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20 pages, 5511 KiB  
Article
Antioxidant Effect of a Plant-Derived Extracellular Vesicles’ Mix on Human Skin Fibroblasts: Induction of a Reparative Process
by Rossella Di Raimo, Davide Mizzoni, Antonella Aloi, Giulia Pietrangelo, Vincenza Dolo, Giuseppina Poppa, Stefano Fais and Mariantonia Logozzi
Antioxidants 2024, 13(11), 1373; https://doi.org/10.3390/antiox13111373 - 9 Nov 2024
Viewed by 902
Abstract
Plant-Derived Extracellular Vesicles extracellular vesicles (PDEVs) from organic agriculture (without the use of pesticides and microbicides) contain high levels of antioxidants. Organic PDEVs have shown an increased antioxidant power compared to PDEVs from single plants, suggesting a synergistic effect of the bioactives constitutively [...] Read more.
Plant-Derived Extracellular Vesicles extracellular vesicles (PDEVs) from organic agriculture (without the use of pesticides and microbicides) contain high levels of antioxidants. Organic PDEVs have shown an increased antioxidant power compared to PDEVs from single plants, suggesting a synergistic effect of the bioactives constitutively expressed in the PDEVs from single fruits. With this study, we wanted to investigate the beneficial effects of a mix of PDEVs on human skin cells. We found detectable levels of citric acid, ascorbic acid, glutathione, catalase, and SOD in a mix of PDEVs deriving from five different fruits (grape, red orange, papaya, pomegranate, and tangerine). We then treated H2O2-conditioned fibroblasts with the mix of PDEVs. The results showed that the PDEVs’ mixture reverted the H2O2-induced redox imbalance, restoring mitochondrial homeostasis, with a strong reduction of mitochondrial anion superoxide and an increase in sirtuin levels. The antioxidant action was consistent with wound repair on a lesion produced in a fibroblast’s monolayer. This result was consistent with an increased level of vimentin and matrix metalloproteinase-9, whose expression is directly related to the efficiency of the reparative processes. These data support a beneficial role of PDEVs in both preventing and treating skin injuries through their potent antioxidant and reparative activities. Full article
(This article belongs to the Special Issue The OxInflammation Process and Tissue Repair)
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<p>PDEVs biophysical characterization. (<b>a</b>) Size and distribution of PDEVs through NTA; (<b>b</b>) Distribution of PDEVs’ zeta potential.</p>
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<p>PDEVs morphological characterization through TEM. (<b>a</b>) Round structure and membrane integrity of PDEVs with sizes between 50 and 80 nm; (<b>b</b>) Round structure and membrane integrity of PDEVs with sizes between 150 and 200 nm; (<b>c</b>) PDEVs plasma membrane visible at 34.000 magnification and (<b>d</b>) 64.000 (insert). The arrow indicates the plasma membrane of vesicles.</p>
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<p>Cellular uptake of Dil-PDEVs in skin fibroblasts after (<b>a</b>) 24 h of treatment; (<b>b</b>) 48 h of treatment; and (<b>c</b>) 72 h of treatment. PDEVs were labeled with Dil (red), and nuclei were counterstained with DAPI (blue).</p>
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<p>PDEVs effect of mitochondrial metabolism. (<b>a</b>) Analysis of mitochondrial membrane potential; (<b>b</b>) analysis of mitochondrial anion superoxide levels. Data are expressed as mean ± SE. * <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. Statistical analysis was performed using one-way ANOVA Bonferroni. M.I.F. (a.u.) = Mean I Intensity of Fluorescence (arbitrary unit).</p>
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<p>Quantification of extracellular sirtuin 1 concentration. Data are expressed as mean ± SE. **** <span class="html-italic">p</span> &lt; 0.0001. Statistical analysis was performed using one-way ANOVA Bonferroni.</p>
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<p>Wound healing assay of skin fibroblasts. Representative images are shown from three independent experiments. Data are expressed as mean ± SE. ns: not significant; **** <span class="html-italic">p</span> &lt; 0.0001. Statistical analysis was performed using unpaired <span class="html-italic">t</span>-test (Student’s <span class="html-italic">t</span>-test). Scale bar = 500 µm.</p>
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<p>Collagen I expression in skin fibroblasts at the wound site. Cells were stained with anti-collagen I primary antibody, subsequently a secondary antibody AlexaFluor<sup>®</sup> 488 conjugated was added (green) and nuclei were counterstained with DAPI (blue). Scale bar = 100 µm.</p>
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<p>MMP-9 expression in skin fibroblasts at the wound site. Cells were stained with anti-MMP-9-FITC (green) and nuclei were counterstained with DAPI (blue). Scale bar = 50 µm.</p>
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<p>Quantification of extracellular vimentin. Data are expressed as mean ± SE. **** <span class="html-italic">p</span> &lt; 0.0001. Statistical analysis was performed using unpaired <span class="html-italic">t</span>-test (Student’s <span class="html-italic">t</span>-test).</p>
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17 pages, 3842 KiB  
Article
Metallurgical Waste for Sustainable Agriculture: Converter Slag and Blast-Furnace Sludge Increase Oat Yield in Acidic Soils
by Olga V. Zakharova, Peter A. Baranchikov, Svetlana P. Chebotaryova, Gregory V. Grigoriev, Nataliya S. Strekalova, Tatiana A. Grodetskaya, Igor N. Burmistrov, Sergey V. Volokhov, Denis V. Kuznetsov and Alexander A. Gusev
Agronomy 2024, 14(11), 2642; https://doi.org/10.3390/agronomy14112642 - 9 Nov 2024
Viewed by 446
Abstract
The study is the first to examine the combined use of blast-furnace sludge as a source of microelements and converter slag as a soil-deoxidizing agent in oat (Avena sativa L.) cultivation in sod-podzolic soils. It has been established that blast-furnace sludge is [...] Read more.
The study is the first to examine the combined use of blast-furnace sludge as a source of microelements and converter slag as a soil-deoxidizing agent in oat (Avena sativa L.) cultivation in sod-podzolic soils. It has been established that blast-furnace sludge is a highly dispersed waste, which contains about 50% iron, 7% zinc, and a small amount of calcium, silicon, magnesium, aluminum, and sulfur. Hazardous components such as lead, arsenic, etc., are not detected. Converter slag comprises porous granules up to 3 mm in size, consisting mainly of calcium compounds (CaO, Ca(CO)3, CaSiO3, CaFe2O4) and a small amount of Mn, Al, and Mg trace elements. In a laboratory experiment, blast-furnace sludge increased the germination of oats by 5–10%, regardless of the addition of a deoxidizer (slag), but at the same time suppressed the growth of stem length by a maximum of 18% at 1 g∙kg−1. The addition of slag raised substrate pH and increased the index by 8% at a sludge concentration of 0.1 g∙kg−1. Root length in deoxidizer-free variants increased by 50–60% and with the addition of slag by 27–47%. Root dry mass also increased under the addition of sludge by 85–98%; however, the addition of slag reduced the indicator to the control level. In a field experiment with the combined application of waste, an increase in yield by more than 30% was shown. When soil was treated with slag and sludge, the height of plants increased by an average of 18%. It should be noted that the introduction of waste did not affect the quality of the grain. The use of slag increased the lead content in the soil, which is probably due to the sorption properties of calcium compounds in the slag, since lead was not found in the analyzed waste. Presumably, lead is sorbed by slag from the lower soil horizons, concentrating and immobilizing it in the upper layer. This version is supported by the absence of lead accumulation in straw and oat grain. The zinc-containing sludge increased the content of this element by 33% in the soil, as well as by 6% in straw and by 14% in grain. Thus, we found that the studied metallurgical wastes can be used as nutrients for agriculture, both individually and jointly. Overall, the proposed approach will contribute both to reducing the amount of accumulated waste and to improving the efficiency and sustainability of agricultural production and CO2 sequestration. However, the features of the accumulation of heavy metals in soil and plants under the influence of the analyzed types of waste require more in-depth study, including within the framework of long-term field experiments. Full article
(This article belongs to the Section Plant-Crop Biology and Biochemistry)
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<p>Experimental field.</p>
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<p>Waste characterization: (<b>a</b>) SEM micrograph with elemental composition and diffraction pattern of blast-furnace sludge sample; (<b>b</b>) SEM micrograph with elemental composition and diffraction pattern of converter slag.</p>
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<p>Morphophysiological parameters of oats: (<b>a</b>) germination; (<b>b</b>) average stem length; (<b>c</b>) average root length; (<b>d</b>) experimental plants. The * symbol marks significant differences with the control at a significance level of <span class="html-italic">p</span> &lt; 0.05. *—differences from the untreated variant, **—differences from the slag-treated variant.</p>
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<p>Weight of spring oat: (<b>a</b>) raw stem weight; (<b>b</b>) dry stem weight; (<b>c</b>) raw root weight; (<b>d</b>) dry root weight. The * symbol marks significant differences with the control at a significance level of <span class="html-italic">p</span> &lt; 0.05. *—differences from the untreated variant, **—differences from the slag-treated variant.</p>
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<p>Elemental analysis of plants: (<b>a</b>) root, control; (<b>b</b>) stem, control; (<b>c</b>) root, sludge; (<b>d</b>) stem, sludge; (<b>e</b>) root, slag + sludge; (<b>f</b>) stem, slag + sludge.</p>
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<p>Productivity and crop quality indicators of spring oats: (<b>a</b>) yield; (<b>b</b>) height of plants; (<b>c</b>) net photosynthesis productivity; (<b>d</b>) protein content in grain. The * symbol marks significant differences with the control at a significance level of <span class="html-italic">p</span> &lt; 0.05. *—differences from the untreated variant, **—differences from the slag-treated variant.</p>
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16 pages, 1926 KiB  
Article
TLR2/TLR5 Signaling and Gut Microbiota Mediate Soybean-Meal-Induced Enteritis and Declined Growth and Antioxidant Capabilities in Large Yellow Croaker (Larimichthys crocea)
by Lei Zheng, Chao Zeng, Wanqin Zhu, Jiaonan Zhang, Lei Wang, Jianchun Shao and Wei Zhao
J. Mar. Sci. Eng. 2024, 12(11), 2016; https://doi.org/10.3390/jmse12112016 - 8 Nov 2024
Viewed by 471
Abstract
Soybean meal, renowned for its high yield, cost efficiency, and protein richness, serves as a pivotal plant-based alternative to fish meal. However, high soybean meal inclusion in Larimichthys crocea diets is linked to enteritis and oxidative damage, with unknown mechanisms. Our study aims [...] Read more.
