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.

Dietary saturated fatty acid and polyunsaturated fatty acid oppositely affect hepatic NOD-like receptor protein 3 inflammasome through regulating nuclear factor-kappa B activation

AIM: To investigate the effect of different dietary fatty acids on hepatic inflammasome activation.METHODS: Wild-type C57BL/6 mice were fed either a high-fat diet or polyunsaturated fatty acid (PUFA)-enriched diet. Primary hepatocytes were treated with either saturated fatty acids (SFAs) or PUFAs as well as combined with lipopolysaccharide (LPS). The expression of NOD-like receptor protein 3 (NLRP3) inflammasome, peroxisome proliferator-activated receptor-γ and nuclear factor-kappa B (NF-κB) was determined by real-time PCR and Western blot. The activity of Caspase-1 and interleukine-1β production were measured.RESULTS: High-fat diet-induced hepatic steatosis was sufficient to induce and activate hepatic NLRP3 inflammasome. SFA palmitic acid (PA) directly activated NLRP3 inflammasome and increased sensitization to LPS-induced inflammasome activation in hepatocytes. In contrast, PUFA docosahexaenoic acid (DHA) had the potential to inhibit NLRP3 inflammasome expression in hepatocytes and partly abolished LPS-induced NLRP3 inflammasome activation. Furthermore, a high-fat diet increased but PUFA-enriched diet decreased sensitization to LPS-induced hepatic NLRP3 inflammasome activation in vivo. Moreover, PA increased but DHA decreased phosphorylated NF-κB p65 protein expression in hepatocytes.CONCLUSION: Hepatic NLRP3 inflammasome activation played an important role in the development of non-alcoholic fatty liver disease. Dietary SFAs and PUFAs oppositely regulated the activity of NLRP3 inflammasome through direct activation or inhibition of NF-κB.

Saturated Fatty Acid Induces Insulin Resistance Partially Through Nucleotide-binding Oligomerization Domain 1 Signaling Pathway in Adipocytes

Objective To investigate the potential role of nucleotide-binding oligomerization domain 1 （NOD1）, a component of the innate immune system, in mediating lipid-induced insulin resistance in adipocytes. Methods Adipocytes from Toll-like receptor 4 deficiency mice were used for stimulation experiments. The effect of oleate/palmitate mixture on nuclear factor-κB （NF-κB） activation was analyzed by reporter plasmid assay. The release of proinflammatory chemokine/cytokines production was determined by using real-time PCR. Insulin-stimulated glucose uptake was measured by 2-deoxy-D-[SH] glucose uptake assay. Chemokine/cytokine expression and glucose uptake in adipocytes transfected with small interfering RNA （siRNA） targeting NOD 1 upon fatty acids treatment were analyzed. Results Oleate/palmitate mixture activated the NF-κB pathway and induced interleukin-6, tumor necrosis factor-R, and monocyte chemoattractant protein-1 mRNA expressions in adipocytes from mice deficient in Toll-like receptor 4, and these effects were blocked by siRNA targeting NOD1. Furthermore, saturated fatty acids decreased the ability of insulin-stimulated glucose uptake. Importantly, siRNA targeting NOD 1 partially reversed saturated fatty acid-induced suppression of insulin-induced glucose uptake. Conclusion NOD1 might play an important role in saturated fatty acid-induced insulin resistance in adipocytes, suggesting a mechanism by which reduced NOD1 activity confers beneficial effects on insulin action.

