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19 pages, 5921 KiB  
Article
Distribution of Trachelospermum jasminoides Under the Influence of Different Environmental Factors
by Huan Yu, Zhihang Zhuo, Zhipeng He, Quanwei Liu, Xinqi Deng and Danping Xu
Agriculture 2025, 15(3), 285; https://doi.org/10.3390/agriculture15030285 (registering DOI) - 28 Jan 2025
Abstract
Trachelospermum jasminoides (Lindl.) Lem. is a well-known herb with important medicinal and economic values. It is widely used in the treatment of inflammations in China. As global climate change intensifies, the ecological niche of plants has correspondingly shifted. Therefore, understanding the distribution of [...] Read more.
Trachelospermum jasminoides (Lindl.) Lem. is a well-known herb with important medicinal and economic values. It is widely used in the treatment of inflammations in China. As global climate change intensifies, the ecological niche of plants has correspondingly shifted. Therefore, understanding the distribution of suitable habitats for T. jasminoides under different climate conditions is of great significance for its cultivation, introduction, and conservation. This research utilizes the MaxEnt model in combination with the Geographic Information System (ArcGIS) to analyze the present and future potential habitat distributions of T. jasminoides. Based on 227 documented occurrence points and 15 ecological variables, the results emphasize that the key environmental limitations influencing the optimal habitats of T. jasminoides are the precipitation during the coldest quarter, the mean temperature of the driest quarter, precipitation in the warmest quarter, temperature seasonality (standard deviation × 100), and the human impact index. At present, the combined area of suitable and highly suitable habitats for T. jasminoides amounts to 15.76 × 104 km2, with the highly suitable habitats predominantly situated in East and Central China. Based on climate scenario forecasts, within the SSP1-2.6 climate scenario, the total suitable habitat area for T. jasminoides is projected to increase relative to the current situation. Nevertheless, in the SSP2-4.5 and SSP5-8.5 climate scenarios, the suitable habitat area is anticipated to initially rise and then decline. The distribution center is mainly concentrated in the provinces of Hunan and Jiangxi, with the centroid shifting southeastward compared to the current situation. The findings of this research offer valuable insights for the effective cultivation, preservation, and sustainable use of T. jasminoides resources. Full article
(This article belongs to the Section Ecosystem, Environment and Climate Change in Agriculture)
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<p>Distribution points of <span class="html-italic">T. jasminoides</span> in China during the current period (green solid circles indicate regions where the species is currently present).</p>
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<p>Correlation heatmap of bioclimatic factors (dark blue and dark red indicate that the absolute value of the correlation coefficient is greater than 0.8. The colors gradually transition from the sides towards the center, with the absolute value of the correlation coefficient also decreasing accordingly).</p>
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<p>ROC curve and AUC value (the AUC value represents the area under the ROC curve; a larger value indicates higher accuracy in the model’s predictions).</p>
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<p>Variable importance determined via the folding Jackknife.</p>
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<p>Response curves of dominant environmental factors ((<b>a</b>) denotes the response curve of the presence probability of <span class="html-italic">T. jasminoides</span> to the precipitation of coldest quarter; (<b>b</b>) denotes the response curve of the presence probability of <span class="html-italic">T. jasminoides</span> to the mean temperature of driest quarter; (<b>c</b>) denotes the response curve of the presence probability of <span class="html-italic">T. jasminoides</span> to the precipitation of warmest quarter; (<b>d</b>) denotes the response curve of the presence probability of <span class="html-italic">T. jasminoides</span> to the temperature seasonality (standard deviation × 100); (<b>e</b>) denotes the response curve of the presence probability of <span class="html-italic">T. jasminoides</span> to the human footprint index).</p>
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<p>Optimal survival areas for <span class="html-italic">T. jasminoides</span> under current climate conditions (yellow represents low suitable habitat distribution, red represents moderate suitable habitat distribution, and purple represents high suitable habitat distribution).</p>
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<p>Optimal survival areas for <span class="html-italic">T. jasminoides</span> under future climate conditions: (<b>a</b>) optimal survival area distribution of <span class="html-italic">T. jasminoides</span> under the SSP1-2.6 scenario in the 2050s; (<b>b</b>) optimal survival area distribution of <span class="html-italic">T. jasminoides</span> under the SSP1-2.6 scenario in the 2090s; (<b>c</b>) optimal survival area distribution of <span class="html-italic">T. jasminoides</span> under the SSP2-4.5 scenario in the 2050s; (<b>d</b>) optimal survival area distribution of <span class="html-italic">T. jasminoides</span> under the SSP2-4.5 scenario in the 2090s; (<b>e</b>) optimal survival area distribution of <span class="html-italic">T. jasminoides</span> under the SSP5-8.5 scenario in the 2050s; (<b>f</b>) optimal survival area distribution of <span class="html-italic">T. jasminoides</span> under the SSP5-8.5 scenario in the 2090s. Yellow indicates areas of low suitability, red indicates areas of moderate suitability, and purple indicates areas of high suitability.</p>
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<p>Migration of the <span class="html-italic">T. jasminoides</span> Centroid Under Future Climate Conditions (the blue arrows indicate the movement direction of the centroid in different years under the SSP5-8.5 emission hypothesis; the red arrows show the centroid movement direction in different years under the SSP2-4.5 emission hypothesis; the green arrows represent the movement direction of the centroid in different years under the SSP1-2.6 emission hypothesis; the solid black dot marks the current position of the centroid).</p>
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18 pages, 3629 KiB  
Article
Isolation, In Vitro Antioxidant Capacity, Hypoglycemic Activity and Immunoactivity Evaluation of Polysaccharides from Coriandrum sativum L.
by Weiwei Jin, Huan Zhou, Haijun Zhao, Yue Pei, Fengxian Su, Yan Li and Tao Luo
Antioxidants 2025, 14(2), 149; https://doi.org/10.3390/antiox14020149 - 27 Jan 2025
Viewed by 230
Abstract
Coriander (Coriandrum sativum) is a classical medicinal and edible herb as well as a spice, but the physicochemical and biological properties of its polysaccharides have not been fully studied. In this study, the polysaccharides were extracted using an ultrasonic-assisted method and [...] Read more.
Coriander (Coriandrum sativum) is a classical medicinal and edible herb as well as a spice, but the physicochemical and biological properties of its polysaccharides have not been fully studied. In this study, the polysaccharides were extracted using an ultrasonic-assisted method and purified from fresh coriander, and then the coriander polysaccharide (CSP) fraction was separated using an agarose gel Q-Sepharose Fast Flow column. The total sugar content, protein content and monosaccharides composition of CSPs were determined using a phenol–sulfuric acid method, Coomassie Brilliant Blue method and HPLC. The structural characterization was detected using ultraviolet spectrophotometry and FT-IR spectroscopy. DPPH and ABTS free radicals were used to explore their antioxidant activities, while the inhibitory abilities of α-amylase and α-glucosidase were used to evaluate their hypoglycemic activity. After that, the immunomodulatory and antitumor activities were investigated using macrophage RAW264.7 and HepG2 cells as the targets. The results showed that the total sugar and protein contents of CSPs were 66.90 ± 1.44% and 1.06 ± 0.32%, respectively. CSPs were mainly composed of fucose, rhamnose, arabinose, galactose, glucose, galacturonic acid and glucuronic acid, with a molar ratio of 1.13:15.11:9.60:25.98:1.55:44.33:2.29, and may be an acidic heteropolysaccharide containing pyran rings, α- and β-glycosidic bonds and glucuronic acid. Results from in vitro experiments of biological activities showed that the IC50 of CSPs for scavenging DPPH and ABTS free radicals were 0.759 mg/mL and 1.758 mg/mL, respectively; the IC50 values for inhibiting the activities of α-amylase and α-glucosidase were 0.634 mg/mL and 2.178 mg/mL, respectively; the CSPs with a concentration of 25~200 μg/mL showed no obvious toxicity to macrophage RAW264.7, and when treated with 100 μg/mL of CSPs, the relative cell phagocytosis capacity and secreted nitric oxide amount of RAW264.7 were 153.75 ± 12.01% and 133.56 ± 5.37%, respectively; CSPs showed a concentration-dependent ability to inhibit the growth of HepG2 cells within the test concentration of 0.25–2.0 mg/mL. Summarizing the results, due to their excellent antioxidant, immunomodulatory and anti-tumor activities, the coriander acid polysaccharides were expected to show good potential in comprehensive development of food and medicine. Full article
(This article belongs to the Special Issue Plant Materials and Their Antioxidant Potential, 2nd Edition)
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<p>Q Sepharose <sup>TM</sup> Fast Flow elution curve of coriander polysaccharides.</p>
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<p>UV–visible spectrum of CSPs.</p>
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<p>FT-IR spectroscopy of CSPs.</p>
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<p>Ion chromatograms of standard monosaccharides (<b>A</b>) and CSPs (<b>B</b>).</p>
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<p>In vitro DPPH and ABTS antioxidant activity of CSPs. Vc was used as a positive control. Different lowercase letters represent significant difference at 0.05 level.</p>
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<p>In vitro hypoglycemic activity of CSPs. Acarbose was used as a reference. Different lowercase letters represent significant difference at 0.05 level.</p>
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<p>Effects of CSPs on RAW264.7 cytotoxicity. Compared with the control group, *** represents <span class="html-italic">p</span> &lt; 0.001.</p>
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<p>Effects of CSPs on phagocytosis of RAW264.7 cells. Compared with the negative control group, * represents <span class="html-italic">p</span> &lt; 0.05 and *** represents <span class="html-italic">p</span> &lt; 0.001.</p>
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<p>Effects of CSPs on NO secretion in RAW264.7 cells. Compared with the blank control group, * represents <span class="html-italic">p</span> &lt; 0.05, ** represents <span class="html-italic">p</span> &lt; 0.01, and *** represents <span class="html-italic">p</span> &lt; 0.001.</p>
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<p>Effects of CSPs on proliferation of HepG2 cells. Compared with the control group, * represents <span class="html-italic">p</span> &lt; 0.05 and *** represents <span class="html-italic">p</span> &lt; 0.001.</p>
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16 pages, 1306 KiB  
Article
Chemical Composition of Essential Oil from Apium graveolens L. and Its Biological Activities Against Sitophilus zeamais Motschulsky (Coleoptera: Dryophthoridae)
by Ruchuon Wanna, Darika Bunphan, Benjapon Kunlanit, Phirayot Khaengkhan, Parinda Khaengkhan and Hakan Bozdoğan
Plants 2025, 14(3), 347; https://doi.org/10.3390/plants14030347 - 24 Jan 2025
Viewed by 359
Abstract
The use of essential oils from certain herbal plants offers a promising alternative to synthetic insecticides for controlling the maize weevil, Sitophilus zeamais Motschulsky (Coleoptera: Dryophthoridae), a major pest that causes significant damage to stored grains. Essential oils, particularly from aromatic herbs in [...] Read more.