Soybean meal, renowned for its high yield, cost efficiency, and protein richness, serves as a pivotal plant-based alternative to fish meal. However, high soybean meal inclusion in Larimichthys crocea diets is linked to enteritis and oxidative damage, with unknown mechanisms. Our study aims to elucidate the molecular basis of soybean-meal-induced enteritis and its impact on intestinal microbiota in L. crocea. To this end, four isonitrogenous and isolipidic diets with varying soybean meal levels (0% FM, 15% SBM15, 30% SBM30, and 45% SBM45) were administered to L. crocea for 8 weeks. The results indicated that the SBM30 and SBM45 treatments significantly hindered fish growth, digestive efficiency, and protein utilization. Furthermore, high soybean meal levels (SBM30 and SBM45) activated intestinal Toll-like receptors (TLR2A, TLR2B, TLR5, and TLR22), stimulating C-Rel and mTOR protein expression and elevating ERK phosphorylation. This led to increased pro-inflammatory cytokine production (IL-1β, IL-6, and TNF-α) and decreased anti-inflammatory cytokine expression (IL-4/13A, IL-4/13B, and TGF-β), suggesting a potential signaling pathway for soybean-meal-induced enteritis. Furthermore, enteritis induced by high soybean meal levels led to oxidative damage, evident from increased MDA levels and decreased antioxidant enzyme activities (SOD and CAT). The SBM30 and SBM45 treatments increased Firmicutes and Bacteroidetes abundance in fish gut microbiota, while Proteobacteria abundance decreased. This microbiota shift may enhance soybean meal nutrient utilization, yet high soybean meal concentrations still impair growth. A soybean-meal-rich diet promotes harmful bacteria like Rhodococcus and depletes probiotics like Ralstonia, increasing disease risks. L. crocea has limited tolerance for soybean meal, necessitating advanced processing to mitigate anti-nutritional factors. Ultimately, exploring alternative protein sources beyond soybean meal for fish meal replacement is optimal for L. crocea. Full article
(This article belongs to the Special Issue Nutrition and Physiology of Marine Fish)
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<p>The mRNA expression levels of specific intestinal inflammation-related factors, including Toll-like receptor genes (<b>A</b>), pro-inflammatory cytokine genes (<b>B</b>), and anti-inflammatory cytokine genes (<b>C</b>), in large yellow croaker (<span class="html-italic">Larimichthys crocea</span>) fed with various experimental diets. Values are shown as mean ± SD (<span class="html-italic">n</span> = 4). Different lowercase letters indicate a significant difference (<span class="html-italic">p</span> &lt; 0.05). Indicators without lowercase letters indicate no significant difference (<span class="html-italic">p</span> &gt; 0.05).</p>
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<p>Protein level of C-Rel and phosphorylation levels of ERK and mTOR in the gut of large yellow croaker (<span class="html-italic">Larimichthys crocea</span>) fed with experimental diets (<b>A</b>). The protein levels were quantified using ImageJ (<b>B</b>,<b>C</b>). The phosphorylation levels were quantified using ImageJ (<b>D</b>). Values are shown as mean ± SD (<span class="html-italic">n</span> = 4). Different lowercase letters indicate a significant difference (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>Composition of gut microflora at phylum level (<b>A</b>) and differentially abundant phyla (<b>B</b>) in large yellow croaker (<span class="html-italic">Larimichthys crocea</span>) fed experimental feeds. Values are shown as mean ± SD (<span class="html-italic">n</span> = 3). The red star represents a statistically significant difference (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>Composition of gut microflora at genus level (<b>A</b>) and differentially abundant genera (<b>B</b>) in large yellow croaker (<span class="html-italic">Larimichthys crocea</span>) fed experimental feeds. Values are shown as mean ± SD (<span class="html-italic">n</span> = 3). The red star represents a statistically significant difference (<span class="html-italic">p</span> &lt; 0.05).</p>
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15 pages, 3144 KiB  
Article
Effect of Inoculation with Rhizosphere Phosphate-Solubilizing Bacteria on the Growth and Physiological Characteristics of Parashorea chinensis
by Zhongheng Hu, Shuang Wei, Wannian Li, Tong Wu, Saif Ullah and Mei Yang
Forests 2024, 15(11), 1932; https://doi.org/10.3390/f15111932 - 1 Nov 2024
Viewed by 848
Abstract
Given the challenges of slow growth and low survival rates in the early stages of Parashorea chinensis cultivation, identifying sustainable methods to enhance seedling performance is critical for successful reforestation and conservation efforts. This study aimed to address these by investigating the growth-promoting [...] Read more.
Given the challenges of slow growth and low survival rates in the early stages of Parashorea chinensis cultivation, identifying sustainable methods to enhance seedling performance is critical for successful reforestation and conservation efforts. This study aimed to address these by investigating the growth-promoting effects of phosphate-solubilizing bacteria (PSB). One-year-old seedlings of P. chinesis were inoculated with PSB strains isolated from the rhizosphere soil of Parashorea chinensis H. Wang plantations Y3, W5, H8, and a mixed strain (Mix), with inoculated seedling as a control (CK). The effects of inoculation on seedling growth, photosynthetic physiology, plant nutrient status, and physiological indicators were comprehensively evaluated. Results showed that PSB inoculation increased seedling height and basal diameter growth of P. chinensis, with an increase of 1.56 cm and 0.53 mm compared to CK, respectively, though the differences were not significant. The Mix treatment significantly improved photosynthesis, with increases in net photosynthetic rate (106.3%), transpiration rate (93.89%), and intercellular CO2 concentration (75.51%) compared to CK. Nutrient levels including total nitrogen, total phosphorus, and total potassium were significantly increased by 15.98%, 25.54%, and 32.12%, respectively, in the Mix treatments compared to CK. Moreover, stress resistance also improved, with higher proline content, soluble sugar, and soluble protein levels. Antioxidant enzyme activities (SOD, CAT, and POD) were increased by 9.83%, 23.66%, and 292.32%, respectively, while MDA content was significantly reduced by 69.01%. The mixed strain treatment also significantly increased acid phosphatase activity by 111.88%. In conclusion, PSB inoculation, particularly with the mixed strain, promoted growth and nutrient uptake photosynthetic efficiency and stress resistance in P. chinensis seedlings, offering a promising biotechnological solution for improving seedling performance. Full article
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<p>The effect of different bacterial inoculation treatments on photosynthetic pigment content of <span class="html-italic">Parashorea Chinensis</span> seedlings. Note: Different lowercase letters on the bars indicate significant differences between different bacterial solutions (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>The effects of different bacterial solution treatments on the photosynthetic characteristics of <span class="html-italic">Parashorea chinensis</span> seedlings. Note: Different lowercase letters on the bars indicate significant differences between different bacterial solutions (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>The effects of different bacterial solution treatments on the nutrient levels of <span class="html-italic">Parashorea chinensis</span> seedlings. Note: Different lowercase letters on the bars indicate significant differences between different bacterial solutions (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>The effects of different bacterial solution treatments on the osmotic regulators of <span class="html-italic">Parashorea chinensis</span> seedlings. Note: Different lowercase letters on the bars indicate significant differences between different bacterial solutions (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>The effects of different bacterial solution treatments on the malondialdehyde (MDA) content of <span class="html-italic">Parashorea chinensis</span> seedlings. Note: Different lowercase letters on the bars indicate significant differences between different bacterial solutions (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>The effects of different bacterial solution treatments on the enzyme activity of <span class="html-italic">Parashorea chinensis</span> seedlings. Note: Different lowercase letters on the bars indicate significant differences between different bacterial solutions (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>Correlation analysis between growth and physiological parameters of <span class="html-italic">Parashorea chinensis</span> seedlings. Note: H: height growth; D: diameter growth; Chl: total chlorophyll; <span class="html-italic">Pn</span>: net photosynthetic rate; <span class="html-italic">Ci</span>: intercellular CO<sub>2</sub> concentration; <span class="html-italic">Gs</span>: stomatal conductance; <span class="html-italic">Tr</span>: transpiration rate; TP: total phosphorus; TK: total potassium; TN: total nitrogen; Pro: proline; SS: soluble sugar; SP: soluble protein; MDA: malondialdehyde; CAT: catalase activity; SOD: superoxide dismutase activity; POD: peroxidase activity; ACP: acid phosphatase. * <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.</p>
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13 pages, 2805 KiB  
Article
A New Mutagenesis Tool for Songpu Mirror Carp (Cyprinus carpio L.) for Selective Breeding: Atmospheric-Pressure Room-Temperature Plasma Mutagenesis Technology
by Xiaona Jiang, Chitao Li, Mei Shang, Xuesong Hu, Yanlong Ge and Zhiying Jia
Fishes 2024, 9(11), 448; https://doi.org/10.3390/fishes9110448 - 1 Nov 2024
Viewed by 532
Abstract
As a new, safe, and efficient method, Atmospheric-Pressure Room-Temperature Plasma (ARTP) mutagenesis has been widely applied in the field of microbial breeding and industrial applications, but it is rarely used in fish. In this study, ARTP mutagenesis technology was applied for the first [...] Read more.