Effects of microalgal polyunsaturated fatty acid oil on body weight and lipid accumulation in the liver of C57BL/6 mice fed a high fat diet

Dietary polyunsaturated fatty acids （PUFAs）, which are abundant in marine fish oils, have recently received global attention for their prominent anti-obesogenic effects. Among PUFAs, eicosapentaenoic acid （EPA; 20：5n-3） and docosahexaenoic acid （DHA; 22：6n-3）, which are n-3 long-chain PUFAs widely referred to as omega-3 oils, were reported to prevent the development of obesity in rodents and humans. In the present study, we evaluated the anti-obesity effects of microalgal oil on high-fat induced obese C57BL/6 mice, compared with commercial omega-3 fish oil and vegetable corn oil. Microalgal oil is an inherent mixture of several PUFAs, including EPA, DHA and other fatty acids produced from a marine microalgal strain of Thraustochytriidae sp. derived mutant. It was found to contain more PUFAs （〉80%） and more omega-3 oils than commercial omega-3 fish oil （PUFAs 〉31%） and corn oil （PUFAs 59%）. All three types of oils induced weight loss in high-fat-induced obese mice, with the loss induced by microalgal oil being most significant at 9 weeks （10% reduction）. However, the oils tested did not improve blood lipid levels, although microalgal oil showed an apparent inhibitory effect on lipid accumulation in the liver. These findings may be attributed to the higher PUFA content, including omega-3 oils of microalgal oil than other oils. Collectively, these findings suggest that microalgal oil, derived from Thraustochytriidae sp. derived mutant, is a prominent candidate for replacement of omega-3 fish oils based on its apparent anti-obesity effect in vivo.

Reproductive performance and gestational effort in relation to dietary fatty acids in guinea pigs

Background： Dietary saturated（SFAs） and polyunsaturated（PUFAs） fatty acids can highly affect reproductive functions by providing additional energy, modulating the biochemical properties of tissues, and hormone secretions. In precocial mammals such as domestic guinea pigs the offspring is born highly developed. Gestation might be the most critical reproductive period in this species and dietary fatty acids may profoundly influence the gestational effort. We therefore determined the hormonal status at conception, the reproductive success, and body mass changes during gestation in guinea pigs maintained on diets high in PUFAs or SFAs, or a control diet.Results： The diets significantly affected the females＇ plasma fatty acid status at conception, while cortisol and estrogen levels did not differ among groups. SFA females exhibited a significantly lower body mass and litter size, while the individual birth mass of pups did not differ among groups and a general higher pup mortality rate in larger litters was diminished by PUFAs and SFAs. The gestational effort, determined by a mother＇s body mass gain during gestation, increased with total litter mass, whereas this increase was lowest in SFA and highest in PUFA individuals. The mother＇s body mass after parturition did not differ among groups and was positively affected by the total litter mass in PUFA females.Conclusions： While SFAs reduce the litter size, but also the gestational effort as a consequence, PUFA supplementation may contribute to an adjustment of energy accumulations to the total litter mass, which may both favor a mother＇s body condition at parturition and perhaps increase the offspring survival at birth.

Oils with different degree of saturation:effects on ileal digestibility of fat and corresponding additivity and bacterial community in growing pigs

Background Oils are important sources of energy in pig diets.The combination of oils with different degree of saturation contributes to improve the utilization efficiency of the mixed oils and may reduce the cost of oil supplemented.An experiment was conducted to evaluate the effects of oils with different degree of saturation on the fat digestibility and corresponding additivity and bacterial community in growing pigs.Methods Eighteen crossbred(Duroc×Landrace×Yorkshire)barrows(initial body weight:29.3±2.8 kg)were surgically fitted with a T-cannula in the distal ileum.The experimental diets included a fat-free basal diet and 5 oil-added diets.The 5 oil-added diets were formulated by adding 6%oil with different ratio of unsaturated to saturated fatty acids(U:S)to the basal diet.The 5 oils were palm oil(U:S=1.2),canola oil(U:S=12.0),and palm oil and canola oil were mixed in different proportions to prepare a combination of U:S of 2.5,3.5 and 4.5,respectively.Results The apparent and standardized ileal digestibility(AID and SID)of fat and fatty acids increased linearly(P<0.05)as the U:S of dietary oils increased except for SID of fat and C18:2.The AID and SID of fat and fatty acids differed among the dietary treatments(P<0.05)except for SID of unsaturated fatty acids(UFA)and C18:2.Fitted one-slope broken-line analyses for the SID of fat,saturated fatty acids(SFA)and UFA indicated that the breakpoint for U:S of oil was 4.14(R^(2)=0.89,P<0.01),2.91(R^(2)=0.98,P<0.01)and 3.84(R^(2)=0.85,P<0.01),respectively.The determined SID of fat,C18:1,C18:2 and UFA in the mixtures was not different from the calculated SID of fat,C18:1,C18:2 and UFA.However,the determined SID of C16:0,C18:0 and SFA in the mixtures were greater than the calculated SID values(P<0.05).The abundance of Romboutsia and Turicibacter in pigs fed diet containing palm oil was greater than that in rapeseed oil treatment group,and the two bacteria were negatively correlated with SID of C16:0,C18:0 and SFA(P<0.05).Conclusions The optimal U:S for improving the utilization efficiency of mixed oil was 4.14.The SID of fat and UFA for palm oil and canola oil were additive in growing pigs,whereas the SID of SFA in the mixture of two oils was greater than the sum of the values of pure oils.Differences in fat digestibility caused by oils differing in degree of saturation has a significant impact on bacterial community in the foregut.