The use of essential oils from certain herbal plants offers a promising alternative to synthetic insecticides for controlling the maize weevil, Sitophilus zeamais Motschulsky (Coleoptera: Dryophthoridae), a major pest that causes significant damage to stored grains. Essential oils, particularly from aromatic herbs in the Apiaceae family, are widely used in medicinal, cosmetic, and food industries and provided insecticidal properties to mitigate the environmental and health hazards associated with synthetic insecticides. This research aimed to investigate the insecticidal and repellent effects of Apium graveolens L. (celery) seed essential oil against S. zeamais. Chemical analysis of the commercially produced essential oil from A. graveolens seeds was conducted using a gas chromatograph–mass spectrometer (GC-MS), and the biological activity of the essential oil was determined by ingestion, contact, fumigation, and repellent tests. The analysis identified D-limonene (64.21%) and α-humulene (17.46%) as primary components of the oil. Toxicity assays revealed an observable contact toxicity, with higher concentrations and prolonged exposure increasing its effectiveness. The contact toxicity assays reported an LC50 of 19.83 nL/adult after 72 h. Additionally, the essential oil displayed repellent effects, effectively deterring weevils at concentrations above 16 µL/L air, but its feeding deterrence was weak. The essential oil’s strong insecticidal and repellent properties, which were concentration- and time-dependent, highlighted its potential as a sustainable alternative to synthetic pesticides for integrated pest management. Full article
(This article belongs to the Special Issue Green Insect Control: The Potential Impact of Plant Essential Oils)
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<p>The adult mortality of <span class="html-italic">S. zeamais</span> after contact with essential oil of <span class="html-italic">A. graveolens</span> seeds. There were no significant differences observed in the mortality within 24 h (<span class="html-italic">p</span> &gt; 0.05). The significant differences were found within 48–168 h (<span class="html-italic">p</span> &lt; 0.01). Means of the same period followed by the same letter were not significantly different (LSD: <span class="html-italic">p</span> &gt; 0.05).</p>
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<p>The adult mortality of <span class="html-italic">S. zeamais</span> after fumigation with essential oil of <span class="html-italic">A. graveolens</span> seeds. The data could not be statistically analyzed at 24 and 168 h. There were significant differences found within 48–144 h (<span class="html-italic">p</span> &lt; 0.05). Means of the same period followed by the same letter were not significantly different (LSD: <span class="html-italic">p</span> &gt; 0.05).</p>
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<p>The adult of <span class="html-italic">S. zeamais</span> was repelled after fumigation with essential oil of <span class="html-italic">A. graveolens</span> seeds. There were no significant differences observed in the mortality within 24–168 h (<span class="html-italic">p</span> &gt; 0.05), except significant differences were found within 48 h (<span class="html-italic">p</span> &lt; 0.05). Means of the same period followed by the same letter were not significantly different (LSD: <span class="html-italic">p</span> &gt; 0.05).</p>
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36 pages, 3075 KiB  
Review
Chromatographic and Spectroscopic Analyses of Cannabinoids: A Narrative Review Focused on Cannabis Herbs and Oily Products
by Céline Duchateau, Caroline Stévigny, Jehan Waeytens and Eric Deconinck
Molecules 2025, 30(3), 490; https://doi.org/10.3390/molecules30030490 - 22 Jan 2025
Viewed by 870
Abstract
Cannabis sativa L. is cultivated nowadays for agricultural, industrial, and medicinal applications and also for recreational use. The latter is due to the presence of delta-9-tetrahydrocannabinol, a psychoactive substance. Recreational cannabis policies vary between different countries, which has led to the lack of [...] Read more.
Cannabis sativa L. is cultivated nowadays for agricultural, industrial, and medicinal applications and also for recreational use. The latter is due to the presence of delta-9-tetrahydrocannabinol, a psychoactive substance. Recreational cannabis policies vary between different countries, which has led to the lack of a clearly defined legal context for cannabis and also a diversity of products derived from or containing cannabis on the (il)legal market. These cannabis-derived products have regained attention, notably because of their cannabinoid content. This review aims to assess and present analytical methods developed to analyze phytocannabinoids with spectroscopic and chromatographic techniques in specific cannabis matrices: herbs and oily products. Published papers from 2018–November 2024 were searched for with precise criteria, analyzed, and summarized. In the studies, liquid and gas chromatographic techniques (>70% reviewed papers) were the most used and have been widely applied using similar methods, and most papers were focused on cannabis herbs (>75%). Techniques were also compared and future challenges were identified. A comparison of different specificities of chromatographic and spectroscopic techniques discussed in this current review has also been established and summarized. Full article
(This article belongs to the Special Issue Review Papers in Analytical Chemistry)
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43 pages, 4236 KiB  
Review
Plant Polyphenols as Heart’s Best Friends: From Health Properties, to Cellular Effects, to Molecular Mechanisms of Action
by Sanja Stankovic, Slavica Mutavdzin Krneta, Dragan Djuric, Verica Milosevic and Dragan Milenkovic
Int. J. Mol. Sci. 2025, 26(3), 915; https://doi.org/10.3390/ijms26030915 - 22 Jan 2025
Viewed by 766
Abstract
Polyphenols are micronutrients found in fruits, vegetables, tea, coffee, cocoa, medicinal herbs, fish, crustaceans, and algae. They can also be synthesized using recombinant microorganisms. Interest in plant-derived natural compounds has grown due to their potential therapeutic effects with minimal side effects. This is [...] Read more.
Polyphenols are micronutrients found in fruits, vegetables, tea, coffee, cocoa, medicinal herbs, fish, crustaceans, and algae. They can also be synthesized using recombinant microorganisms. Interest in plant-derived natural compounds has grown due to their potential therapeutic effects with minimal side effects. This is particularly important as the aging population faces increasing rates of chronic diseases such as cancer, diabetes, arthritis, cardiovascular, and neurological disorders. Studies have highlighted polyphenols’ capacity to reduce risk factors linked to the onset of chronic illnesses. This narrative review discusses polyphenol families and their metabolism, and the cardioprotective effects of polyphenols evidenced from in vitro studies, as well as from in vivo studies, on different animal models of cardiac disease. This study also explores the molecular mechanisms underlying these benefits. Current research suggests that polyphenols may protect against ischemia, hypertension, cardiac hypertrophy, heart failure, and myocardial injury through complex mechanisms, including epigenetic and genomic modulation. However, further studies under nutritionally and physiologically relevant conditions, using untargeted multigenomic approaches, are needed to more comprehensively elucidate these mechanisms and firmly prove the cardioprotective effects of polyphenols. Full article
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<p>Mechanisms of action of resveratrol in cell models.</p>
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<p>Mechanisms of action of quercetin in cell models.</p>
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<p>Mechanisms of action of gallic acid in cell models.</p>
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<p>Mechanisms of action of resveratrol in animal models.</p>
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<p>The standardized mean differences (SMD) and 95% confidence intervals (95% CI) for (<b>a</b>) the myocardial injury and cardiac function parameters and (<b>b</b>) the oxidative stress and inflammation parameters in the meta-analysis of analyses of resveratrol’s cardioprotective effects by Zhang et al., 2024 [<a href="#B108-ijms-26-00915" class="html-bibr">108</a>]. CK—creatine kinase; CK-MB—creatine kinase–myocardial band; LVEDP—left ventricular end-diastolic pressure; HR—heart rate; ROS—reactive oxygen species; MDA—malondialdehyde; SOD—superoxide dismutase; TNF-α—tumor necrosis factor-alpha; IL-6—interleukin-6.</p>
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<p>Mechanisms of action of quercetin in animal models.</p>
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<p>Mechanisms of action of gallic acid in animal models.</p>
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<p>Mechanisms of action of curcumin in animal models.</p>
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<p>The standardized mean differences (SMD) and 95% confidence intervals (95% CI) according to data from meta-analysis of epigallocatechin gallate’s cardioprotective effects in preclinical models by Wei et al. 2023 [<a href="#B122-ijms-26-00915" class="html-bibr">122</a>]. LVDP—left ventricular developed pressure; LVEDP—left ventricular end-diastolic pressure; LVSP—left ventricular systolic pressure; +dP/dt max—maximum rate of rise in left ventricular pressure; −dP/dt max—maximum rate of decline in left ventricular pressure; MDA—malondialdehyde; SOD—superoxide dismutase; CAT—catalase; CK—creatine kinase; CK-MB—creatine kinase–myocardial band; LDH—lactate dehydrogenase; TnT—troponin T.</p>
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<p>The standardized mean differences (SMD) and 95% confidence intervals (95% CI) according to data from two meta-analyses of curcumin’s cardioprotective effects in preclinical models by Zeng et al. 2023 [<a href="#B117-ijms-26-00915" class="html-bibr">117</a>]—blue and Li et al. 2023 [<a href="#B118-ijms-26-00915" class="html-bibr">118</a>]—red. LVEF—left ventricular ejection fraction; LVFS—left ventricular fractional shortening; LDH— lactate dehydrogenase; MDA—malondialdehyde; SOD—superoxide dismutase; CAT—catalase; TNF-α—tumor necrosis factor-alpha.</p>
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<p>A summary of health effect of polyphenols on the heart from preclinical studies, and heart cells from in vitro studies, together with potential underlying molecular mechanisms of action.</p>
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23 pages, 9197 KiB  
Article
Comparative Analysis of Metabolites of Wild and Cultivated Notopterygium incisum from Different Origins and Evaluation of Their Anti-Inflammatory Activity
by Fukang Kong, Yannan Kou, Xu Zhang, Yue Tian, Bin Yang and Weihao Wang
Molecules 2025, 30(3), 468; https://doi.org/10.3390/molecules30030468 - 22 Jan 2025
Viewed by 331
Abstract
The dried rhizome of Notopterygium incisum (NI) from the Umbelliferae family, genuinely produced in Sichuan, China, is a classic traditional Chinese medicinal herb for treating wind-dampness arthralgia. Due to scarce natural resources, wild NI is gradually being replaced by cultivated types. However, knowledge [...] Read more.
