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Keywords = fatty acid synthase system

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17 pages, 4823 KiB  
Article
Genome-Wide Analysis of KCS Gene Family in Ginkgo biloba L. and Functional Identification of KCS7 in Oleic Acid Synthesis
by Xingyu Zhang, Kaifang Fan, Zhi Feng, Zhi Yao, Jinyuan Li, Shuguang Zhang, Xiaoqin Mi, Fuwen Wu, Yiqiang Wang and Meng Li
Genes 2025, 16(7), 773; https://doi.org/10.3390/genes16070773 - 30 Jun 2025
Viewed by 345
Abstract
Background: β-ketolipoyl coenzyme A synthase (KCS) is an essential limiting catalyst involved in carbon chain elongation during fatty acid biosynthesis, characterized by strict substrate specificity. C18:1 (oleic acid) plays a vital role in cell membranes and is essential for nutrient storage and stress [...] Read more.
Background: β-ketolipoyl coenzyme A synthase (KCS) is an essential limiting catalyst involved in carbon chain elongation during fatty acid biosynthesis, characterized by strict substrate specificity. C18:1 (oleic acid) plays a vital role in cell membranes and is essential for nutrient storage and stress defense. There are indications of significant accumulation and rapid synthesis of C18:1 during the early growth stages of Ginkgo biloba L. episperm. The KCS gene family in G. biloba has yet to be analyzed, and the role of KCS in oleic acid synthesis remains unexplored. Methods: In this study, this issue was investigated using transcriptomic and metabolomic data, bioinformatics analysis to screen a key gene from the KCS gene family, and dual validation using yeast and Arabidopsis thaliana expression systems to probe its function. Results: A total of 11 members of the GbKCS gene family were identified, and the dynamics of these genes were analyzed during exocarp development in the G. biloba genome. Among them, the gene designated GbKCS7 showed a highly direct association with the content of C18:1. Heterologous expression of GbKCS7 in yeast increased C18:1N12 and C18:1 content by 3.18-fold and 2.07-fold, respectively. Overexpression of GbKCS7 in Arabidopsis showed that C18:1 was increased by 27.70% and 31.43% in GbKCS7-OE-1 and GbKCS7-OE-2 strains, correspondingly, in juxtaposition to the non-transgenic plants. In addition, the content of VLCFAs increased to varying degrees. Conclusions: These outcomes offer important insights for investigating the role of KCS genes in fatty acid synthesis to further improve G.biloba resistance. Full article
(This article belongs to the Section Plant Genetics and Genomics)
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13 pages, 4529 KiB  
Article
Variation in Intramuscular Fat Deposition of Goats and Sheep and Its Correlation with Gut Microbiota
by Lei Yang, Shaobin Li, Jiagong Hou, Zhisheng Tang, Bingang Shi, Yuzhu Luo, Jiqing Wang and Fangfang Zhao
Foods 2025, 14(11), 1885; https://doi.org/10.3390/foods14111885 - 26 May 2025
Viewed by 581
Abstract
The meat quality of sheep and goats differs even within the same age, gender, and farming systems. Intramuscular fat (IMF) content is an important factor affecting the quality of livestock meat because it affects muscle color, tenderness, juiciness, water-holding capacity, and flavor. This [...] Read more.
The meat quality of sheep and goats differs even within the same age, gender, and farming systems. Intramuscular fat (IMF) content is an important factor affecting the quality of livestock meat because it affects muscle color, tenderness, juiciness, water-holding capacity, and flavor. This study evaluates the differences in IMF deposition characteristics between Longdong cashmere goats and Tan sheep, and also explores the correlations between these variations and the gut microbiota. The results revealed that the IMF contents in shoulder and rump meat, as well as the blood lipid levels, of Longdong cashmere goats were higher than those of Tan sheep (p < 0.05). The content of fatty acid synthase (FAS) in the duodenum of the goats was lower, but the content of hormone-sensitive lipase (HSL) in both the pancreas and duodenum was greater (p < 0.05). The Chao1 and β diversity showed differences between the two breeds, observed not only in the abomasum but also in the colon. The specific microbiota identified from the goats were involved in the lipid metabolism pathway. The concentrations of acetic acid and propionic acid in the colonic and abomasal chyme were decreased in the goats when compared to the sheep (p < 0.05). The contents of FAS in the colonic chyme of the goats were significantly lower, while HSL in the abomasal chyme was significantly higher than that of the sheep. The correlation analysis of IMF deposition with gut microbiota showed that Acetobacter and UBA1711 in the abomasum, as well as Faecousia, WQUU01, UBA5905, and GCA-900066495 in the colon, were positively correlated with the IMF content in shoulder meat and the level of LDL (except for UBA1711), but negatively associated with the content of propionic acid (|r| > 0.45, p < 0.05). This preliminary study has demonstrated that some specific bacteria in the abomasum and colon were associated with IMF deposition, while also providing an indicative reference range for further investigation into the effects of microbes on IMF deposition. Full article
(This article belongs to the Section Meat)
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13 pages, 1492 KiB  
Article
Effects of Nitrogen Fertilizer Spraying Time on Source–Sink Nitrogen Metabolism and Seed Oil Quality of Paeonia ostii ‘Fengdan’
by Nannan Zhang, Xingqiao Liu, Xiaolei Ma, Yabing Zhang, Duoduo Wang, Dingding Zuo, Chengwei Song and Xiaogai Hou
Agronomy 2025, 15(4), 892; https://doi.org/10.3390/agronomy15040892 - 3 Apr 2025
Viewed by 577
Abstract
The spraying time of nitrogen fertilizer is a key factor to consider when fertilizing with an intelligent micro-sprinkler irrigation system. This study aims to investigate the impact of nitrogen fertilizer spraying time on the seed oil quality of tree peony, with the expectation [...] Read more.
