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17 pages, 2001 KiB  
Article
Farnesol Improves Endoplasmic Reticulum Stress and Hepatic Metabolic Dysfunction Induced by Tunicamycin in Mice
by Naqash Goswami, Lionel Kinkpe, Lun Hua, Yong Zhuo, Zhengfeng Fang, Lianqiang Che, Yan Lin, Shengyu Xu, Xuemei Jiang, Bin Feng and De Wu
Biology 2025, 14(2), 213; https://doi.org/10.3390/biology14020213 - 18 Feb 2025
Viewed by 342
Abstract
Endoplasmic reticulum (ER) stress significantly affects liver metabolism, often leading to disorders such as hepatic steatosis. Tunicamycin (TM), a known ER stress inducer, is frequently used to model metabolic stress, but its specific effects on liver energy homeostasis remain unclear. This study investigates [...] Read more.
Endoplasmic reticulum (ER) stress significantly affects liver metabolism, often leading to disorders such as hepatic steatosis. Tunicamycin (TM), a known ER stress inducer, is frequently used to model metabolic stress, but its specific effects on liver energy homeostasis remain unclear. This study investigates how farnesol (FOH), a natural compound with antioxidant and anti-inflammatory properties, counteracts TM-induced ER stress and its associated metabolic disruptions in the liver. Using both primary hepatocytes and a mouse model, this study demonstrates that TM treatment caused upregulation of ER stress markers, including ATF4, and disrupted genes related to lipid metabolism and gluconeogenesis. Co-treatment with FOH reduced these stress markers and restored the expression of metabolic genes. In vivo, FOH treatment alleviated oxidative stress, reduced lipid accumulation, and restored normal glycogen and lipid metabolism. Histological analysis further confirmed that FOH preserved liver architecture and minimized cellular damage. FOH also stabilized serum lipid profiles and modulated key metabolic biomarkers, suggesting its protective role against TM-induced liver injury. These findings suggest that FOH has therapeutic potential in mitigating ER stress-related metabolic dysfunctions, offering promising insights for the treatment of liver diseases linked to metabolic stress. Full article
(This article belongs to the Special Issue Molecular Basis of Metabolic Homeostasis)
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Figure 1
<p>Ameliorative action of farnesol against tunicamycin-induced changes in relative expression of the endoplasmic reticulum stress-related genes Chop, Grp78, and Atf4 in primary hepatocytes. Values represent the mean ± SEM from three independent biological replicates. Statistical significance was determined by one-way ANOVA followed by Tukey’s multiple comparison test. Veh: Control (vehicle); TM: Tunicamycin; FOH+TM: Farnesol plus tunicamycin. * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, vs. Veh, # <span class="html-italic">p</span> &lt; 0.05 vs. TM.</p>
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<p>Ameliorative effect of farnesol on tunicamycin-induced changes in the relative expression of hepatic energy homeostasis-related genes. (<b>A</b>) Gene expression of Srebp1c; (<b>B</b>) Gene expression of Cpt1a; (<b>C</b>) Gene expression of Pepck; (<b>D</b>) Gene expression of G6pase; (<b>E</b>) Gene expression of Scd1; (<b>F</b>) Gene expression of Fas in primary hepatocytes. Values represent the mean ± SEM from three independent biological replicates. Veh: control (vehicle); TM: tunicamycin; FOH+TM: farnesol plus tunicamycin. * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001, **** <span class="html-italic">p</span> &lt; 0.0001 vs. Veh; # <span class="html-italic">p</span> &lt; 0.05, ## <span class="html-italic">p</span> &lt; 0.01, ### <span class="html-italic">p</span> &lt; 0.001 vs. TM.</p>
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<p>Ameliorative action of farnesol against tunicamycin-induced alterations in the relative expression of endoplasmic reticulum stress (ERS)-related genes. (<b>A</b>) Gene expression of Chop; (<b>B</b>) Gene expression of Grp78; (<b>C</b>) Gene expression of Atf4 in the liver tissue of mice. Values represent the mean ± SEM from eight biological replicates per group. Veh: control (vehicle); TM: tunicamycin; FOH+TM: farnesol plus tunicamycin. *<span class="html-italic">p</span> &lt; 0.05, *** <span class="html-italic">p</span> &lt; 0.001 vs. Veh; **** <span class="html-italic">p</span> &lt; 0.0001 vs. Veh; ### <span class="html-italic">p</span> &lt; 0.001 vs. TM.</p>
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<p>Ameliorative effect of farnesol on tunicamycin-induced alterations in the relative expression of metabolic genes. (<b>A</b>) Gene expression of Srebp1c; (<b>B</b>) Gene expression of Cpt1a; (<b>C</b>) Gene expression of Pepck; (<b>D</b>) Gene expression of G6pase; (<b>E</b>) Gene expression of Scd1; (<b>F</b>) Gene expression of Fas in liver tissue. Values represent the mean ± SEM from eight biological replicates per group. Veh: Control (vehicle); TM: Tunicamycin; FOH+TM: Farnesol plus tunicamycin. * <span class="html-italic">p</span> &lt; 0.05, *** <span class="html-italic">p</span> &lt; 0.001, **** <span class="html-italic">p</span> &lt; 0.0001 vs. Veh; # <span class="html-italic">p</span> &lt; 0.05, ## <span class="html-italic">p</span> &lt; 0.01, ### <span class="html-italic">p</span> &lt; 0.001 vs. TM.</p>
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<p>Protective effect of farnesol against tunicamycin-induced oxidative stress in the liver. (<b>A</b>) MDA levels; (<b>B</b>) SOD activity; (<b>C</b>) GSH levels. Values represent the mean ± SEM from eight biological replicates per group. Statistical significance was determined using one-way ANOVA followed by Tukey’s multiple comparison test. Veh: Control (vehicle); TM: Tunicamycin; FOH+TM: Farnesol plus tunicamycin. * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, **** <span class="html-italic">p</span> &lt; 0.0001 vs. Veh; # <span class="html-italic">p</span> &lt; 0.05, ## <span class="html-italic">p</span> &lt; 0.01, ### <span class="html-italic">p</span> &lt; 0.001 vs. TM.</p>
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<p>Histopathological and ultrastructural changes demonstrated by liver tissue of mice in hematoxylin and eosin staining after exposure to tunicamycin. (<b>A</b>) Control (Veh) group histological examination of H&amp;E-stained. (<b>B</b>) Tunicamycin (TM) group histological examination of H&amp;E-stained. (<b>C</b>) Farnesol plus tunicamycin (FOH+TM) group histological examination of H&amp;E-stained. Note: The farnesol-treated group significantly improves the health of liver tissue of mice when compared with the tunicamycin-treated and control groups of experimental mice.</p>
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<p>The ameliorative action of farnesol against tunicamycin-induced alterations. (<b>A</b>) Liver glycogen levels; (<b>B</b>) Blood glucose levels; (<b>C</b>) Non-esterified fatty acid levels; (<b>D</b>) Liver TAG levels; (<b>E</b>) Serum TAG levels. Values represent the mean ± SEM from eight independent biological replicates. Control (Veh), tunicamycin (TM), and farnesol plus tunicamycin (FOH+TM) treatment groups. * <span class="html-italic">p</span> &lt; 0.05, *** <span class="html-italic">p</span> &lt; 0.001, **** <span class="html-italic">p</span> &lt; 0.0001 vs. Veh; # <span class="html-italic">p</span> &lt; 0.05, ## <span class="html-italic">p</span> &lt; 0.01, #### <span class="html-italic">p</span> &lt; 0.0001 vs. TM.</p>
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<p>Ameliorative action of farnesol against tunicamycin-induced alterations in blood serum markers. (<b>A</b>) Serum ALT levels; (<b>B</b>) Serum AST levels; (<b>C</b>) ALT/AST ratio; (<b>D</b>) NEFA levels; (<b>E</b>) Serum TC levels; (<b>F</b>) Serum HDL-C levels; (<b>G</b>) Serum LDL-C levels. Values represent the mean ± SEM from eight independent biological replicates. Control (Veh), tunicamycin (TM), and farnesol plus tunicamycin (FOH+TM) treatment groups. * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001 vs. Veh.</p>
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<p>Ameliorative action of farnesol against tunicamycin-induced alterations in liver weight parameters. (<b>A</b>) Relative weight of liver; (<b>B</b>) Liver weight/BW ratio in experimental mice. Values represent the mean ± SEM from eight independent biological replicates. Control (Veh), tunicamycin (TM), and farnesol plus tunicamycin (FOH+TM) treatment groups. *** <span class="html-italic">p</span> &lt; 0.001, **** <span class="html-italic">p</span> &lt; 0.0001 vs. Veh.</p>
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12 pages, 3194 KiB  
Article
Preventing Oral Dual Biofilm Development with Innovative Bioactive Varnishes
by Tainá de Lima Costa, Regina Maria Puppin-Rontani and Aline Rogéria Freire de Castilho
J. Funct. Biomater. 2025, 16(2), 70; https://doi.org/10.3390/jfb16020070 - 18 Feb 2025
Viewed by 379
Abstract
This study introduces innovative varnishes incorporating natural bioactive compounds to inhibit the formation of oral dual biofilms, a critical contributor to dental caries and other oral diseases. The purpose of this study was to evaluate the effectiveness of bioactive varnishes containing tt-farnesol, [...] Read more.
This study introduces innovative varnishes incorporating natural bioactive compounds to inhibit the formation of oral dual biofilms, a critical contributor to dental caries and other oral diseases. The purpose of this study was to evaluate the effectiveness of bioactive varnishes containing tt-farnesol, quercetin, and theobromine in inhibiting the formation of mixed Streptococcus mutans and Candida albicans biofilms. Mixed biofilms of Streptococcus mutans UA159 and Candida albicans SC5314 were grown in 96-well plates containing a specialized culture medium. Approximately 0.2 mL of experimental varnishes with A—1.5% or B—4.5% concentrations of tt-farnesol, quercetin, and theobromine were separately added to the wells using a disposable applicator, with a vehicle varnish (lacking bioactives) serving as the control. Biofilms were incubated at 37 °C with 5% CO2 for 24 h. Microbial viability was determined in terms of colony-forming units per milliliter (CFU/mL), and biofilm morphology was evaluated qualitatively via scanning electron microscopy (SEM). Statistical analyses were performed using ANOVA/Tukey tests at a 5% significance level. Varnishes A and B achieved significant reductions in microbial populations within the biofilms (p < 0.05) compared to the vehicle control (C). SEM imaging revealed marked structural disruptions in the biofilms, validating the quantitative results. Higher bioactive concentrations demonstrated enhanced inhibitory effects. Bioactive varnishes enriched with theobromine, quercetin, and tt-farnesol represent a novel and effective strategy for inhibiting oral dual biofilm development, offering a promising advancement in preventive dentistry. Full article
(This article belongs to the Special Issue Role of Dental Biomaterials in Promoting Oral Health)
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<p>Schematic diagram of the assays performed in this study.</p>
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<p>Box plot (mean and standard deviation) of the different varnish formulations (A, B, and C—control) for <span class="html-italic">S. mutans</span> (SM) and <span class="html-italic">C. albicans</span> (CA) biofilms (CFU/mL) (<span class="html-italic">n</span> = 6).</p>
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<p>Scanning electron microscopy of <span class="html-italic">S. mutans</span> and <span class="html-italic">C. albicans</span> biofilms. Images show the typical architecture of a dual biofilm (SM + CA), with fewer cells after treatment with different concentrations (<b>A</b>,<b>B</b>) and no biofilm reduction ((<b>C</b>)-vehicle) at 1000× magnification. Reprinted from Ref. [<a href="#B31-jfb-16-00070" class="html-bibr">31</a>].</p>
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42 pages, 23331 KiB  
Article
Exploratory Analysis of Free and Glycosidically Bound Volatile Compounds in Australian-Grown and Imported Dragon Fruit: Implications for Industry Standard Development
by Si-Yuan Chen, Cheng-Yuan Xu, Muhammad Sohail Mazhar and Mani Naiker
Chemosensors 2025, 13(2), 70; https://doi.org/10.3390/chemosensors13020070 - 15 Feb 2025
Viewed by 410
Abstract
Dragon fruit (Hylocereus spp.), a globally popular tropical fruit, is highly regarded for its unique sensory attributes and potential health benefits. However, the volatile organic compound (VOC) profiles that define its aroma and flavor are underexplored. This exploratory study investigates the VOC [...] Read more.