The dried rhizome of Notopterygium incisum (NI) from the Umbelliferae family, genuinely produced in Sichuan, China, is a classic traditional Chinese medicinal herb for treating wind-dampness arthralgia. Due to scarce natural resources, wild NI is gradually being replaced by cultivated types. However, knowledge is limited regarding the differences in chemical composition and pharmacological effects between wild and cultivated NI and between Sichuan-grown and other-region-grown NI. In this study, a plant metabolomics strategy, based on GC–MS and UHPLC-Orbitrap MS, was employed to compare metabolic profiles between wild and cultivated NI and between cultivated NI from Sichuan and cultivated NI from Gansu and Qinghai. In total, 195 metabolites were identified, and the biosynthetic pathways of coumarins and phenolic acids, which were the most abundant secondary metabolites in NI, were summarized. Additionally, seven key metabolic intermediates were uncovered, revealing the reasons for the differences in metabolic profiles between wild and cultivated NI. The anti-inflammatory effect of wild and cultivated NI was verified by inflammatory gene expression and neutrophil count using a zebrafish yolk sac inflammation model. Overall, this study presents information on the types and synthesis of pharmacodynamic substances in NI and provides a basis for its cultivation and applications. Full article
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<p>Morphological features of wild and cultivated <span class="html-italic">N. incisum</span> originating from different places. G: Gansu Province; H: Qinghai Province; S: Sichuan Province; W: wild; C: cultivated.</p>
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<p>Multivariate statistical analyses of volatile components in nine NI samples (<b>A</b>) GC–MS (gas chromatography–mass spectrometry) chromatogram of wild NI from Sichuan Province. (<b>B</b>) PCA (principal component analysis) plot showing separation of samples based on their volatile component profiles. (<b>C</b>) Clustering heat map of NI samples based on their volatile component profiles. (<b>D</b>) OPLS-DA score plot and permutation test diagram of wild vs. cultivated NI based on their volatile component profiles. 1.0029*t[1] represents the scaling factor (1.0029) multiplied by t[1]; 1.10457*to[1] represents the scaling factor (1.10457) multiplied by to[1] (<b>E</b>) OPLS-DA score plots and permutation test plots of cultivated NI from Sichuan vs. cultivated NI from Qinghai and Gansu based on volatile oil components. 1.0002*t[1] represents the scaling factor (1.0002) multiplied by t[1]; 1.09302*to[1] represents the scaling factor (1.09302) multiplied by to[1] (<b>F</b>) Volcano plot showing differential volatile components between wild NI and cultivated NI. (<b>G</b>) Volcano plot showing differential volatile components between NI cultivated in Sichuan and NI cultivated in Qinghai and Gansu. G: Gansu Province; H: Qinghai Province; S: Sichuan Province; W: wild; C: cultivated.</p>
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<p>TIC (total ion chromatogram) of QC sample acquired by UHPLC-Orbitrap MS. (<b>A</b>) TIC in positive ion mode; (<b>B</b>) TIC in negative ion mode.</p>
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<p>Multivariate statistical analysis of non-volatile metabolites in different batches of NI. (<b>A</b>) Sector chart of the proportion of various components in wild and cultivated NIs from different growing areas. (<b>B</b>) PCA score diagram of samples. (<b>C</b>) Clustering heat map based on contents of constituents in nine batches of NI samples. (<b>D</b>) OPLS-DA score diagram and permutation test result for wild NI vs. cultivated NI. (<b>E</b>) OPLS-DA score diagram and permutation test result for cultivated NI from Sichuan vs. cultivated NI from Qinghai and Gansu. (<b>F</b>) Volcano plot of DAMs between wild and cultivated NI. (<b>G</b>) Volcano diagram of DAMs between cultivated NI from Sichuan and cultivated NI from Qinghai and Gansu. G: Gansu Province; H: Qinghai Province; S: Sichuan Province; W: wild; C: cultivated.</p>
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<p>Pathway enrichment analysis of DAMs. (<b>A</b>) Enrichment analysis of metabolic pathways enriched with DAMs in W vs. C. (<b>B</b>) Enrichment analysis of metabolic pathways enriched with DAMs in SC vs. HC and GC.</p>
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<p>Phenolic acid and coumarin biosynthesis pathways construct in nine batches of NI. G: Gansu Province; H: Qinghai Province; S: Sichuan Province; W: wild; C: cultivated.</p>
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<p>Chemical structure of seven important intermediates.</p>
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<p>Correlation analysis between amino acids and key phenolic acid and coumarin components in NI. Asterisks indicate significance of correlation (* significant at <span class="html-italic">p</span> &lt; 0.05; ** significant at <span class="html-italic">p</span> &lt; 0.01). Oval in red color: positive correlation; oval in blue color: negative correlation.</p>
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<p>Neutrophil visualization and counts as indexes of anti-inflammatory effects of SW and SC-2 in the zebrafish yolk sac system. (<b>A</b>) Recruitment of neutrophils to zebrafish yolk sac in control, model, and positive control groups, as well as in groups treated with Sichuan wild NI (SW) and Sichuan cultivated NI (SC-2) at different doses. (<b>B</b>) Histogram of neutrophil counts in zebrafish yolk sac treated with different doses of SW. (<b>C</b>) Histogram of neutrophil counts in zebrafish yolk sac treated with different doses of SC-2. Yellow dashed area in (<b>A</b>) represents the location of zebrafish yolk sac analysis; green fluorescent particles represent neutrophils. Compared with model group, *, **, and *** indicate significant differences at <span class="html-italic">p</span> &lt; 0.05, <span class="html-italic">p</span> &lt; 0.01, and <span class="html-italic">p</span> &lt; 0.001, respectively.</p>
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<p>qRT-PCR results showing transcript levels of genes encoding inflammatory factors in the zebrafish inflammation model. (<b>A</b>) Histogram of transcript levels of genes encoding inflammation markers in zebrafish after SW administration. (<b>B</b>) Histogram of transcript levels of genes encoding inflammation markers in zebrafish after SC-2 administration. Compared with model group, * and ** indicate significant difference at <span class="html-italic">p</span> &lt; 0.05 and <span class="html-italic">p</span> &lt; 0.01, respectively.</p>
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27 pages, 9904 KiB  
Article
Phenolic Compounds and Pharmacological Potential of Lavandula angustifolia Extracts for the Treatment of Neurodegenerative Diseases
by Olha Mykhailenko, Viktoriia Hurina, Nataliia Herbina, Yuliia Maslii, Liudas Ivanauskas, Inna Vladymyrova, Dmytro Lytkin, Zigmantas Gudžinskas, Hanna Severina, Olena Ruban and Victoriya Georgiyants
Plants 2025, 14(2), 289; https://doi.org/10.3390/plants14020289 - 20 Jan 2025
Viewed by 666
Abstract
The search for neuroprotective compounds in lavender is driven by its traditional use for brain health, with antioxidant activity serving as a key mechanism in reducing oxidative stress and supporting cognitive function. Lavender’s potential to protect neurons is based on its calming, anti-stress [...] Read more.
The search for neuroprotective compounds in lavender is driven by its traditional use for brain health, with antioxidant activity serving as a key mechanism in reducing oxidative stress and supporting cognitive function. Lavender’s potential to protect neurons is based on its calming, anti-stress properties, which increase the brain’s resistance to neurodegeneration. Although lavender is not a traditional medicinal plant in Ukraine, it is increasingly recognised for its medicinal properties and is widely cultivated in the country. Lavender use in Ukraine is influenced by both global herbal practices and local medical traditions. The aim of this study was to optimise the preparation of lavender herb extracts, perform chemical profiling and evaluate their antioxidant and neuroprotective activities. The study focused on Lavandula angustifolia cultivated in Lviv, Ukraine. Modern analytical methods were used, including HPLC, spectrophotometry, molecular docking, lyophilisation and pharmacological testing. The selection of the optimal conditions for obtaining lavender herb extracts was determined on the basis of the results of the total yield of phenolic compounds in each extract, where it was found that the raw material–solvent ratio (1:10) in water and 50% ethanol gave the highest yield of substances; the preferred extraction time was 20 min, and the temperature was 60–70 °C, especially for water extraction. Further HPLC analysis identified marker compounds including rosmarinic acid (28.31 mg/g), chlorogenic acid (1.64 mg/g) and luteolin (0.23 mg/g) in the lyophilised ethanol extract, which were previously recognised as neuroprotective markers by molecular docking. The water extract showed higher antioxidant (total 50.85 mg/g) and neuroprotective activity, probably due to synergistic interactions among the components. Behavioural tests further demonstrated the neuroprotective potential of lavender herb. These results demonstrate the potential neuroprotective activity of lavender herb and open new possibilities for its use in the treatment of various neurodegenerative diseases. Full article
(This article belongs to the Section Phytochemistry)
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<p>The design of the experiment.</p>
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<p><span class="html-italic">Lavandula angustifolia</span> and its raw material: (<b>a</b>) general view of plants cultivated in the Lviv Botanical Garden of National University named after Ivan Franko (Lviv, Ukraine, 2022), photo by O. Mykhailenko; (<b>b</b>) general appearance of dry lavender herb; (<b>c</b>) separated flowers, leaves and stems of lavender.</p>
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<p>Quantity of phenolic compounds extracted from herb, flowers and stems of lavender. Whiskers represent the standard deviation. Different letters above the bars indicate significant differences (<span class="html-italic">p</span> &lt; 0.05) between variants.</p>
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<p>Effect of extraction time, type of extractant and ratio of HRM to extractant on total content of phenolic compounds from lavender herb. Whiskers represent standard deviation.</p>
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<p>Effect of temperature on quantity of phenolic compounds extracted from lavender herb. Different letters above bars indicate significant differences (<span class="html-italic">p</span> &lt; 0.05) between variants. Whiskers represent standard deviation.</p>
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<p>Combined conformational arrangement of the reference inhibitor donepezil (grey) and (<b>a</b>) apigenin (green) and 6,7-dihydroxyisoflavone (blue), and (<b>b</b>) chlorogenic (purple), neochlorogenic (blue) and rosmarinic acids (yellow) in the active site of AChE.</p>
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<p>Interaction of luteolin (<b>a</b>) and rosmarinic acid (<b>b</b>) with amino acid residues of the NMDAR inhibitor site.</p>
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<p>Typical HPLC chromatogram of lavender herb detected at 325 nm: 1—chlorogenic acid (RT 11.64 min); 2—rosmarinic acid (RT 37.18 min); 3—luteolin (RT 43.37 min).</p>
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<p>Typical HPLC-ABTS chromatogram of lavender herb extract (LLEE, 50% ethanol) detected at 325 nm (HPLC, green) and 650 nm (ABTS, red): 1—unknown compound; 2—chlorogenic acid; 3—rosmarinic acid; 4—luteolin.</p>
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21 pages, 3263 KiB  
Article
Effects of Echinacea purpurea and Alkylamides on Respiratory Virus Replication and IL-8 Expression In Vitro
by Keely Puchalski, Johanne A. Gerstel, Abiola Jimoh, Yalda Shokoohinia and Jeffrey Langland
Molecules 2025, 30(2), 386; https://doi.org/10.3390/molecules30020386 - 17 Jan 2025
Viewed by 359
Abstract
Echinacea purpurea is a perennial medicinal herb with important immunomodulatory and anti-inflammatory properties, especially purported for the alleviation of cold and flu symptoms. Different classes of secondary metabolites of the plant, such as alkylamides, caffeic acid derivatives, polysaccharides, flavonoids, and glycoproteins, are believed [...] Read more.