As a new, safe, and efficient method, Atmospheric-Pressure Room-Temperature Plasma (ARTP) mutagenesis has been widely applied in the field of microbial breeding and industrial applications, but it is rarely used in fish. In this study, ARTP mutagenesis technology was applied for the first time to a common carp strain, Songpu mirror carp (Cyprinus carpio L.), to increase genetic variation in this species. The appropriate experimental conditions were determined to include a radio frequency output power of 160 W and the processing of fertilized eggs for 360 s. The ARTP treatment group had a lower survival rate than the control group. The CV of morphological characters in the ARTP treatment group was significantly higher than that in the control group, and the CV of body weight was the highest (p < 0.05). In addition, the deformity rate in the ARTP treatment group was significantly higher than in the control group (p < 0.05). Individuals with high weight and no deformities were screened within the selection pressure of 1:15 of ARTP treatment group and fed in the same pool with the control group of the same age. The measurement of serum indices showed that, in the ARTP treatment group, TP, ALP, ALB, T-CHO, LDL levels were significantly higher than those in the control group (p < 0.05). Furthermore, the relative expressions of SOD, growth-related genes GH, IGF-I, protein synthesis-related genes TOR and 4EBP1 were significantly higher in the ARTP treatment group than in the control group (p < 0.05). In summary, Songpu mirror carp subjected to ARTP treatment showed a higher growth potential and antioxidant capacity. Full article
(This article belongs to the Special Issue Genetics and Breeding in Aquaculture)
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<p>Hatching rates and fry survival rates for Songpu mirror carp under different conditions. (<b>a</b>) Survival rates of fertilized Songpu mirror carp eggs under different treatment times and output power levels. (<b>b</b>) Hatchability rates of fertilized Songpu mirror carp eggs under different treatment times and output powers. The treatment time was 0 s and the output power was 0 W in the control group, and the other groups constituted the ARTP treatment group. Lowercase letters in the column chart indicate significant differences determined by using the Bonferroni <span class="html-italic">t</span> test in SAS 9.1 (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>Box plots of W and phenotypic traits in Songpu mirror carp. Box plots of the W, SL, H, BW and HL of Sonpu mirror carp in three periods in the ARTP treatment and control groups initiated at 5 months after fertilization. The black dots in the figure indicate values greater than 1.5 times the interquartile range.</p>
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<p>Malformation types in Songpu mirror carp, arranged from large to small according to total length. Types of deformities include lack of fin (<b>A</b>,<b>B</b>,<b>E</b>–<b>G</b>), deformity of mouth and skull (<b>C</b>), lack of operculum (<b>D</b>,<b>H</b>,<b>I</b>), scales on the back (<b>C</b>,<b>J</b>), and high back (<b>F</b>,<b>I</b>–<b>K</b>). Bar indicates 50 mm.</p>
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<p>Morphological parameter ratios of different stages in Songpu mirror carp. The ratios of morphological parameters of Songpu mirror carp at 5 months and 14 months after fertilization, including W/SL, BH/SL, BW/SL and HL/SL Statistically significant differences were defined at <span class="html-italic">p</span> &lt; 0.05 (** <span class="html-italic">p</span> &lt; 0.01).</p>
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<p>Relative expression levels of protein synthesis-related genes in the dorsal muscles (<b>a</b>) and intestine (<b>b</b>) in the ARTP treatment and control groups. Relative mRNA expression of <span class="html-italic">S6K</span> compared to that in the ARTP treatment group. Statistically significant differences were defined at <span class="html-italic">p</span> &lt; 0.05 (* <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).</p>
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<p>Relative expression levels of antioxidant-related genes in the liver (<b>a</b>) and growth-related genes in dorsal muscles (<b>b</b>) in the ARTP treatment and control groups, respectively. Relative mRNA expression levels of <span class="html-italic">SOD</span> compared to those in the ARTP treatment group and <span class="html-italic">GH</span> compared to those in the control group. Statistically significant differences were defined at <span class="html-italic">p</span> &lt; 0.05 (* <span class="html-italic">p</span> &lt; 0.05; ** <span class="html-italic">p</span> &lt; 0.01).</p>
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16 pages, 3148 KiB  
Article
Evaluation of the Dietary Arginine Supplementation on Yellow Catfish: From a Low-Temperature Farming Perspective
by Quanquan Cao, Mohamed S. Kisha, Alkhateib Gaafar, Abdelgayed Metwaly Younes, Haifeng Liu and Jun Jiang
Biology 2024, 13(11), 881; https://doi.org/10.3390/biology13110881 - 29 Oct 2024
Viewed by 562
Abstract
The yellow catfish is an economically significant freshwater fish with increasing importance in aquaculture. However, the low temperature environments prevalent in certain regions pose challenges to its growth, development, and overall health. This study aimed to explore the impact of dietary arginine (Arg) [...] Read more.
The yellow catfish is an economically significant freshwater fish with increasing importance in aquaculture. However, the low temperature environments prevalent in certain regions pose challenges to its growth, development, and overall health. This study aimed to explore the impact of dietary arginine (Arg) addition on the growth, digestive capacity, and intestinal antioxidant response in fish under low temperature acclimation (18 °C). Total 720 fish were randomly distributed into six groups, each containing 120 fish. Over the course of eight weeks, each group was fed with diets about varying Arg concentrations (1.79–3.26 g/kg). The results indicated that Arg supplementation resulted in an increase in specific growth rate (SGR), feed intake (FI), feed efficiency (FE), as well as pancreatic enzyme activities in both pancreas and intestine. Conversely, malondialdehyde (MDA) and protein carbonyl (PC) contents initially decreased but increased with higher Arg concentrations. Glutathione peroxidase 1a (GPX1a) showed a positive correlation with nuclear factor-erythroid 2-related factor-2 (Nrf2), showing its role in antioxidative capacity. Furthermore, this study revealed that Arg significantly enhanced the activities of anti-superoxide anion, anti-hydroxyl radical, and anti-oxidative enzymes, along with the relative mRNA abundance of Copper-Zinc superoxide dismutase (CuZnSOD), catalase, GPX1a, glutamate-cysteine ligase catalytic subunit (GCLC), and Nrf2 in the intestine. It was determined that yellow catfish weighing between 61.0 g and 89.0 g require an intake of 26.8 g of Arg per kilogram of diet based on polynomial regression analysis of specific growth rate (SGR), which is equivalent to 37.0 g of dietary protein, under sub-low temperature conditions of 18 °C. Full article
(This article belongs to the Special Issue Nutrition, Environment, and Fish Physiology)
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<p>Quadratic regression analysis of specific growth rate (SGR) for yellow catfish (<span class="html-italic">Pelteobagrus fulvidraco</span>) fed diets containing graded levels of Arg (g/kg) for 8 weeks.</p>
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<p>Relative expression of Copper-Zinc superoxide dismutase (CuZnSOD), catalase (CAT) and glutathione peroxidase 1a (GPx1a) and glutamate-cysteine ligase catalytic subunit (GCLC) in the intestine of yellow catfish fed diets containing graded levels of Arg (g/kg). Data represents the means of three replicate groups of yellow catfish with 12 fish per group; error bars indicate standard error. Different letters above a bar denote significant difference (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>Relative expression of NF-E2-related factor 2 (Nrf2) in the intestine of yellow catfish fed diets containing graded levels of Arg (g/kg). Data represents means of three replicate groups of yellow catfish with 12 fish per group; error bars indicate standard error. Different letters above a bar denote significant difference (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>Loading plot of principal component analysis (PCA) in intestine representing measured variables and the individuals based on all the gene expressions of six graded Arg levels (g/kg) in the (PC1 × PC2) coordination plane. Ellipses in group from the six conditions (1.79 g/kg diet in red, 2.11 g/kg in yellow, 2.36 g/kg diet in green, 2.68 g/kg diet in light blue, 2.95 g/kg diet in blue and 3.26 g/kg diets in pink).</p>
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<p>Role of Nrf2 in the regulation of intestinal metabolism in yellow catfish under Arg feed. Nrf2 can induce the downstream pathway genes including GPX, CuZnSOD, CAT, GCLC to increase the anti-oxidative capacity.</p>
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17 pages, 2784 KiB  
Article
Pathological Alterations in Heart Mitochondria in a Rat Model of Isoprenaline-Induced Myocardial Injury and Their Correction with Water-Soluble Taxifolin
by Natalia V. Belosludtseva, Tatyana A. Uryupina, Lyubov L. Pavlik, Irina B. Mikheeva, Eugeny Yu. Talanov, Natalya I. Venediktova, Dmitriy A. Serov, Mikhail R. Stepanov, Mikhail A. Ananyan and Galina D. Mironova
Int. J. Mol. Sci. 2024, 25(21), 11596; https://doi.org/10.3390/ijms252111596 - 29 Oct 2024
Viewed by 438
Abstract
Mitochondrial damage and associated oxidative stress are considered to be major contributory factors in cardiac pathology. One of the most potent naturally occurring antioxidants is taxifolin, especially in its water-soluble form. Herein, the effect of a 14-day course of the peroral application of [...] Read more.
