Long-chain Unsaturated Fatty Acids Research Articles

Effect of Amorphous Halomonas-PHB on Growth, Body Composition, Immune-Related Gene Expression and Vibrio anguillarum Resistance of Hybrid Grouper (Epinephelus fuscoguttatus ♀ × E. lanceolatu ♂) Juveniles

Poly-β-hydroxybutyrate (PHB) is a bacterial metabolite produced by bacteria such as Halomonas sp. that serves as a carbon and energy storage compound for bacteria under nutrient-limited conditions. Two experiments were conducted to investigate the effects of dietary supplementation with Halomonas-PHB on hybrid grouper (Epinephelus fuscoguttatus ♀ × E. lanceolatu ♂). In experiment I, juvenile groupers were fed basal diets supplemented with 3% Halomonas-PHB (3% HM-PHB) containing 1.4% PHB and 3% Halomonas (3% HM) without PHB, as well as a control diet, for seven weeks. The results showed no significant difference in survival rate, weight gain, and crude fat content between the 3% HM-PHB group and the control group; however, the crude protein of the 3% HM-PHB group was significantly lower than that of the control group. Furthermore, supplementation with 3% HM-PHB increased the fatty acids content in fish muscles, including long-chain unsaturated fatty acids C18:1n9, EPA, and DHA. In experiment II, groupers were fed a basal diet supplemented with 6.5% Halomonas-PHB (6.5% HM-PHB) containing 3% PHB and 6.5% Halomonas (6.5% HM) containing no PHB, as well as a basal diet (Control). After seven weeks of rearing, the fish were challenged with Vibrio anguillarum for 48 h. Although no significant difference in survival rate and growth was observed among different groups, the dietary supplement of 6.5% Halomonas-PHB improved the survival rate of V. anguillarum challenged grouper and significantly increased the gene expressions of catalase (CAT) and superoxide dismutase (SOD) in blood, interleukin 1 (IL1) and interleukin 10 (IL10) in the liver, spleen, head kidney, and blood (p < 0.05). In conclusion, dietary supplementation of Halomonas-PHB had no significantly positive effect on fish growth performance but increased the content of fatty acids, including long-chain unsaturated fatty acids C18:1n9, EPA, and DHA in fish muscle; it also improved the V. anguillarum resistance, possibly through increasing immune-related gene expression in different tissues and organs. Our findings offer compelling evidence that Halomonas-PHB can be utilized as a feed additive in intensive grouper farming to enhance the groupers’ resistance to Vibrio.

Clotrimazole Identified as a Selective UGT2B4 Inhibitor Using Canagliflozin-2'-O-Glucuronide Formation as a Selective UGT2B4 Probe Reaction.

UGT2B4 is a highly expressed drug-metabolizing enzyme in the liver contributing to the glucuronidation of several drugs. To enable quantitatively assessing UGT2B4 contribution toward metabolic clearance, a potent and selective UGT2B4 inhibitor that can be used for reaction phenotyping was sought. Initially, a canagliflozin-2'-O-glucuronyl transferase activity assay was developed in recombinant UGT2B4 and human liver microsomes (HLM) [±2% bovine serum albumin (BSA)]. Canagliflozin-2'-O-glucuronidation (C2OG) substrate concentration at half-maximal velocity value in recombinant UGT2B4 and HLM were similar. C2OG formation intrinsic clearance was five- to seven-fold higher in incubations containing 2% BSA, suggesting UGT2B4 susceptibility to the inhibitory unsaturated long-chain fatty acids released during the incubation. Monitoring for C2OG formation, 180 compounds were evaluated for UGT2B4 inhibition potency in the presence and absence of 2% BSA. Compounds that exhibited an apparent UGT2B4 IC50 of < 1 μM in HLM with 2% BSA were evaluated for inhibition of UGT1A1, UGT1A3, UGT1A4, UGT1A6, UGT1A9, UGT2B7, UGT2B10, UGT2B15, and UGT2B17 catalytic activities to establish selectivity suitable for supporting UGT reaction phenotyping. In this study, clotrimazole was identified as a potent UGT2B4 inhibitor (HLM apparent IC50 of 11 to 35 nM ± 2% BSA). Moreover, clotrimazole exhibited selectivity for UGT2B4 inhibition (>24-fold) over the other UGT enzymes evaluated. Additionally, during this study it was discovered that the previously described UGT2B7 inhibitors 16α- and 16β-phenyllongifolol also inhibit UGT2B4. Clotrimazole, a potent and selective UGT2B4 inhibitor, will prove essential during UGT reaction phenotyping. SIGNIFICANCE STATEMENT: To mechanistically evaluate drug interactions, it is essential to understand the contribution of individual enzymes to the metabolic clearance of a drug. The present study describes the development of a UGT2B4 activity assay that enabled the discovery of the highly selective and potent UGT2B4 inhibitor clotrimazole. Clotrimazole can be used in UGT reaction phenotyping studies to estimate fractional contribution of UGT2B4.