The spraying time of nitrogen fertilizer is a key factor to consider when fertilizing with an intelligent micro-sprinkler irrigation system. This study aims to investigate the impact of nitrogen fertilizer spraying time on the seed oil quality of tree peony, with the expectation of providing theoretical support for the application of intelligent micro-sprinkler irrigation systems in the production of tree peony. In 2022 and 2023, foliar nitrogen application was conducted on Paeonia ostii ‘Fengdan’ utilizing an intelligent micro-spray irrigation system, with four distinct nitrogen fertilizer spraying times (3:00–4:00, 7:00–8:00, 14:00–15:00, and 19:00–20:00). Based on this, the study assessed nitrogen metabolism indicators in leaves and seeds at various growth stages and the fatty acid composition of seed oil in Paeonia ostii ‘Fengdan’. The results revealed that foliar nitrogen application between 14:00 and 15:00 significantly enhanced the levels of free amino acids (FAA), nitrate reductase (NR), glutamine synthetase (GS), and glutamate synthase (GOGAT) activity in both leaves and seeds. Furthermore, the ratio of α-linolenic acid in the seed oil was significantly increased. Correlation analysis demonstrated a positive or highly significant positive correlation between the levels of nitrogen metabolism indicators and the ratio of unsaturated fatty acids. In conclusion, foliar nitrogen application between 14:00 and 15:00 significantly enhances the FAA content and the activity of nitrogen metabolism enzymes within the leaves and seeds and promotes the synthesis of unsaturated fatty acids in seed oil. This study contributes to the efficient and high-quality cultivation of tree peony. Full article
(This article belongs to the Section Soil and Plant Nutrition)
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19 pages, 1190 KiB  
Article
FASN, SCD, and PLAG1 Gene Polymorphism and Association with Carcass Traits and Fatty Acid Profile in Hanwoo Cattle
by Jia Yu, Sajida Naseem, Sungkwon Park, Sunjin Hur, Yoonbin Choi, Teahyung Lee, Xiangzi Li and Seongho Choi
Animals 2025, 15(6), 897; https://doi.org/10.3390/ani15060897 - 20 Mar 2025
Viewed by 713
Abstract
Genetic polymorphisms have a great impact on enhancing quantitative traits in cattle. In this study, Fatty acid synthase (FASN) g. 16024 (A>G), Stearoyl-CoA desaturase (SCD) g. 10329 (C>T), and pleomorphic adenoma gene (PLAG1) g. 25003338 (C>G) genotypic [...] Read more.
Genetic polymorphisms have a great impact on enhancing quantitative traits in cattle. In this study, Fatty acid synthase (FASN) g. 16024 (A>G), Stearoyl-CoA desaturase (SCD) g. 10329 (C>T), and pleomorphic adenoma gene (PLAG1) g. 25003338 (C>G) genotypic and allelic polymorphisms were evaluated, along with their associations with fatty acid composition, adipogenic gene expression, and carcass characteristics (carcass weight, yield grade, backfat thickness, and marbling score) in Hanwoo steers. A total of 128 Hanwoo steers were selected for this study and the Polymerase chain reaction–restriction fragment length polymorphism (PCR-RFLP) method was used to identify polymorphism of these genes. The AG genotype and G allele in FASN g. 16024 (A>G), CT genotype and T allele in SCD g. 10329 (C>T), and GG genotype and G allele in PLAG1 g. 25003338 (C>G) showed higher frequency and positively correlated with carcass traits, yield, and quality grades. Fatty acid composition results indicate that C18:3n-6, C20:1, and C20:2n-6 were significantly higher in the AA genotype of FASN gene, C14:1 and C18:3n-6 in the CC genotype, and C16:1 in the TT genotype of SCD gene. C12:0, C14:0, C16:1, C18:0, and C20:0 were higher in the CC genotype of PLAG1 gene. Furthermore, RT-qPCR analysis of adipogenesis-related genes (AMP-activated protein kinase-α (AMPKα), Carnitine palmitoyl transferase-1β (CPT1), G-coupled protein receptor-43 (GPR43), and SCD) across different SNP genotypes suggests a systemic interaction between genetic factors and adipogenesis in beef cattle. This study emphasizes the significance of FASN g. 16024 (A>G), SCD g. 10329 (C>T), and PLAG1 g. 25003338 (C>G) SNPs for genetic selection to enhance beef quality and elucidate lipid metabolic pathways in Hanwoo cattle. Full article
(This article belongs to the Section Animal Genetics and Genomics)
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17 pages, 4522 KiB  
Article
Poly-γ-Glutamic Acid from a Novel Bacillus subtilis Strain: Strengthening the Skin Barrier and Improving Moisture Retention in Keratinocytes and a Reconstructed Skin Model
by Hyun-Ju Ko, SeoA Park, Eunjin Shin, Jinhwa Kim, Geun Soo Lee, Ye-Jin Lee, Sung Min Park, Jungno Lee and Chang-Gu Hyun
Int. J. Mol. Sci. 2025, 26(3), 983; https://doi.org/10.3390/ijms26030983 - 24 Jan 2025
Viewed by 3176
Abstract
A novel Bacillus subtilis HB-31 strain was isolated from Gotjawal Wetland in Jeju Island, Republic of Korea. A mucus substance produced by this strain was identified as high-molecular-weight poly-γ-glutamic acid (γ-PGA) using NMR, Fourier transform infrared spectroscopy, and size-exclusion chromatography/multi-angle light scattering analyses. [...] Read more.