Dragon fruit (Hylocereus spp.), a globally popular tropical fruit, is highly regarded for its unique sensory attributes and potential health benefits. However, the volatile organic compound (VOC) profiles that define its aroma and flavor are underexplored. This exploratory study investigates the VOC profiles of Australian-grown and imported dragon fruit, focusing on free volatiles and glycosidically bound volatiles released through acid and enzymatic hydrolysis. The analysis aims to uncover the competitive advantages of Australian-grown dragon fruit, providing a scientific foundation for establishing industry standards in Australia, where such standards are currently absent. Using gas chromatography–mass spectrometry (GC-MS) and statistical analysis such as principal component analysis (PCA), this study revealed significant differences in VOC profiles influenced by variety and growing region. Northern Territory-grown white-fleshed dragon fruit (NTW) displayed the most diverse and unique volatile profile, with key unique contributors such as acetoin, phenylethyl alcohol, and prenol, highlighting its potential as a premium product. Despite regional similarities, Queensland-grown white- (QLDW) and red-fleshed (QLDR) dragon fruit exhibited distinct profiles, with compounds such as farnesol and linoleic acid ethyl ester serving as distinguishing markers. Overseas white- (OverseasW) and red-fleshed (OverseasR) samples had less complex profiles, likely related to earlier harvesting and postharvest treatments, emphasizing the impact of such practices on volatile complexity. Glycosidically bound volatiles were identified as latent precursors that enhance aroma during ripening and processing. This research underscores the importance of VOC profiling in potentially assisting with establishing industry standards for Australian dragon fruit, enabling the differentiation of domestic varieties from imports and enhancing market competitiveness. As this is a novel and exploratory study, future research should prioritize the identification of unknown compounds and refine methodologies to better understand the dynamic changes in VOCs during storage and ripening. These findings provide valuable insights for optimizing postharvest practices and developing standards that support the Australian dragon fruit industry’s growth and global positioning. Full article
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<p>Hydrolysis pathways of glycosidically bound volatiles [<a href="#B4-chemosensors-13-00070" class="html-bibr">4</a>]. (<b>A</b>) Terpene polyol glycosides release terpene polyols and, subsequently, aromatic compounds such as hotrienol and neroloxide through enzymatic and chemical hydrolysis. (<b>B</b>) Norisoprenoid glycosides yield β-damascenone precursors via enzymatic hydrolysis, followed by acid-catalyzed rearrangements to produce β-damascenone.</p>
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<p>Total ion chromatograms of free volatile compounds extracted from dragon fruit samples. Peak number corresponds to <a href="#chemosensors-13-00070-t002" class="html-table">Table 2</a>. (<b>a</b>) QLDW; (<b>b</b>) QLDR; (<b>c</b>) NTW; (<b>d</b>) OverseasW; (<b>e</b>) OverseasR. Note: X axis—retention time (mins); Y axis:—peak intensity counts.</p>
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<p>Total ion chromatograms of free volatile compounds extracted from dragon fruit samples. Peak number corresponds to <a href="#chemosensors-13-00070-t002" class="html-table">Table 2</a>. (<b>a</b>) QLDW; (<b>b</b>) QLDR; (<b>c</b>) NTW; (<b>d</b>) OverseasW; (<b>e</b>) OverseasR. Note: X axis—retention time (mins); Y axis:—peak intensity counts.</p>
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<p>Total ion chromatograms of volatile compounds released by acid hydrolysis from dragon fruit samples. Peak number corresponds to <a href="#chemosensors-13-00070-t003" class="html-table">Table 3</a>. (<b>a</b>) QLDW; (<b>b</b>) QLDR; (<b>c</b>) NTW; (<b>d</b>) OverseasW; (<b>e</b>) OverseasR. Note: X axis—retention time (mins); Y axis—peak intensity counts.</p>
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<p>Total ion chromatograms of volatile compounds released by acid hydrolysis from dragon fruit samples. Peak number corresponds to <a href="#chemosensors-13-00070-t003" class="html-table">Table 3</a>. (<b>a</b>) QLDW; (<b>b</b>) QLDR; (<b>c</b>) NTW; (<b>d</b>) OverseasW; (<b>e</b>) OverseasR. Note: X axis—retention time (mins); Y axis—peak intensity counts.</p>
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<p>Total ion chromatograms of volatile compounds from dragon fruit samples released by enzymatic hydrolysis. Peak number corresponds to <a href="#chemosensors-13-00070-t004" class="html-table">Table 4</a>. (<b>a</b>) QLDW; (<b>b</b>) QLDR; (<b>c</b>) NTW; (<b>d</b>) OverseasW; (<b>e</b>) OverseasR. Note: X axis—retention time (mins); Y axis—peak intensity counts.</p>
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<p>Total ion chromatograms of volatile compounds from dragon fruit samples released by enzymatic hydrolysis. Peak number corresponds to <a href="#chemosensors-13-00070-t004" class="html-table">Table 4</a>. (<b>a</b>) QLDW; (<b>b</b>) QLDR; (<b>c</b>) NTW; (<b>d</b>) OverseasW; (<b>e</b>) OverseasR. Note: X axis—retention time (mins); Y axis—peak intensity counts.</p>
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<p>Venn diagrams of volatile compounds present in different dragon fruit types (QLDW, QLDR, NTW, OverseasW, and OverseasR). (<b>A</b>) shows free volatile compounds present in dragon fruits. (<b>B</b>) shows volatile compounds released by acid hydrolysis present in dragon fruit. (<b>C</b>) shows volatile compounds released by enzymatic hydrolysis present in dragon fruit.</p>
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<p>PCA biplot of volatile compounds and dragon fruit samples (QLDW, QLDR, NTW, OverseasW, and OverseasR). Volatile compounds are labeled as X1, X2, …, Xn, corresponding to their respective compounds listed in <a href="#chemosensors-13-00070-t0A1" class="html-table">Table A1</a>. High-contribution compounds are positioned farther from the origin and aligned with specific principal components. The arrows represent individual volatile compounds, with their length indicating contribution and direction reflecting alignment with Dim1 (39.2% variance) and Dim2 (30.3% variance). Sample differentiation reflects unique volatile profiles, with NTW showing clear separation and Overseas samples clustering near the origin.</p>
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<p>TIC of blank for free volatile compound analysis. Note: X axis—retention time (mins); Y axis—peak intensity counts.</p>
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<p>TIC of blank for analysis of volatile compounds released by acid hydrolysis. Peak i: 2,4-Decadienal; ii: Hexanoic acid; iii: 2-Heptaflurobutyroxypentadecane; iv: 2-Nonen-1-ol; v: Triethylene glycol monododecyl ether; vi: Octaethylene glycol monododecyl ether. Note: X axis—retention time (mins); Y axis—peak intensity counts.</p>
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<p>TIC of blank for analysis of volatile compounds released by enzymatic hydrolysis. Peak I: Acetic acid; II: Cerasynt; III: Octaethylene glycol monododecyl ether. Note: X axis—retention time (mins); Y axis—peak intensity counts.</p>
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22 pages, 3962 KiB  
Review
Compounds Involved in the Invasive Characteristics of Lantana camara
by Hisashi Kato-Noguchi and Midori Kato
Molecules 2025, 30(2), 411; https://doi.org/10.3390/molecules30020411 - 19 Jan 2025
Viewed by 708
Abstract
Lantana camara L. is native to tropical America and has naturalized in many other tropical, subtropical, and temperate regions in Asia, Africa, Oceania, North and South America, and Europe. L. camara infests diverse habitats with a wide range of climatic factors, and its [...] Read more.
Lantana camara L. is native to tropical America and has naturalized in many other tropical, subtropical, and temperate regions in Asia, Africa, Oceania, North and South America, and Europe. L. camara infests diverse habitats with a wide range of climatic factors, and its population increases aggressively as one of the world’s 100 worst invasive alien species. Its infestation reduces species diversity and abundance in the natural ecosystems and reduces agricultural production. The life history characteristics of L. camara, such as its high reproductive ability and high adaptive ability to various environmental conditions, may contribute to its ability to infest and increase its population. Possible evidence of the compounds involved in the defense functions of L. camara against natural enemies, such as herbivore mammals and insects, parasitic nematodes, pathogenic fungi and bacteria, and the allelochemicals involved in its allelopathy against neighboring competitive plant species, have accumulated in the literature over three decades. Lantadenes A and B, oleanonic acid, and icterogenin are highly toxic to herbivore mammals, and β-humulene, isoledene, α-copaene thymol, and hexadecanoic acid have high insecticidal activity. β-Caryophyllene and cis-3-hexen-1-ol may function as herbivore-induced plant volatiles which are involved in sending warning signals to undamaged tissues and the next plants of the same species. Farnesol and farnesal may interrupt insect juvenile hormone biosynthesis and cause abnormal metamorphosis of insects. Several triterpenes, such as lantanolic acid, lantoic acid, pomolic acid, camarin, lantacin, camarinin, ursolic acid, and oleanonic acid, have demonstrated nematocidal activity. Lantadene A, β-caryophyllene, germacrene-D, β-curcumene, eicosapentaenoic acid, and loliolide may possess antimicrobial activity. Allelochemicals, such as caffeic acid, ferulic acid, salicylic acid, α-resorcylic acid, p-hydroxybenzoic acid, vanillic acid, unbelliferone, and quercetin, including lantadenes A and B and β-caryophyllene, suppress the germination and growth of neighboring plant species. These compounds may be involved in the defense functions and allelopathy and may contribute to L. camara’s ability to infest and to expand its population as an invasive plant species in new habitats. This is the first review to focus on how compounds enhance the invasive characteristics of L. camara. Full article
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Graphical abstract

Graphical abstract
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<p>Stand and leaves of <span class="html-italic">L. camara</span>.</p>
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<p>Flowers and fruits of <span class="html-italic">L. camara</span>.</p>
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<p>The compounds involved in the defense function against herbivore mammals.</p>
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<p>The compounds involved in the defense function against herbivorous insects.</p>
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<p>The compounds involved in the defense function against parasitic nematodes.</p>
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<p>The compounds involved in the defense function against pathogenic fungi and bacteria.</p>
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<p>The compounds involved in the allelopathy.</p>
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<p>Action mechanisms of the compounds discussed in this paper. These compounds are involved in the hepathoxic, insecticidal, nematocidal, fungicidal, and allelopathic activity of <span class="html-italic">L. camara</span>. Purple arrow: direct action; blue arrow: secondary and tertiary action.</p>
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10 pages, 646 KiB  
Article
Influence of Quercetin and tt-Farnesol Enrichment on Physicochemical Properties of a Universal Adhesive System
by Roberta Janaína Soares Mendes, Regina Maria Puppin-Rontani and Aline Rogéria Freire de Castilho
Adhesives 2025, 1(1), 3; https://doi.org/10.3390/adhesives1010003 - 6 Jan 2025
Viewed by 521
Abstract
This study investigated the impact of natural compound enrichment, specifically quercetin and trans, trans-farnesol (tt-farnesol), on the physicochemical properties of a universal adhesive system. A preliminary DPPH assay was conducted to determine the optimal concentrations of quercetin (0.24 mg/mL) and tt [...] Read more.