Food for Thought: Have We Been Giving the Wrong Dietary Advice?

Background: Since 1984 UK citizens have been advised to reduce total dietary fat intake to 30% of total energy and saturated fat intake to 10%. The National Institute of Clinical Excellence [NICE] suggests a further benefit for Coronary Heart Disease [CHD] prevention by reducing saturated fat [SFA] intake to 6% - 7% of total energy and that 30,000 lives could be saved by replacing SFAs with Polyunsaturated fats [PUFAs]. Methods: 20 volumes of the Seven Countries Study, the seminal work behind the 1984 nutritional guidelines, were assessed. The evidence upon which the NICE guidance was based was reviewed. Nutritional facts about fat and the UK intake of fat are presented and the impact of macronutrient confusion on public health dietary advice is discussed. Findings: The Seven Countries study classified processed foods, primarily carbohydrates, as saturated fats. The UK government and NICE do the same, listing biscuits, cakes, pastries and savoury snacks as saturated fats. Processed foods should be the target of public health advice but not natural fats, in which the UK diet is deficient. With reference to the macro and micro nutrient composition of meat, fish, eggs, and dairy foods the article demonstrates that dietary trials cannot change one type of fat for another in a controlled study. Interpretation: The evidence suggests that processed food is strongly associated with the increase in obesity, diabetes, CHD, and other modern illness in our society. The macro and micro nutrients found in meat, fish, eggs and dairy products, are vital for human health and consumption of these nutritious foods should be encouraged.

Lipid Metabolic Profile Interrelationships between the Silkworm(Bombyx mori L.)and Its Host Plant(Morus)Varieties

The lipid content and composition in tender, medium and coarse leaves of 3 mulberry varieties were analyzed. Tender leaf contained more lipid than medium and coarse leaves. Fatty acid composition of lipids also varied with leaf maturity. Tender leaf contained more precursor fatty acids, medium leaf more unsaturated fatty acids and coarse leaf more saturated fatty acids. V-1 mulberry variety had more foliar lipid con- tent than S-36 and M-5. Lipid composition also varied among the 3 mulberry varieties. V-1 and S-36 leaves contained more unsaturated fatty acids and M-5 more saturated fatty acids. V instar silkworm was fed with the medium leaf of the 3 mulberry varieties, and the total lipid content and fatty acid composition of the haemolymph, fatbody and excreta were analyzed. Lauric acid （12：0）, undetectable in the haemolymph, was recovered in significant quantity in the excreta. Saturated fatty acids （16：0 and 18：0） were higher in the haemolymph of larvae fed on M-5 while unsaturated fatty acids （18：2 and 18：3） were higher in those fed on S-36 and V-1 leaves. But, the percentage levels of 18：1 and 18：2 fatty acids in the leaf and haemolymph were swapped. While the leaf of M-5 mulberry variety had 18：1 and 18：2 fatty acid components at 1.93% and 16.8%, the larvae feeding on the leaf had 18：1 and 18：2 at 21.5% and 6.95% in the haemolymph. Similar correction in the percentages of the two unsaturated fatty acids was observed in the larvae feeding on the leaves of S-36 and V-1. Saturation of co -6 double bond of linoleic acid by a reductase in the lower end of the midgut was proposed.