Effective complete replacement of fish oil by linseed oil in diets for thick-lipped grey mullet (Chelon labrosus) juveniles reared at three environmental salinities

One of the major challenges for fish oil replacement by alternative oils is the limited capacity of marine fish species to convert 18C fatty acids into long-chain poly unsaturated fatty acids (LC-PUFA). Since salinity markedly affects this conversion capacity, euryhaline species, such as thick-lipped grey mullet (Chelon labrosus), may have a good potential to use diets without inclusion of fish oil. To test this hypothesis, thick-lipped grey mullet juveniles were fed two different diets based on fish oil (FO diet) or linseed oil (LO diet) at three different salinities: 46 ppt, 35 ppt, and 16 ppt. At the end of the trial, the biochemical and fatty acid composition of whole-body, liver, and muscle, as well as the relative expression of the genes codifying for the elongase Elovl5 and the desaturase Fads2 in liver, were determined. The results showed the good capacity of mullet juveniles to convert 18C fatty acids into LC-PUFA, which was enhanced by the replacement of FO by LO and the reduction of salinity to an isosmotic level (16 ppt). Besides, these juveniles efficiently used diets with complete replacement of FO by LO with growth performance, body composition, and LC-PUFA fatty acid profiles similar to those of fish fed FO, particularly when reared at 16 ppt.

Lipase-catalyzed synthesis and characterization of medium-long-medium (MLM) type structured triacylglycerols from peony seed oil

Structured triacylglycerols (STAGs) with medium-chain fatty acids (FAs) at sn-1,3 positions and long-chain FAs at the sn-2 position are known as medium-long-medium (MLM) type STAGs. They can not only provide energy immediately and control weight effectively, but may also provide essential or beneficial FAs to meet the human needs. In this study, MLM type STAGs were synthesized from lipase-catalyzed acidolysis of peony seed oil (PSO) with octanoic acid (OcA) and decanoic acid (DA) as medium-chain FAs in solvent-free systems. Optimal reaction conditions were 1:8 (PSO to OcA/DA), 40 °C and 24 h. The OcA content and DA content at sn-1,3 of two STAGs were 81.1% and 82.5%. The total essential FA (α-linolenic acid and linoleic acid) contents at sn-2 reached 61.5% and 57.2%, and long-chain unsaturated FA contents reached 93.3% and 87.4%. Infrared absorptions of two STAGs were weaker than those of PSO at 723 cm−1 [-(CH2)n-rocking] and 3011 cm−1 [ = CH (cis) stretching]. The end melting temperature and maximum weight loss temperature of STAG rich in DA were higher than those of STAG rich in OcA. This study is conducive to the deep processing of PSO and provides a new and promising substrate for MLM type STAG synthesis.