Mitochondrial damage and associated oxidative stress are considered to be major contributory factors in cardiac pathology. One of the most potent naturally occurring antioxidants is taxifolin, especially in its water-soluble form. Herein, the effect of a 14-day course of the peroral application of the water-soluble taxifolin (aqTAX, 15 mg/kg of body weight) on the progression of ultrastructural and functional disorders in mitochondria and the heart’s electrical activity in a rat model of myocardial injury induced with isoprenaline (ISO, 150 mg/kg/day for two consecutive days, subcut) was studied. The delayed ISO-induced myocardial damage was accompanied by an increase in the duration of RR and QT intervals, and long-term application of aqTAX partially restored the disturbed intraventricular conduction. It was shown that the injections of ISO lead to profound ultrastructural alterations of myofibrils and mitochondria in cardiomyocytes in the left ventricle myocardium, including the impairment of the ordered arrangement of mitochondria between myofibrils as well as a decrease in the size and the number of these organelles per unit area. In addition, a reduction in the protein level of the subunits of the respiratory chain complexes I-V and the activity of the antioxidant enzymes catalase, glutathione peroxidase, and Mn-SOD in mitochondria was observed. The application of aqTAX caused an increase in the efficiency of oxidation phosphorylation and a partial restoration of the morphometric parameters of mitochondria in the heart tissue of animals with the experimental pathology. These beneficial effects of aqTAX are associated with the inhibition of lipid peroxidation and the normalization of the enzymatic activities of glutathione peroxidase and Mn-SOD in rat cardiac mitochondria, which may reduce the oxidative damage to the organelles. Taken together, these data allow one to consider this compound as a promising cardioprotector in the complex therapy of heart failure. Full article
(This article belongs to the Section Biochemistry)
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<p>Experimental study design. Male Wistar rats were randomly assigned to the following groups: the control (Cntl) group was given two subcutaneous (SC) injections of normal saline (NS); the control + aqTAX (Cntl + aqTAX) group was watered with aqTAX (aqTAX, 150 mg/kg/day body weight, <span class="html-italic">per os</span>); the isoprenaline (ISO) group was treated with isoprenaline hydrochloride (ISO, 150 mg/kg body weight, s.c.); the isoprenaline + aqTAX (ISO + aqTAX) group received two injections of ISO and were given a freshly prepared mixture of aqTAX and drinking water for the next 14 days. Rats (<span class="html-italic">n</span> = 10 <span class="html-italic">per</span> group) were sacrificed 15 days after the first injection, and the above analyses were carried out.</p>
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<p>Electrophysiological characteristics of the heart of experimental animals. (<b>A</b>) Representative lead II ECG records. (<b>B</b>) Heart rate (beats per minute) and the most common ECG intervals: RR and QT (in seconds) in the studied animal groups (control (Cntl), the control + aqTAX (Cntl + aqTAX) group, the isoprenaline hydrochloride (ISO) group, and the isoprenaline hydrochloride + aqTAX (ISO + aqTAX) group. Data are presented as mean values ± SEM from a group of five rats. Statistical significance was analyzed using one-way analysis of variance and Tukey’s post hoc 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.</p>
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<p>Electron micrographs of mitochondria in the subsarcolemmal region of a rat cardiomyocyte from the left ventricle of the heart: (<b>A</b>) the control group (Cntl); (<b>B</b>) control rats after a course of treatment with aqTAX (Cntl + aqTAX); (<b>C</b>) isoprenaline-induced myocardial injury (ISO); (<b>D</b>) ISO-injured rats after a course of treatment with aqTAX (ISO + aqTAX). Representative micrographs from three independent experiments are shown. Lead citrate and uranyl acetate × 5000. Scale bars, 1 µm.</p>
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<p>Morphometric analysis of rat heart subsarcolemmal mitochondria in the studied animal groups (control (Cntl), control + aqTAX (Cntl + aqTAX), isoprenaline hydrochloride (ISO), and isoprenaline hydrochloride + aqTAX (ISO + aqTAX)). (<b>A</b>) Average number of the organelles in rat cardiomyocytes, normalized to the area (mm<sup>2</sup>) of the TEM images analyzed. (<b>B</b>) Bar graph summarizing the number of damaged (vacuolated) mitochondria (% of the control) between conditions. (<b>C</b>) Relative distribution of mitochondrial size (perimeter, µm) in rat cardiomyocytes under different treatments. The assays were carried out from at least three rat left ventricles in each group. At least 30 fields of view per group were analyzed. Data are presented as mean values ± standard derivations. * <span class="html-italic">p</span> &lt; 0.05; *** <span class="html-italic">p</span> &lt; 0.001.</p>
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<p>Content of the subunits of five respiratory chain complexes (CI-NDUFB8, CII-SDHB, CIII-UQCRC2, CIV-MTCO1, and CV-vATP5A) in rat heart mitochondria from the experimental groups. (<b>A</b>) Representative Western blot of typical oxidative phosphorylation (OXPHOS) subunits of complexes I–V in the mitochondria isolated from the organs of experimental animals. The analysis was performed using the Total OXPHOS Rodent WB Antibody Cocktail and anti-VDAC1 antibodies (Abcam, #ab110413 and #ab15895, Cambridge, UK; 10 μg of mitochondrial protein per lane). PC–positive control (Abcam, #ab110341, Cambridge, UK). (<b>B</b>) Densitometric analysis of the level of the OXPHOS subunits in the cardiac mitochondria of experimental animals. Data are expressed as mean values ± standard errors. Control values are taken as 100%. Significant differences tested with a one-way analysis of variance and Tukey’s post hoc test. * <span class="html-italic">p</span> &lt; 0.05.</p>
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<p>Oxidative stress-related makers of the blood plasma and heart mitochondria of rats from the experimental groups: control (Cntl), control + aqTAX (Cntl + aqTAX), isoprenaline hydrochloride (ISO), and isoprenaline hydrochloride + aqTAX (ISO + aqTAX). (<b>A</b>) Plasma levels of thiobarbituric acid reactive substances (TBARS). (<b>B</b>) TBARS levels in isolated rat heart mitochondria. (<b>C</b>) Enzymatic activities of the superoxide scavenging enzyme manganese superoxide dismutase (MnSOD) in rat heart mitochondria. (<b>D</b>) Glutathione peroxidase (GPx) enzyme activities in rat heart mitochondria. (<b>E</b>) Enzymatic activities of catalase in rat heart mitochondria. Values are mean ± SEM from five animals in each group. Significant differences tested with a one-way analysis of variance and Tukey’s post hoc 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.</p>
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18 pages, 11849 KiB  
Article
Preparation Process Optimization of Glycolipids from Dendrobium officinale and the Difference in Antioxidant Effects Compared with Ascorbic Acid
by Yan Long, Jiajing Yang, Hongfei Ji, Xiao Han, Yuting Fan, Keyao Dai, Haiyu Ji and Juan Yu
Nutrients 2024, 16(21), 3664; https://doi.org/10.3390/nu16213664 - 28 Oct 2024
Viewed by 585
Abstract
Background:Dendrobium officinale glycolipids (DOG), often left as residues after hot water extraction for polysaccharide production, are often discarded. Methods: This study investigates the optimal extraction of DOG using response surface methodology, focusing on liquid–solid ratios, ethanol concentrations, extraction temperatures, and extraction [...] Read more.