Effects of the ratio of unsaturated fatty acid to saturated fatty acid on the growth performance,carcass and meat quality of finishing pigs

The effects on finishing pigs(80-100 kg BW) fed diets supplemented with oil sources containing different ratios of unsaturated to saturated fatty acids(UFA:SFA ratio) were evaluated in 15 barrows and15 gilts(Duroc × Large White × Landrace). Three experimental diets were evaluated using a randomized complete block design, with broken rice, soybean meal and rice bran as the main feedstuffs in the control diet. Diets 2 and 3 consisted of the control diet supplemented with 3% oil, with UFA:SFA ratios of 2.5:1 and 5:1, respectively. Overall, there was no significant difference(P> 0.05) found in the average daily gain(ADG) of the pigs fed the treatment diets; however, the pigs fed the control diet and diet 3 had better(P < 0.05) feed conversion ratios(FCR) than the pigs fed diet 2. The pigs fed diets 2 and 3, which were supplemented with oil at UFA:SFA ratios of 2.5:1 and 5:1, had greater(P < 0.05) average daily feed intakes(ADFI) than the pigs in the control group. Additionally, it was found that the gender of the pigs had an effect(P < 0.05) on the FCR. Interaction effects between the experimental diets and the gender of the pigs(P < 0.05) were found in the ADFI and FCR. There were no significance differences(P > 0.05)among the treatment groups with regard to the carcass quality of the pigs; however, it was found that the gilts had greater(P < 0.01) loin eye areas than the barrows fed diets 2 and 3 and the loin eye area of pig fed diet 2 was the largest(P < 0.05); In the case of the meat quality parameters, it was clearly found that the pigs fed the control diet had a greater(P < 0.05) lightness(L~*) in the meat colour, and the lowest cooking loss was found in the pigs fed the diet supplemented with fat containing the UFA:SFA ratio of 5:1.Overall, the dietary treatment did not significantly affect the drip loss, thawing loss and shear force of the pork. In conclusion, the supplementation of oil with UFA:SFA ratios of 2.5:1 and 5:1 has the potential to improve pork quality.

Comparative studies on phosphate ore flotation collectors prepared by hogwash oil from different regions

In order to compare the differences between variations of phosphate ore flotation collectors prepared by hogwash oils, JZQ-F collectors were prepared by the hydrolysis method using four types of hogwash oils from different regions in China. The components of the hogwash oils were determined using Fourier transform infrared spectroscopy(FIIR) spectra analysis, while the components of the JZQ-F were determined through gas chromatography–mass spectrometry(GC–MS). The flotation effects of the JZQ-F collectors were investigated by flotation experiments and the adsorption characteristics were determined through the quartz crystal microbalance(QCM-D). Results show that the JZQ-F collector from the Ordos area has 27.43% unsaturated fatty acids. JZQ-F collectors from Beijing, Qingdao, and Dongguan areas contain over 62% of unsaturated fatty acids, which can acquire phosphorous concentrates with a the grade of P_2O_5 above 31.96% and the recovery higher than 91.52%. These three collectors have a larger adsorption capacity, faster adsorption rate, and stronger adsorption structure on apatite surface, which contributed to a good adsorption performance and a favorable flotation effect.