This study investigated the impact of natural compound enrichment, specifically quercetin and trans, trans-farnesol (tt-farnesol), on the physicochemical properties of a universal adhesive system. A preliminary DPPH assay was conducted to determine the optimal concentrations of quercetin (0.24 mg/mL) and tt-farnesol (1.43 mg/mL) based on their radical scavenging abilities. These compounds were then incorporated into the adhesive system. Specimens (n = 5; 7 mm × 1 mm) of the adhesive system, both with and without the added compounds, were prepared and tested for water sorption, solubility, Knoop hardness, and softening percentage. Water sorption and solubility were measured after immersion in deionized water for 7 days, and Knoop hardness was measured before and after immersion in 75% ethanol. Softening percentage was calculated based on changes in hardness. Data on water sorption, solubility, and percentage of softening were submitted to the Student’s t-test (α = 5%) while Knoop hardness values were submitted to the Mann–Whitney test (α = 5%). Both quercetin and tt-farnesol exhibited important antioxidant activity (85.5% and 82%, respectively). Water sorption was similar for both groups (p > 0.05) but the experimental adhesive had a significantly higher solubility, lower hardness, and higher softening. The incorporation of quercetin and tt-farnesol into a universal adhesive system detrimentally affects its essential physicochemical properties, compromising its performance. Full article
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Figure 1
<p>Flowchart of the experimental design used in this study. (<b>A</b>)—Preliminary study: Natural compounds that were submitted to the DPPH assay; (<b>B</b>)—Physicochemical characterization: Groups and assays. Source: Authors.</p>
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<p>Hardness and softening percentage of control (CG) and experimental (EG) groups.</p>
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20 pages, 5931 KiB  
Article
Trans, Trans-Farnesol Enhances the Anti-Bacterial and Anti-Biofilm Effect of Arachidonic Acid on the Cariogenic Bacteria Streptococcus mutans and Streptococcus sobrinus
by Farah Haj-Yahya, Doron Steinberg and Ronit Vogt Sionov
Int. J. Mol. Sci. 2024, 25(21), 11770; https://doi.org/10.3390/ijms252111770 - 1 Nov 2024
Viewed by 1406
Abstract
Background: Streptococcus mutans and Streptococcus sobrinus are Gram-positive bacteria involved in the development of dental caries, as they are able to form biofilms on tooth enamel, ferment sugars into acids, and survive under acidic conditions. This ultimately leads to a local lowering of [...] Read more.
Background: Streptococcus mutans and Streptococcus sobrinus are Gram-positive bacteria involved in the development of dental caries, as they are able to form biofilms on tooth enamel, ferment sugars into acids, and survive under acidic conditions. This ultimately leads to a local lowering of the pH value on the tooth surface, which causes enamel cavities. Hypothesis: One measure to reduce caries is to limit the growth of cariogenic bacteria by using two anti-bacterial agents with different mechanisms of action. The hypothesis of this study was that the anti-bacterial activity of ω-6 polyunsaturated arachidonic acid (AA) against S. mutans and S. sobrinus can be enhanced by the sesquiterpene alcohol trans, trans-farnesol (t,t-farnesol). Methods: The anti-bacterial activity of single and combined treatment was determined by the checkerboard assay. Bacterial viability was assessed by live/dead SYTO 9/propidium iodide (PI) staining on flow cytometry. Anti-biofilm activity was determined by MTT metabolic assay, crystal violet staining of biofilm biomass, SYTO 9/PI staining by spinning disk confocal microscopy (SDCM) and high-resolution scanning electron microscopy (HR-SEM). Results: t,t-Farnesol lowered the minimum inhibitory concentration (MIC) and the minimum biofilm inhibitory concentration (MBIC) of AA at sub-MICs. AA reduced the metabolic activity of preformed mature biofilms, while t,t-farnesol had no significant effect. The enhanced anti-bacterial effect of the combined t,t-farnesol/AA treatment was further evidenced by increased PI uptake, indicating membrane perforation. The enhanced anti-biofilm effect was further verified by SDCM and HR-SEM. Gene expression studies showed reduced expression of some biofilm-related genes. Conclusions: Altogether, our study suggests a potential use of the two naturally occurring compounds arachidonic acid and t,t-farnesol for preventing biofilm formation by the cariogenic bacteria S. mutans and S. sobrinus. These findings have implications for caries prevention. Full article
(This article belongs to the Special Issue Natural Compounds: Advances in Antimicrobial Activity)
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<p>The chemical structures of arachidonic acid (AA) and trans, trans-farnesol (t,t-farnesol).</p>
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<p>Increased anti-bacterial effect of arachidonic acid and t,t-farnesol against <span class="html-italic">S. mutans</span> (<b>A</b>) and <span class="html-italic">S. sobrinus</span> (<b>B</b>) when combined together. The graphs present the percentage turbidity of planktonic growing bacteria after a 24 h incubation with different concentrations of AA and farnesol. N = 3. a: <span class="html-italic">p</span> &lt; 0.05 when compared to control bacteria. b: <span class="html-italic">p</span> &lt; 0.05 when compared to single treatments.</p>
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<p>Membrane perforation caused by arachidonic acid (AA) is aggravated when <span class="html-italic">S. mutans</span> is co-treated with t,t-farnesol. (<b>A</b>). Density dot plots of PI versus SYTO 9 fluorescence intensities of <span class="html-italic">S. mutans</span> that was treated with the indicated compounds for 2 h. The samples were analyzed by flow cytometry. The different colors represent the density of the events at a given position. (<b>B</b>). Percentage of live (SYTO 9<sup>high</sup>PI<sup>low</sup>) and dead (SYTO 9<sup>high</sup>PI<sup>high</sup>) bacteria, respectively. N = 3. a: <span class="html-italic">p</span> &lt; 0.05 when compared to control bacteria. b: <span class="html-italic">p</span> &lt; 0.05 when compared to single treatments.</p>
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<p>Increased membrane perforation when <span class="html-italic">S. sobrinus</span> is co-treated with arachidonic acid (AA) and t,t-farnesol. (<b>A</b>). Density dot plots of PI versus SYTO 9 fluorescence intensities of <span class="html-italic">S. sobrinus</span> that was treated with the indicated compounds for 2 h. The samples were analyzed by flow cytometry. The different colors represent the density of the events at a given position. (<b>B</b>). Percentages of live (SYTO 9<sup>high</sup>PI<sup>low</sup>) and dead (SYTO 9<sup>high</sup>PI<sup>high</sup>; SYTO 9<sup>low</sup>PI<sup>high</sup>; SYTO 9<sup>neg</sup>PI<sup>neg</sup>) bacteria, respectively. SYTO 9<sup>high</sup>PI<sup>high</sup>, SYTO 9<sup>low</sup>PI<sup>high</sup> and SYTO 9<sup>neg</sup>PI<sup>neg</sup> represent three different stages of bacterial cell death: SYTO 9<sup>high</sup>PI<sup>high</sup> represents bacteria with membrane perforation; SYTO 9<sup>low</sup>PI<sup>high</sup> represents bacteria with membrane perforation with initial cytoplasmic leakage; while SYTO 9<sup>neg</sup>PI<sup>neg</sup> represents dead bacteria, which have lost nucleic acids (neg = negative). N = 3. a: <span class="html-italic">p</span> &lt; 0.05 when compared to control bacteria. b: <span class="html-italic">p</span> &lt; 0.05 when compared to single treatments.</p>
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<p>Membrane hyperpolarization induced by arachidonic acid (AA) is intensified when <span class="html-italic">S. sobrinus</span> is co-treated with t,t-farnesol. (<b>A</b>,<b>B</b>). Histograms of DiOC2(3) green fluorescence (<b>A</b>) and red fluorescence (<b>B</b>) of bacteria exposed to 12.5 µg/mL t,t-Farnesol and/or 12.5 µg/mL AA for 15 min. (<b>C</b>). Summary of the relative DiOC2(3) green and red fluorescence intensities of <span class="html-italic">S. sobrinus</span> exposed to the indicated concentrations of the two compounds. RFI = Relative fluorescence intensities calculated according to the geometric mean on the De Novo FCS Express 7.12.0007 software. DiOC2(3) is a potentiometric drug whose red fluorescence is intensified upon membrane hyperpolarization, while the green fluorescence intensity is unaffected by the ΔΨ [<a href="#B50-ijms-25-11770" class="html-bibr">50</a>,<a href="#B51-ijms-25-11770" class="html-bibr">51</a>]. Thus, an increase in the red fluorescence relative to the green fluorescence is an indication of membrane hyperpolarization. N = 3. a: <span class="html-italic">p</span> &lt; 0.05 when compared to control bacteria. b: <span class="html-italic">p</span> &lt; 0.05 when compared to single treatments.</p>
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<p>Increased anti-biofilm effects against <span class="html-italic">S. mutans</span> (<b>A</b>,<b>B</b>) and <span class="html-italic">S. sobrinus</span> (<b>C</b>,<b>D</b>) when arachidonic acid (AA) is combined with t,t-farnesol. (<b>A</b>,<b>C</b>) The metabolic activity of the biofilms after a 24 h incubation with different concentrations of AA and t,t-farnesol, as measured by the MTT assay. (<b>B</b>,<b>D</b>) The biofilm biomass after a 24 h incubation with different concentrations of AA and t,t-farnesol, as measured by crystal violet (CV) staining. N = 3. a: <span class="html-italic">p</span> &lt; 0.05 when compared to control bacteria. b: <span class="html-italic">p</span> &lt; 0.05 when compared to single treatments.</p>
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<p>Spinning disk confocal microscopy (SDCM) of <span class="html-italic">S. mutans</span> biofilms formed in the absence or presence of 3.125 µg/mL AA and 12.5 µg/mL t,t-farnesol for 24 h. (<b>A</b>). The individual and merged fluorescence images of the three stains: SYTO 9 that enters both live and dead bacteria and emits green fluorescence; PI that can only enter bacteria with damaged membrane and emits red fluorescence; and AlexaFluor<sup>647</sup> Dextran 10,000 that shows the EPS, emits far-red fluorescence and is presented here in blue. The images cover an area of 1497.6 µm × 1497.6 µm. (<b>B</b>–<b>D</b>) The relative fluorescence intensities (RFI) of SYTO 9 (<b>B</b>), PI (<b>C</b>) and Dextran 10,000 (<b>D</b>) in each of the biofilm layers. The numbers on the Y-axis represent the RFI values calculated by NIS element × 10<sup>−8</sup>. (<b>E</b>) The area under the curve (AUC) of the graphs in B-D. Here the total RFI values were multiplied by 10<sup>−9</sup>. N = 3–4. a: <span class="html-italic">p</span> &lt; 0.05 when compared to control bacteria. b: <span class="html-italic">p</span> &lt; 0.05 when compared to single treatments.</p>
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<p>HR-SEM images of <span class="html-italic">S. mutans</span> biofilms formed in the absence or presence of 3.125 µg/mL AA and 12.5 µg/mL t,t-farnesol for 24 h. (<b>A</b>–<b>D</b>) ×5000 magnifications. The bar represents 20 µm. (<b>E</b>–<b>H</b>) ×20,000 magnifications. The bar represents 5 µm.</p>
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<p>Effect of arachidonic acid (AA) and t,t-farnesol on preformed <span class="html-italic">S. mutans</span> biofilms. <span class="html-italic">S. mutans</span> was allowed to form biofilms in BHI supplemented with 1% sucrose for 24 h. The mature biofilms were then washed in PBS and exposed to different concentrations of AA and/or t,t-farnesol for 24 h, and the metabolic activity of the remaining biofilms was measured using the MTT assay. N = 3. a: <span class="html-italic">p</span> &lt; 0.05 when compared to control biofilms.</p>
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<p>Effect of arachidonic acid (AA) and t,t-farnesol on the expression of biofilm-related genes. <span class="html-italic">S. mutans</span> was exposed to 3.125 µg/mL AA and/or 12.5 µg/mL t,t-farnesol for 2 h prior to RNA isolation. The gene expression was determined by semiquantitative real-time PCR using <span class="html-italic">gltA</span> and <span class="html-italic">glnA</span> as internal standards, and the relative expression was calculated against control bacteria that were set to 1. N = 3. a: <span class="html-italic">p</span> &lt; 0.05 when compared to control bacteria.</p>
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15 pages, 638 KiB  
Article
The Comparison and Brewing Value of Saaz Hop Pedigree
by Jana Olšovská, Lenka Straková, Vladimír Nesvadba, Tomáš Vrzal and Jaroslav Přikryl
Beverages 2024, 10(4), 101; https://doi.org/10.3390/beverages10040101 - 22 Oct 2024
Cited by 2 | Viewed by 1065
Abstract
The well-known hop variety, Saaz, which gives the Pilsner lager beer its characteristic hop aroma, may be threatened by climate change in the future. Therefore, new Saaz-related hop varieties, including Saaz Late, Saaz Brilliant, Saaz Comfort, and Saaz Shine, were recently bred. A [...] Read more.