A novel Bacillus subtilis HB-31 strain was isolated from Gotjawal Wetland in Jeju Island, Republic of Korea. A mucus substance produced by this strain was identified as high-molecular-weight poly-γ-glutamic acid (γ-PGA) using NMR, Fourier transform infrared spectroscopy, and size-exclusion chromatography/multi-angle light scattering analyses. We evaluated whether γ-PGA strengthened the skin barrier using keratinocytes and a reconstructed skin model. In keratinocytes, γ-PGA treatment dose-dependently increased the mRNA expression of skin barrier markers, including filaggrin, involucrin, loricrin, serine palmitoyl transferase, fatty acid synthase, and 3-hydroxy-3-methylglutaryl coenzyme A reductase. γ-PGA also enhanced hyaluronic acid synthesis by upregulating hyaluronic acid synthase-1, -2, and -3 mRNA levels and promoted aquaporin 3 expression, which is involved in skin hydration. In the reconstructed skin model, topical application of 1% γ-PGA elevated filaggrin, involucrin, CD44, and aquaporin 3 expression, compared to the control. These results suggest that the newly isolated HB-31 can be used as a commercial production system of high-molecular-weight γ-PGA, which can serve as an effective ingredient for strengthening the skin barrier and improving moisture retention. Further research is needed to explore the long-term effects of γ-PGA on skin health and its application in treating skin conditions. Full article
(This article belongs to the Section Molecular Pathology, Diagnostics, and Therapeutics)
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15 pages, 588 KiB  
Review
Metabolite-Sensing Receptors: Emerging Targets for Modulating Chronic Pain Pathways
by Ciprian Pușcașu, Corina Andrei, Octavian Tudorel Olaru and Anca Zanfirescu
Curr. Issues Mol. Biol. 2025, 47(1), 63; https://doi.org/10.3390/cimb47010063 - 17 Jan 2025
Cited by 2 | Viewed by 1720
Abstract
Chronic pain is a debilitating condition affecting millions worldwide, often resulting from complex interactions between the nervous and immune systems. Recent advances highlight the critical role of metabolite-sensing G protein-coupled receptors (GPCRs) in various chronic pain types. These receptors link metabolic changes with [...] Read more.
Chronic pain is a debilitating condition affecting millions worldwide, often resulting from complex interactions between the nervous and immune systems. Recent advances highlight the critical role of metabolite-sensing G protein-coupled receptors (GPCRs) in various chronic pain types. These receptors link metabolic changes with cellular responses, influencing inflammatory and degenerative processes. Receptors such as free fatty acid receptor 1 (FFAR1/GPR40), free fatty acid receptor 4 (FFAR4/GPR120), free fatty acid receptor 2 (FFAR2/GPR43), and Takeda G protein-coupled receptor 5 (TGR5/GPR131/GPBAR1) are key modulators of nociceptive signaling. GPR40, activated by long-chain fatty acids, exhibits strong anti-inflammatory effects by reducing cytokine expression. Butyrate-activated GPR43 inhibits inflammatory mediators like nitric oxide synthase-2 and cyclooxygenase-2, mitigating inflammation. TGR5, activated by bile acids, regulates inflammation and cellular senescence through pathways like NF-κB and p38. These receptors are promising therapeutic targets in chronic pain, addressing the metabolic and inflammatory factors underlying nociceptive sensitization and tissue degeneration. This review explores the molecular mechanisms of metabolite-sensing receptors in chronic pain, their therapeutic potential, and challenges in clinical application. By uncovering these mechanisms, metabolite-sensing receptors could lead to safer, more effective pain management strategies. Full article
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25 pages, 8725 KiB  
Article
The Altered Lipid Composition and Key Lipid Metabolic Enzymes in Thiacloprid-Resistant Myzus persicae, with Special Attention Paid to the Function of MpTHEM6a
by Jinfeng Hu, Wenhua Rao, Feng Chen, Xianzhi Zhou, Jun Wang, Lei Lin and Guocheng Fan
Int. J. Mol. Sci. 2024, 25(22), 12112; https://doi.org/10.3390/ijms252212112 - 11 Nov 2024
Cited by 1 | Viewed by 1156
Abstract
Neonicotinoid resistance is increasingly prevalent in the agricultural pest Myzus persicae. Lipids play a critical role in insect defense systems, but their contribution to insect neonicotinoid resistance is disregarded. We conducted metabolomics and transcriptomics studies on M. persicae thiacloprid-resistant (THG-R) and -susceptible [...] Read more.