Echinacea purpurea is a perennial medicinal herb with important immunomodulatory and anti-inflammatory properties, especially purported for the alleviation of cold and flu symptoms. Different classes of secondary metabolites of the plant, such as alkylamides, caffeic acid derivatives, polysaccharides, flavonoids, and glycoproteins, are believed to be biologically and pharmacologically active. Although previous research suggests that the alkylamides present in Echinacea may be responsible for reducing the symptoms associated with the common cold or flu through their immunomodulatory activity, the roles of specific alkylamides and their targets (i.e., immune and/or antiviral) have not been well-elucidated or established. This study tested the antiviral and cytokine regulatory activity of various specific alkylamides that are present predominantly in Echinacea root extracts and found that one specific alkylamide, Dodeca-2E,4E-Dienoic acid isobutylamide, had potent antiviral activity against rhinovirus (the causative agent of most common colds) and influenza virus, as well as potent inhibition of IL-8 cytokine production. IL-8 is responsible for many of the symptoms associated with the common cold and is upregulated in other common respiratory infections. The broad activity and low cytotoxicity of this specific alkylamide support its potential use for treating rhinovirus and influenza virus infections. Full article
(This article belongs to the Special Issue Bioactive Compounds: Applications and Benefits for Human Health)
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<p>Viral plaque assay showing inhibitory effect of <span class="html-italic">Echinacea purpurea</span> extract on rhinovirus and influenza virus replication. HeLaH1 or MDCK cells were infected with either rhinovirus or influenza virus, respectively, in the presence of increasing concentrations (0–125 μg/mL) of <span class="html-italic">Echinacea purpurea</span> 70% ethanol root extract. After 72 h of infection, cells were fixed and stained with crystal violet to visualize viral plaques. Representative images of plaques from each treatment group are shown. Viral plaques were counted and normalized to untreated controls (0 µg). Vehicle controls (70% ethanol) were tested at the same volumes, corresponding to the treatment doses, and the results for the maximum volume used are graphed in <a href="#molecules-30-00386-f002" class="html-fig">Figure 2</a>. Results did not differ from untreated controls and had no effect on the treatment outcomes.</p>
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<p>Rhinovirus plaque reduction assay with <span class="html-italic">Echinacea</span> extract and phenolics. (<b>A</b>) Quantification of plaque numbers from rhinovirus plaque assay treated with <span class="html-italic">Echinacea</span> crude extract from <a href="#molecules-30-00386-f001" class="html-fig">Figure 1</a>. (<b>B</b>) HeLaH1 cells were infected with rhinovirus in the presence of increasing concentrations (0–16 μg/mL) of five phenolic compounds: echinacoside, cafteric acid, chlorogenic acid, cichoric acid, and dicaffeoylquinic acid. After 72 h of infection, cells were fixed and stained with crystal violet to visualize plaques. Viral plaques were counted and normalized to untreated controls (0 µg). Statistical significance was determined using a one-sample <span class="html-italic">t</span>-test. Significant differences are indicated by <span class="html-italic">p</span> &lt; 0.001 (***). Error bars represent the mean ± SD (<span class="html-italic">n</span> = 3). Vehicle controls (70% ethanol for the crude extract) were tested at the same volumes, and the results for the maximum volume used are shown in (<b>A</b>). For phenolic compounds (<b>B</b>), DMSO vehicle doses corresponding to the tested concentrations are shown to account for any potential dose-dependent effects of the solvent. These controls did not differ from untreated controls and had no effect on the treatment outcomes.</p>
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<p>Rhinovirus plaque reduction assay with <span class="html-italic">Echinacea</span> extract and alkylamides. (<b>A</b>) Quantification of plaque numbers from rhinovirus plaque assay treated with <span class="html-italic">Echinacea</span> crude extract from <a href="#molecules-30-00386-f001" class="html-fig">Figure 1</a>. (<b>B</b>) HeLaH1 cells were infected with rhinovirus in the presence of increasing concentrations (0–16 μg/mL) of various alkylamide compounds: A, B, C, and D (see key for compound names). After 72 h of infection, cells were fixed and stained with crystal violet to visualize plaques. Viral plaques were counted and normalized to untreated controls (0 µg). Statistical significance was determined using a one-sample <span class="html-italic">t</span>-test. Significant differences are indicated by <span class="html-italic">p</span> &lt; 0.01 (**), and <span class="html-italic">p</span> &lt; 0.001 (***). Error bars represent the mean ± SD (<span class="html-italic">n</span> = 3). (<b>C</b>) LC-MS analysis of the crude ethanolic extract was performed using UV detection at 330 nm, with the four alkylamide peaks (A–D) measured as absorbance values in milli-absorbance units (mAU). Alkylamides A1 and A2 represent <span class="html-italic">E</span>/<span class="html-italic">Z</span> isomers of alkylamide A (Dodeca-2<span class="html-italic">E</span>,4<span class="html-italic">E</span>,8<span class="html-italic">Z</span>,10(<span class="html-italic">E</span>/<span class="html-italic">Z</span>)-TE acid isobutylamide). When the activity of the alkylamides (<b>B</b>) was compared to the concentrations of alkylamides quantified in the crude <span class="html-italic">Echinacea</span> extract ((<b>A</b>), alk/mL), comparable concentrations of total alkylamides present in the crude extract had similar levels of activity relative to the pure alkylamides (with a viral IC80 around 2 µg/mL for both the crude extract and alkylamides C and D). Vehicle controls (70% ethanol for the crude extract) were tested at the same volumes, and the results for the maximum volume used are shown in (<b>A</b>). For alkylamide compounds (<b>B</b>), DMSO vehicle doses corresponding to the tested concentrations are shown to account for any potential dose-dependent effects of the solvent. These controls did not differ from untreated controls and had no effect on the treatment outcomes.</p>
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<p>Influenza plaque reduction assay with <span class="html-italic">Echinacea</span> extract and phenolics. (<b>A</b>) Quantification of plaque numbers from influenza virus plaque assay treated with <span class="html-italic">Echinacea</span> crude extract in <a href="#molecules-30-00386-f001" class="html-fig">Figure 1</a>. (<b>B</b>) MDCK cells were infected with influenza virus in the presence of increasing concentrations (0–16 µg/mL) of various phenolic compounds: echinacoside, cafteric acid, chlorogenic acid, cichoric acid, and dicaffeoylquinic acid. After 72 h of infection, cells were fixed and stained with crystal violet to visualize plaques. Viral plaques were counted and normalized to untreated controls (0 µg). Statistical significance was determined using a one-sample <span class="html-italic">t</span>-test. Significant differences are indicated by <span class="html-italic">p</span> &lt; 0.01 (**), and <span class="html-italic">p</span> &lt; 0.001 (***). Error bars represent the mean ± SD (<span class="html-italic">n</span> = 3). Vehicle controls (70% ethanol for the crude extract) were tested at the same volumes, and the results for the maximum volume used are shown in (<b>A</b>). For phenolic compounds (<b>B</b>), DMSO vehicle doses corresponding to the tested concentrations are shown to account for any potential dose-dependent effects of the solvent. These controls did not differ from untreated controls and had no effect on the treatment outcomes.</p>
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<p>Influenza plaque reduction assay with <span class="html-italic">Echinacea</span> extract and alkylamides. (<b>A</b>) Quantification of plaque numbers from influenza virus plaque assay treated with <span class="html-italic">Echinacea</span> crude extract in <a href="#molecules-30-00386-f001" class="html-fig">Figure 1</a>. (<b>B</b>): MDCK cells were infected with influenza virus in the presence of increasing concentrations (0–16 µg/mL) of various alkylamide compounds: A, B, C, and D (see Key for actual compound names). After 72 h of infection, cells were fixed and stained with crystal violet to visualize plaques. Viral plaques were counted and normalized to untreated controls (0 µg). Statistical significance was determined using a one-sample <span class="html-italic">t</span>-test. Significant differences are indicated by <span class="html-italic">p</span> &lt; 0.05 (*), <span class="html-italic">p</span> &lt; 0.01 (**), and <span class="html-italic">p</span> &lt; 0.001 (***). Error bars represent the mean ± SD (<span class="html-italic">n</span> = 3). Vehicle controls (70% ethanol for the crude extract) were tested at the same volumes, and the results for the maximum volume used are shown in (<b>A</b>). For alkylamide compounds (<b>B</b>), DMSO vehicle doses corresponding to the tested concentrations are shown to account for any potential dose-dependent effects of the solvent. These controls did not differ from untreated controls and had no effect on the treatment outcomes.</p>
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<p>Regulation of LPS-induced IL-8 secretion with <span class="html-italic">Echinacea</span> extract and alkylamides. Interleukin-8 (IL-8) levels were measured by ELISA in human THP-1 cells treated with varying concentrations of <span class="html-italic">Echinacea</span> root 70% ethanol extract (50 µg/mL) or the various alkylamides (1 µg/mL) in the presence or absence of LPS (1 µg/mL). Statistical significance was determined using a one-sample <span class="html-italic">t</span>-test. Significant differences are indicated by <span class="html-italic">p</span> &lt; 0.05 (*), <span class="html-italic">p</span> &lt; 0.01 (**), and <span class="html-italic">p</span> &lt; 0.001 (***). Error bars represent the mean ± SD (<span class="html-italic">n</span> = 3). Vehicle controls (70% ethanol and DMSO) were tested at the same volumes corresponding to the treatment doses, both with and without LPS, and were found to have no significant effect on IL-8 production compared to mock controls.</p>
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<p>CC<sub>50</sub> and IC<sub>50</sub> of <span class="html-italic">E. purpurea</span> root extract and effective alkylamides. (<b>A</b>) To determine the CC<sub>50</sub> of alkylamides C (Dodeca-2<span class="html-italic">E</span>-ENE-8,10-Diynoic acid isobutylamide) and D (Dodeca-2<span class="html-italic">E</span>,4<span class="html-italic">E</span>-Dienoic acid isobutylamide) and the <span class="html-italic">E. purpurea</span> EtOH extract, MTS assays were performed. HeLaH1 or MDCK cells were dosed with increasing concentrations (10–1000 µg/mL) of either <span class="html-italic">E. purpurea</span> extract or individual alkylamides and were incubated for 24 h, followed by addition of the MTS reagent. Absorbance was measured using a microplate reader. (<b>B</b>) The CC<sub>50</sub> was determined by plotting the treatment concentration (x axis) vs. cell viability (y axis) dose–response curve and analyzing viability as a percentage of the control. The IC<sub>50</sub> was determined from the viral plaque assay results in <a href="#molecules-30-00386-f003" class="html-fig">Figure 3</a> and <a href="#molecules-30-00386-f005" class="html-fig">Figure 5</a>. The selectivity index (SI) was then calculated.</p>
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17 pages, 295 KiB  
Review
A Review of Chemical Variability and Metal Contamination of Herbaceous Plants in Terms of Health Safety—A Case Study of Tanacetum vulgare L.