The well-known hop variety, Saaz, which gives the Pilsner lager beer its characteristic hop aroma, may be threatened by climate change in the future. Therefore, new Saaz-related hop varieties, including Saaz Late, Saaz Brilliant, Saaz Comfort, and Saaz Shine, were recently bred. A comparison study was carried out to evaluate whether these varieties are acceptable for traditional lagers. For this purpose, sensorial and chemical analyses of hops and related beers, namely, an analysis of hop resins and oils, were performed. Sensory profiles of Saaz varieties are very similar (fine, hoppy aroma; floral; herbal), except for Saaz Comfort, which has a slightly higher aroma intensity, and Saaz Shine, which has the most noticeable fruity scent, with traces of citrus. The chemical profiles are also very similar, with α-humulene, β-pinene, (E)-β-farnesene, β-caryophyllene, and myrcene being the most abundant. Decoction mashing and kettle hopping technology with bottom fermentation show that the compared varieties result in very similar lager beers with hoppy, floral, herbal, fruity, and spicy aromas. Typical hop oils include farnesol, linalool, methyl geranate, β-pinene, and limonene. The high concentration of farnesol in beer correlates with the concentrations of (E)-β-farnesene and farnesol in hops. New Saaz varieties are widely used to produce Pilsner lager without affecting the traditional sensory aroma of this widespread style. Varieties have a higher yield of approximately 25% and bitter acid concentrations of approximately 15%, with Saaz Comfort comprising approximately 100%. Furthermore, the concentration of hop oils is approximately 40% higher in Saaz Shine than a traditional Saaz variety. Moreover, Saaz Shine and Saaz Comfort have very good resistance to drought, which is an important property from a climate change perspective. Full article
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<p>Principal component analysis of Saaz varieties and their hop oil content.</p>
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<p>Principal component analysis of single-hopped beers by relative Saaz varieties and their hop oil content.</p>
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13 pages, 4284 KiB  
Article
Total Iridoid Glycosides from Swertia mussotii Franch. Alleviate Cholestasis Induced by α-Naphthyl Isothiocyanate through Activating the Farnesoid X Receptor and Inhibiting Oxidative Stress
by Qi Dong, Zhenhua Wang, Na Hu, Fangfang Tie, Zenggen Liu, Ying Sun, Yue Wang, Nixia Tan and Honglun Wang
Int. J. Mol. Sci. 2024, 25(19), 10607; https://doi.org/10.3390/ijms251910607 - 2 Oct 2024
Viewed by 925
Abstract
Cholestasis refers to a physiological and pathological process caused by bile acid (BA) overaccumulation inside the circulatory system and liver, leading to systemic and hepatocellular damage. Activating the farnesol X receptor (FXR) to restore BA homeostasis is a promising strategy for treating cholestasis. [...] Read more.
Cholestasis refers to a physiological and pathological process caused by bile acid (BA) overaccumulation inside the circulatory system and liver, leading to systemic and hepatocellular damage. Activating the farnesol X receptor (FXR) to restore BA homeostasis is a promising strategy for treating cholestasis. The objective of this research is to reveal solid evidence for the fact that the total iridoid glycosides from Swertia mussotii Franch. (IGSM) alleviate cholestasis. In this research, the whole plant of S. mussotii was extracted with 70% ethanol and separated by macroporous adsorption resin. A rat cholestasis model was established by the injection of α-naphthyl isothiocyanate (ANIT) at a dose of 75 mg/kg. Biochemical and oxidative stress indicators were determined using commercial assay kits. The mRNA abundance of FXR and target proteins was assessed using RT-qPCR. In addition, the effects of main compounds with FXR were evaluated by molecular docking after IGSM analysis using UPLC. The results indicated that IGSM alleviated ANIT-induced cholestasis through reducing serum ALT, AST, AKP, and TBA levels; increasing the mRNA levels of Fxr, Besp, Ntcp, and Mep2; and reducing oxidative stress. The proportion of iridoid compounds in IGSM exceeded 50%, which may be the active substance basis of IGSM. This study provides a theoretical reference for IGSM in the treatment of cholestasis, and future studies may delve more deeply into the FXR regulatory pathway. Full article
(This article belongs to the Section Biochemistry)
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<p>IGSM alleviated liver injury and cholestasis by reducing the serum levels of AKP, ALT, AST, and TBA in ANIT-induced cholestasis rats. ** <span class="html-italic">p</span> &lt; 0.01 when compared to the control group; # <span class="html-italic">p</span> &lt; 0.05 when compared to the ANIT model group.</p>
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<p>IGSM improved the levels of SOD, MDA, CAT, and GSH-Px in the liver tissues in the ANIT-induced cholestasis rats. * <span class="html-italic">p</span> &lt; 0.05 when compared to the control group; # <span class="html-italic">p</span> &lt; 0.05 when compared to the ANIT model group.</p>
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<p>IGSM improved liver pathology in ANIT-induced cholestasis rats (HE stained, 200× magnification). (<b>A</b>) Control, (<b>B</b>) model, (<b>C</b>) high-dose group, (<b>D</b>) medium-dose group, and (<b>E</b>) low-dose group.</p>
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<p>IGSM affected the mRNA levels of FXR and related proteins in the liver tissues of ANIT-induced cholestasis rats. ** <span class="html-italic">p</span> &lt; 0.01 when compared to the control group; # <span class="html-italic">p</span> &lt; 0.05 and ## <span class="html-italic">p</span> &lt; 0.01 when compared to the ANIT model group.</p>
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<p>Chromatograms of the four reference substances (<b>A</b>) and IGSM (<b>B</b>). 1—sweroside, 2—gentiopicroside, 3—swertiamarin, and 4—mangiferin.</p>
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<p>The binding of FXR and compounds: sweroside (<b>1</b>), gentiopicroside (<b>2</b>), swertiamarin (<b>3</b>), mangiferin (<b>4</b>), and obeticholic acid (<b>5</b>).</p>
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<p>The interactions between FXR and compounds: sweroside (<b>1</b>), gentiopicroside (<b>2</b>), swertiamarin (<b>3</b>), mangiferin (<b>4</b>), and obeticholic acid (<b>5</b>) by molecular docking analysis.</p>
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12 pages, 1008 KiB  
Article
Repellent Effects of Coconut Fatty Acid Methyl Esters and Their Blends with Bioactive Volatiles on Winged Myzus persicae (Sulzer) Aphids (Hemiptera: Aphididae)
by Félix Martín, Pedro Guirao and María Jesús Pascual-Villalobos
Insects 2024, 15(9), 731; https://doi.org/10.3390/insects15090731 - 23 Sep 2024
Viewed by 1076
Abstract
Myzus persicae (Sulzer) (Hemiptera: Aphididae) is one of the most important aphid crop pests, due to its direct damage and its ability to transmit viral diseases in crops. The objective is to test whether spraying nanoemulsions of botanical products repels winged individuals of [...] Read more.
Myzus persicae (Sulzer) (Hemiptera: Aphididae) is one of the most important aphid crop pests, due to its direct damage and its ability to transmit viral diseases in crops. The objective is to test whether spraying nanoemulsions of botanical products repels winged individuals of M. persicae in a bioassay in culture chambers. The bioactive volatiles were applied on pepper plants at a dose of 0.2% alone or at 0.1% of each component in blends. A treated plant and a control plant were placed at each side of an entomological cage inside a growth chamber. The winged individuals were released between the plants, in a black-painted Petri dish suspended by wires in the upper half of the cage. The most repellent products were farnesol (repellency index, RI = 40.24%), (E)-anethole (RI = 30.85%) and coconut fatty acid methyl ester (coconut FAME) (RI = 28.93%), alone or in the following blends: farnesol + (E)-anethole + distilled lemon oil (RI = 36.55%) or (E)-anethole + distilled lemon oil + coconut FAME (RI = 30.63%). The observed effect of coconut FAME on aphids is the first report of this product having a repellent effect on a crop pest. Repellent substances for viral disease vectors should be further investigated to develop new strategies for plant protection. Full article
(This article belongs to the Special Issue Chemical Toxicology and Insecticide Resistance on Insect Pests)
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<p>Entomological cage with choice bioassay (control plant and treated plant) and the winged aphid shuttle.</p>
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<p>Distribution of choice percentage of aphids (<span class="html-italic">M. persicae</span> winged) 24 h after release in cages (<span class="html-italic">n</span> = 24) with treated and control pepper plants. The bioactive volatiles were formulated as nanoemulsions at 0.2% or 0.1% in blends of more than one product, with Tween 80. Statistical significance according to an exact two-sided binomial test (* <span class="html-italic">p</span> &lt; 0.05; ** <span class="html-italic">p</span> &lt; 0.01; *** <span class="html-italic">p</span> &lt; 0.001; ns <span class="html-italic">p</span> &gt; 0.05). When the confidence interval crosses the red dashed line at 50%, there is no significant repellency or attraction. Error bars correspond to the Clopper–Pearson 95% confidence intervals.</p>
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14 pages, 3754 KiB  
Article
Effects of Sucrose and Farnesol on Biofilm Formation by Streptococcus mutans and Candida albicans
by Wit Yee Wint, Mayu Miyanohara, Chika Terada-Ito, Hidenori Yamada, Koufuchi Ryo and Takatoshi Murata
Microorganisms 2024, 12(8), 1737; https://doi.org/10.3390/microorganisms12081737 - 22 Aug 2024
Cited by 2 | Viewed by 1236
Abstract
Candida albicans (C. albicans) and Streptococcus mutans (S. mutans) are frequently detected in the plaque biofilms of children with early childhood caries. This study investigated the effects of sucrose and farnesol on biofilm formation by the oral pathogens S. [...] Read more.