Neonicotinoid resistance is increasingly prevalent in the agricultural pest Myzus persicae. Lipids play a critical role in insect defense systems, but their contribution to insect neonicotinoid resistance is disregarded. We conducted metabolomics and transcriptomics studies on M. persicae thiacloprid-resistant (THG-R) and -susceptible (FFJ-S) populations. A total of 149 lipid metabolites were identified, with 90 upregulated and 59 downregulated in THG-R compared to in FFJ-S. Metabolites in the arachidonic acid (AA) pathway substantially varied between THG-R and FFJ-S. For example, arachidonic acid, (±)11-HETE, and prostaglandin B1 were significantly upregulated, while prostaglandin A1, tetranor-PGDM, 8,15-diHETE, and (±)11(12)-EET were significantly decreased in THG-R. Transcriptomics profiles and qPCR indicated that lipid metabolic enzymes, including fatty acid synthase (FAS), the elongase of very-long-chain fatty acids (ELO), fatty acid desaturase (FAD), and phospholipase (PL) genes, were not overexpressed in THG-R. Among the twelve thioesterase genes, only MpTHEM6a was significantly upregulated in THG-R. Knocking down the expression of MpTHEM6a in THG-R significantly increased the toxicity of the three neonicotinoids, reduced the lifespan of adults, and decreased the number of nonviable nymphs produced by female adults. The metabolites AA, (±)11-HETE, and prostaglandin B1 are potential biomarkers in neonicotinoid-resistant M. persicae. MpTHEM6a may become a potential target for combating neonicotinoid-resistant M. persicae. Full article
(This article belongs to the Section Molecular Endocrinology and Metabolism)
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11 pages, 1431 KiB  
Article
Efficient Catalytic Conversion of Acetate to Citric Acid and Itaconic Acid by Engineered Yarrowia lipolytica
by Yuchen Ning, Renwei Zhang, Huan Liu, Yue Yu, Li Deng and Fang Wang
Catalysts 2024, 14(10), 710; https://doi.org/10.3390/catal14100710 - 10 Oct 2024
Cited by 1 | Viewed by 1792
Abstract
The bioconversion of agricultural and industrial wastes is considered a green and sustainable alternative method for producing high-value biochemicals. As a major catalytic product of greenhouse gases and a by-product in the fermentation and lignocellulose processing industries, acetate is a promising bioconversion raw [...] Read more.
The bioconversion of agricultural and industrial wastes is considered a green and sustainable alternative method for producing high-value biochemicals. As a major catalytic product of greenhouse gases and a by-product in the fermentation and lignocellulose processing industries, acetate is a promising bioconversion raw material. In this work, endogenous and heterologous enzymes were manipulated in Yarrowia lipolytica to achieve the conversion of acetate to high-value citric acid and itaconic acid, respectively. After the combinational expression of the key enzymes in the acetate metabolic pathway, the citric acid synthesis pathway, and the mitochondrial transport system, acetate could be efficiently converted to citric acid. Coupled with the down-regulation of fatty acid synthase expression in the competitive pathway, more acetyl-CoA flowed into the synthesis of citric acid, and the titer reached 15.11 g/L with a productivity of 0.51 g/g acetate by the engineered Y. lipolytica, which is comparable to the results using glucose as the substrate. On this basis, the heterologous cis-aconitate decarboxylase from Aspergillus terreus was introduced into the engineered Y. lipolytica to achieve the catalytic synthesis of itaconic acid from acetate. Combined with investigating the effects of multiple enzymes in the synthesis pathway, the titer of itaconic acid reached 1.87 g/L with a yield of 0.43 g/g DCW by the final engineered strain, which is the highest reported titer of itaconic acid derived from acetate by engineered microbes in shake flasks. It is demonstrated that acetate has the potential to replace traditional starch-based raw materials for the synthesis of high-value organic acids and our work lays a foundation for the rational utilization of industrial wastes and the catalytic products of greenhouse gases. Full article
(This article belongs to the Special Issue Recent Advances in Biocatalysis and Enzyme Engineering)
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38 pages, 7020 KiB  
Article
Antidiabetic and Antihyperlipidemic Activities and Molecular Mechanisms of Phyllanthus emblica L. Extract in Mice on a High-Fat Diet
by Hsing-Yi Lin, Cheng-Hsiu Lin, Yueh-Hsiung Kuo and Chun-Ching Shih
Curr. Issues Mol. Biol. 2024, 46(9), 10492-10529; https://doi.org/10.3390/cimb46090623 - 20 Sep 2024
Viewed by 2068
Abstract
We planned to explore the protective activities of extract of Phyllanthus emblica L. (EPE) on insulin resistance and metabolic disorders including hyperlipidemia, visceral obesity, and renal dysfunction in high-fat diet (HFD)-progressed T2DM mice. Mice treatments included 7 weeks of HFD induction followed by [...] Read more.