Quantity and Quality Evaluation of Girardinia Diversifolia and Girardinia Bullosa Seed Oil

Kenya imports more than 65% of its edible oil requirements. This is due to low oil production and limited sources that cannot adequately meet the high demand by the ever-increasing population. This study aimed to determine the fatty acid profile and assess the quantity and quality of Girardinia diversifolia and Girardinia bullosa seed oil. Its quality was assessed in terms of iodine value (IV), saponification value (SV), acid value(AV), peroxide value(PV), unsaponifiable matter, refractive index (RI), specific gravity(SG) and moisture content and volatile matter. The percentage oil content (18.4 to 21.2/100g dry) was low relative to most common edible oil plants. The highest moisture content recorded was 39.60 with the lowest being 31.24g/ 100g air-dried seeds. The RI ranged from 1.4662-1.4704. The IV ranged from 166-192. The PV was in the range of 8.68 to 13.6. The SV (164-179) was comparable to that of corn oil (187-147). The AV varied from 4.58-7.22. The percent unsaponifiable matter of the oil varied from 2.14-4.10. The fatty acids detected in the oil were decanoic acid, hexadecanoic acid, octadecanoic acid, 9-octadecaenoic acid, 9,12-octadecadienoic acid, eicosanoic acid and heneicosanoic acid. The results showed that Girardinia oil is stable in the first two hours of heating. The results indicated that the oil had a high content of long-chain unsaturated fatty acids, and this guarantees the possibility of using the oil in the food industry.

Course-based intra-articular injection of medical chitosan mitigates excessive deposition of triacylglycerides in the synovial tissue of the knee osteoarthritis.

This study aimed to investigate the clinical efficacy of intra-articular injections of medical chitosan for treating knee osteoarthritis (KOA) and measure the lipid metabolism profiles of the synovial tissue. Sixty patients with KOA undergoing conservative treatment were recruited and randomized into two groups: one without pharmacological intervention (OA group) and the other receiving course-based intra-articular medical chitosan injections (CSI group). Quantitative lipidomic profile of synovial tissue was analyzed. Functional scores, including Kellgren-Lawrence rating (K-L), Visual Analog Scale (VAS), Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) scoring, and American Knee Society (AKS) scoring were conducted. Survival from the initial conservative treatment to final knee arthroplasty was significantly longer in the CSI group compared to the OA group. Except for the presurgery VAS score, no statistically significant differences were observed in the other scores, including K-L, initial VAS, WOMAC, and AKS. However, the CSI group experienced more reductions in AKS-Knee subscores compared to the OA group. Compared to the CSI group, the OA group exhibited a significant upregulation in most differential lipids, particularly triacylglycerides (TAGs, 77%). The OA group had notably higher levels of long-chain unsaturated fatty acids. Intra-articular injection of medical chitosan significantly prolongs the survival period before knee arthroplasty and reduces the deposition of TAGs metabolites.

Transcriptome Analysis Reveals the Potential Key Genes in Nutritional Deposition in the Common Carp (Cyprinus carpio).

The common carp (Cyprinus carpio) is one of the most important aquaculture species in China, known for its remarkable adaptability and nutritional profile. However, the specific molecular response mechanisms regulating the nutritional deposition of carp remain inadequately elucidated. This study conducted a comprehensive analysis of muscle nutritional content and transcriptome data from liver and muscle tissues of three distinct carp varieties. The aim was to elucidate the key genes and signaling pathways that regulate muscle nutritional composition in carp. The findings revealed that FFRC carp (FFRC) exhibited significantly higher levels of crude fat, total n-3 polyunsaturated fatty acids, and total n-6 polyunsaturated fatty acids in muscle tissue compared to Ying carp (YC) and Huanghe carp (HC) (p < 0.05). Transcriptomic analyses correlated these elevated levels with a marked upregulation of genes involved in the activation and transportation of fatty acid (fabp7, acsl5, acsbg2) as well as biosynthesis and elongation of long-chain unsaturated fatty acids (elovl2, fads2) within the liver. Furthermore, the flavor amino acid, essential amino acids, and crude protein content in the muscle of HC were significantly higher than in FFRC and YC (p < 0.05). Transcriptomic analyses indicated that this was associated with significant changes in the expression of genes related to amino acid metabolism (asns, alt, ldha, glul, setd, prodh, l3hypdh, hoga1) within their muscle tissue. This research provides a theoretical foundation for the precise modulation of the muscle nutritional composition in carp.