Background:Dendrobium officinale glycolipids (DOG), often left as residues after hot water extraction for polysaccharide production, are often discarded. Methods: This study investigates the optimal extraction of DOG using response surface methodology, focusing on liquid–solid ratios, ethanol concentrations, extraction temperatures, and extraction times, while preliminarily analyzing DOG’s structural properties. Additionally, the differences in antioxidant effects between DOG and ascorbic acid based on intestinal flora metabolism were further evaluated. Results: The optimal parameters for DOG extraction were determined as follows: liquid–solid ratio of 20 mL/g, ethanol concentration of 70%, extraction temperature of 70 °C, and extraction time of 2.5 h, yielding 2.64 ± 0.18%. In addition, DOG was identified as a diglyceride, mainly composed of glucose, mannose, linoleic acid, 9,12,15-octadecatrienoic acid, and presented certain direct free radicals scavenging effects. In animal experiments, unlike the direct free scavenging effects of ascorbic acid, DOG increased intestinal Bacteroides acidifaciens abundance in mice, up-regulated piceatannol expression, and down-regulated 1-naphthol expression, which contributed to antioxidant effects by enhancing the activities of SOD and GSH-Px while reducing MDA content. Conclusions: DOG was a diglyceride isolated from D. officinale residues after hot water extraction, and presented strong antioxidant effects by regulating intestinal flora metabolism. These findings could promote the efficient utilization of D. officinale and support further development of DOG in functional food applications. Full article
(This article belongs to the Section Nutrition and Metabolism)
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<p>Response surface (<b>a</b>–<b>c</b>,<b>g</b>–<b>i</b>) and contour plots (<b>d</b>–<b>f</b>,<b>j</b>–<b>l</b>) of the DOG extraction with various variables interaction including liquid–solid ratios, ethanol concentrations, extraction temperatures, extraction time. The colors transition from blue to red indicated the increasing DOG yields.</p>
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<p>UV full wavelength scanning (<b>a</b>) and FTIR (<b>b</b>) spectra of DOG.</p>
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<p>Scavenging rates of DOG and ascorbic acid on ABTS (<b>a</b>) and DPPH (<b>b</b>) free radicals.</p>
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<p>Antioxidant effects of DOG on oxidative damaged mice sera. (<b>a</b>), SOD activities; (<b>b</b>), GSH-Px activities; (<b>c</b>), MDA contents; (<b>d</b>), total antioxidant capacity. Note: **, <span class="html-italic">p</span> &lt; 0.01 compared with blank group; <sup>##</sup>, <span class="html-italic">p</span> &lt; 0.01 compared with model group.</p>
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<p>Effects of ascorbic acid and DOG on intestinal microbial diversity. (<b>a</b>), Venn diagram; (<b>b</b>), relative contents at species level; (<b>c</b>), ternary plot among blank, model and ascorbic acid groups; (<b>d</b>), ternary plot among blank, model and DOG groups.</p>
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<p>Differential metabolites analysis in ascorbic acid group compared with model group. (<b>a</b>), Heatmap with metabolites ID; (<b>b</b>), model overfitting analysis; (<b>c</b>), volcano map; (<b>d</b>), KEGG enrichment plot.</p>
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<p>Differential metabolites analysis in DOG-H group compared with model group: (<b>a</b>) Heatmap with metabolites ID; (<b>b</b>) model overfitting analysis; (<b>c</b>) volcano map, and (<b>d</b>) KEGG enrichment plot.</p>
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25 pages, 31111 KiB  
Article
Experimental Analysis of Cavitation Erosion: Parameter Sensitivity and Testing Protocols
by SeyedMehdi Mohammadizadeh, José Gilberto Dalfré Filho, Cassiano Sampaio Descovi, Ana Inés Borri Genovez and Thomaz Eduardo Teixeira Buttignol
Coatings 2024, 14(10), 1288; https://doi.org/10.3390/coatings14101288 - 9 Oct 2024
Cited by 1 | Viewed by 840
Abstract
The scientific goal of this study was to investigate the effects of various parameters on cavitation-induced erosion, with the aim to enhance the understanding and assessment of cavitation resistance in hydraulic systems. Cavitation erosion poses significant challenges to the durability and efficiency of [...] Read more.
The scientific goal of this study was to investigate the effects of various parameters on cavitation-induced erosion, with the aim to enhance the understanding and assessment of cavitation resistance in hydraulic systems. Cavitation erosion poses significant challenges to the durability and efficiency of hydraulic components, such as those found in hydropower plants and pumping stations. Prompted by the need to improve the reliability of cavitation testing and material assessment, this research conducted a comprehensive sensitivity analysis of a cavitation jet apparatus (CJA). This study employed an experimental platform that consisted of a vertical cylindrical test tank, a submerged nozzle, and an aluminum sample. By examining a range of orifice diameters, this research identified that smaller diameters led to increased erosion intensity, with the most pronounced effects observed at a diameter of 2 mm. Furthermore, various standoff distances (SoDs) were tested, which revealed that shorter distances resulted in greater erosion, with the highest impact noted at an SoD of 5 cm. This study also evaluated different nozzle geometries, where it was found that a 132° conical sharped edges nozzle, combined with an orifice diameter of 2 mm and an SoD of 5 cm, produced the most severe erosion. Conversely, chamfered edges nozzles and a commercial nozzle (MEG2510) with an SoD of 10 cm or greater showed reduced erosion. These results highlight that by standardizing the testing duration to 1200 s, the CJA could reliably assess the cavitation resistance of materials. This study established a clear relationship between increased pressure and higher impact forces, which led to more severe erosion. The findings underscore the effectiveness of the CJA in evaluating material resistance under various cavitation conditions, thus addressing a critical need for reliable cavitation testing tools. Full article
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<p>Francis Turbine Damaged by Cavitation [<a href="#B18-coatings-14-01288" class="html-bibr">18</a>].</p>
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<p>FESEM Micrographs Depicting Cavitation Erosion Pits on Copper (Cu, <b>Top</b>), 17-4PH Stainless Steel (<b>Middle</b>), and Tungsten (W, <b>Bottom</b>) Subjected to Hydrostatic Pressures of <math display="inline"><semantics> <mrow> <mn>0.1</mn> </mrow> </semantics></math>, 3, 6, and 10 MPa (From <b>Left</b> to <b>Right</b>). The Magnification Scales are <math display="inline"><semantics> <mrow> <mn>60</mn> </mrow> </semantics></math>, <math display="inline"><semantics> <mrow> <mn>500</mn> </mrow> </semantics></math>, and <math display="inline"><semantics> <mrow> <mn>5000</mn> </mrow> </semantics></math>, Which Correspond to 200, 40, and 4 μm Scale Bars, Respectively [<a href="#B8-coatings-14-01288" class="html-bibr">8</a>].</p>
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<p>Mind Map Showing the Sequence of Steps in the Materials and Methods Section.</p>
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<p>CJA: (<b>a</b>) Scheme of Cavitating Jet Apparatus, (<b>b</b>) Cylindrical Test Tank Detail, (<b>c</b>) Submerged Jet, and (<b>d</b>) Photo of the Experimental System (Dimensions Not to Scale).</p>
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<p>Schematic Representation of Jet Amplitude (Dimensions Not to Scale).</p>
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<p>Nozzle Schemes Used in Cavitation Jet Apparatuses: (<b>a</b>) 20° Conical Nozzle, (<b>b</b>) 132° Conical Nozzle, (<b>c</b>) Circular Nozzle With a Radius of 12.5 mm, (<b>d</b>) <span class="html-small-caps">MEG 2510</span> WashJet<sup>®</sup> Spray Nozzle, (<b>e</b>) Sharped Edges, and (<b>f</b>) Chamfered Edges (Dimensions Not to Scale).</p>
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<p>SEM Image of Al 6351-T6 Sample Surface.</p>
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<p>Schematic Representation of the Jet Amplitude Using a Circular Sharped Edges Nozzle With an Orifice Diameter of 2 mm and an SoD of 5 cm.</p>
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<p>Pressure Versus Orifice Diameter (132° Sharped Edges Nozzle, SoD of 5 cm).</p>
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<p>Impact Force and Upstream Pressure Versus Time Periods: (<b>a</b>) 10 s, (<b>b</b>) 5 s, (<b>c</b>) 3 s, (<b>d</b>) 1 s, (<b>e</b>) 0.5 s, and (<b>f</b>) 0.1 s.</p>
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<p>Effects of SoD on Cavitation Erosion Over the Exposure Time. The Tests Used a Nozzle Orifice Diameter of 2 mm and a 20° Conical Sharped Edges Nozzle.</p>
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<p>Comparison of Cavitation Erosion Quantities Versus Standoff Distances at <span class="html-italic">t</span> = 1200 s. The Tests Used a Nozzle Orifice Diameter of 2 mm and a 20° Conical Sharped Edges Nozzle.</p>
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<p>Overview of Erosion Patterns for Various SoDs With a 2 mm Orifice Diameter and a 20° Conical Sharped Edges Nozzle, as Measured at <span class="html-italic">t</span> = 1200 s.</p>
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<p>Evolution of the Cavitation Erosion on an Aluminum Sample Over Time, Up to <span class="html-italic">t</span> = 1200 s. The Test uUtilized a 5 cm SoD, a 2 mm Orifice Diameter, and an Intensively Cavitation-Impacted Diameter of 4 cm With a 20° Conical Sharped Edges Nozzle.</p>
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<p>Effects of Various Nozzle Geometries on Cavitation Erosion Over Different Exposure Times (Nozzle Orifice Diameter = 2 mm, SoD = 5 cm, Intensively Cavitation-Impacted Diameter of Specimen = 4 cm).</p>
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<p>Comparison of Erosion Intensity for Different Nozzle Geometries at <span class="html-italic">t</span> = 1200 s (Orifice Diameter = 2 mm, SoD = 5 cm, Intensively Cavitation-Impacted Diameter = 4 cm).</p>
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<p>Cumulative Mass Loss Versus Time (132° Conical Sharped Edges Nozzle, SoD = 5 cm, and Nozzle Orifice Diameter = 2 mm).</p>
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<p>Cavitation Erosion (pits/cm<sup>2</sup>) Versus Time (132° Conical Sharped Edges Nozzle, <span class="html-italic">t</span> = 1200 s, SoD = 0.05 m, Intensively Cavitation-Impacted Diameter of Sample = 4 cm, and Nozzle Orifice Diameter = 2 mm).</p>
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19 pages, 7566 KiB  
Article
Cytotoxicity, Proapoptotic Activity and Drug-like Potential of Quercetin and Kaempferol in Glioblastoma Cells: Preclinical Insights
by Magdalena Kusaczuk, Elena Tovar-Ambel, Paola Martín-Cabrera, Mar Lorente, Nélida Salvador-Tormo, Agnieszka Mikłosz, Adrian Chabowski, Guillermo Velasco and Monika Naumowicz
Int. J. Mol. Sci. 2024, 25(19), 10740; https://doi.org/10.3390/ijms251910740 - 5 Oct 2024
Viewed by 1247
Abstract
Despite the increasing understanding of the pathogenesis of glioblastoma (GBM), treatment options for this tumor remain limited. Recently, the therapeutic potential of natural compounds has attracted great interest. Thus, dietary flavonoids quercetin (QCT) and kaempferol (KMF) were investigated as potential cytostatic agents in [...] Read more.