Candida albicans (C. albicans) and Streptococcus mutans (S. mutans) are frequently detected in the plaque biofilms of children with early childhood caries. This study investigated the effects of sucrose and farnesol on biofilm formation by the oral pathogens S. mutans and C. albicans, including their synergistic interactions. Biofilm formation dynamics were monitored using the Cell Index (CI). The CI for S. mutans increased in the brain–heart infusion medium, peaking at 10 h; however, the addition of sucrose reduced the CI. For C. albicans yeast cells, the CI increased at sucrose concentrations > 0.5%, peaking at 2 h. Mixed cultures of S. mutans and C. albicans yeast cells showed significantly higher CI values in the presence of sucrose, suggesting a synergistic effect on biofilm formation. Farnesol consistently suppressed biofilm formation by C. albicans yeast cells, even in the presence of sucrose, and higher farnesol concentrations resulted in greater inhibition. Regarding C. albicans hyphal cells, sucrose did not enhance biofilm formation, whereas farnesol significantly reduced biofilm formation at all concentrations tested. These findings elucidate the complex roles of sucrose and farnesol in biofilm formation by S. mutans and C. albicans and emphasize the potential of farnesol as an effective oral biofilm inhibitor. Full article
(This article belongs to the Special Issue Oral Microbiota and Candidiasis)
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<p>Effect of sucrose on the CIs at different time points. (<b>a</b>) Comparison of CIs between BHI, BHI + sucrose (0.5% and 1.0%), and BHI + <span class="html-italic">S. mutans</span> (BHI + Sm), BHI + sucrose + <span class="html-italic">S. mutans</span> (BHI + Suc + Sm). (<b>b</b>) Comparison of CIs between BHI, BHI + sucrose (0.5, 1.0, and 2.0%), and BHI + <span class="html-italic">C. albicans</span> yeast cells (BHI + Ca). BHI + Suc + Ca (BHI + Suc + Ca). Abbreviations: CI, Cell index; BHI, brain−heart infusion medium; PBS; phosphate-buffered saline, Suc, sucrose; Sm, <span class="html-italic">S. mutans</span>; Ca, <span class="html-italic">C. albicans</span>.</p>
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<p>Effect of sucrose on CIs after 5 h of incubation. (<b>a</b>) Comparison of CIs between BHI + Sm, BHI + 0.5 Suc + Sm, and BHI + 1.0 Suc + Sm. (<b>b</b>) Comparison of CIs between BHI + Ca, BHI + 0.5 Suc + Ca, BHI + 1.0 Suc + Ca, and BHI + 2.0 Suc + Ca. Data represent the means of three replicates, and error bars indicate standard errors. Means with the same letters are not significantly different (Tukey HSD, <span class="html-italic">p</span> &lt; 0.05).</p>
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<p>Effect of sucrose on the CIs of mixed cultures of <span class="html-italic">S. mutans</span> and <span class="html-italic">C. albicans</span>. (<b>a</b>) Effect of sucrose on the CIs of mixed cultures of <span class="html-italic">S. mutans</span> and <span class="html-italic">C. albicans</span> at different time points. (<b>b</b>) Effect of sucrose on the CI of the mixed culture of <span class="html-italic">S. mutans</span> and <span class="html-italic">C. albicans</span> after 5 h of incubation. Data represent the means of three replicates, and error bars indicate standard errors. Means with the same letters are not significantly different (Tukey HSD, <span class="html-italic">p</span> &lt; 0.05).</p>
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<p>Effect of farnesol on the CIs of <span class="html-italic">C. albicans</span> yeast cells at different time points. (<b>a</b>) Comparison of CIs between BHI + Ca, and BHI + Ca + Far (1, 2, 5, and 10 mM). (<b>b</b>) Comparison of CIs between BHI + Ca, BHI + Ca + 1.0 Suc, and BHI + Ca + 1.0 Suc + Far (1, 2, 5, and 10 mM). Abbreviations: Far, farnesol.</p>
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<p>Effect of farnesol CIs of <span class="html-italic">C. albicans</span> yeast cells after 5 h of incubation. (<b>a</b>) Comparison of CIs between BHI + Ca and BHI + Ca + Far (1, 2, 5, and 10 mM). (<b>b</b>) Comparison of CIs between BHI + Ca, BHI + Ca + 1.0 Suc, and BHI + Ca + 1.0 Suc + Far (1, 2, 5, and 10 mM). Data represent the means of three replicates, and error bars indicate standard errors. Means with the same letters are not significantly different (Tukey HSD, <span class="html-italic">p</span> &lt; 0.05).</p>
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<p>The individual and combined effects of sucrose and farnesol on biofilm formation by <span class="html-italic">C. albicans</span> hyphal cells at different time points. (<b>a</b>) Comparison of the effects of sucrose on the CIs of <span class="html-italic">C. albicans</span> between BHI + Suc (0.5% and 1.0%), and BHI + <span class="html-italic">C. albicans</span> (BHI + Ca). (<b>b</b>) Comparison of the effect of farnesol on CIs of <span class="html-italic">C. albicans</span> between BHI + <span class="html-italic">C. albicans</span> (BHI + Ca), and BHI + <span class="html-italic">C. albicans</span> with varying concentrations of farnesol (1, 2, 5, and 10 mM). (<b>c</b>) Comparison of the combined effects of farnesol and sucrose on CIs of <span class="html-italic">C. albicans</span> between BHI + <span class="html-italic">C. albicans</span> (BHI + Ca), and BHI + <span class="html-italic">C. albicans</span> with sucrose and varying concentrations of farnesol (1, 2, 5, and 10 mM).</p>
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<p>The individual and combined effects of sucrose and farnesol on biofilm formation by <span class="html-italic">C. albicans</span> hyphal cells after 5 h of incubation. (<b>a</b>) Comparison of the effect of Suc on CIs of <span class="html-italic">C. albicans</span> after 5 h of incubation between BHI + Ca, BHI + 0.5% Suc + Ca, and BHI + 1.0% Suc + Ca. (<b>b</b>) Comparison of the effect of farnesol on CIs of <span class="html-italic">C. albicans</span> after 5 h of incubation between BHI + Ca and BHI + C. albicans with varying concentrations of farnesol (1, 2, and 5 mM). (<b>c</b>) Comparison of the combined effect of farnesol and Suc on CIs of <span class="html-italic">C. albicans</span> after 5 h of incubation between BHI + Ca, BHI + 1.0 Suc + Ca, BHI + 1.0 Suc + Ca + 1 Far, and BHI + 1.0 Suc + Ca + 2 Far. Data represent the means of three replicates and error bars indicate standard errors. Means with the same letters are not significantly different (Tukey HSD, <span class="html-italic">p</span> &lt; 0.05).</p>
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21 pages, 4502 KiB  
Article
Farnesol Emulsion as an Effective Broad-Spectrum Agent against ESKAPE Biofilms
by Li Tan, Rong Ma, Adam J. Katz and Nicole Levi
Antibiotics 2024, 13(8), 778; https://doi.org/10.3390/antibiotics13080778 - 17 Aug 2024
Viewed by 1347
Abstract
The family of ESKAPE pathogens is comprised of Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter. Together they are the main contributors of nosocomial infections and are well established for their ability to “escape” antibiotics. Farnesol is [...] Read more.
The family of ESKAPE pathogens is comprised of Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter. Together they are the main contributors of nosocomial infections and are well established for their ability to “escape” antibiotics. Farnesol is an FDA-approved cosmetic and flavoring agent with significant anti-biofilm properties. In a proprietary emulsion, farnesol has been shown to be capable of disrupting S. aureus, P. aeruginosa, and A. baumannii biofilms. The current work demonstrates that this farnesol emulsion reduces the number of viable bacteria, while also leading to reductions in biomass, of the other three ESKAPE pathogens: Enterococcus faecium, Klebsiella pneumoniae, and Enterobacter, both in vitro and in an ex vivo human skin model. A concentration of 0.5 mg/mL was effective for impeding biofilm development of all three bacteria, while 1 mg/mL for E. faecium and K. pneumoniae, or 0.2 mg/mL for E. cloacae, was able to kill bacteria in established biofilms. Contrary to antibiotics, no resistance to farnesol was observed for E. faecium or K. pneumoniae. The results indicate that farnesol is effective for direct cell killing and also has the ability to induce biofilm detachment from surfaces, as confirmed using Live/Dead image analysis. Our findings confirm that farnesol emulsion is an effective broad-spectrum agent to impede ESKAPE biofilms. Full article
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Figure 1
<p>Inhibition of <span class="html-italic">E. faecium</span> biofilm formation. (<b>A</b>) <span class="html-italic">E. faecium</span> biofilm development is inhibited with exposure to farnesol for 24 h. Data are represented as the mean ± standard deviation (SD) (n = 3). (<b>B</b>) Flattened views of three-dimensional stacked Live/Dead images of <span class="html-italic">E. faecium</span> biofilms after 24 h exposure to farnesol. Green indicates live cells (SYTO<sup>®</sup> 9 staining), while red indicates dead cells (propidium iodide staining). Scale bars are 20 μm. (<b>C</b>) Quantitative evaluation of fluorescence intensity by Photoshop<sup>®</sup> from the images in (<b>B</b>), with data representing the mean ± SD (n = 3). (<b>D</b>) Quantitative evaluation of fluorescence intensity by Comstat2 from the images in (<b>B</b>) (biomass (µm<sup>3</sup>/µm<sup>2</sup>) and average thickness (µm)), using Otsu thresholding [<a href="#B31-antibiotics-13-00778" class="html-bibr">31</a>]. Ctrl indicates the ethanol control corresponding to the same volume of ethanol used to carry farnesol. Ctrl_0.5 = 1.7%, Ctrl_1 = 3.3%, and Ctrl_2 = 6.7% of ethanol. * <span class="html-italic">p</span> &lt; 0.0001 against each of other groups.</p>
Full article ">Figure 2
<p>Inhibition of <span class="html-italic">K. pneumoniae</span> biofilm formation. (<b>A</b>) <span class="html-italic">K. pneumoniae</span> biofilm development is inhibited with exposure to farnesol for 24 h. Data are represented as the mean ± SD (n = 3). (<b>B</b>) Flattened views of three-dimensional stacked Live/Dead images of <span class="html-italic">K. pneumoniae</span> biofilms after 24 h exposure to farnesol. Green indicates live cells (SYTO<sup>®</sup> 9 staining), while red indicates dead cells (propidium iodide staining). Scale bars are 20 μm. (<b>C</b>) Quantitative evaluation of fluorescence intensity by Photoshop<sup>®</sup> from the images in (<b>B</b>), with data representing the mean ± SD (n = 3). (<b>D</b>) Quantitative evaluation of fluorescence intensity by Comstat2 from the images in (<b>B</b>) (biomass (µm<sup>3</sup>/µm<sup>2</sup>) and average thickness (µm)), using Otsu thresholding [<a href="#B31-antibiotics-13-00778" class="html-bibr">31</a>]. Ctrl indicates the ethanol control corresponding to the same volume of ethanol used to carry farnesol. Ctrl_0.5 = 1.7%, Ctrl_1 = 3.3%, and Ctrl_3 = 10% of ethanol. * <span class="html-italic">p</span> &lt; 0.0001 against each of other groups.</p>
Full article ">Figure 3
<p>Inhibition of <span class="html-italic">E. cloacae</span> biofilm formation. (<b>A</b>) <span class="html-italic">E. cloacae</span> biofilm development is inhibited with exposure to farnesol for 24 h. Data are represented as the mean ± SD (n = 3). (<b>B</b>) Flattened views of three-dimensional stacked Live/Dead images of <span class="html-italic">E. cloacae</span> biofilms after 24 h exposure to farnesol. Green indicates live cells (SYTO<sup>®</sup> 9 staining), while red indicates dead cells (propidium iodide staining). Scale bars are 20 μm. (<b>C</b>) Quantitative evaluation of fluorescence intensity by Photoshop<sup>®</sup> from the images in (<b>B</b>), with data representing the mean ± SD (n = 3). (<b>D</b>) Quantitative evaluation of fluorescence intensity by Comstat2 from the images in (<b>B</b>) (biomass (µm<sup>3</sup>/µm<sup>2</sup>) and average thickness (µm)), using Otsu thresholding [<a href="#B31-antibiotics-13-00778" class="html-bibr">31</a>]. Ctrl indicates the ethanol control corresponding to the same volume of ethanol used to carry farnesol. Ctrl_0.2 = 0.67%, Ctrl_0.5 = 1.7%, and Ctrl_1 = 3.3% of ethanol. * <span class="html-italic">p</span> &lt; 0.0001 against each of other groups.</p>