We planned to explore the protective activities of extract of Phyllanthus emblica L. (EPE) on insulin resistance and metabolic disorders including hyperlipidemia, visceral obesity, and renal dysfunction in high-fat diet (HFD)-progressed T2DM mice. Mice treatments included 7 weeks of HFD induction followed by EPE, fenofibrate (Feno), or metformin (Metf) treatment daily for another 4-week HFD in HFD-fed mice. Finally, we harvested blood to analyze some tests on circulating glycemia and blood lipid levels. Western blotting analysis was performed on target gene expressions in peripheral tissues. The present findings indicated that EPE treatment reversed the HFD-induced increases in blood glucose, glycosylated HbA1C, and insulin levels. Our findings proved that treatment with EPE in HFD mice effectively controls hyperglycemia and hyperinsulinemia. Our results showed that EPE reduced blood lipid levels, including a reduction in blood triglyceride (TG), total cholesterol (TC), and free fatty acid (FFA); moreover, EPE reduced blood leptin levels and enhanced adiponectin concentrations. EPE treatment in HFD mice reduced BUN and creatinine in both blood and urine and lowered albumin levels in urine; moreover, EPE decreased circulating concentrations of inflammatory NLR family pyrin domain containing 3 (NLRP3) and kidney injury molecule-1 (KIM-1). These results indicated that EPE displayed antihyperglycemic and antihyperlipidemic activities but alleviated renal dysfunction in HFD mice. The histology examinations indicated that EPE treatment decreased adipose hypertrophy and hepatic ballooning, thus contributing to amelioration of lipid accumulation. EPE treatment decreased visceral fat amounts and led to improved systemic insulin resistance. For target gene expression levels, EPE enhanced AMP-activated protein kinase (AMPK) phosphorylation expressions both in livers and skeletal muscles and elevated the muscular membrane glucose transporter 4 (GLUT4) expressions. Treatment with EPE reduced hepatic glucose-6-phosphatase (G6Pase) and phosphoenolpyruvate carboxykinase (PEPCK) expressions to suppress glucose production in the livers and decreased phosphorylation of glycogen synthase kinase 3β (GSK3β) expressions to affect hepatic glycogen synthesis, thus convergently contributing to an antidiabetic effect and improving insulin resistance. The mechanism of the antihyperlipidemic activity of EPE involved a decrease in the hepatic phosphorylation of mammalian target of rapamycin complex C1 (mTORC1) and p70 S6 kinase 1 (S6K1) expressions to improve insulin resistance but also a reduction in hepatic sterol regulatory element binding protein (SREBP)-1c expressions, and suppression of ACC activity, thus resulting in the decreased fatty acid synthesis but elevated hepatic peroxisome proliferator-activated receptor (PPAR) α and SREBP-2 expressions, resulting in lowering TG and TC concentrations. Our results demonstrated that EPE improves insulin resistance and ameliorates hyperlipidemia in HFD mice. Full article
(This article belongs to the Section Molecular Pharmacology)
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17 pages, 4567 KiB  
Article
Intestinal Ketogenesis and Permeability
by Anna Casselbrant, Erik Elias, Peter Hallersund, Erik Elebring, Jakob Cervin, Lars Fändriks and Ville Wallenius
Int. J. Mol. Sci. 2024, 25(12), 6555; https://doi.org/10.3390/ijms25126555 - 14 Jun 2024
Cited by 1 | Viewed by 2332
Abstract
Consumption of a high-fat diet (HFD) has been suggested as a contributing factor behind increased intestinal permeability in obesity, leading to increased plasma levels of microbial endotoxins and, thereby, increased systemic inflammation. We and others have shown that HFD can induce jejunal expression [...] Read more.