Palmitate induces fat accumulation by activating C/EBPβ-mediated G0S2 expression in HepG2 cells

AIM To determine the role of G0/G1 switch gene 2(G0 S2) and its transcriptional regulation in palmitate-induced hepatic lipid accumulation.METHODS Hep G2 cells were treated with palmitate,or palmitate in combination with CCAAT/enhancer binding protein(C/EBP)β si RNA or G0 S2 si RNA. The m RNA expression of C/EBPβ,peroxisome proliferator-activated receptor(PPAR)γ and PPARγ target genes(G0 S2,GPR81,GPR109 A and Adipoq) was examined by q PCR. The protein expression of C/EBPβ,PPARγ,and G0 S2 was determined by Western blotting. Lipid accumulation was detected with Oil Red O staining and quantified by absorbance value of the extracted Oil Red O dye. Lipolysis was evaluated by measuring the amount of glycerol released into the medium. RESULTS Palmitate caused a dose-dependent increase in lipid accumulation and a dose-dependent decrease in lipolysis in Hep G2 cells. In addition,palmitate increased the m RNA expression of C/EBPβ,PPARγ,and PPARγ target genes(G0 S2,GPR81,GPR109 A,and Adipoq) and the protein expression of C/EBPβ,PPARγ,and G0 S2 in a dose-dependent manner. Knockdown of C/EBPβ decreased palmitate-induced PPARγ and its target genes(G0 S2,GPR81,GPR109 A,and Adipoq) m RNA expression and palmitate-induced PPARγ and G0 S2 protein expression in Hep G2 cells. Knockdown of C/EBPβ also attenuated lipid accumulation and augmented lipolysis in palmitate-treated Hep G2 cells. G0 S2 knockdown attenuated lipid accumulation and augmented lipolysis,while G0 S2 knockdown had no effects on the m RNA expression of C/EBPβ,PPARγ,and PPARγ target genes(GPR81,GPR109 A and Adipoq) in palmitate-treated Hep G2 cells. CONCLUSION Palmitate can induce lipid accumulation in Hep G2 cells by activating C/EBPβ-mediated G0 S2 expression.

Long-term high fructose and saturated fat diet affects plasma fatty acid profile in rats

As the consumption of fructose and saturated fatty acids(FAs) has greatly increased in western diets and is linked with an increased risk of metabolic syndrome,the aim of this study was to investigate the effects of a mod-erate(10 weeks) and a prolonged(30 weeks) high fructose and saturated fatty acid(HFS) diet on plasma FA com-position in rats.The effects of a few weeks of HFS diet had already been described,but in this paper we tried to es-tablish whether these effects persist or if they are modified after 10 or 30 weeks.We hypothesized that the plasma FA profile would be altered between 10 and 30 weeks of the HFS diet.Rats fed with either the HFS or a standard diet were tested after 10 weeks and again after 30 weeks.After 10 weeks of feeding,HFS-fed rats developed the metabolic syndrome,as manifested by an increase in fasting insulinemia,total cholesterol and triglyceride levels,as well as by impaired glucose tolerance.Furthermore,the plasma FA profile of the HFS group showed higher proportions of monounsaturated FAs like palmitoleic acid [16:1(n-7)] and oleic acid [18:1(n-9)],whereas the proportions of some polyunsaturated n-6 FAs,such as linoleic acid [18:2(n-6)] and arachidonic acid [20:4(n-6)],were lower than those in the control group.After 30 weeks of the HFS diet,we observed changes mainly in the levels of 16:1(n-7)(decreased) and 20:4(n-6)(increased).Together,our results suggest that an HFS diet could lead to an adaptive response of the plasma FA profile over time,in association with the development of the metabolic syndrome.

Mechanisms of Steatosis-Derived Hepatocarcinogenesis:Lessons from HCV Core Gene Transgenic Mice