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<p>Farnesol disrupts established <span class="html-italic">E. faecium</span> biofilms. (<b>A</b>) The 24 h old established biofilms exposed to farnesol for 24 h have reduced cell viability, indicative of biofilm disruption and cell killing. Data are represented as the mean ± SD (n = 3). (<b>B</b>) Flattened views of three-dimensional stacked Live/Dead images of 24 h old established <span class="html-italic">E. faecium</span> biofilms after 24 h exposure to farnesol. Green indicates live cells (SYTO<sup>®</sup> 9 staining), while red indicates dead cells (propidium iodide staining). Scale bars are 20 μm. (<b>C</b>) Quantitative evaluation of fluorescence intensity by Photoshop<sup>®</sup> from the images in (<b>B</b>), with data representing the mean ± SD (n = 3). (<b>D</b>) Quantitative evaluation of fluorescence intensity by Comstat2 from the images in (<b>B</b>) (biomass (µm<sup>3</sup>/µm<sup>2</sup>) and average thickness (µm)), using Otsu thresholding [<a href="#B31-antibiotics-13-00778" class="html-bibr">31</a>]. Ctrl indicates the ethanol control corresponding to the same volume of ethanol used to carry farnesol. Ctrl_3 = 10%, Ctrl_6 = 20%, and Ctrl_10 = 33.3% of ethanol. * <span class="html-italic">p</span> &lt; 0.0001 against each of other groups.</p>
Full article ">Figure 5
<p>Farnesol disrupts established <span class="html-italic">K. pneumoniae</span> biofilms. (<b>A</b>) The 24 h old established biofilms exposed to farnesol for 24 h have reduced cell viability, indicative of biofilm disruption and cell killing. Data are represented as the mean ± SD (n = 3). (<b>B</b>) Flattened views of three-dimensional stacked Live/Dead images of 24 h old established <span class="html-italic">K. pneumoniae</span> biofilms after 24 h exposure to farnesol. Green indicates live cells (SYTO<sup>®</sup> 9 staining), while red indicates dead cells (propidium iodide staining). Scale bars are 20 μm. (<b>C</b>) Quantitative evaluation of fluorescence intensity by Photoshop<sup>®</sup> from the images in (<b>B</b>), with data representing the mean ± SD (n = 3). (<b>D</b>) Quantitative evaluation of fluorescence intensity by Comstat2 from the images in (<b>B</b>) (biomass (µm<sup>3</sup>/µm<sup>2</sup>) and average thickness (µm), using Otsu thresholding [<a href="#B31-antibiotics-13-00778" class="html-bibr">31</a>]. Ctrl indicates the ethanol control corresponding to the same volume of ethanol used to carry farnesol. Ctrl_1 = 3.3%, Ctrl_3 = 10%, and Ctrl_6 = 20% of ethanol. * <span class="html-italic">p</span> &lt; 0.0001 against each of other groups.</p>
Full article ">Figure 6
<p>Farnesol disrupts established <span class="html-italic">E. cloacae</span> biofilms. (<b>A</b>) The 24 h old established biofilms exposed to farnesol for 24 h have reduced cell viability, indicative of biofilm disruption and cell killing. Data are represented as the mean ± SD (n = 3). (<b>B</b>) Flattened views of three-dimensional stacked Live/Dead images of 24 h old established <span class="html-italic">E. cloacae</span> biofilms after 24 h exposure to farnesol. Green indicates live cells (SYTO<sup>®</sup> 9 staining), while red indicates dead cells (propidium iodide staining). Scale bars are 20 μm. (<b>C</b>) Quantitative evaluation of fluorescence intensity by Photoshop<sup>®</sup> from the images in (<b>B</b>), with data representing the mean ± SD (n = 3). (<b>D</b>) Quantitative evaluation of fluorescence intensity by Comstat2 from the images in (<b>B</b>) (biomass (µm<sup>3</sup>/µm<sup>2</sup>) and average thickness (µm)), using Otsu thresholding [<a href="#B31-antibiotics-13-00778" class="html-bibr">31</a>]. Ctrl indicates the ethanol control corresponding to the same volume of ethanol used to carry farnesol. Ctrl_0.2 = 0.67%, Ctrl_0.5 = 1.7%, and Ctrl_1 = 3.3% of ethanol. * <span class="html-italic">p</span> ≤ 0.0005 against each of other groups.</p>
Full article ">Figure 7
<p>Farnesol kills <span class="html-italic">E. faecium</span> cells and facilitates biofilm detachment without inducing resistance. (<b>A</b>) <span class="html-italic">E. faecium</span> development of resistance to rifampicin or farnesol with serial passaging at sub-MIC doses. (<b>B</b>) Propidium iodide (PI) influx into <span class="html-italic">E. faecium</span> indicates cell killing. Data include the mean of three replicates. (<b>C</b>) The ethanol vehicle does not detach or kill <span class="html-italic">E. faecium</span> cells following 24 h exposure of established biofilms to the ethanol control, as indicated by no green or red fluorescence. Supernatants recovered from the biofilms indicate that live (green) biomass is detached from the surface at a low dose of farnesol. Increasing farnesol doses disrupt the live biomass and further kill the <span class="html-italic">E. faecium</span> cells (red floating material in the supernatant). Scale bars, 20 μm. Ctrl_1 = 3.3%, and Ctrl_3 = 10% of ethanol.</p>
Full article ">Figure 8
<p>Farnesol kills <span class="html-italic">K. pneumoniae</span> cells and facilitates biofilm detachment without inducing resistance. (<b>A</b>) <span class="html-italic">K. pneumoniae</span> development of resistance to rifampicin or farnesol with serial passaging at sub-MIC doses. (<b>B</b>) Propidium iodide (PI) influx into <span class="html-italic">K. pneumoniae</span> indicates cell killing. Data include the mean of three replicates. (<b>C</b>) The ethanol vehicle does not detach or kill <span class="html-italic">K. pneumoniae</span> cells following 24 h exposure of established biofilms to the ethanol control, as indicated by no green or red fluorescence. Supernatants recovered from the biofilms indicate that live (green) biomass is detached from the surface at a low dose of farnesol. Increasing farnesol doses disrupt the live biomass and further kill the <span class="html-italic">K. pneumoniae</span> cells (red floating material in the supernatant). Scale bars, 20 μm. Ctrl_1 = 3.3% of ethanol.</p>
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<p>Farnesol prevents biofilm development and disrupts established <span class="html-italic">E. faecium</span> biofilm<span class="html-italic">s</span> on ex vivo intact or burned human skin. (<b>A</b>,<b>B</b>) <span class="html-italic">E. faecium</span> biofilm development on ex vivo human skin is inhibited by exposure to farnesol at 1 mg/mL for 24 h, as visualized by H&amp;E-stained images (<b>A</b>), and quantification of viable bacteria in log<sub>10</sub> CFU per square centimeter (cm<sup>2</sup>) of skin (<b>B</b>). (<b>C</b>,<b>D</b>) The same concentration of farnesol (1 mg/mL) is also effective for inhibiting <span class="html-italic">E. faecium</span> biofilm development on burned ex vivo human skin treated for 24 h, as visualized by H&amp;E-stained images (<b>C</b>), and quantification of viable bacteria in log<sub>10</sub> CFU per cm<sup>2</sup> of skin (<b>D</b>). (<b>E</b>,<b>F</b>) Established <span class="html-italic">E. faecium</span> biofilms developed for 24 h on ex vivo human skin, and then exposed to farnesol for 24 h (15 mg/mL), have reductions in biofilm development, as visualized by H&amp;E-stained images (<b>E</b>), and quantification of viable bacteria in log<sub>10</sub> CFU per cm<sup>2</sup> of skin (<b>F</b>). (<b>G</b>,<b>H</b>) The same concentration of farnesol (15 mg/mL) is also effective for inhibiting <span class="html-italic">E. faecium</span> biofilm development on burned ex vivo human skin treated for 24 h, as visualized by H&amp;E-stained images (<b>G</b>), and quantification of viable bacteria in log<sub>10</sub> CFU per cm<sup>2</sup> of skin (<b>H</b>). The presence of biofilm is indicated by arrowheads in (<b>A</b>,<b>C</b>,<b>E</b>,<b>G</b>). Quantitative data in (<b>B</b>,<b>D</b>,<b>F</b>,<b>H</b>) represent the mean ± SD (n = 3 separate pieces of skin) from two independent donors. Ctrl_1 = 3.3 and Ctrl_15 = 50% of ethanol.</p>
Full article ">Figure 10
<p>Farnesol prevents biofilm development and disrupts established <span class="html-italic">K. pneumoniae</span> biofilm<span class="html-italic">s</span> on ex vivo intact or burned human skin. (<b>A</b>,<b>B</b>) <span class="html-italic">K. pneumoniae</span> biofilm development on ex vivo human skin is inhibited by exposure to farnesol at 1 mg/mL for 24 h, as visualized by H&amp;E-stained images (<b>A</b>), and quantification of viable bacteria in log<sub>10</sub> CFU per cm<sup>2</sup> of skin (<b>B</b>). (<b>C</b>,<b>D</b>) The same concentration of farnesol (1 mg/mL) is also effective for inhibiting <span class="html-italic">K. pneumoniae</span> biofilm development on burned ex vivo human skin treated for 24 h, as visualized by H&amp;E-stained images (<b>C</b>), and quantification of viable bacteria in log<sub>10</sub> CFU per cm<sup>2</sup> of skin (<b>D</b>). (<b>E</b>,<b>F</b>) Established <span class="html-italic">K. pneumoniae</span> biofilms developed for 24 h on ex vivo human skin, and then exposed to farnesol for 24 h (15 mg/mL), have reductions in biofilm development, as visualized by H&amp;E-stained images (<b>E</b>), and quantification of viable bacteria in log<sub>10</sub> CFU per cm<sup>2</sup> of skin (<b>F</b>). (<b>G</b>,<b>H</b>) The same concentration of farnesol (15 mg/mL) is also effective for inhibiting <span class="html-italic">K. pneumoniae</span> biofilm development on burned ex vivo human skin treated for 24 h, as visualized by H&amp;E-stained images (<b>G</b>), and quantification of viable bacteria in log<sub>10</sub> CFU per cm<sup>2</sup> of skin (<b>H</b>). The presence of biofilm is indicated by arrowheads in (<b>A</b>,<b>C</b>,<b>E</b>,<b>G</b>). Quantitative data in (<b>B</b>,<b>D</b>,<b>F</b>,<b>H</b>) represent the mean ± SD (n = 3 separate pieces of skin) from two independent donors. Ctrl_6 = 20% of ethanol; Ctrl_15 = 50% of ethanol.</p>
Full article ">Figure 11
<p>Farnesol prevents biofilm development and disrupts established <span class="html-italic">E. cloacae</span> biofilm<span class="html-italic">s</span> on ex vivo intact or burned human skin. (<b>A</b>,<b>B</b>) <span class="html-italic">E. cloacae</span> biofilm development on ex vivo human skin is inhibited by exposure to farnesol at 1 mg/mL for 24 h, as visualized by H&amp;E-stained images (<b>A</b>), and quantification of viable bacteria in log<sub>10</sub> CFU per cm<sup>2</sup> of skin (<b>B</b>). (<b>C</b>,<b>D</b>) The same concentration of farnesol (1 mg/mL) is also effective for inhibiting <span class="html-italic">E. cloacae</span> biofilm development on burned ex vivo human skin treated for 24 h, as visualized by H&amp;E-stained images (<b>C</b>), and quantification of viable bacteria in log<sub>10</sub> CFU per cm<sup>2</sup> of skin (<b>D</b>). (<b>E</b>,<b>F</b>) Established <span class="html-italic">E. cloacae</span> biofilms developed for 24 h on ex vivo human skin, and then exposed to farnesol for 24 h (15 mg/mL), have reductions in biofilm development, as visualized by H&amp;E-stained images (<b>E</b>), and quantification of viable bacteria in log<sub>10</sub> CFU per cm<sup>2</sup> of skin (<b>F</b>). (<b>G</b>,<b>H</b>) The same concentration of farnesol (15 mg/mL) is also effective for <span class="html-italic">E. cloacae</span> biofilm development on burned ex vivo human skin treated for 24 h, as visualized by H&amp;E-stained images (<b>G</b>), and quantification of viable bacteria in log<sub>10</sub> CFU per cm<sup>2</sup> of skin (<b>H</b>). The presence of biofilm is indicated by black arrowheads in (<b>A</b>,<b>C</b>,<b>E</b>,<b>G</b>). The gray arrows in (<b>C,G</b>) indicate broken stratum corneum of burned epidermis, which was penetrated by <span class="html-italic">E. cloacae</span> to develop biofilm. Quantitative data in (<b>B</b>,<b>D</b>,<b>F</b>,<b>H</b>) represent the mean ± SD (n = 3 separate pieces of skin) from two independent donors. Ctrl_1 = 3.3% of ethanol; Ctrl_15 = 50% of ethanol.</p>
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13 pages, 2269 KiB  
Article
Antifungal Effect of Poly(methyl methacrylate) with Farnesol and Undecylenic Acid against Candida albicans Biofilm Formation
by Milica Išljamović, Debora Bonvin, Milena Milojević, Simona Stojanović, Milan Spasić, Branislava Stojković, Predrag Janošević, Suzana Otašević and Marijana Mionić Ebersold
Materials 2024, 17(16), 3936; https://doi.org/10.3390/ma17163936 - 8 Aug 2024
Viewed by 1321
Abstract
The control of Candida albicans biofilm formation on dentures made of poly(methyl methacrylate) (PMMA) is an important challenge due to the high resistance to antifungal drugs. Interestingly, the natural compounds undecylenic acid (UDA) and farnesol (FAR) both prevent C. albicans biofilm formation and [...] Read more.