Essential Omega-3 (n-3PUFA) Nutritional Content of Ptilinopus melanosphila and Chalcophaps indicia Meat

Ptilinopus melanosphila and Chalcophaps indica, known as Puna birds among local people of the Sula Islands, North Maluku, Indonesia, while Treron aromaticus, known as Puna Giu, belong to the Columbidae family. These birds are frequently hunted for consumption due to the delectable taste of their meat. This study aims to examine the essential Omega-3 (n-3PUFA) nutritional content, utilizing these two avian species (Ptilinopus melanosphila and Chalcophaps indicia) as samples. Local perceptions consider these birds as identical, hence this study also provides insights into the disparities between the two species based on meat quality analysis. The analysis encompasses proximate analysis, fatty acids, and amino acids, conducted on 6 samples, each encompassing 3 breast meats. These samples were mixed and blended, followed by chemical analysis at the integrated laboratory of Universitas IPB Baranangsiang, with proximate analysis executed at PAU IPB. The results reveal that Ptilinopus melanosphila meat contains crucial nutrients for human consumption, specifically long-chain omega-3 unsaturated fatty acids (n-3PUFA), which are more abundant compared to Chalcophaps indica and several other meat products. The myriad of significant benefits offered by this avian species underscores the importance of recommending its preservation to parties engaged in ex-situ conservation efforts. Furthermore, it is advocated to stimulate the establishment of regional culinary businesses specializing in nutrient-rich Black-Naped Fruit Dove meat.

Differentiation of Cis/trans-geometrical isomers in long-chain unsaturated fatty acids based on ion mobility and theoretical calculations

In recent years, fatty acids containing conjugated CCs have attracted extensive research attention due to their biological activities against human diseases. However, their differentiation is challenging. This study developed a comprehensive analytical solution to accurately differentiate cis/trans-fatty acid isomers using ion mobility mass spectrometry (IM-MS) and theoretical calculations. Cis/trans-fatty acids were mobility-differentiated via simple complexation with 1,5,9-triazacyclododecane (9C3N) or 1,4,8,11-tetraazacyclotetradecane (10C4N) and metal ions, obtaining baseline separation with a peak-to-peak resolution of 0.35–0.92. Moreover, the conformation of the complexes was optimized theoretically, revealing different binding modes between the cis/trans-fatty acid-9C3N/10C4N–metal ion systems, yielding in-depth structural data on the complexes and elucidating the principles of mobility separation. Furthermore, the proposed method was assessed in terms of quantification, accuracy, and precision repeatability. Finally, the method was applied to analyze oil samples. Given its simplicity, speed, and lack of chemical derivatization or chromatographic separation, this technique has potential applications in food analysis.

Effects of food waste hydrolysate as an external carbon source on defoaming in wastewater treatment with activated sludge process

The activated sludge process is the most widely used technology for treating municipal wastewater. However, thick foam often occurs in activated sludge process. Here, we reported for the first time the effect of food waste hydrolysate (FWH) as an external carbon source on defoaming in activated sludge process. The study found that FWH was effective in defoaming at a wide dose range of 50–1600 mg/L total solids, as exhibiting that the foaming tendency of FWH-added foam mixed liquor was reduced to 0 mL-foam/mL-air·min from initial 0.171 mL-foam/mL-air·min in the control without adding FWH with 100 % of defoaming efficiency. Fatty acids, oils, and solid particles in FWH jointly contributed to the deformation. Among these factors, the concentration of long-chain unsaturated fatty acids was mainly responsible for the defoaming. This work provides a cost-effective strategy to solve the foaming problem in activated sludge process as well as providing external carbon sources.

Long-chain unsaturated fatty acids sensor controlling the type III/VI secretion system is essential for Edwardsiella piscicida infection

Edwardsiella piscicida is an acute marine pathogen that causes severe damage to the aquaculture industry worldwide. The pathogenesis of E. piscicida is dependent mainly on the type III secretion system (T3SS) and type VI secretion system (T6SS), both of which are critically regulated by EsrB and EsrC. In this study, we revealed that fatty acids influence T3SS expression. Unsaturated fatty acids (UFAs), but not saturated fatty acids (SFAs), directly interact with EsrC, which abolishes the function of EsrC and results in the turn-off of T3/T6SS. Moreover, during the in vivo colonization of E. piscicida, host fatty acids were observed to be transported into E. piscicida through FadL and to modulate the expression of T3/T6SS. Furthermore, the esrCR38G mutant blocked the interaction between EsrC and UFAs, leading to dramatic growth defects in DMEM and impaired colonization in HeLa cells and zebrafish. In conclusion, this study revealed that the interaction between UFAs and EsrC to turn off T3/T6SS expression is essential for E. piscicida infection.