by Anna Turek
Appl. Sci. 2025, 15(2), 911; https://doi.org/10.3390/app15020911 - 17 Jan 2025
Viewed by 479
Abstract
The article presents a review of studies on the chemical diversity of essential oils and extracts from Tanacetum vulgare L. under the influence of environmental factors and the bioaccumulation of heavy metals in the plant. These issues are crucial for consumer safety, especially [...] Read more.
The article presents a review of studies on the chemical diversity of essential oils and extracts from Tanacetum vulgare L. under the influence of environmental factors and the bioaccumulation of heavy metals in the plant. These issues are crucial for consumer safety, especially when they concern plants widely used in alternative medicine. Stress factors to which plants are subjected may result in a decrease in their biological activity or toxic effects (as in the case of Tanacetum vulgare L.). The issue is particularly important in the context of increasing environmental pollution, e.g., with heavy metals, and climate change, which is also a stress factor. The presented review clearly shows that detailed research is required both to identify the components of T. vulgare with specific therapeutic potential and to assess their quantitative changes under the influence of a specific stressor. It was found that there is a need to standardize analytical procedures used in research on the bioaccumulation of metals by medicinal plants. The importance of environmental monitoring in herb-growing areas, as an important element of health policy, is also emphasized. Full article
21 pages, 8794 KiB  
Article
Preparation of Ethosome Gel with Total Flavonoids from Vernonia anthelmintica (L.) Willd. for the Treatment of Vitiligo
by Dongmei Qin, Yongjie Cui, Mengyue Zheng, Zhiguo Yang and Xinbing Wang
Gels 2025, 11(1), 73; https://doi.org/10.3390/gels11010073 - 17 Jan 2025
Viewed by 486
Abstract
Vernonia anthelmintica (L.) Willd. is a traditional medicinal herb in Chinese medicine, extensively used by various ethnic groups due to the numerous advantages derived from its total flavonoids. These benefits encompass anti-inflammatory and antioxidant effects, and the promotion of melanin production, showcasing its [...] Read more.
Vernonia anthelmintica (L.) Willd. is a traditional medicinal herb in Chinese medicine, extensively used by various ethnic groups due to the numerous advantages derived from its total flavonoids. These benefits encompass anti-inflammatory and antioxidant effects, and the promotion of melanin production, showcasing its significant efficacy in addressing vitiligo. To improve transdermal absorption and enhance the antioxidant effectiveness of the treatment, ethosome containing total flavonoids were prepared utilizing the ultrasound injection technique. The resulting ethosome was then carefully mixed with 0.7% Carbomer 934 gel in equal parts, yielding a gel concentration of 0.302 mg/g. This formulation produced small, consistent ethosome that exhibited high encapsulation efficiency and notable stability. In vitro analyses demonstrated sustained release characteristics of the gel and considerable therapeutic effectiveness against vitiligo resulting from hydroquinone exposure. Histological examinations performed through hematoxylin and eosin (H&E) staining of mouse skin revealed increased melanin production and increased activities of tyrosinase (TYR), cholinesterase (CHE), and mouse monoamine oxidase (MAO), while levels of superoxide dismutase (SOD) and malondialdehyde (MDA) were reduced. These findings underscore the promising effectiveness of this treatment strategy and validate the efficacy of the dosage form. Full article
(This article belongs to the Section Gel Applications)
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<p>Appearance and characterization of three ethosomes. (<b>a</b>) Particle size distribution of three ethosomes. (<b>b</b>) Appearance and zeta potential distribution of ethanol ethosome. (<b>c</b>) Appearance and zeta potential distribution of 1,2-propanediol ethosome. (<b>d</b>) Appearance and zeta potential distribution of mixture ethosome.</p>
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<p>Effect of cholesterol/lecithin ratio on total flavonoid ethosome. (<b>a</b>) Effect of cholesterol/lecithin ratio on size of total flavonoid ethosome. (<b>b</b>) Effect of cholesterol/lecithin ratio on PDI of total flavonoid ethosome. (<b>c</b>) Effect of cholesterol/lecithin ratio on zeta potential of total flavonoid ethosome. (<b>d</b>) Effect of cholesterol/lecithin ratio on encapsulation efficiency of total flavonoid ethosome. (<b>e</b>) Appearance of total flavonoid ethosome with different cholesterol/lecithin ratio.</p>
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<p>Effect of alcohol ratio on the total flavonoid ethosome of <span class="html-italic">Vernonia anthelmintica</span>. (<b>a</b>) Effect of alcohol ratio on size of total flavonoid ethosome. (<b>b</b>) Effect of alcohol ratio on PDI of total flavonoid ethosome. (<b>c</b>) Effect of alcohol ratio on zeta potential of total flavonoid ethosome. (<b>d</b>) Effect of alcohol ratio on encapsulation efficiency of total flavonoid ethosome. (<b>e</b>) Appearance of total flavonoid ethosome with different alcohol ratio.</p>
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<p>Effect of ethanol/1,2-propanediol ratio on the total flavonoid ethosome. (<b>a</b>) Effect of ethanol/1,2-propanediol ratio on size of total flavonoid ethosome. (<b>b</b>) Effect of ethanol/1,2-propanediol ratio on PDI of total flavonoid ethosome. (<b>c</b>) Effect of ethanol/1,2-propanediol ratio on zeta potential of total flavonoid ethosome. (<b>d</b>) Effect of ethanol/1,2-propanediol ratio on encapsulation efficiency of total flavonoid ethosome. (<b>e</b>) Appearance of total flavonoid ethosome with different ethanol/1,2-propanediol ratios.</p>
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<p>Effect of soy lecithin concentration on the total flavonoid ethosome of <span class="html-italic">Vernonia anthelmintica</span>. (<b>a</b>) Effect of soy lecithin concentration on size of total flavonoid ethosome. (<b>b</b>) Effect of soy lecithin concentration on PDI of total flavonoid ethosome. (<b>c</b>) Effect of soy lecithin concentration on zeta potential of total flavonoid ethosome. (<b>d</b>) Effect of soy lecithin concentration on encapsulation efficiency of total flavonoid ethosome. (<b>e</b>) Appearance of total flavonoid ethosome with different soy lecithin concentrations.</p>
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<p>Appearance and particle size potential distribution of total flavonoid ethosome of <span class="html-italic">Vernonia anthelmintica</span>.</p>
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<p>Transmission electron microscope (TEM) image of total flavonoid ethosome from <span class="html-italic">Vernonia anthelmintica</span> (×100 k).</p>
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<p>Stability study of the total flavonoid ethosome.</p>
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<p>Appearance of ethosome gel with different drug/matrix ratios: (<b>A</b>) 1:1; (<b>B</b>) 1:3; (<b>C</b>) 1:6.</p>
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<p>SEM characterization of ethosome gel (×100 k).</p>
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<p>Visualization of mouse skin tissue. (<b>A</b>) Control group. (<b>B</b>) Model group. (<b>C</b>) Ethosome gel group. (<b>D</b>) Solution group. (<b>E</b>) KL Group.</p>
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<p>Staining results of mouse skin tissue sections and quantification of melanin. Data were evaluated using one-way ANOVA (mean ± SD, <span class="html-italic">n</span> = 6). <sup>##</sup> <span class="html-italic">p</span> &lt; 0.01 vs. control; ** <span class="html-italic">p</span> &lt; 0.01 vs. model.</p>
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<p>Effects of ethosome gel on serum index in model mice. (<b>A</b>) Effects of ethosome gel on TYR. (<b>B</b>) Effects of ethosome gel on CHE. (<b>C</b>) Effects of ethosome gel on MAO. (<b>D</b>) Effects of ethosome gel on SOD. (<b>E</b>) Effects of ethosome gel on MDA. Data were evaluated using one-way ANOVA (mean ± SD, <span class="html-italic">n</span> = 6). <sup>##</sup> <span class="html-italic">p</span> &lt; 0.01 vs. control. * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01 vs. model.</p>
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25 pages, 7816 KiB  
Article
Genome-Wide Identification of CONSTANS-like (COL) Gene Family and the Potential Function of ApCOL08 Under Salt Stress in Andrographis paniculata
by Yizhu Zhao, Jiahao Xu, Xinyi Xu, Hui Liu, Qinxiang Chang, Ling Xu and Zongsuo Liang
Int. J. Mol. Sci. 2025, 26(2), 724; https://doi.org/10.3390/ijms26020724 - 16 Jan 2025
Viewed by 321
Abstract
Andrographis paniculata is an important medicinal herb known as a “natural antibiotic”, which has been used in Southeast Asia for thousands of years. The CONSTANS-like (COL) gene is an important regulatory factor for plant photoperiod flowering and stress response. However, there [...] Read more.