Hepatitis C virus(HCV)is a major cause of chronic hepatitis,liver cirrhosis,and hepatocellular carcinoma(HCC)worldwide.Among the structural proteins of HCV,the HCV core protein has the ability to regulate gene transcription,lipid metabolism,cell proliferation,apoptosis,and autophagy,all of which are closely related to the development of HCC.Transgenic mice carrying the HCV core gene exhibited age-dependent insulin resistance,hepatic steatosis,and HCC that resembled the clinical characteristics of chronic hepatitis C patients.Several dietary modifications,including calorie restriction and diets rich in saturated fatty acids,trans fatty acids,or cholesterol,were found to influence hepatic steatogenesis and tumorigenesis in HCV core gene transgenic mice.These strategies modulated hepatocellular stress and proliferation,in addition to hepatic fibrotic processes and the microenvironment,thereby corroborating a close interconnection between dietary habits and steatosis-related hepatocarcinogenesis.In this review,we summarize the findings obtained from mouse models transgenic for the HCV genome,with a special focus on HCV core gene transgenic mice,and discuss the mechanisms of steatogenesis and hepatocarcinogenesis induced by the HCV core protein and the impact of dietary habits on steatosis-derived HCC development.

Coconut oil:what do we really know about it so far?

In recent years,coconut oil has emerged as a potential‘miracle’food.Some media vehicles and health specialists assure that this fat is capable of promoting health benefits,such as weight reduction,cholesterol lowering,prevention of cardiovascular diseases,and anti-inflammatory effect,among others.These claims are used to market the product and boost its sales by coconut oil companies.However,governmental regulatory agencies in many countries are still sceptical about the benefits obtained by the consumption of coconut oil due to its high-saturated fatty acid content.In light of such controversy,this review focused on analysing the published literature on the alleged health claims,in order to investigate if there is enough scientific evidence to support them.It was verified that the metabolism of lauric acid,the major fatty acid in coconut oil,remains unclear.Many studies reported that the product was not efficient in weight loss.Also,it has been reported that the consumption of coconut oil increased low-density lipoprotein cholesterol,consequently increasing the risk of cardiovascular diseases.In general,the studies present conflicting results and there is a lack of long-term human-based clinical trials.Therefore,as a saturated fat,coconut oil should be consumed with moderation and the health allegations should not be used to market the product,once they are not scientifically proven so far.

Dietary butyrate,lauric acid and stearic acid improve gut morphology and epithelial cell turnover in weaned piglets

This study was to evaluate the effects of the supplementation of saturated fatty acids with different chain lengths on growth performance,intestinal morphology,epithelial cell proliferation,differentiation and apoptosis in weaned piglets.Thirty-two weaned piglets(Duroc×Landrace×Yorkshire,BW=7.81±0.26 kg)were weaned at 21 d and randomly assigned to 1 of 4 experimental treatments:(1)a basal diet(control);(2)control+0.3% butyrate(BT);(3)control+0.3%lauric acid(LA);(4)control+0.3% stearic acid(SA).All piglets were then slaughtered for tissue sampling after having been fed experimental diets for 28 d after weaning.Supplementation of BT increased the gain-to-feed ratio(G:F)(P<0.05)compared to piglets fed the control diet from 14 to 28 d.In addition,the villus height(VH)to crypt depth(CD)ratio(VH:CD ratio)of the ileum were higher in the BT and LA diets than that of the control diet(P<0.05).The SA-supplemented diet increased ileal VH(P<0.05),whereas the BT-supplemented diet increased jejunal CD(P<0.05).Compared to the control,diets supplemented with BT,LA,or SA all tended to increase jejunal proliferation(Ki67/crypt positive cells)(P=0.190);diets supplemented with BT or SA significantly increased the number of ki67-positive cells in the ileal crypt(P<0.05).Furthermore,in the jejunum,the protein expression of activated caspase 3 and villin were increased in piglets fed BT,LA,or SA diets compared to those on the control diet(P<0.05).In the ileum,compared with the control diet,the BT diet tended to increase the protein level of mammalian phosphorylation target of rapamycin(p-m TOR,P<0.10);LA or SA diets significantly increased p-m TOR protein expression(P<0.05).These results show that dietary supplementation of BT,LA,or SA promotes jejunal cell renewal in weaned piglets.At the same time,increased proliferation of ileal crypt cells by promoting p-m TOR expression has beneficial effects on ileal morphology in weaned piglets.