Lipidomic Assessment of the Inhibitory Effect of Standardized Water Extract of Hydrangea serrata (Thunb.) Ser. Leaves during Adipogenesis.

Obesity is primarily exacerbated by excessive lipid accumulation during adipogenesis, with triacylglycerol (TG) as a major lipid marker. However, as the association between numerous lipid markers and various health conditions has recently been revealed, investigating the lipid metabolism in detail has become necessary. This study investigates the lipid metabolic effects of Hydrangea serrata (Thunb.) Ser. hot water leaf extract (WHS) on adipogenesis using LC-MS-based lipidomics analysis of undifferentiated, differentiated, and WHS-treated differentiated 3T3-L1 cells. WHS treatment effectively suppressed the elevation of glycerolipids, including TG and DG, and prevented a molecular shift in fatty acyl composition towards long-chain unsaturated fatty acids. This shift also impacted glycerophospholipid metabolism. Additionally, WHS stabilized significant lipid markers such as the PC/PE and LPC/PE ratios, SM, and Cer, which are associated with obesity and related comorbidities. This study suggests that WHS could reduce obesity-related risk factors by regulating lipid markers during adipogenesis. This study is the first to assess the underlying lipidomic mechanisms of the adipogenesis-inhibitory effect of WHS, highlighting its potential in developing natural products for treating obesity and related conditions. Our study provides a new strategy for the development of natural products for the treatment of obesity and related diseases.

Abstract 1122: Endothelial Cd36 Mediates Lipid-induced Stiffening Of Aortic Endothelium And Western Diet-induced Barrier Disruption

Introduction: Western diet (WD) associates with elevations in blood lipids. Endothelial cells (EC) uptake lipids through scavenger receptors such as CD36. In our studies we demonstrated that exposure to lipids induces a stiffening response in ECs, with in vivo studies showing that this stiffening is abrogated in mice with a global CD36 knockout. Hypothesis: WD associates with aortic barrier disruption. in vitro studies have demonstrated that EC stiffening correlates with barrier disruption. It is our hypothesis that EC stiffening mediates WD-induced aortic barrier disruption in an endothelial CD36-dependent manner. Methods: To generate a model of endothelial-specific CD36 deficiency, we crossed tamoxifen-inducible Cre mice with an EC-promoter with CD36 fl/fl mice. Cre only mice are employed as controls. At the end of the study window, aortas are excised for assessment of EC stiffening via atomic force microscopy. Separate cohorts are assessed for barrier disruption via quantification of Evans blue dye (EBD) leakage or VE-cadherin junctional thickness. Inhibition of CD36 in vitro was accomplished with SSO. For cell experiments we employed long-chain fatty acids (LCFAs), lipid ligands for CD36. Results: WD induced significant stiffening of the aortic endothelium [p&lt;0.05]. This phenomenon was only observed in male (n=5-6) and not female (n=3-10) mice. EC stiffening associated with aortic barrier disruption as indicated by increased EBD permeation (n=3-8) and increased VE-cadherin thickness (n=3-4) [p&lt;0.05]. Use of an endothelial-specific CD36 deficiency significantly abrogated stiffening and barrier disruption [p&lt;0.05]. Controls demonstrated no effects by tamoxifen (n=2-3). Exposure to 5 μM of palmitic and stearic acids (saturated LCFAs) results in stiffening while exposure to oleic acid (unsaturated LCFA) does not (n=5-12) [p&lt;0.05]. Palmitic acid-induced EC stiffening was shown to be dependent on CD36 (n=5-6) and activation of RhoA protein kinase (n=3-8) [p&lt;0.05]. Conclusions: Our results demonstrate that endothelial CD36 is required for WD-induced EC stiffening and subsequent disruption of the aortic barrier. Furthermore, our results suggest that CD36 mediates EC stiffening through its role as scavenger receptor to saturated LCFAs.