Andrographis paniculata is an important medicinal herb known as a “natural antibiotic”, which has been used in Southeast Asia for thousands of years. The CONSTANS-like (COL) gene is an important regulatory factor for plant photoperiod flowering and stress response. However, there is currently no detailed research on the COL genes of A. paniculata. In our study, we performed a genome-wide analysis of A. paniculata COL (ApCOL) family members using bioinformatics tools and identified nine ApCOL genes. Based on phylogenetic analysis, ApCOLs were categorized into three groups, with members of the same group having similar structures. Gene duplication events indicated that only one pair of duplicated genes was identified, possibly caused by segmental duplication. In terms of evolutionary relationships, the COL proteins of A. paniculata and Sesamum indicum were closely related, showing that they are highly similar in the phylogenetic tree. In addition, ApCOL genes showed tissue specificity and were specifically highly expressed mainly in leaves and flowers. Based on the cis-regulatory element prediction results, we examined the expression levels of ApCOLs under hormone and salt stress, and ApCOL08 was significantly induced. With subcellular localization results consistent with the prediction, we transformed ApCOL08 into yeast and showed significant resistance to salt stress. Our study suggests that ApCOL genes have important roles in response to abiotic stress and plant development and initially identifies key genes for future molecular regulation studies. Full article
(This article belongs to the Section Molecular Plant Sciences)
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<p>Interchromosomal homology analysis of <span class="html-italic">ApCOL</span> gene in the <span class="html-italic">A. paniculata</span> genome. The purple line represents the replication events of the <span class="html-italic">ApCOL</span> genes, and the gray line represents the syntenic blocks in the <span class="html-italic">A. paniculata</span> genome.</p>
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<p>NJ Phylogenetic tree of eight plant COL proteins. The phylogenetic tree is divided into three groups and six subgroups, each marked with different colors. Circular nodes represent Bootstrap values above 50%.</p>
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<p>The Nightingale Rose Chart calculates the number of <span class="html-italic">COL</span> members in eight species and shows the number of members in the three groups. The eight plants include <span class="html-italic">A. paniculata</span>, Arabidopsis (<span class="html-italic">A.thaliana</span>), soybean (<span class="html-italic">G. max</span>), sesame (<span class="html-italic">S. indicum</span>), tomato (<span class="html-italic">S. lycopersicum</span>), grape (<span class="html-italic">V. vinifera</span>), rice (<span class="html-italic">O. sativa</span>), and maize (<span class="html-italic">Z. mays</span>).</p>
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<p>Phylogenetic tree, conserved motifs, and gene structure analyses of <span class="html-italic">A. paniculata ApCOL</span> genes. (<b>A</b>) Evolutionary relationships of the nine <span class="html-italic">ApCOL</span> genes. (<b>B</b>) Conserved motif composition of ApCOL proteins. (<b>C</b>) Distribution of conserved protein structural domains of <span class="html-italic">ApCOL</span>s. (<b>D</b>) Exon–intron structure analysis of <span class="html-italic">ApCOL</span>s. Exons and introns are represented by yellow boxes and gray lines, respectively.</p>
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<p>Distribution of cis-regulatory elements in the 2000 bp sequence of the promoter region of the <span class="html-italic">ApCOL</span> gene. Each cis-regulatory element is indicated by a different color and is positioned in the same position as the corresponding position of the promoter.</p>
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<p>Synteny relationship analysis of <span class="html-italic">COL</span> genes between <span class="html-italic">A. paniculata</span> and other plants. Gray lines in the background represent collinear blocks in the <span class="html-italic">A. paniculata</span> and other plant genomes, and green lines indicate synthetic <span class="html-italic">COL</span> gene pairs.</p>
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<p>Tissue-specific expression profiles of <span class="html-italic">ApCOL</span> genes in different tissues of <span class="html-italic">A. paniculata</span>. The logarithm of FPKM values was normalized for creating heatmaps. Different colors represent gene expression levels, with high expression levels approaching red and low expression levels approaching green. Genes with relative expression values &gt; 0.3 in each tissue are listed in the four squares of the right-hand panel, and no genes were expressed in seeds. L: Leaves; S: Stem; B: Branch; F: Flowers; SD: Seed.</p>
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<p>The expression profile of <span class="html-italic">ApCOL</span> genes in the leaves of <span class="html-italic">A. paniculata</span> under three PGR treatments (6-BA, SA, and NAA) was analyzed by qRT-PCR. The relative expression levels of genes were calculated by using the 2<sup>−ΔΔCt</sup> method. The UBC gene serves as an internal reference gene. The results were visualized as log<sub>2</sub>-fold changes. 6-BA-5-5/6-BA-5-10: <span class="html-italic">A. paniculata</span> treated with 5 μM 6-BA for 5 and 10 days; 6-BA-10-5/6-BA-10-10: <span class="html-italic">A. paniculata</span> treated with 10 μM 6-BA for 5 and 10 days; SA-5-5/SA-5-10: <span class="html-italic">A. paniculata</span> treated with 5 μM SA for 5 and 10 days; SA-10-5/SA-10-10: <span class="html-italic">A. paniculata</span> treated with 10 μM SA for 5 and 10 days; NAA-5-5/NAA-5-10: <span class="html-italic">A. paniculata</span> treated with 5 μM NAA for 5 and 10 days; NAA-10-5/NAA-10-10: <span class="html-italic">A. paniculata</span> treated with 10 μM NAA for 5 and 10 days.</p>
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<p>Expression analysis of <span class="html-italic">COL</span> genes in <span class="html-italic">A. paniculata</span> leaves under salt treatment. Expression analysis of <span class="html-italic">ApCOL</span>s in the leaves of <span class="html-italic">A. paniculata</span> under salt stress. (<b>A</b>) The expression profile of <span class="html-italic">ApCOL</span> genes under salt treatment was analyzed by qRT-PCR. The relative expression levels of genes were calculated by using the 2<sup>−ΔΔCt</sup> method. The UBC gene serves as an internal reference gene. The results were visualized as log<sub>2</sub>-fold changes. (<b>B</b>) Relative expression of nine <span class="html-italic">ApCOL</span>s in response to salt stress. The histogram was presented by the mean and standard error of the data. Lowercase letters “a, b, c, d” indicate a significant level of 0.05, and different letters indicate significant differences between groups (<span class="html-italic">p</span> &lt; 0.05). NaCl-50-5/NaCl-50-10: <span class="html-italic">A. paniculata</span> treated with 50 mM NaCl for 5 and 10 days; NaCl-100-5/NaCl-100-10: <span class="html-italic">A. paniculata</span> treated with 100 mM NaCl for 5 and 10 days.</p>
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<p>Expression analysis of <span class="html-italic">COL</span> genes in <span class="html-italic">A. paniculata</span> leaves under salt treatment. Expression analysis of <span class="html-italic">ApCOL</span>s in the leaves of <span class="html-italic">A. paniculata</span> under salt stress. (<b>A</b>) The expression profile of <span class="html-italic">ApCOL</span> genes under salt treatment was analyzed by qRT-PCR. The relative expression levels of genes were calculated by using the 2<sup>−ΔΔCt</sup> method. The UBC gene serves as an internal reference gene. The results were visualized as log<sub>2</sub>-fold changes. (<b>B</b>) Relative expression of nine <span class="html-italic">ApCOL</span>s in response to salt stress. The histogram was presented by the mean and standard error of the data. Lowercase letters “a, b, c, d” indicate a significant level of 0.05, and different letters indicate significant differences between groups (<span class="html-italic">p</span> &lt; 0.05). NaCl-50-5/NaCl-50-10: <span class="html-italic">A. paniculata</span> treated with 50 mM NaCl for 5 and 10 days; NaCl-100-5/NaCl-100-10: <span class="html-italic">A. paniculata</span> treated with 100 mM NaCl for 5 and 10 days.</p>
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<p>Subcellular localization of the ApCOL08 and ApCOL09 proteins in <span class="html-italic">A. paniculata</span>.</p>
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<p>Comparison of growth between INVSc1 (pYES2-<span class="html-italic">ApCOL08</span>) and INVSc1 (pYES2) under treatment with six concentrations of NaCl (0 M, 0.5 M, 1 M, 1.3 M, 1.5 M, 2 M). Photos were taken after incubating at 30 °C for 4 d and 7 d, respectively.</p>
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19 pages, 1749 KiB  
Article
In Vitro Determination of Antimicrobial, Antioxidant and Antiviral Properties of Greek Plant Extracts
by Aliki Tsakni, Eirini Kyriakopoulou, Sophia Letsiou, Panagiotis Halvatsiotis, Haralambos Rigopoulos, Niki Vassilaki and Dimitra Houhoula
Microorganisms 2025, 13(1), 177; https://doi.org/10.3390/microorganisms13010177 - 16 Jan 2025
Viewed by 674
Abstract
The medicinal potential of plant extracts, especially their antimicrobial, antioxidant, antiviral and cytotoxic properties, has gained significant attention in recent years. This study examined the in vitro bioactivities of several selected Greek medicinal plants, like Eucalyptus globulus L., Thymus vulgaris L., Salvia rosmarinus [...] Read more.