Despite the increasing understanding of the pathogenesis of glioblastoma (GBM), treatment options for this tumor remain limited. Recently, the therapeutic potential of natural compounds has attracted great interest. Thus, dietary flavonoids quercetin (QCT) and kaempferol (KMF) were investigated as potential cytostatic agents in GBM. Moreover, the physicochemical properties of QCT and KMF, determining their bioavailability and therapeutic efficiency, were evaluated. We proved that both polyphenols significantly reduced the viability of GBM cells. We also demonstrated that both QCT and KMF evoked the cytotoxic effect in T98G cells via induction of apoptotic cell death as shown by increased activity of caspase 3/7 and caspase 9 together with an overexpression of the cleaved form of PARP. Apoptosis was additionally accompanied by the activation of stress responses in QCT- and KMF-treated cells. Both polyphenols caused oxidative stress and endoplasmic reticulum (ER) stress, as demonstrated by the increased generation of reactive oxygen species (ROS), deregulated expressions of superoxide dismutases (SOD2 and Sod1 on protein and transcriptomic levels, respectively), as well as an overexpression of ERO1α, GRP78, p-JNK, and an up-regulation of Chop, Atf4 and Atf6α genes. The antitumor effect of QCT and KMF was also confirmed in vivo, showing reduced growth of tumor xenografts in the chick chorioallantoic membrane (CAM) experiment. Moreover, electrophoretic light scattering (ELS) was used to measure the zeta potential of cell membranes upon exposition to QCT and KMF. Additionally, on the basis of existing physicochemical data, the drug-likeness score of QCT and KMF was evaluated. Analyses showed that both compounds accomplish Lipinski’s Rule of 5, and they both fit into the criteria of good central nervous system (CNS) drugs. Altogether, our data support the idea that QCT and KMF might be plausible candidates for evaluation as therapeutic agents in preclinical models of glioblastoma. Full article
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<p>The viability of glioblastoma cells treated with QCT and KMF. Results of the MTT analysis after 24 and 48 h of exposure to QCT and KMF are shown for (<b>A</b>) T98G; (<b>B</b>) U118MG; (<b>C</b>) U87MG cells. (<b>D</b>) Cell viability results plotted against logarithmic values of drug concentrations together with a tabulated summary of calculated IC<sub>50</sub> values are shown for T98G, U118MG, and U87MG cells treated for 24 and 48 h. The results represent means for pooled triplicate values from three independent experiments. Significant alterations are expressed relative to controls and marked with asterisks. Statistical significance was * <span class="html-italic">p</span> ≤ 0.05; ** <span class="html-italic">p</span> ≤ 0.005; *** <span class="html-italic">p</span> ≤ 0.0005; **** <span class="html-italic">p</span> ≤ 0.0001.</p>
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<p>Typical pH-dependence of the zeta potential of human glioblastoma T98G cells treated with (<b>A</b>) QCT; (<b>B</b>) KMF for 24 and 48 h.</p>
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<p>The effect of QCT and KMF on proliferation and apoptosis of GBM cells. (<b>A</b>) The ATP production in T98G cells upon treatment with QCT and KMF for 48 h; the immunofluorescence microscopy of Ki67-immunostained T98G cells. The Ki67-staining and the DNA counterstain with DAPI was performed for cells cultured for 48 h. (<b>B</b>) Representative images visualized under ZAISS-Axioplan 2 fluorescence microscope are shown (magnification ×20). (<b>C</b>) Bar graph illustrating the number of Ki67-immunolabeled T98G cells. (<b>D</b>) Representative Western blot images showing expressions of apoptosis-related proteins in T98G cells treated with QCT and KMF for 48 h; (<b>E</b>) Caspase 3/7 activity in T98G cells exposed to QCT and KMF for 48 h. (<b>F</b>) Caspase 9 activity in T98G cells exposed to QCT and KMF for 48 h. Significant alterations are expressed relative to controls and marked with asterisks. Statistical significance was * <span class="html-italic">p</span> ≤ 0.05; ** <span class="html-italic">p</span> ≤ 0.005; *** <span class="html-italic">p</span> ≤ 0.0005; **** <span class="html-italic">p</span> ≤ 0.0001.</p>
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<p>(<b>A</b>) The effect of QCT and KMF on stress responses in glioblastoma T98G cells. ROS levels in cells treated with QCT and KMF for 48 h. (<b>B</b>) Representative Western blot images showing expression levels of proteins connected with ER stress and oxidative stress in cells treated with QCT and KMF for 48 h. (<b>C</b>) Real-time qPCR analysis of <span class="html-italic">Atf6α</span>, <span class="html-italic">Atf4</span>, <span class="html-italic">Chop</span>, and <span class="html-italic">Sod1</span> gene expression in cells treated with QCT and KMF for 48 h. Results are shown as relative fold change in mRNA expression in comparison to control, where the expression level was set as 1. Significant alterations are expressed relative to controls and marked with asterisks. Statistical significance was * <span class="html-italic">p</span> ≤ 0.05; *** <span class="html-italic">p</span> ≤ 0.0005; **** <span class="html-italic">p</span> ≤ 0.0001.</p>
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<p>(<b>A</b>) Chicken embryo in vivo assay. Time schedule of the chicken embryo in vivo experiment. (<b>B</b>) Weight of control tumors vs. tumors treated with 200 μM QCT and 200 μM KMF. Significant alterations are expressed relative to controls and marked with asterisks. Statistical significance was * <span class="html-italic">p</span> ≤ 0.05; ** <span class="html-italic">p</span> ≤ 0.005.</p>
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<p>Tentative model of QCT and KMF mode of action in glioblastoma cells. Continuous arrows symbolize well-established molecular pathways. Dashed arrows show possible indirect interactions.</p>
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12 pages, 2273 KiB  
Article
Effects of Water-Deficit Stress on the Growth and Physiological Characteristics of Chloranthus spicatus Seedlings
by Yangjuan Shang, Zhibing Wan, Changheng Zhao, Xing Su and Xinyi Liu
Horticulturae 2024, 10(10), 1054; https://doi.org/10.3390/horticulturae10101054 - 3 Oct 2024
Viewed by 656
Abstract
Chloranthus spicatus is one of the main scented tea varieties cultivated mainly in the Huangshan region, and dried flowers of these plants are mainly used for imparting a characteristic aroma to the tea. However, climatic variations in Huangshan limit its cultivation, with water [...] Read more.
Chloranthus spicatus is one of the main scented tea varieties cultivated mainly in the Huangshan region, and dried flowers of these plants are mainly used for imparting a characteristic aroma to the tea. However, climatic variations in Huangshan limit its cultivation, with water deficit (WD) being the main limiting factor. The present study evaluated the effects of different WD intensities on the growth and physiological parameters of C. spicatus seedlings to determine the optimal soil moisture content for their large-scale cultivation. The experimental design comprised a control group (95–100%) and three treatment groups, namely mild WD (75–80%), moderate WD (55–60%), and severe WD (35–40%). Each treatment lasted 45 days and was given to 10 potted C. spicatus seedlings, with 3 replicates. Measurements were conducted for the shoot length and diameter; biomass; photosynthesis parameter; activities of antioxidant enzymes, namely superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT); and soluble protein (SP) and malonylaldehyde (MDA) contents of the seedlings. The results indicated that mild and moderate WD positively affected C. spicatus seedlings in terms of shoot length, diameter, biomass, root/shoot ratio, photosynthesis rate, intercellular CO2 concentration, water use efficiency, and carboxylation efficiency. Moreover, the growth and photosynthesis were increased in the mild and moderate WD seedlings but decreased in the severe WD seedlings. Mild and moderate WD also led to a significant decrease in the antioxidant enzyme activities and the MDA content of seedlings (p < 0.05), all of which exhibited higher levels in severe WD seedlings. These results indicated that severe water stress restricted the growth of seedlings, while mild and moderate stress proved beneficial. SP content of the seedlings increased under mild and moderate WD but decreased under severe WD. We infer that the strong hydrophilicity of SPs in the seedlings results in the accumulation of water in plant cells, thus conferring resistance to drought stress. In conclusion, reducing the soil moisture content by 55–80% in the Huangshan region may be a promising strategy for boosting its cultivation. Full article
(This article belongs to the Section Biotic and Abiotic Stress)
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<p>Phenotypic changes in <span class="html-italic">Chloranthus spicatus</span> seedlings under different water-deficit stress intensities after 45 days.</p>
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<p>Biomass of <span class="html-italic">Chloranthus spicatus</span> seedlings under water-deficit (WD) stress after 45 days. Columns with different small letters indicate significant differences (Duncan’s multiple range test, <span class="html-italic">p</span> &lt; 0.05).</p>
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<p>Variations in photosynthesis parameters (<span class="html-italic">P<sub>n</sub></span>, <span class="html-italic">T<sub>r</sub></span>, <span class="html-italic">C<sub>i</sub></span>, <span class="html-italic">G<sub>s</sub></span>, <span class="html-italic">WUE</span>, and <span class="html-italic">CE</span>) in the leaves of <span class="html-italic">Chloranthus spicatus</span> seedlings under water-deficit (WD) stress after 45 days. Dots with different small letters indicate significant differences (Duncan’s multiple range test, <span class="html-italic">p</span> &lt; 0.05).</p>
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<p>Changes in superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) activities in the leaves of <span class="html-italic">Chloranthus spicatus</span> seedlings under different water-deficit (WD) treatments after 45 days. Dots with different small letters indicate significant differences (Duncan’s multiple range test, <span class="html-italic">p</span> &lt; 0.05).</p>
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<p>Soluble protein and malonylaldehyde contents in the leaves of <span class="html-italic">Chloranthus spicatus</span> seedlings under different water-deficit (WD) treatments after 45 days. Dots with different small letters indicate significant differences (Duncan’s multiple range test, <span class="html-italic">p</span> &lt; 0.05).</p>
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16 pages, 12018 KiB  
Article
Optimizing Sugarcane Clonal Propagation In Vitro by Using Calcium Ammonium Nitrate and Ammonium Sulfate
by Yuanli Wu, Faisal Mehdi, Zhengying Cao, Yimei Gan, Shuting Jiang, Limei Zan, Shuzhen Zhang and Benpeng Yang
Plants 2024, 13(19), 2767; https://doi.org/10.3390/plants13192767 - 2 Oct 2024
Viewed by 745
Abstract
To replace explosive nitrate-based chemicals in MS medium, this study developed a new, safer, and more cost-effective method using fertilizer-grade calcium ammonium nitrate and ammonium sulfate. This approach replaces ammonium nitrate and potassium nitrate, ensuring both safety and cost efficiency for sugarcane propagation. [...] Read more.