Analysis of heterologous expression of phaCBA promotes the acetoin stress response mechanism in Bacillus subtilis using transcriptomics and metabolomics approaches.

Acetoin, a versatile platform chemical and popular food additive, poses a challenge to the biosafety strain Bacillus subtilis when produced in high concentrations due to its intrinsic toxicity. Incorporating the PHB synthesis pathway into Bacillus subtilis 168 has been shown to significantly enhance the strain's acetoin tolerance. This study aims to elucidate the molecular mechanisms underlying the response of B. subtilis 168-phaCBA to acetoin stress, employing transcriptomic and metabolomic analyses. Acetoin stress induces fatty acid degradation and disrupts amino acid synthesis. In response, B. subtilis 168-phaCBA down-regulates genes associated with flagellum assembly and bacterial chemotaxis, while up-regulating genes related to the ABC transport system encoding amino acid transport proteins. Notably, genes coding for cysteine and D-methionine transport proteins (tcyB, tcyC and metQ) and the biotin transporter protein bioY, are up-regulated, enhancing cellular tolerance. Our findings highlight that the expression of phaCBA significantly increases the ratio of long-chain unsaturated fatty acids and modulates intracellular concentrations of amino acids, including L-tryptophan, L-tyrosine, L-leucine, L-threonine, L-methionine, L-glutamic acid, L-proline, D-phenylalanine, L-arginine, and membrane fatty acids, thereby imparting acetoin tolerance. Furthermore, the supplementation with specific exogenous amino acids (L-alanine, L-proline, L-cysteine, L-arginine, L-glutamic acid, and L-isoleucine) alleviates acetoin's detrimental effects on the bacterium. Simultaneously, the introduction of phaCBA into the acetoin-producing strain BS03 addressed the issue of insufficient intracellular cofactors in the fermentation strain, resulting in the successful production of 70.14g/L of acetoin through fed-batch fermentation. This study enhances our understanding of Bacillus's cellular response to acetoin-induced stress and provides valuable insights for the development of acetoin-resistant Bacillus strains.

Human lipocalins bind and export fatty acids through the secretory pathway of yeast cells.

The activation of fatty acids to their acyl-CoA derivatives is a crucial step for their integration into more complex lipids or their degradation via beta-oxidation. Yeast cells employ five distinct acyl-CoA synthases to facilitate this ATP-dependent activation of acyl chains. Notably, mutant cells that are deficient in two of these fatty acid-activating (FAA) enzymes, namely, Faa1 and Faa4, do not take up free fatty acids but rather export them out of the cell. This unique fatty acid export pathway depends on small, secreted pathogenesis-related yeast proteins (Pry). In this study, we investigate whether the expression of human fatty acid-binding proteins, including Albumin, fatty acid-binding protein 4 (Fabp4), and three distinct lipocalins (ApoD, Lcn1, and Obp2a), could promote fatty acid secretion in yeast. To optimize the expression and secretion of these proteins, we systematically examined various signal sequences in both low-copy and high-copy number plasmids. Our findings reveal that directing these fatty-acid binding proteins into the secretory pathway effectively promotes fatty acid secretion from a sensitized quadruple mutant model strain (faa1∆ faa4∆ pry1∆ pry3∆). Furthermore, the level of fatty acid secretion exhibited a positive correlation with the efficiency of protein secretion. Importantly, the expression of all human lipid-binding proteins rescued Pry-dependent fatty acid secretion, resulting in the secretion of both long-chain saturated and unsaturated fatty acids. These results not only affirm the in vitro binding capabilities of lipocalins to fatty acids but also present a novel avenue for enhancing the secretion of valuable lipidic compounds. Given the growing interest in utilizing yeast as a cellular factory for producing poorly soluble compounds and the potential of lipocalins as platforms for engineering substrate-binding specificity, our model is considered as a powerful tool for promoting the secretion of high-value lipid-based molecules.