The medicinal potential of plant extracts, especially their antimicrobial, antioxidant, antiviral and cytotoxic properties, has gained significant attention in recent years. This study examined the in vitro bioactivities of several selected Greek medicinal plants, like Eucalyptus globulus L., Thymus vulgaris L., Salvia rosmarinus L. and Ocimum basilicum L., are well-known for their traditional therapeutic use. Minimum inhibitory concentration (MIC) assays were used to evaluate the antimicrobial activity of the extracts against pathogenic bacteria. The antioxidant activity was carried out using the DPPH method, while the cytotoxicity of the plants was determined using the Alamar Blue method. In addition, the antiviral efficacy of the samples was tested against DENV in different cell lines. The majority of medicinal herbs demonstrated significant antimicrobial action (MIC = 30–3000 μg∙mL−1). The extracts showed great antioxidant activity, while the Salvia rosmarinus L. extract turned out to be the most effective (IC50 = 12.89 ± 0.11 μg∙mL−1). In contrast, the extract of Eucalyptus globulus L. had the lowest antioxidant action (IC50 = 71.02 ± 0.42 μg∙mL−1). The results of the Alamar Blue method were presented with CC50 values, and it was shown that Eucalyptus globulus L. extract exhibited the highest cytotoxicity (CC50 = 5.94% v/v ± 0.04). Similarly, the results of the antiviral potential of extracts were expressed as EC50 values, and Eucalyptus globulus L. was characterized as the most effective sample against dengue virus infection, with EC50 values estimated at 2.37% v/v ± 0.6 (HuhD-2 cells infected with DENV-2) and 0.36% v/v ± 0.004 (Huh7.5 cells infected with DVR2A). These findings provide a foundation for further studies in order to combat infectious diseases and promote human health. Full article
(This article belongs to the Section Antimicrobial Agents and Resistance)
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<p>Capacity of the natural extracts to scavenge DPPH radicals.</p>
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<p>Comparison of antioxidant activity of natural extracts.</p>
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<p>Cytotoxicity of the plant extracts. * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01 and *** <span class="html-italic">p</span> &lt; 0.001 vs. control (Student’s <span class="html-italic">t</span>-test). # <span class="html-italic">p</span> &lt; 0.05, ## <span class="html-italic">p</span> &lt; 0.01 and ### <span class="html-italic">p</span> &lt; 0.001 across extracts (one-way ANOVA).</p>
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<p>Effect of plant extracts on DENV serotype 2 replication. Values from control cells were set to one hundred. Bars represent mean values from three independent experiments in triplicate. Error bars indicate standard deviations. * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01 and *** <span class="html-italic">p</span> &lt; 0.001 vs. control (Student’s <span class="html-italic">t</span>-test).</p>
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<p>Effect of plant extracts on DENV infection. Values from control cells were set to one hundred. Bars represent mean values from three independent experiments in triplicate. Error bars indicate standard deviations. * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01 and *** <span class="html-italic">p</span> &lt; 0.001 vs. control (Student’s <span class="html-italic">t</span>-test).</p>
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12 pages, 1283 KiB  
Article
Preliminary Exploration of Natural Polyphenols and/or Herbal Mix Replacing Sodium Nitrate to Improve the Storage Quality of Pork Sausage
by Angeliki-Niki Kaloudi, Agori Karageorgou, Michael Goliomytis and Panagiotis Simitzis
Appl. Sci. 2025, 15(2), 789; https://doi.org/10.3390/app15020789 - 15 Jan 2025
Viewed by 392
Abstract
Synthetic agents are regularly used as preservatives in pork meat products such as sausages. However, these compounds can be harmful to human health, due to their carcinogenic potential. As a result, natural antioxidants derived from herbs and medicinal plants are continuously and thoroughly [...] Read more.
Synthetic agents are regularly used as preservatives in pork meat products such as sausages. However, these compounds can be harmful to human health, due to their carcinogenic potential. As a result, natural antioxidants derived from herbs and medicinal plants are continuously and thoroughly assessed to replace/minimize the application of synthetic agents during meat processing and storage. The current study was consequently implemented to highlight the effects of natural polyphenols and/or an herbal mix on the quality characteristics of pork sausages. Five batches of sausages were manufactured using the same recipe and raw materials; the control, without sodium nitrate and natural preservatives (CON), and four groups supplemented with sodium nitrate at 0.015% (SN), olive polyphenols at 0.2% (POL), an herbal mix containing rosemary, savory, camelia, thyme, lemon balm and turmeric at 0.2% (NM), or the concomitant addition of olive polyphenols (0.1%) and the herbal mix (0.1%) (POL + NM). Assessment of pH, color, cooking loss, tenderness and oxidative stability was implemented immediately after the manufacture of the sausages (day 0) and at 1, 2 and 3 months after refrigerated storage. It was illustrated that the oxidative stability of sausages was ameliorated as a consequence of POL or/and NM addition, since the values of malondialdehyde (MDA) were significantly decreased compared to the control (p < 0.001) at months 1, 2 and 3, and the values were similar to those observed in the SN group that served as a positive control. Values for the lightness (L), shear force and cooking loss were similar among the experimental treatments (p > 0.05). On the other hand, pH values were greater in the SN and NM groups than in the other groups (p < 0.05). Redness (a*) was significantly higher in the SN group and yellowness (b*) in the NM and POL + NM groups compared to the other groups (p < 0.001). It can be concluded that the examined polyphenols and/or herbal mix could be used as an alternative to nitrates for the amelioration of oxidative stability in pork sausages. Full article
(This article belongs to the Special Issue Recent Processing Technologies for Improving Meat Quality)
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<p>Technological flow diagram for preparing the pork sausages.</p>
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<p>Effect of polyphenols and/or herbal mix on the (<b>A</b>) pH, (<b>B</b>) cooking loss (%) and (<b>C</b>) shear force (N) values of pork sausages on month 0, 1, 2 and 3 after refrigerated storage. The treatment groups were as follows: control (CON), with the addition of sodium nitrite (SN), polyphenols (POL), herbal mix (NM) and polyphenols and herbal mix (POL + NM). <sup>a,b</sup> Values with different letters within a parameter in a specific storage period are significantly different (<span class="html-italic">p</span> &lt; 0.05). <sup>A–C</sup> Values with different letters within the same group among different storage periods are significantly different (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>Effect of polyphenols and/or herbal mix on the (<b>A</b>) pH, (<b>B</b>) cooking loss (%) and (<b>C</b>) shear force (N) values of pork sausages on month 0, 1, 2 and 3 after refrigerated storage. The treatment groups were as follows: control (CON), with the addition of sodium nitrite (SN), polyphenols (POL), herbal mix (NM) and polyphenols and herbal mix (POL + NM). <sup>a,b</sup> Values with different letters within a parameter in a specific storage period are significantly different (<span class="html-italic">p</span> &lt; 0.05). <sup>A–C</sup> Values with different letters within the same group among different storage periods are significantly different (<span class="html-italic">p</span> &lt; 0.05).</p>
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25 pages, 2950 KiB  
Article
Ajuga reptans L. Herb Extracts: Phytochemical Composition and Pharmacological Activity Screening
by Svitlana Maliuvanchuk, Andriy Grytsyk, Oksana Popadynets, Taras Kotyk, Ain Raal and Oleh Koshovyi
Plants 2025, 14(2), 219; https://doi.org/10.3390/plants14020219 - 14 Jan 2025
Viewed by 480
Abstract
The genus Ajuga (Lamiaceae family) comprises approximately 300 species, which are widely used in traditional medicine for their diaphoretic, antiseptic, hemostatic, and anti-inflammatory properties, but scarcely in official ones. Therefore, the study of Ajuga reptans holds promise for developing new medicinal products. In [...] Read more.
The genus Ajuga (Lamiaceae family) comprises approximately 300 species, which are widely used in traditional medicine for their diaphoretic, antiseptic, hemostatic, and anti-inflammatory properties, but scarcely in official ones. Therefore, the study of Ajuga reptans holds promise for developing new medicinal products. In aqueous and aqueous-alcoholic soft extracts of the A. reptans herb, 16 amino acids, 20 phenolics, and 10 volatile substances were identified by HPLC and GC/MS. The assays of the main substances’ groups were also determined by spectrophotometry (vitamin K1, polyphenols, tannins, flavonoids, and hydroxycinnamic acids) and titrometry (ascorbic and organic acids). A. reptans herb extracts are practically non-toxic, exhibit hepatoprotective activity (dose 25 mg/kg) in experimental carbon tetrachloride-induced hepatitis, moderate anti-inflammatory activity (dose 100 mg/kg) in carrageenan-induced edema models, and possess significant local hemostatic (reducing bleeding time by 40.6%) and wound-healing properties (complete wound healing after 9 days). The aqueous-ethanolic soft A. reptans extract (extractant 50% ethanol) demonstrated the most pronounced hepatoprotective and anti-inflammatory effects. A. reptans extracts are capable of inhibiting the growth of microorganisms and showing higher activity against Gram-positive bacteria. A. reptans herb extracts are promising agents for implementation in official medicine as wound healing and hepatoprotective remedies after further preclinical and clinical studies. Full article
(This article belongs to the Special Issue Medicinal Plants: Phytochemistry and Pharmacology Studies)
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<p><span class="html-italic">Ajuga reptans</span> L.</p>
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<p>TLC chromatogram of phenolic compounds in <span class="html-italic">Ajuga reptans</span> L. herb extracts: 1–3—extracts of <span class="html-italic">A. reptans</span>; 4—<span class="html-italic">A. reptans</span> extract (extraction agent: 50% ethanol); 5—<span class="html-italic">A. reptans</span> extract (extraction agent: 70% ethanol); 6—comparison solution (rutin (R<sub>f</sub> = 0.05), apigenin-7-glucoside (R<sub>f</sub> = 0.07), chlorogenic acid (R<sub>f</sub> = 0.22), isoquercitrin (R<sub>f</sub> = 0.24), hyperoside (R<sub>f</sub> = 0.26), apigenin (R<sub>f</sub> = 0.35), luteolin (R<sub>f</sub> = 0.42), caffeic acid (R<sub>f</sub> = 0.45), ferulic acid (R<sub>f</sub> = 0.77), quercetin (R<sub>f</sub> = 0.79)).</p>
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<p>Structural features of liver lobules under the influence of the toxicant. 1—destructured hepatocytes, 2—lymphocytes. Staining: hematoxylin and eosin. Magnification: ×200.</p>
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<p>Structural features of liver lobules in animals administered AR2: 1—hepatic plates, 2—sinusoids, 3—blood vessels. Staining: hematoxylin and eosin. Magnification: ×200.</p>
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<p>Structural features of liver lobules in animals that received the “Silibor” preparation. 1—hepatic plates, 2—sinusoid, 3—central vein. Staining: hematoxylin and eosin. Magnification: ×200.</p>
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<p>Antimicrobial activity of <span class="html-italic">Ajuga reptans</span> L. herb extracts prepared using AR1 (extraction agent: purified water) (1), AR2 (extraction agent: 50% ethanol) (2), AR3 (extraction agent: 70% ethanol) (3), and gentamicin (4) against cultures of <span class="html-italic">Enterococcus faecalis</span> (<b>A</b>) and β-hemolytic <span class="html-italic">Streptococcus</span> group G (<b>B</b>).</p>
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19 pages, 2933 KiB  
Article
Therapeutic Potential of Lappula patula Extracts on Germline Development and DNA Damage Responses in C. elegans
by Qinghao Meng, Anna Hu, Weiyu Xiao, Robert P. Borris and Hyun-Min Kim
Pharmaceuticals 2025, 18(1), 89; https://doi.org/10.3390/ph18010089 - 13 Jan 2025
Viewed by 455
Abstract
Background: Lappula patula (L. patula) is a plant with known medicinal properties, and its extracts have shown promise as potential anti-cancer agents. This study aimed to evaluate the nematocidal effects of L. patula extracts and investigate their impact on germline development, DNA [...] Read more.