To replace explosive nitrate-based chemicals in MS medium, this study developed a new, safer, and more cost-effective method using fertilizer-grade calcium ammonium nitrate and ammonium sulfate. This approach replaces ammonium nitrate and potassium nitrate, ensuring both safety and cost efficiency for sugarcane propagation. Six local sugarcane varieties—Zhongtang1 (ZT1), Zhongtang3 (ZT3), Zhongtang6 (ZT6), Guitang42 (GT42), Guitang44 (GT44), and Guiliu 07150 (GT07150)—were used. In the control group (Ck), nitrate ions (NO3) were 39.28 mM, and ammonium ions (NH4+) were 20.49 mM, with a 2:1 ratio. In the treatment groups, the concentrations of nitrate ions (NO3) and ammonium ions (NH4+) included treatment 1 (19.69 mM NO3 and 10.3 mM NH4+), treatment 2 (29.54 mM and 15.44 mM), treatment 3 (39.38 mM and 20.59 mM), treatment 4 (49.225 mM and 25.74 mM), treatment 5 (59.07 mM and 30.89 mM), and treatment 6 (68.915 mM and 36.03 mM), respectively, all with the same 2:1 ratio. Fifty bottles per treatment, with three replicates, were used for each sugarcane plantlets treatment. After five subcultures, the optimal ratio was determined by assessing morphological and physiological parameters, nitrogen levels, and SOD enzyme activity. The results indicated that treatment 3 (39.38 mM and 20.59 mM) and treatment 4 (49.225 mM and 25.74 mM) had the best morphological and physiological indicators. The optimal doses of calcium ammonium nitrate and ammonium sulfate were found in treatments 3 and 4, as well as in the control, with no significant difference among them. However, treatment 3, due to its lower dose, was more cost effective. To improve cost efficiency in practical production, it is recommended to use the lower concentration ratio of treatment 3 for plant tissue culture plantlets. Full article
(This article belongs to the Section Plant Development and Morphogenesis)
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Figure 1
<p>The growth response of six sugarcane plantlets varieties subjected to six different gradient ratios of ammonium nitrogen (NH<sub>4</sub><sup>+</sup>-N) and nitrate nitrogen (NO<sub>3</sub><sup>−</sup>-N). (<b>A</b>) GT 42, (<b>B</b>) GT 44, (<b>C</b>) GL 07150, (<b>D</b>) ZT 1, (<b>E</b>) ZT 3, and (<b>F</b>) ZT6. Note: CK (Control) contains ammonium nitrate and potassium nitrate.</p>
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<p>Illustrates the growth rate of sugarcane plantlets of six varieties under seven gradient ratios of ammonium nitrogen (NH<sub>4</sub><sup>+</sup>-N) and nitrate nitrogen (NO<sub>3</sub><sup>−</sup>-N). Different lowercase letters indicate significant differences (<span class="html-italic">p</span> &lt; 0.05). Note: CK (Control) contains ammonium nitrate and potassium nitrate. Abbreviations: Zhongtang 1 (ZT1), Zhongtang 3 (ZT3), Zhongtang 6 (ZT6), Guitang 42 (GT142), Guitang 44 (GT144), and Guiliu 07150 (GL 07150).</p>
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<p>Root growth of six varieties of sugarcane original plantlets under six gradient ratios of ammonium nitrogen (NH<sub>4</sub><sup>+</sup>-N) and nitrate nitrogen (NO<sub>3</sub><sup>−</sup>-N). (<b>A</b>) GT 42, (<b>B</b>) GT 44, (<b>C</b>) GL 07150, (<b>D</b>) ZT 1, (<b>E</b>) ZT 3, and (<b>F</b>) ZT 6. Note: CK (Control) contains ammonium nitrate and potassium nitrate.</p>
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<p>The average root length of six sugarcane original plantlets under seven gradient ratios of ammonium nitrogen (NH<sub>4</sub><sup>+</sup>-N) and nitrate nitrogen (NO<sub>3</sub><sup>−</sup>-N). Different lowercase letters indicate significant differences (<span class="html-italic">p</span> &lt; 0.05). Note: CK (Control) contains ammonium nitrate and potassium nitrate.</p>
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<p>This figure illustrates the average number of roots in six sugarcane original plantlets under seven gradient ratios of ammonium nitrogen (NH<sub>4</sub><sup>+</sup>-N) and nitrate nitrogen (NO<sub>3</sub><sup>−</sup>-N). Different lowercase letters indicate significant differences (<span class="html-italic">p</span> &lt; 0.05). Note: CK (Control) contains ammonium nitrate and potassium nitrate.</p>
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<p>This figure represents the SOD activity of six sugarcane original plantlets under seven gradient ratios of ammonium nitrogen (NH<sub>4</sub><sup>+</sup>-N) and nitrate nitrogen (NO<sub>3</sub><sup>−</sup>-N). Different lowercase letters indicate significant differences (<span class="html-italic">p</span> &lt; 0.05). Note: CK (Control) contains ammonium nitrate and potassium nitrate.</p>
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<p>Chlorophyll content of six sugarcane original plantlets under seven gradient ratios of ammonium nitrogen (NH<sub>4</sub><sup>+</sup>-N) and nitrate nitrogen (NO<sub>3</sub><sup>−</sup>-N). Different lowercase letters indicate significant differences (<span class="html-italic">p</span> &lt; 0.05). Note: CK (Control) contains ammonium nitrate and potassium nitrate.</p>
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<p>Soluble protein content in six sugarcane plantlets under seven gradient ratios of ammonium nitrogen (NH<sub>4</sub><sup>+</sup>-N) and nitrate nitrogen (NO<sub>3</sub><sup>−</sup>-N). Different lowercase letters indicate significant differences (<span class="html-italic">p</span> &lt; 0.05). Note: CK (Control) contains ammonium nitrate and potassium nitrate.</p>
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<p>Ammonium nitrogen content in six sugarcane original plantlets under seven gradient ratios of ammonium nitrogen (NH<sub>4</sub><sup>+</sup>-N) and nitrate nitrogen (NO<sub>3</sub><sup>−</sup>-N). Different lowercase letters indicate significant differences (<span class="html-italic">p</span> &lt; 0.05). Note: CK (Control) contains ammonium nitrate and potassium nitrate.</p>
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<p>Nitrate nitrogen content in six sugarcane original plantlets under seven gradient ratios of ammonium nitrogen (NH<sub>4</sub><sup>+</sup>-N) and nitrate nitrogen (NO<sub>3</sub><sup>−</sup>-N). Different lowercase letters indicate significant differences (<span class="html-italic">p</span> &lt; 0.05). Note: CK (Control) contains ammonium nitrate and potassium nitrate.</p>
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21 pages, 8570 KiB  
Article
Transcriptome and Metabolome Analyses Reveal the Molecular Mechanisms of Albizia odoratissima’s Response to Drought Stress
by Shuoxing Wei, Feng Gao, Zhihui Wang, Guoping Yin, Shizhi Wen, Hanbiao Ou and Zhiming Liu
Plants 2024, 13(19), 2732; https://doi.org/10.3390/plants13192732 - 29 Sep 2024
Viewed by 644
Abstract
Albizia odoratissima is a deciduous tree species belonging to the family Leguminosae. It is widely distributed in the southern subtropical and tropical areas of China and has important ecological and economic value. The growth and metabolic processes of A. odoratissima are affected [...] Read more.
