成是由脂肪酸延长酶催化的 ,在一些植物中 ,该酶系统的有关基因已被克隆研究。人们还从草地泡沫等植物中获得特异性催化芥酸进入Sn - 2位的酶的目的基因。

超长链脂肪酸的研究进展

对超长链脂肪酸的合成代谢途径、生物学功能、分离纯化方法、分析检测技术以及应用等进行综述,并对其研究及应用前景进行了展望。

The Benefit Risk Assessment of Consumption of Marine Species Based on Benefit-Risk Analysis for Foods(BRAFO)-tiered Approach

Objective To assess the net health effect caused by the consumption of specific marine species based on Benefit-Risk Analysis for Foods （BRAFO）-tiered approach. Methods Twenty species were collected from the Zhoushan Archipelago, China. Concentrations of n-3 long-chain polyunsaturated fatty acids, methyl mercury （MeHg）, and dioxin-like compounds （DLCs） in the samples were analyzed for benefit risk assessment based on BRAFO-tiered approach. Results Based on the BRAFO-tiered approach, reference scenario （no intake） and alternative scenario （intake of specific species of 200 g/week） were determined. The exposure to MeHg/DLCs via alternative scenario of all studied species did not exceed provisional tolerable weekly/monthly intake. However, the adult population with high DLCs exposure in China would significantly exceed the upper limit of DLCs via an additional alternative scenario of some species such as Auxis thazard. The results of deterministic computation showed that alternative scenario of all studied species generated clear net beneficial effects on death prevention and child IQ gain. Conclusion The alternative scenario of all studied species could be recommended to population with average DLCs exposure, and the reference scenario of species with relatively high DLCs concentration could be recommended to population exposed to high DLCs.

Modifier-concept of colorectal carcinogenesis: Lipidomics as a technical tool in pathway analysis

In the modifier concept of intestinal carcinogenesis, lipids have been established as important variables and one focus is given to long-chain fatty acids. Increased consumption of long-chain fatty acids is in discussion to modify the development of colorectal carcinoma in humans. Saturated long-chain fatty acids, in particular, are assumed to promote carcinogenesis, whereas poly-unsaturated forms are likely to act in the opposite way. At present, the molecular mechanisms behind these effects are not well understood. Recently, it has been demonstrated by lipidomics and associated molecular techniques, that activation and metabolic channeling of long-chain fatty acids are important mechanisms to modify colorectal carcinogenesis. In this Editorial, an overview about the present concept of long-chain fatty acids and its derivatives in colorectal carcinogenesis as well as technical algorithms in lipid analysis is given.

Cloning,Functional Characterization and Expression Analysis of the elovl4a Gene in the Large Yellow Croaker(Larimichthys crocea)

Elovl4 is a fatty acyl elongase which participates in the biosynthesis of long-chain polyunsaturated fatty acids(LCPUFA).Herein we isolated and functionally characterized the elovl4a gene in the large yellow croaker Larimichthys crocea,and investigated the regulatory effects of transcription factors Hnf4α,Lxrα,Pparαon the activity of elovl4a promoter.Tissue expression pattern revealed that elovl4a was widely expressed in several tissues and predominantly in brain.Heterologous expression in yeast showed that L.crocea Elovl4a could effectively elongate both C18 and C20 PUFA substrates to C22 fatty acid.In addition,the affinity of large yellow croaker Elovl4a to n-6 series fatty acids is weak and cannot prolong 18:2n-6 and 18:3n-6.Moreover,L.crocea elovl4a reporter activities were elevated by 1.31-,1.39-and 1.48-fold via over-expression of Lxrα,Pparαand Hnf4α,respectively.The above findings can enrich the knowledge of biosynthesis pathway of LC-PUFA in the large yellow croaker and contribute to elucidate the LC-PUFA anabolism mechanism in fish.

Tumor cell membrane remodeling with universal ligand for CAR-T cells to inhibit solid tumors

Heterogeneity is a major obstacle to the success of CAR-T therapy in treating solid tumors. The complex tumor micro-environment and varying phenotypes of tumor cells might result in antigen escape, drug resistance, and tumor recurrence. To address this issue, we proposed to use lipid-modified fluorescein isothiocyanate(Lip-FITC) as an artificial ligand to normalize the phenotypes of solid tumor cells. In murine cutaneous melanoma and colon adenocarcinoma that prefer to utilize exogenous long-chain fatty acids, we observed much more uptake of Lip-FITC and significantly increased FITC fluorescence on tumor cell membranes than normal cells. This specific exogenous labeling with FITC enhanced the recognition and selectivity of CART cells in solid tumors, bypassing the limitations derived from antigen expression differences in adoptive cell therapies. Lipid metabolism analysis and in vitro experiments demonstrated the sufficient uptake of long-chain fatty acid(LCFAs)-modified Lip-FITC by solid tumor cells, as well as satisfactory ligand assembly on cell membranes. In solid tumor model, the treatment induced the recognition and initiation of CART cells and successfully suppressed tumor progression through T-cell immunity.