Consumption of a high-fat diet (HFD) has been suggested as a contributing factor behind increased intestinal permeability in obesity, leading to increased plasma levels of microbial endotoxins and, thereby, increased systemic inflammation. We and others have shown that HFD can induce jejunal expression of the ketogenic rate-limiting enzyme mitochondrial 3-hydroxy-3-methylglutaryl-CoA synthase (HMGCS). HMGCS is activated via the free fatty acid binding nuclear receptor PPAR-α, and it is a key enzyme in ketone body synthesis that was earlier believed to be expressed exclusively in the liver. The function of intestinal ketogenesis is unknown but has been described in suckling rats and mice pups, possibly in order to allow large molecules, such as immunoglobulins, to pass over the intestinal barrier. Therefore, we hypothesized that ketone bodies could regulate intestinal barrier function, e.g., via regulation of tight junction proteins. The primary aim was to compare the effects of HFD that can induce intestinal ketogenesis to an equicaloric carbohydrate diet on inflammatory responses, nutrition sensing, and intestinal permeability in human jejunal mucosa. Fifteen healthy volunteers receiving a 2-week HFD diet compared to a high-carbohydrate diet were compared. Blood samples and mixed meal tests were performed at the end of each dietary period to examine inflammation markers and postprandial endotoxemia. Jejunal biopsies were assessed for protein expression using Western blotting, immunohistochemistry, and morphometric characteristics of tight junctions by electron microscopy. Functional analyses of permeability and ketogenesis were performed in Caco-2 cells, mice, and human enteroids. Ussing chambers were used to analyze permeability. CRP and ALP values were within normal ranges and postprandial endotoxemia levels were low and did not differ between the two diets. The PPARα receptor was ketone body-dependently reduced after HFD. None of the tight junction proteins studied, nor the basal electrical parameters, were different between the two diets. However, the ketone body inhibitor hymeglusin increased resistance in mucosal biopsies. In addition, the tight junction protein claudin-3 was increased by ketone inhibition in human enteroids. The ketone body β-Hydroxybutyrate (βHB) did not, however, change the mucosal transition of the large-size molecular FD4-probe or LPS in Caco-2 and mouse experiments. We found that PPARα expression was inhibited by the ketone body βHB. As PPARα regulates HMGCS expression, the ketone bodies thus exert negative feedback signaling on their own production. Furthermore, ketone bodies were involved in the regulation of permeability on intestinal mucosal cells in vitro and ex vivo. We were not, however, able to reproduce these effects on intestinal permeability in vivo in humans when comparing two weeks of high-fat with high-carbohydrate diet in healthy volunteers. Further, neither the expression of inflammation markers nor the aggregate tight junction proteins were changed. Thus, it seems that not only HFD but also other factors are needed to permit increased intestinal permeability in vivo. This indicates that the healthy gut can adapt to extremes of macro-nutrients and increased levels of intestinally produced ketone bodies, at least during a shorter dietary challenge. Full article
(This article belongs to the Special Issue The Role of Tight Junction Proteins in Health and Disease)
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18 pages, 1992 KiB  
Article
Effect of Dietary Eicosapentaenoic and Docosahexaenoic Fatty Acid Supplementation during the Last Month of Gestation on Fatty Acid Metabolism and Oxidative Status in Charolais Cows and Calves
by Diana Brozić, Kristina Starčević, Marina Vranić, Krešimir Bošnjak, Maja Maurić Maljković and Tomislav Mašek
Animals 2024, 14(9), 1273; https://doi.org/10.3390/ani14091273 - 24 Apr 2024
Cited by 2 | Viewed by 1700
Abstract
Fatty acids (FAs) are of utmost importance in the peripartal period for the development of the central nervous and immune systems of the newborn. The transport of polyunsaturated fatty acids (PUFAs) through the placenta is considered to be minimal in ruminants. Nevertheless, the [...] Read more.
Fatty acids (FAs) are of utmost importance in the peripartal period for the development of the central nervous and immune systems of the newborn. The transport of polyunsaturated fatty acids (PUFAs) through the placenta is considered to be minimal in ruminants. Nevertheless, the cow’s FAs are the main source of FAs for the calf during gestation. This research aimed to investigate the influence of low-dose eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) supplementation during late gestation on the FA metabolism of cows and their calves. A total of 20 Charolais cows during the last month of their gestation were included in the feeding trial and were divided into a control group (CON) and an experimental group (EPA + DHA). The latter received a supplement in the amount of 100 g/day (9.1 and 7.8 g/cow/day of EPA and DHA, respectively). Supplementation of low-dose EPA and DHA alters colostrum and milk fatty acid composition through the elevation of n-3 long-chain polyunsaturated fatty acids (LC-PUFAs) without affecting milk fat and protein concentrations and oxidative status. Plasma composition in cows was significantly altered, while the same effect was not detected in calf plasma. No significant change in mRNA expression was detected for the genes fatty acid synthase (FASN) and acetyl-CoA carboxylase alpha (ACACA). Full article
(This article belongs to the Special Issue Advances in the Nutrition and Management of Transition Dairy Cows)
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24 pages, 8214 KiB  
Article
Differences in Fatty Acid and Central Carbon Metabolite Distribution among Different Tissues of Alfalfa–Rhizobia Symbiotic System
by Bao-Fu Lu, Wen-Juan Kang, Shang-Li Shi, Jian Guan, Fang Jing and Bei Wu
Agronomy 2024, 14(3), 511; https://doi.org/10.3390/agronomy14030511 - 1 Mar 2024
Cited by 2 | Viewed by 1753
Abstract
Fatty acid and central carbon metabolism are crucial energy metabolism reactions. However, to date, few studies have examined their distribution characteristics within the alfalfa–rhizobia symbiotic system. To clarify the distributional differences and accumulation rates of fatty acids and central carbon with this system, [...] Read more.