Background: Lappula patula (L. patula) is a plant with known medicinal properties, and its extracts have shown promise as potential anti-cancer agents. This study aimed to evaluate the nematocidal effects of L. patula extracts and investigate their impact on germline development, DNA damage responses, and apoptosis in Caenorhabditis elegans (C. elegans), a model organism for studying these processes. Methods: C. elegans was exposed to L. patula extracts to assess survival, development, and incidence of male phenotype. Germline abnormalities were examined using microscopy at different developmental stages. The DNA damage response was evaluated through the expression of the atm-1, atl-1 and pCHK-1. Apoptosis was quantified by monitoring cell death during the pachytene stage. LC-MS was used to identify bioactive compounds within the extracts. Results: Exposure to L. patula extracts resulted in a dose-dependent reduction in worm survival and larval developmental progress, with no significant impact on the male incidence. Germline defects were observed, including increased nuclear spacing at premeiotic and pachytene stages, altered number of bivalents during diakinesis. These defects correlated with a significant decrease in brood size. Also, L. patula extracts activated the DNA damage response pathway, marked by increased expression of atm-1 and atl-1. Moreover, the extracts induced apoptosis in the germline in a pCHK-1-independent manner. LC-MS analysis revealed 31 potential anti-tumor compounds, supporting the extract’s cytotoxic properties. Conclusions: Lappula patula extracts exhibit potent nematocidal and cytotoxic properties, suggesting their potential for cancer therapy. The observed DNA damage and apoptosis in C. elegans emphasize the extract’s promising role in anti-cancer drug development. Further studies are needed to explore the therapeutic potential of these compounds in clinical settings. Full article
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<p>Dose-dependent nematocidal effects of <span class="html-italic">Lappula patula</span> extracts on <span class="html-italic">C. elegans</span> survival and development, with no impact on OP50 growth. (<b>A</b>) <span class="html-italic">Lappula patula</span> extracts significantly reduced the survival and development of <span class="html-italic">C. elegans</span>. The effects were evaluated by treating worms with different concentrations of <span class="html-italic">L. patula</span> extracts (0.03, 0.3, and 3 µg/mL, indicated by brown, orange, and gray colors, respectively) and monitoring their survival, adult formation, and male (HIM) phenotype over a 48 h period. A clear inverse relationship was observed between the dose of the herbal extract and the survival and adult worm percentages, indicating that higher concentrations of <span class="html-italic">L. patula</span> extract led to a marked decrease in worm viability and maturation. However, the percentage of males did not exhibit a dose-dependent trend, suggesting that the disruption in sex chromosome segregation may not be directly influenced by the dosage of the extract. Statistical significance was assessed using a two-tailed <span class="html-italic">t</span>-test, with ** <span class="html-italic">p</span> &lt; 0.01; *** <span class="html-italic">p</span> &lt; 0.001; and **** <span class="html-italic">p</span> &lt; 0.0001, comparing the control (+DMSO) with the treated samples. (<b>B</b>) To evaluate whether the nematocidal effects of <span class="html-italic">Lappula patula</span> could be attributed to an inhibition of bacterial growth, we assessed the growth of OP50 in the presence of <span class="html-italic">L. patula</span> extract. Over a 24 h incubation period, no significant inhibition of bacterial growth was observed at 0.03 μg/mL of <span class="html-italic">L. patula</span> extract.</p>
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<p>Disruption of germline development caused by <span class="html-italic">Lappula patula</span> extracts. (<b>A</b>) DAPI-stained nuclei during germline development. Exposure to <span class="html-italic">Lappula patula</span> extracts led to an increase in the gaps between nuclei in the PMT and pachytene, as indicated by the arrows. The distances between adjacent nuclei were greater in the worms treated with the herb extract compared with the control (DMSO). Worms exposed to the herbal extract often exhibited fewer DAPI-stained bivalent bodies during diakinesis, suggesting impaired DNA recombination. Scale bar: 2 µm. (<b>B</b>) Quantification of the increased nuclear spacing in the premeiotic tip (PMT) and pachytene stages shown in the panel. (<b>C</b>) Quantification of crescent-shaped nuclei in the germline. The number of crescent-shaped nuclei per gonad arm is indicated. (<b>D</b>) Quantification of DAPI-stained bivalents in the germline. The percentage of bivalent is indicated. (<b>E</b>) Quantification of germline size. Germline size, as indicated by the length of the PMT, TZ, and pachytene stages, was measured in worms treated with <span class="html-italic">L. patula</span> or <span class="html-italic">O. cornuta</span> extracts. (<b>F</b>) Brood size of <span class="html-italic">Lappula patula</span>-exposed worms. Treatment with <span class="html-italic">Lappula patula</span> extracts led to a notable decrease in the number of offspring produced by hermaphrodite worms over a span of four days. Statistical significance was determined using a two-tailed Mann–Whitney test, as indicated by asterisks. All experiments were performed with <span class="html-italic">C. elegans</span> hermaphrodites, and the data are presented as the mean ± SEM.</p>
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<p><span class="html-italic">Lappula patula</span> extracts activated the DNA damage response, leading to upregulation of <span class="html-italic">ATM-1</span> and <span class="html-italic">ATL-1</span>, and increased apoptosis but did not induce pCHK-1 foci. (<b>A</b>) Exposure to <span class="html-italic">Lappula patula</span> extracts significantly enhanced the mRNA expression of <span class="html-italic">atm-1</span> and <span class="html-italic">atl-1</span>, two essential proteins involved in the DNA damage response pathway, confirming the activation of this cellular defense mechanism. Both <span class="html-italic">atm-1</span> and <span class="html-italic">atl-1</span> are critical for activating repair pathways such as homologous recombination and cell cycle arrest, and their upregulation in response to extract exposure provides evidence of the activation of these protective mechanisms. This upregulation suggests that the extracts may trigger a cellular reaction to DNA damage, which is necessary to repair the compromised genome. (<b>B</b>) No distinct increase in pCHK-1 foci (arrows) was observed in the germline cells of worms treated with <span class="html-italic">Lappula patula</span> extracts. Despite the elevation of <span class="html-italic">atm-1</span> and <span class="html-italic">atl-1</span> levels, no significant increase in pCHK-1 levels was observed, implying that while the DNA damage response was triggered, the downstream signaling associated with checkpoint activation, particularly the phosphorylation of CHK-1, did not occur as expected. The extract from <span class="html-italic">O.c.</span> was used as a positive control for these experiments, as it has been previously shown to activate the DNA damage response pathway robustly (<span class="html-italic">p</span> = 0.825 in control and <span class="html-italic">L.p.</span>, <span class="html-italic">p</span> = 0.0021 in control and <span class="html-italic">O.c.</span>; scale bar = 2 µm. (<b>C</b>) When examined during the pachytene stage, a significant increase in apoptosis (arrow) was detected in the germline cells exposed to <span class="html-italic">Lappula patula</span> extracts. Statistical significance was determined by a two-tailed Mann–Whitney test, with asterisks denoting <span class="html-italic">p</span> values indicating significant differences between the control and experimental groups. Scale bar = 20 µm.</p>
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<p>Fragmentation patterns of 31 potential anti-tumor components identified in <span class="html-italic">Lappula patula</span> extracts. Out of 112 identified substances, 31 were found to be potential anti-tumor components. The peaks (hollow arrows) shown represent the breakdown patterns of these 31 components, analyzed using PeakView Analyst TF 1.6 software. The <span class="html-italic">x</span> axis shows the mass-to-charge ratio (<span class="html-italic">m</span>/<span class="html-italic">z</span>), and the <span class="html-italic">y</span> axis shows the intensity.</p>
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<p><span class="html-italic">Lappula patula</span> extract triggered a DNA damage response, which resulted in disturbances in germline development, enhanced apoptotic cell death, and disruption of the normal meiotic process. These findings suggest that the extract has cytotoxic effects. Furthermore, <span class="html-italic">Lappula patula</span> contains at least 31 compounds known for their potential anti-tumor properties. This presence of bioactive compounds underscores the potential of <span class="html-italic">Lappula patula</span> as a source of natural anti-cancer agents. Given the growing interest in plant-based treatments for cancer, further investigation into the specific compounds and their mechanisms in <span class="html-italic">Lappula patula</span> is essential to fully explore its therapeutic potential.</p>
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