Long-Chain Polyunsaturated Fatty Acids Accelerate the Rate of Insulin Aggregation and Enhance Toxicity of Insulin Aggregates.

Long-chain polyunsaturated fatty acids (LCPUFAs) are essential components of a human diet. These molecules are critically important for cognitive attention and memory, mood states, coronary circulation, and cirrhosis. However, recently reported findings demonstrated that docosahexaenoic (DHA) and arachidonic acids (ARA), ω-3 and ω-6 LCPUFAs, accelerated the aggregation rates of insulin and α-synuclein, proteins that are directly linked to diabetes type 2 and Parkinson's disease, respectively. Furthermore, both DHA and ARA uniquely altered the structure and toxicity of the corresponding protein aggregates. Our objective is to ascertain whether other LCPUFAs, alongside long-chain unsaturated fatty acid (LCUFA) proteins, exhibit similar effects on amyloidogenic proteins. To explore this matter, we investigated the effect of 10 different LCPUFAs and LCUFAs on the rate of insulin aggregation. We found that all of the analyzed fatty acids strongly accelerated insulin aggregation. Moreover, we found that protein aggregates that were formed in the presence of these fatty acids exerted significantly higher cell toxicity compared with insulin fibrils grown in the lipid-free environment. These findings show that interactions between amyloid-associated proteins and LCPUFAs can be the underlying molecular cause of neurodegenerative diseases.

Cistanche deserticola improves ovariectomized-induced osteoporosis mainly by regulating lipid metabolism: Insights from serum metabolomics using UPLC/Q-TOF-MS

Ethnopharmacological relevanceCistanche deserticola (C. deserticola) is an edible and traditional medicine widely used in China, which has been confirmed to be effective in the treatment of postmenopausal osteoporosis (PMOP). Despite its proven efficacy, the exact role of C. deserticola in bone metabolism and its underlying mechanism has remained unclear. Aim of the studyIn this research, we employed an in vivo model utilizing ovariectomized (OVX) rats to characterize the anti-osteoporotic activity and metabolic mechanism of the ethanol extract of C. deserticola (CHE). Materials and methodsFifty female Sprague-Dawley (SD) rats were randomly divided into five groups including sham operation group, model group, 0.1 g/kg estradiol valerate (EV) group as the positive control, low (0.6 g/kg) and high (1.2 g/kg) dosage CHE groups. Biochemical parameter analyses and histopathological experiments were conducted to assess the pharmacodynamic effects. Metabolomic analysis was conducted on serum samples to examine the metabolic profiles, identify potential biomarkers, and elucidate the metabolic pathways associated with CHE in OVX rats. ResultsCHE treatment demonstrated significant anti-osteoporosis activity by regulating serum biochemical markers of bone turnover, improving cancellous bone structure, and reversing the decrease in bone mineral density. Furthermore, the clinical equivalent dose group (CHL) achieved superior overall outcomes. The main interventions of CHE on OVX rats involved the modulation of several key pathways, including steroid hormone biosynthesis, arachidonic acid metabolism, tyrosine and tryptophan metabolism, biotin metabolism, regulation of TRP channels by inflammatory mediators, primary bile acid biosynthesis, regulation of lipolysis in adipocytes, and bile secretion. 23 potential efficacy-related biomarkers within the metabolic network were identified. Among them, long-chain unsaturated fatty acids (eg. DHA and docosapentaenoic acid), steroid hormones, amino acids and carbohydrates were strongly correlated with bone resorption and formation markers. Additionally, it was observed four pathways (nucleotide, carbon, amino acid, and lipid metabolism) were implicated in the effects of CHE. ConclusionThis study demonstrates that CHE improves bone loss in PMOP mainly through regulating lipid metabolism pathways, which provides an evidence base for CHE treatment of PMOP.

NMR and Docking Calculations Reveal Novel Atomistic Selectivity of a Synthetic High-Affinity Free Fatty Acid vs. Free Fatty Acids in Sudlow's Drug Binding Sites in Human Serum Albumin.