The control of Candida albicans biofilm formation on dentures made of poly(methyl methacrylate) (PMMA) is an important challenge due to the high resistance to antifungal drugs. Interestingly, the natural compounds undecylenic acid (UDA) and farnesol (FAR) both prevent C. albicans biofilm formation and could have a synergetic effect. We therefore modified PMMA with a combination of UDA and FAR (UDA+FAR), aiming to obtain the antifungal PMMA_UDA+FAR composites. Equal concentrations of FAR and UDA were added to PMMA to reach 3%, 6%, and 9% in total of both compounds in composites. The physico-chemical properties of the composites were characterized by Fourier-transform infrared spectroscopy and water contact angle measurement. The antifungal activity of the composites was tested on both biofilm and planktonic cells with an XTT test 0 and 6 days after the composites’ preparation. The effect of the UDA+FAR combination on C. albicans filamentation was studied in agar containing 0.0125% and 0.4% UDA+FAR after 24 h and 48 h of incubation. The results showed the presence of UDA and FAR on the composite and decreases in the water contact angle and metabolic activity of both the biofilm and planktonic cells at both time points at non-toxic UDA+FAR concentrations. Thus, the modification of PMMA with a combination of UDA+FAR reduces C. albicans biofilm formation on dentures and could be a promising anti-Candida strategy. Full article
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Figure 1

Figure 1
<p>Fourier-transform infrared spectra of undecylenic acid (UDA), farnesol (FAR), poly(methyl methacrylate) (0% UDA+FAR), and composites with 3%, 6%, and 9% UDA+FAR.</p>
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<p>Water contact angle on the surface of the composites with 0%, 3%, 6%, and 9% of the total concentration of both antimicrobial compounds undecylenic acid (UDA) and farnesol (FAR). Asterisks above the columns show significant differences among the groups and compared to the controls (<span class="html-italic">p</span> &lt; 0.05, Tukey’s test). The results are presented as means ± SDs.</p>
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<p>Comparison of a metabolically active <span class="html-italic">Candida</span> (<span class="html-italic">C.</span>) <span class="html-italic">albicans</span> biofilm (<b>a</b>) and planktonic (<b>b</b>) cells after incubation with poly(methyl methacrylate) (PMMA) modified with the combination of equal concentrations of tundecylenic acid (UDA) and farnesol (FAR) (0%, 3%, 6%, and 9% UDA+FAR in total) in two points: 0 days (T0) and 6 days (T6) after PMMA_UDA+FAR composite preparation. The asterisks denote significant differences (<span class="html-italic">p</span> &lt; 0.05) compared to the PMMA control.</p>
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<p>The effect of the combination of undecylenic acid (UDA) and farnesol (FAR) on <span class="html-italic">C. albicans</span> growth after 24 h of incubation on the agar surface loaded with different UDA+FAR concentrations. The results are given as a mean ± standard deviation of optical density reading at a wavelength of 620 nm.</p>
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<p>Representative optical micrographs showing morphologic changes of <span class="html-italic">C. albicans</span> cells embedded in the agar without the combination of undecylenic acid (UDA) + farnesol (FAR) (control, 0% UDA+FAR, (<b>a</b>,<b>b</b>)), with 0.0125% UDA+FAR (<b>c</b>,<b>d</b>) and 0.4% UDA+FAR (<b>e</b>,<b>f</b>) after 24 h (<b>a</b>,<b>c</b>,<b>e</b>) and 48 h (<b>b</b>,<b>d</b>,<b>f</b>) of incubation. The arrows show spindle-shaped colonies with sporadic hyphal/pseudohyphal forms on the periphery and lateral yeasts in agar loaded with 0.0125% UDA+FAR in total (<b>c</b>,<b>d</b>). The images were taken with a magnification of 40×.</p>
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<p>The viability of human A549 cells incubated with media after the immersion of PMMA with the combination of undecylenic acid (UDA) and farnesol (FAR) in different concentrations, (in total, 3%, 6%, and 9% <span class="html-italic">w</span>/<span class="html-italic">w</span> UDA+FAR, more precisely per 1.5%, 3%, and 4.5% <span class="html-italic">w</span>/<span class="html-italic">w</span> of UDA and FAR, respectively), measured by the MTS test in two studied time points, T0—0 days—and T6—6 days after the PMMA_UDA+FAR disc preparation. The PMMA_UDA+FAR combinations were immersed for 24 h and six days longer for the T0 and T6 time points.</p>
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34 pages, 2751 KiB  
Article
Characterisation of the Volatile Compounds and Key Odourants in Japanese Mandarins by Gas Chromatography–Mass Spectrometry and Gas Chromatography–Olfactometry
by Lingyi Li, Rui Min Vivian Goh, Yunle Huang, Kim-Huey Ee, Aileen Pua, Daphne Tan, Shanbo Zhang, Lionel Jublot, Shao Quan Liu and Bin Yu
Separations 2024, 11(8), 237; https://doi.org/10.3390/separations11080237 - 1 Aug 2024
Viewed by 1580
Abstract
Japanese mandarins are becoming increasingly popular due to their pleasant aroma. The volatiles in four varieties of Japanese mandarins (Iyokan, Ponkan, Shiranui, and Unshiu mikan) were extracted by headspace solid-phase microextraction (HS-SPME) and solvent extraction, then analysed by gas chromatography–mass spectrometry (GC-MS). Principal [...] Read more.
Japanese mandarins are becoming increasingly popular due to their pleasant aroma. The volatiles in four varieties of Japanese mandarins (Iyokan, Ponkan, Shiranui, and Unshiu mikan) were extracted by headspace solid-phase microextraction (HS-SPME) and solvent extraction, then analysed by gas chromatography–mass spectrometry (GC-MS). Principal component analysis (PCA) of the GC-MS data demonstrated distinct segregation of all four Japanese mandarin varieties. Esters, such as neryl acetate, distinguished Iyokan. Methylthymol uniquely characterised Ponkan, valencene was exclusive to Shiranui, and acids like hexanoic acid and heptanoic acid differentiated Unshiu mikan from the other three varieties. Aroma extract dilution analysis (AEDA) revealed 131 key odourants across four Japanese mandarins, including myrcene (peppery, terpenic), perillyl alcohol (green, spicy, floral), trans-nerolidol (sweet, floral), and trans-farnesol (woody, floral, green). Finally, sensory evaluation was conducted on the four Japanese mandarin peel extracts to describe the distinct aroma profile of each variety of Japanese mandarin: Iyokan had higher floral and juicy notes, Ponkan showed higher sulphury notes, Shiranui was perceived to have more albedo notes, and Unshiu mikan exhibited higher peely, green, and woody notes. Full article
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Figure 1

Figure 1
<p>PCA scores and loadings plots of volatile compounds in four varieties of Japanese Mandarin (Iyokan (<span style="color:#ef8c31">▲</span>), Ponkan (<span style="color:#4ab5d6">■</span>), Shiranui (<span style="color:#9463ad">●</span>), and Unshiu mikan (<span style="color:#6bbe8c">★</span>)): (<b>a</b>) Scores plot of volatiles in juices extracted by HS-SPME; (<b>b</b>) Loadings plot of volatiles in juices extracted by HS-SPME; (<b>c</b>) Scores plot of volatiles in peels extracted by HS-SPME; (<b>d</b>) Loadings plot of volatiles in peels extracted by HS-SPME; (<b>e</b>) Scores plot of volatiles in peels extracted by solvent extraction; (<b>f</b>) Loadings plot of volatiles in peels extracted by solvent extraction. The numbers denote the corresponding volatiles reported in <a href="#separations-11-00237-t001" class="html-table">Table 1</a> (juice HS-SPME plots), <a href="#separations-11-00237-t002" class="html-table">Table 2</a> (peel HS-SPME plots), and <a href="#separations-11-00237-t003" class="html-table">Table 3</a> (solvent extraction plots). The black squares indicate the contribution magnitude and direction of variables to the principal components, with their position reflecting the loading value.</p>
Full article ">Figure 1 Cont.
<p>PCA scores and loadings plots of volatile compounds in four varieties of Japanese Mandarin (Iyokan (<span style="color:#ef8c31">▲</span>), Ponkan (<span style="color:#4ab5d6">■</span>), Shiranui (<span style="color:#9463ad">●</span>), and Unshiu mikan (<span style="color:#6bbe8c">★</span>)): (<b>a</b>) Scores plot of volatiles in juices extracted by HS-SPME; (<b>b</b>) Loadings plot of volatiles in juices extracted by HS-SPME; (<b>c</b>) Scores plot of volatiles in peels extracted by HS-SPME; (<b>d</b>) Loadings plot of volatiles in peels extracted by HS-SPME; (<b>e</b>) Scores plot of volatiles in peels extracted by solvent extraction; (<b>f</b>) Loadings plot of volatiles in peels extracted by solvent extraction. The numbers denote the corresponding volatiles reported in <a href="#separations-11-00237-t001" class="html-table">Table 1</a> (juice HS-SPME plots), <a href="#separations-11-00237-t002" class="html-table">Table 2</a> (peel HS-SPME plots), and <a href="#separations-11-00237-t003" class="html-table">Table 3</a> (solvent extraction plots). The black squares indicate the contribution magnitude and direction of variables to the principal components, with their position reflecting the loading value.</p>
Full article ">Figure 1 Cont.
<p>PCA scores and loadings plots of volatile compounds in four varieties of Japanese Mandarin (Iyokan (<span style="color:#ef8c31">▲</span>), Ponkan (<span style="color:#4ab5d6">■</span>), Shiranui (<span style="color:#9463ad">●</span>), and Unshiu mikan (<span style="color:#6bbe8c">★</span>)): (<b>a</b>) Scores plot of volatiles in juices extracted by HS-SPME; (<b>b</b>) Loadings plot of volatiles in juices extracted by HS-SPME; (<b>c</b>) Scores plot of volatiles in peels extracted by HS-SPME; (<b>d</b>) Loadings plot of volatiles in peels extracted by HS-SPME; (<b>e</b>) Scores plot of volatiles in peels extracted by solvent extraction; (<b>f</b>) Loadings plot of volatiles in peels extracted by solvent extraction. The numbers denote the corresponding volatiles reported in <a href="#separations-11-00237-t001" class="html-table">Table 1</a> (juice HS-SPME plots), <a href="#separations-11-00237-t002" class="html-table">Table 2</a> (peel HS-SPME plots), and <a href="#separations-11-00237-t003" class="html-table">Table 3</a> (solvent extraction plots). The black squares indicate the contribution magnitude and direction of variables to the principal components, with their position reflecting the loading value.</p>
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<p>PCA scores and loadings plots of volatile compounds in four varieties of Japanese Mandarin (Iyokan (<span style="color:#ef8c31">▲</span>), Ponkan (<span style="color:#4ab5d6">■</span>), Shiranui (<span style="color:#9463ad">●</span>), and Unshiu mikan (<span style="color:#6bbe8c">★</span>)): (<b>a</b>) Scores plot of volatiles in juices extracted by HS-SPME; (<b>b</b>) Loadings plot of volatiles in juices extracted by HS-SPME; (<b>c</b>) Scores plot of volatiles in peels extracted by HS-SPME; (<b>d</b>) Loadings plot of volatiles in peels extracted by HS-SPME; (<b>e</b>) Scores plot of volatiles in peels extracted by solvent extraction; (<b>f</b>) Loadings plot of volatiles in peels extracted by solvent extraction. The numbers denote the corresponding volatiles reported in <a href="#separations-11-00237-t001" class="html-table">Table 1</a> (juice HS-SPME plots), <a href="#separations-11-00237-t002" class="html-table">Table 2</a> (peel HS-SPME plots), and <a href="#separations-11-00237-t003" class="html-table">Table 3</a> (solvent extraction plots). The black squares indicate the contribution magnitude and direction of variables to the principal components, with their position reflecting the loading value.</p>
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<p>Heatmap of the concentrations and FD factors of the key odourants of four Japanese mandarin peel extracts. “NA” means the odourant was not detected by AEDA via GC-O/MS. For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.</p>
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<p>Sensory profiles of four varieties of Japanese mandarin (Iyokan, Ponkan, Shiranui, and Unshiu mikan) peel extracts.</p>
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19 pages, 4874 KiB  
Article
Comparison of Aroma Profiles of Whiskeys Fermented from Different Grain Ingredients
by Siqian Guo, Dan Wang, Yanting Li, Jingming Li and Jinkun Du
Foods 2024, 13(13), 2031; https://doi.org/10.3390/foods13132031 - 26 Jun 2024
Cited by 1 | Viewed by 1695
Abstract
Different grain sources of whiskey have great potential for aroma expression. In this paper, four whiskeys fermented from different raw materials (barley, wheat, highland barley, and sorghum) were compared. Gas chromatography–mass spectrometry (GC-MS) and sensory evaluation were used to determine the composition of [...] Read more.