Fatty acid and central carbon metabolism are crucial energy metabolism reactions. However, to date, few studies have examined their distribution characteristics within the alfalfa–rhizobia symbiotic system. To clarify the distributional differences and accumulation rates of fatty acids and central carbon with this system, we measured the plant phenotype, nodule formation, nitrogen fixation capacity, and key nitrogen metabolism enzyme activities of Medicago sativa ‘Gannong No. 9’ 35 days post-inoculation (dpi) with Sinorhizobia meliloti LL11. Additionally, we employed targeted metabolomics to analyze central carbon and fatty acid metabolites in various tissue samples of symbiotic and control (C.K.) plants, as well as in S. meliloti LL11. We found that plant height; root length; aboveground fresh and dry weights; underground fresh and dry weights; and nitrate reductase, nitrogen reductase, glutamine synthetase, and glutamate synthase activities were significantly higher in the leaves and roots of symbiotic plants than in those of C.K. plants. Compared to symbiotic plants, C.K. plants exhibited higher total central carbon and fatty acid metabolite content, accounting for 38.61% and 48.17% of C.K. plants, respectively. We detected 32 central carbon and 40 fatty acid metabolites in S. meliloti LL11, with succinate (343,180.8603 ng·mL−1) and hexadecanoic acid (4889.7783 ng·mL−1) being the most. In both symbiotic and C.K. plants, central carbon metabolite was considerably higher than the fatty acid metabolite central. Moreover, the carbon metabolites found in symbiotic plants were primarily distributed in pink nodule roots (PNRs), with malate exhibiting the highest content (4,800,612.3450 ng·g−1), accounting for 53.09% of total central carbon metabolite content. Fatty acid metabolites were mainly found in pink root nodules (P.N.s), which are sites of nitrogen fixation. Trans-10-nonadecenoic acid and hexadecanoic acid exhibited the highest contents, comprising >15% of the total fatty acid metabolite content. We found that petroselaidic acid is only present in P.N., which seems to be closely related to the nitrogen fixation reaction in P.N. In general, symbiotic plants transfer central carbon metabolites to nodules via PNRs to drive nitrogen fixation. However, in P.N.s, these metabolites are limited, leading to accumulation in PNRs. Fatty acid metabolites, crucial for nitrogen fixation, are prevalent in P.N.s. Conversely, C.K. plants without nitrogen fixation distribute these metabolites primarily to the stems, emphasizing growth. This study provides new insights into the energy metabolism of symbiotic nitrogen fixation. Full article
(This article belongs to the Section Grassland and Pasture Science)
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16 pages, 5497 KiB  
Article
The Complement Component 4 Binding Protein α Gene: A Versatile Immune Gene That Influences Lipid Metabolism in Bovine Mammary Epithelial Cell Lines
by Xuanxu Chen, Zhihui Zhao, Xinyi Jiang, Jing Li, Fengshuai Miao, Haibin Yu, Ziwei Lin and Ping Jiang
Int. J. Mol. Sci. 2024, 25(4), 2375; https://doi.org/10.3390/ijms25042375 - 17 Feb 2024
Cited by 4 | Viewed by 2157
Abstract
Complement component 4 binding protein α (C4BPA) is an immune gene which is responsible for the complement regulation function of C4BP by binding and inactivating the Complement component C4b (C4b) component of the classical Complement 3 (C3) [...] Read more.
Complement component 4 binding protein α (C4BPA) is an immune gene which is responsible for the complement regulation function of C4BP by binding and inactivating the Complement component C4b (C4b) component of the classical Complement 3 (C3) invertase pathway. Our previous findings revealed that C4BPA was differentially expressed by comparing the transcriptome in high-fat and low-fat bovine mammary epithelial cell lines (BMECs) from Chinese Holstein dairy cows. In this study, a C4BPA gene knockout BMECs line model was constructed via using a CRISPR/Cas9 system to investigate the function of C4BPA in lipid metabolism. The results showed that levels of triglyceride (TG) were increased, while levels of cholesterol (CHOL) and free fatty acid (FFA) were decreased (p < 0.05) after knocking out C4BPA in BMECs. Additionally, most kinds of fatty acids were found to be mainly enriched in the pathway of the biosynthesis of unsaturated fatty acids, linoleic acid metabolism, fatty acid biosynthesis, and regulation of lipolysis in adipocyte. Meanwhile, the RNA-seq showed that most of the differentially expressed genes (DEGs) are related to PI3K-Akt signaling pathway. The expressions of 3-Hydroxy-3-Methylglutaryl-CoA Synthase 1 (HMGCS1), Carnitine Palmitoyltransferase 1A (CPT1A), Fatty Acid Desaturase 1 (FADS1), and Stearoyl-Coenzyme A desaturase 1 (SCD1) significantly changed when the C4BPA gene was knocked out. Collectively, C4BPA gene, which is an immune gene, played an important role in lipid metabolism in BMECs. These findings provide a new avenue for animal breeders: this gene, with multiple functions, should be reasonably utilized. Full article
(This article belongs to the Section Molecular Endocrinology and Metabolism)
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14 pages, 2275 KiB  
Article
Impact of Difluoromethylornithine and AMXT 1501 on Gene Expression and Capsule Regulation in Streptococcus pneumoniae
by Moses B. Ayoola, Leslie A. Shack, Otto Phanstiel and Bindu Nanduri
Biomolecules 2024, 14(2), 178; https://doi.org/10.3390/biom14020178 - 2 Feb 2024
Cited by 2 | Viewed by 2137
Abstract
Streptococcus pneumoniae (Spn), a Gram-positive bacterium, poses a significant threat to human health, causing mild respiratory infections to severe invasive conditions. Despite the availability of vaccines, challenges persist due to serotype replacement and antibiotic resistance, emphasizing the need for alternative therapeutic strategies. This [...] Read more.