Saturation transfer difference (STD), inter-ligand NOEs (INPHARMA NMR), and docking calculations are reported for investigating specific binding sites of the high-affinity synthetic 7-nitrobenz-2-oxa-1,3-diazoyl-4-C12 fatty acid (NBD-C12 FA) with non-labeled human serum albumin (HSA) and in competition with the drugs warfarin and ibuprofen. A limited number of negative interligand NOEs between NBD-C12 FA and warfarin were interpreted in terms of a short-range allosteric competitive binding in the wide Sudlow's binding site II (FA7) of NBD-C12 FA with Ser-202, Lys-199, and Trp-214 and warfarin with Arg-218 and Arg-222. In contrast, the significant number of interligand NOEs between NBD-C12 FA and ibuprofen were interpreted in terms of a competitive binding mode in Sudlow's binding site I (FA3 and FA4) with Ser-342, Arg-348, Arg-485, Arg-410, and Tyr-411. NBD-C12 FA has the unique structural properties, compared to short-, medium-, and long-chain saturated and unsaturated natural free fatty acids, of interacting with well-defined structures with amino acids of both the internal and external polar anchor sites in Sudlow's binding site I and with amino acids in both FA3 and FA4 in Sudlow's binding site II. The NBD-C12 FA, therefore, interacts with novel structural characteristics in the drug binding sites I and II and can be regarded as a prototype molecule for drug development.

Revealing the synergy mechanisms of organic components anaerobic co-digestion from the prevailing tendency of endogenous inhibitors

The anaerobic co-digestion (AcoD) test among three organic components (model compounds) under high feeding load was carried out in this study, then the dynamic change of intermediate metabolic inhibitors was detected and analyzed to identify the interaction mechanism among organic components from the prevailing tendency of endogenous inhibitors. Carbohydrate and protein show significant two-way synergistic effects for methane production. The addition of protein can supplement additional alkalinity to alleviate the sharp drop of pH (from 4.56 to 6.73) and avoid the acidification caused by high concentration of carbohydrate. Additionally, volatile fatty acids (VFAs) produced by carbohydrate can transform free ammonia to ammonium ion with the largest drop of 54%, thus reducing ammonia toxicity. Carbohydrate and lipid show one-way synergistic effects, namely, lipid can reduce VFAs accumulation by 49% through long chain fatty acids (LCFAs) inhibiting action, and then avoid complete rancidity of carbohydrate fermentation at high concentration. However, protein and lipid show serious two-way antagonistic effects, especially under high load digestion. On one hand, LCFAs would weaken ammonia volatilization and thus aggravate ammonia inhibition; On the other hand, protein would not only accelerate the formation of LCFAs inhibition by promoting lipid hydrolysis, but also transform the degradation pathway of oleic acid (unsaturated LCFAs) and then increase the proportion of stearic acid from 5.17% to 40.33%, which is of higher toxicity than palmitic acid.

Stabilization of Hydroxy-α-Sanshool by Antioxidants Present in the Genus Zanthoxylum.

Japanese pepper (sansho, Zanthoxylum piperitum) contains several types of sanshools belonging to N-alkylamides. Because of the long-chain unsaturated fatty acids present in their structure, sanshools are prone to oxidative deterioration, which poses problems in processing. In this paper, we evaluated the effects of antioxidants from the genus Zanthoxylum in preventing sanshool degradation using accelerated tests. An ethanolic extract of segment membranes of the sansho fruit pericarp was incubated at 70 °C for 7 days with different antioxidants to determine the residual amount of hydroxy-α-sanshool (HαS) in the extract. α-Tocopherol (α-Toc) showed excellent HαS-stabilizing activity at low concentrations. Among phenolic acids, we noted that the HαS-stabilizing activity increased with the number of hydroxy groups per molecule. For example, gallic acid and its derivatives exhibited excellent sanshool-stabilizing activity. Quercetin was found to be a superior HαS stabilizer compared with hesperetin and naringenin. However, the effective concentration was much higher for phenolic compounds than for α-Toc. These substances are believed to play a role in preventing the decomposition of sanshools in the pericarp of sansho. These sanshool stabilizers should be useful in the development of new beverages, foods, cosmetics, and pharmaceuticals that take advantage of the taste and flavor of sansho.