Different grain sources of whiskey have great potential for aroma expression. In this paper, four whiskeys fermented from different raw materials (barley, wheat, highland barley, and sorghum) were compared. Gas chromatography–mass spectrometry (GC-MS) and sensory evaluation were used to determine the composition of the aromatic compounds. A correlation analysis was further conducted between the aromatic compounds and sensory evaluations. Barley whiskey and wheat whiskey had more pronounced fruity, floral, and grain aromas, attributed to esters and terpenes. Barley whiskey had the most compounds (55), followed by highland barley whiskey (54). Highland barley whiskey had the greatest number of unique aroma compounds (seven). It exhibited a unique cocoa aroma related to concentrations of trans-2-nonenal, γ-nonanolactone, 1-nonanol, isoamyl lactate, 2-butanol, and 6-methyl-5-hepten-2-one. Sorghum whiskey had a specific leather and mushroom aroma attributed to 6-methyl-5-hepten-2-one, ethyl lactate, ethyl caprate, phenethyl octanoate, farnesol, α-terpineol, 3-methyl-1-pentanol, and methyleugenol. Alcohols were the main aroma components of grain whiskeys. Isoamyl alcohol (231.59~281.39 mg/L), phenylethyl alcohol (5.755~9.158 mg/L), citronellol (0.224~4.103 mg/L), β-damascenone (0.021~2.431 mg/L), geraniol (0.286~1.416 mg/L), isoamyl acetate (0.157~0.918 mg/L), phenylacetaldehyde (0.162~0.470 mg/L), linalool (0.024~0.148 mg/L), 1-octen-3-ol (0.016~0.145 mg/L), trans-2-nonenal (0.027~0.105 mg/L), and trans-2-octen-1-ol (0.011~0.054 mg/L) were all important aroma compounds in the whiskeys. Full article
(This article belongs to the Section Grain)
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Graphical abstract

Graphical abstract
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<p>The heat map of the aroma composition content (<b>A</b>), PCA score plot (<b>B</b>), and loading plot (<b>C</b>) of the four whiskeys. The numbers in the loading plot correspond to the compound numbers in <a href="#foods-13-02031-t001" class="html-table">Table 1</a>.</p>
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<p>Aroma composition ratio of four whiskeys (<b>A</b>–<b>D</b>, barley whiskey, wheat whiskey, highland barley whiskey, and sorghum whiskey, respectively) and comparison of important aroma components (<b>E</b>–<b>H</b>, alcohols, esters, aldehydes, and terpenes, respectively). Different letters indicate statistically significant differences (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>Flavor radar plots for the four whiskeys.</p>
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<p>Scatter plot of correlation coefficients between aroma sensory attributes. The size of the circle corresponds to the correlation coefficient, and the color represents the positive/negative correlation (red = positive correlation, blue = negative correlation). * indicates a significant association at <span class="html-italic">p</span>-value = 0.05.</p>
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<p>Correlation heat map between aroma sensory attributes and GC-MS analysis.</p>
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<p>Correlation network diagram between aroma sensory attributes and GC-MS analysis. The size of the dot represents the strength of the correlations with other aroma sensory attributes or aroma substances, the thickness of the line represents the strength of the correlations between the two connected attributes or compounds, and the color of the line represents the correlations of different aroma sensory attributes.</p>
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16 pages, 4223 KiB  
Article
Impacts of Dietary Standardized Ileal Digestible Lysine to Net Energy Ratio on Lipid Metabolism in Finishing Pigs Fed High-Wheat Diets
by Jiguang Wang, Haojie Li, He Zhu, Shuangshuang Xia, Fang Zhang, Hui Zhang, Chunxue Liu, Weijiang Zheng and Wen Yao
Animals 2024, 14(12), 1824; https://doi.org/10.3390/ani14121824 - 19 Jun 2024
Viewed by 1206
Abstract
The present study aimed to investigate the impacts of dietary standardized ileal digestible lysine to net energy (SID Lys:NE) ratio on lipid metabolism in pigs fed high-wheat diets. Thirty-six crossbred growing barrows (65.20 ± 0.38 kg) were blocked into two treatment groups, fed [...] Read more.
The present study aimed to investigate the impacts of dietary standardized ileal digestible lysine to net energy (SID Lys:NE) ratio on lipid metabolism in pigs fed high-wheat diets. Thirty-six crossbred growing barrows (65.20 ± 0.38 kg) were blocked into two treatment groups, fed high-wheat diets with either a high SID Lys:NE ratio (HR) or a low SID Lys:NE ratio (LR). Each treatment group consisted of three replicates, with six pigs per pen in each replicate. The diminishing dietary SID Lys:NE ratio exhibited no adverse impacts on the carcass trait (p > 0.05) but increased the marbling score of the longissimus dorsi muscle (p < 0.05). Meanwhile, LR diets tended to increase the serum triglyceride concentration (p < 0.1). LR diets upregulated fatty acid transport protein 4 and acetyl-coA carboxylase α expression levels and downregulated the expression level of adipose triglyceride lipase (p < 0.05). LR diets improved energy metabolism via decreasing the expression levels of AMP-activated protein kinase (AMPK) α1, sirtuin 1 (SIRT1), and peroxisome proliferator-activated receptor-γ coactivator-1α (PGC-1α) (p < 0.05). Additionally, LR diets stimulated hepatic bile acid synthesis via upregulating the expression levels of cytochrome P450 family 7 subfamily A member 1 and cytochrome P450 family 27 subfamily A member 1, and downregulating farnesol X receptor (FXR) and small heterodimer partner (SHP) expression levels (p < 0.05). A lowered SID Lys:NE ratio affected the colonic microbial composition, characterized by increased relative abundances of YRC22, Parabacteroides, Sphaerochaeta, and Bacteroides, alongside a decreased in the proportion of Roseburia, f_Lachnospiraceae_g_Clostridium, Enterococcus, Shuttleworthia, Exiguobacterium, Corynebacterium, Subdoligranulum, Sulfurospirillum, and Marinobacter (p < 0.05). The alterations in microbial composition were accompanied by a decrease in colonic butyrate concentration (p < 0.1). The metabolomic analysis revealed that LR diets affected primary bile acid synthesis and AMPK signaling pathway (p < 0.05). And the mantel analysis indicated that Parabacteroides, Sphaerochaeta, f_Lachnospiraceae_g_Clostridium, Shuttleworthia, and Marinobacter contributed to the alterations in body metabolism. A reduced dietary SID Lys:NE ratio improves energy metabolism, stimulates lipogenesis, and inhibits lipolysis in finishing pigs by regulating the AMPKα/SIRT1/PGC-1α pathway and the FXR/SHP pathway. Parabacteroides and Sphaerochaeta benefited bile acids synthesis, whereas f_Lachnospiraceae_g_Clostridium, Shuttleworthia, and Marinobacter may contribute to the activation of the AMPK signaling pathway. Overall, body metabolism and colonic microbiota collectively controlled the lipid metabolism in finishing pigs. Full article
(This article belongs to the Special Issue Exploration of Sustainable Feed Resources and Pig Dietary Strategies)
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Figure 1

Figure 1
<p>Effects of dietary standardized ileal digestible lysine to net energy ratio on hepatic gene expression levels of energy metabolism (<b>A</b>), bile acid synthesis (<b>B</b>), lipogenesis (<b>C</b>), and lipolysis (<b>D</b>). AMPKα1: AMP-activated protein kinase α1; SIRT1: sirtuin 1; PGC-1α: peroxisome proliferator-activated receptor-γ coactivator-1α; FXR: farnesol X receptor; SHP: small heterodimer partner; CYP7A1: cytochrome P450 family 7 subfamily A member 1; CYP27A1: cytochrome P450 family 27 subfamily A member 1; CYP8B1: cytochrome P450 family 8 subfamily B member 1; CYP7B1: cytochrome P450 family 7 subfamily B member 1; FABP4: fatty acid transport protein 4; CD36: fatty acid translocase; SCD: stearoyl-CoA desaturase; ACCα: acetyl-coA carboxylase α; ACCβ: acetyl-coA carboxylase β; FATP2: fatty acid transport protein 2; HSL: hormone-sensitive lipase; ATGL: adipose triglyceride lipase; CPT: carnitine palmitoyl transferase; GAPDH: glyceraldehyde-3-phosphate dehydrogenase. Data are expressed as the means ± SEM, * indicates significant difference (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>Effects of dietary standardized ileal digestible lysine to net energy ratio on the diversity and structure of colonic microbiota in finishing pigs. (<b>A</b>) A Veen diagram based on ASVs level. (<b>B</b>) Chao 1 index. (<b>C</b>) Principal coordinate analysis (PCoA) based on the Jaccard distance. (<b>D</b>) The top 4 dominant phyla and (<b>E</b>) top 30 genera abundance of bacteria.</p>
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<p>Effects of dietary standardized ileal digestible lysine to net energy ratio on the composition of colonic microbiota in finishing pigs. (<b>A</b>) Linear discriminant analysis effect size of microbiota. (<b>B</b>–<b>N</b>) The statistical analysis of genera. Data are expressed as the means ± SEM; * indicates significant difference (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>Effects of dietary standardized ileal digestible lysine to net energy ratio on the concentration of short-chain fatty acids (SCFAs) and the correlation of differential genera with SCFAs. (<b>A</b>) Concentration of SCFAs in colon. Data are expressed as the means ± SEM; * indicates significant difference (<span class="html-italic">p</span> &lt; 0.05). (<b>B</b>) The correlation heatmap of differential genera with SCFAs. Cells are shaded with color exclusively when <span class="html-italic">p</span> &lt; 0.05; the orange and blue colors represent positive and negative relationship, respectively.</p>
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<p>Effects of dietary standardized ileal digestible lysine to net energy ratio on urine metabolic profiles of finishing pigs. (<b>A</b>) Discriminant analysis of orthogonal partial least squares analysis (OPLS−DA) based on the urine compounds data. (<b>B</b>) The volcano plot of metabolites in LR vs. HR groups, metabolites from distinct classes were presented in separate graph. Red dots represent upregulated metabolites in the LR group, and the blue dots represent downregulated metabolites. (<b>C</b>) Heatmap of 29 differentially expressed metabolites: rows, metabolites; columns, samples. (<b>D</b>) Overview of pathway analysis based on metabolite alterations (top 20 pathways were shown).</p>
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<p>Microbiota–metabolites correlation. Pairwise comparisons of differential metabolites are demonstrated with a color gradient denoting Spearman’s correlation. Correlation between microbiota and metabolic profiles were analyzed with mantel test. Edge width corresponds to the Mantel’s r statistic for the corresponding distance correlation, and red color and bule color denote positive and negative correlation, respectively.</p>
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<p>Potential mechanisms regulating lipid metabolism in finishing pigs fed diets with low standardized ileal digestible lysine to net energy ratio.</p>
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