Streptococcus pneumoniae (Spn), a Gram-positive bacterium, poses a significant threat to human health, causing mild respiratory infections to severe invasive conditions. Despite the availability of vaccines, challenges persist due to serotype replacement and antibiotic resistance, emphasizing the need for alternative therapeutic strategies. This study explores the intriguing role of polyamines, ubiquitous, small organic cations, in modulating virulence factors, especially the capsule, a crucial determinant of Spn’s pathogenicity. Using chemical inhibitors, difluoromethylornithine (DFMO) and AMXT 1501, this research unveils distinct regulatory effects on the gene expression of the Spn D39 serotype in response to altered polyamine homeostasis. DFMO inhibits polyamine biosynthesis, disrupting pathways associated with glucose import and the interconversion of sugars. In contrast, AMXT 1501, targeting polyamine transport, enhances the expression of polyamine and glucose biosynthesis genes, presenting a novel avenue for regulating the capsule independent of glucose availability. Despite ample glucose availability, AMXT 1501 treatment downregulates the glycolytic pathway, fatty acid synthesis, and ATP synthase, crucial for energy production, while upregulating two-component systems responsible for stress management. This suggests a potential shutdown of energy production and capsule biosynthesis, redirecting resources towards stress management. Following DFMO and AMXT 1501 treatments, countermeasures, such as upregulation of stress response genes and ribosomal protein, were observed but appear to be insufficient to overcome the deleterious effects on capsule production. This study highlights the complexity of polyamine-mediated regulation in S. pneumoniae, particularly capsule biosynthesis. Our findings offer valuable insights into potential therapeutic targets for modulating capsules in a polyamine-dependent manner, a promising avenue for intervention against S. pneumoniae infections. Full article
(This article belongs to the Special Issue Polyamine Metabolism and Function)
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25 pages, 2489 KiB  
Review
Intestinal Inflammation and Regeneration–Interdigitating Processes Controlled by Dietary Lipids in Inflammatory Bowel Disease
by Soon Jae Kwon, Muhammad Sohaib Khan and Sang Geon Kim
Int. J. Mol. Sci. 2024, 25(2), 1311; https://doi.org/10.3390/ijms25021311 - 21 Jan 2024
Cited by 7 | Viewed by 5126
Abstract
Inflammatory bowel disease (IBD), including Crohn’s disease and ulcerative colitis, is a disease of chronic inflammatory conditions of the intestinal tract due to disturbance of the inflammation and immune system. Symptoms of IBD include abdominal pain, diarrhea, bleeding, reduced weight, and fatigue. In [...] Read more.
Inflammatory bowel disease (IBD), including Crohn’s disease and ulcerative colitis, is a disease of chronic inflammatory conditions of the intestinal tract due to disturbance of the inflammation and immune system. Symptoms of IBD include abdominal pain, diarrhea, bleeding, reduced weight, and fatigue. In IBD, the immune system attacks the intestinal tract’s inner wall, causing chronic inflammation and tissue damage. In particular, interlukin-6 and interlukin-17 act on immune cells, including T cells and macrophages, to amplify the immune responses so that tissue damage and morphological changes occur. Of note, excessive calorie intake and obesity also affect the immune system due to inflammation caused by lipotoxicity and changes in lipids supply. Similarly, individuals with IBD have alterations in liver function after sustained high-fat diet feeding. In addition, excess dietary fat intake, along with alterations in primary and secondary bile acids in the colon, can affect the onset and progression of IBD because inflammatory cytokines contribute to insulin resistance; the factors include the release of inflammatory cytokines, oxidative stress, and changes in intestinal microflora, which may also contribute to disease progression. However, interfering with de novo fatty acid synthase by deleting the enzyme acetyl-CoA-carboxylase 1 in intestinal epithelial cells (IEC) leads to the deficiency of epithelial crypt structures and tissue regeneration, which seems to be due to Lgr5+ intestinal stem cell function. Thus, conflicting reports exist regarding high-fat diet effects on IBD animal models. This review will focus on the pathological basis of the link between dietary lipids intake and IBD and will cover the currently available pharmacological approaches. Full article
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