Albizia odoratissima is a deciduous tree species belonging to the family Leguminosae. It is widely distributed in the southern subtropical and tropical areas of China and has important ecological and economic value. The growth and metabolic processes of A. odoratissima are affected by drought stress, but the molecular mechanisms remain unknown. Therefore, this study investigated the physicochemical properties, gene expression, and metabolites of A. odoratissima seedlings under drought stress. The results show that, in leaves of A. odoratissima seedlings, drought stress reduced the moisture content, chlorophyll content, photosynthetic efficiency, superoxide dismutase (SOD) activity, and gibberellin (GA) and indoleacetic acid (IAA) contents while increasing the catalase (CAT) and peroxidase (POD) activities and malondialdehyde (MDA), proline, soluble sugar, and soluble protein contents. Within the CK5 (Day 5 of control group) vs. T5 (Day 5 of drought treatment), CK10 vs. T10, CK15 vs. T15, and CK20 vs. T20 groups (CK: control group; T: drought treatment), a total of 676 differentially expressed genes (DEGs) were upregulated and 518 DEGs were downregulated, and a total of 228 and 143 differential accumulation metabolites (DAMs) were identified in the CK10 vs. T10 and CK20 vs. T20 groups. These were mainly involved in the amino acid and alkaloid metabolism pathways in the leaves of the A. odoratissima seedlings. In the amino acid and alkaloid biosynthesis pathways, the relative expression levels of the AoproA (Aod04G002740, ORTHODONTIC APPLIANCE), AoOAT (Aod07G015970, ORNITHINE-OXO-ACID TRANSAMINASE), and AoAOC3 (Aod12G005010/08G003360/05G023920/08G003000/08G003010, AMINE OXIDASE COPPER CONTAINING 3) genes increased, which concurrently promoted the accumulation of arginine, proline, piperine, cadaverine, and lysine. Furthermore, some key transcription factors in the response to drought were identified in the leaves using the weighted gene co-expression network analyses (WGCNA) method. These findings reveal that A. odoratissima seedlings respond to drought stress by improving the capacities of the antioxidant system and secondary metabolism. Full article
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<p>Structural and physiological biochemical changes in leaves of <span class="html-italic">A. odoratissima</span> seedlings under drought stress. (<b>a</b>) Changes in leaf shape on Days 0, 5, 10, 15, and 20. (<b>b</b>) Cross-sections of leaves (×100) (U-ep = upper epidermis, Pal = palisade, L-ep = lower epidermis). (<b>c</b>) Changes in SOD activity. (<b>d</b>) Changes in CAT activity in the leaves of <span class="html-italic">A. odoratissima</span> under drought stress. (<b>e</b>) Changes in POD activity. (<b>f</b>) Changes in MDA content. (<b>g</b>) Changes in proline content. (<b>h</b>) Changes in soluble sugar content. (<b>i</b>) Changes in soluble protein. (<b>j</b>) Changes in chlorophyll content. (<b>k</b>) Changes in intercellular carbon dioxide concentration. (<b>l</b>) Changes in transpiration rate. (<b>m</b>) Changes in net photosynthetic rate. (<b>n</b>) Changes in stomatal conductance. (<b>o</b>) Changes in gibberellin content. (<b>p</b>) Changes in indoleacetic acid content. (<b>q</b>) Changes in trans-Zeatin-riboside. (<b>r</b>) Changes in moisture content. *: significant difference between control and treatment. **: highly significant difference between control and treatment. T5, T10, T15, and T20: Days 5, 10, 15, and 20 of drought treatment.</p>
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<p>Analysis of transcriptional differences between the leaves and roots of <span class="html-italic">A. odoratissima</span> under drought stress. (<b>a</b>) Transcriptome correlation analysis under control. (<b>b</b>) Transcriptome correlation analysis under drought stress. (<b>c</b>) Changes in DEGs in the leaves and roots of <span class="html-italic">A. odoratissima</span> under drought stress. Up represents upregulated genes, and down represents downregulated genes. (<b>d</b>–<b>g</b>) Venn analysis ((<b>d</b>) leaf upregulated differential genes; (<b>e</b>) leaf downregulated differential genes; (<b>f</b>) root upregulated differential genes; (<b>g</b>) root downregulated differential genes). (<b>h</b>–<b>k</b>) KEGG pathways of differentially expressed genes ((<b>h</b>) leaf upregulated differential genes; (<b>i</b>) leaf downregulated differential genes; (<b>j</b>) root upregulated differential genes; (<b>k</b>) root downregulated differential genes). CK5, CK10, CK15, and CK20: Days 5, 10, 15, and 20 in the control groups. T5, T10, T15, and T20: Days 5, 10, 15, and 20 of drought treatment groups.</p>
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<p>Differential analysis of metabolites in leaves of <span class="html-italic">A. odoratissima</span> under drought stress. (<b>a</b>) PCA of differential metabolites. (<b>b</b>) Correlation analysis of differential metabolites. (<b>c</b>) Differential metabolites in leaves. (<b>d</b>) Characterization of temporal changes in differential metabolites in leaves. (<b>e</b>) KEGG pathways of differential metabolites of <span class="html-italic">A. odoratissima</span> after 10 days of drought stress. (<b>f</b>) KEGG pathways of differential metabolites of <span class="html-italic">A. odoratissima</span> after 20 days of drought stress. Orange font indicates the same KEGG pathways of differential metabolites in both after 10 and 20 days.</p>
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<p>The arginine and proline metabolic pathways of <span class="html-italic">A. odoratissima</span> under drought stress. The colors indicate significances shown in a color scale. The solid lines with arrows indicate directions of the processes. Dashed lines represent the two-step and multi-step reactions.</p>
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<p>The tropane, piperidine, and pyridine alkaloid biosynthetic pathways of <span class="html-italic">A. odoratissima</span> under drought stress.</p>
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<p>WGCNA of physiological indicators and transcriptional differences in <span class="html-italic">A. odoratissima</span> under drought stress. (<b>a</b>) Co-expression network of blue module and drought-stress-related genes (ko00620: pyruvate metabolism; ko00260: threonine metabolism; ko00250: glutamate metabolism; ko00230: purine metabolism; ko00195: photosynthesis; ko01230: amino acid biosynthesis). (<b>b</b>) Co-expression network of cyan module and drought-stress-related genes (ko01230: amino acid biosynthesis; ko00280: isoleucine degradation; ko00561: glycerolipid metabolism; ko00620: pyruvate metabolism; ko01212: fatty acid metabolism; ko01040: unsaturated fatty acids).</p>
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<p>A conceptual model of the drought response mechanism in <span class="html-italic">A. odoratissima</span>. Red colour indicates increased content, enzyme activity or up-regulated gene expression, green colour indicates decreased content, enzyme activity or down-regulated gene expression.</p>
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17 pages, 6947 KiB  
Article
Toxic Effect of Methyl-Thiophanate on Bombyx mori Based on Physiological and Transcriptomic Analysis
by Zhen He, Yang Fang, Fengchao Zhang, Yang Liu, Xiaoli Wen, Cui Yu, Xinkai Cheng, Dechen Li, Liang Huang, Hui Ai and Fan Wu
Genes 2024, 15(10), 1279; https://doi.org/10.3390/genes15101279 - 29 Sep 2024
Viewed by 669
Abstract
Background/Objectives: The utilization of methyl-thiophanate (MT) in vegetables and fruits is widespread due to its broad efficiency, yet its potential impact on silkworm growth remains uncertain. This study aims to examine the effects of MT on the growth of silkworms. Specifically, we assessed [...] Read more.
Background/Objectives: The utilization of methyl-thiophanate (MT) in vegetables and fruits is widespread due to its broad efficiency, yet its potential impact on silkworm growth remains uncertain. This study aims to examine the effects of MT on the growth of silkworms. Specifically, we assessed the weights of fifth-instar larvae that were fed mulberry leaves saturated with three concentrations (2.5, 5, and 10 mg/mL) of MT, as well as the weights of a control group. Methods: TEM was used to show the status of the silkworm midgut after MT supplementation. Oxidative stress was evaluated in the presence of MT. Furthermore, a transcriptomic sequencing experiment was conducted to investigate the mechanism through which the development of silkworms is induced by MT. Results: Our findings indicate that the supplementation of MT hindered larval growth compared to the control group, suggesting a toxic effect of MT on silkworms. The transmission electron microscopy (TEM) results show that MT supplementation induced autophagy in the silkworm midgut. MT was also found to induce oxidative stress in silkworms through the activation of reactive oxygen (ROS), superoxide dismutase (SOD), catalase (CAT), and peroxidase (POD) activities. Subsequent transcriptomic analysis revealed 1265 significantly differentially expressed genes (DEGs) in response to MT. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis indicated that these DEGs were associated with antioxidant defense, detoxification processes, lysosome biogenesis, and metabolic pathways. Conclusions: These findings suggest that MT toxicity in silkworm larvae is mediated through the induction of oxidative stress and alterations in metabolism. This study contributes to our understanding of the impacts of MT exposure on silkworms and provides insights into potential pesticides for use in mulberry gardens. Full article
(This article belongs to the Special Issue Genetics, Development and Functional Genes of Insects)
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<p>Content of MT in mulberry leaves. Data are presented as means ± SDs (<span class="html-italic">n</span> = 3). ***: <span class="html-italic">p</span> &lt; 0.001. ND: Not detected.</p>
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<p>(<b>a</b>) Daily body weights of fifth-instar larvae. The data are presented as the means ± SDs (<span class="html-italic">n</span> = 3). **: <span class="html-italic">p</span> &lt; 0.01; ***: <span class="html-italic">p</span> &lt; 0.001. (<b>b</b>) The morphology of <span class="html-italic">Bombyx mori</span> larvae on the fourth day of the fifth instar stage after MT treatment.</p>
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<p>The general morphology of <span class="html-italic">Bombyx mori</span> larvae midgut on the fourth day of the fifth instar stage. Scale bar: 1 μm (<b>a</b>,<b>b</b>).</p>
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<p>MT supplementation induced oxidative stress in silkworm larvae. (<b>a</b>) ROS levels were significantly increased by MT. (<b>b</b>) SOD activities were significantly increased by MT. (<b>c</b>) CAT activities were significantly increased by MT. (<b>d</b>) POD activities were significantly increased by MT. The data are presented as the means ± SDs (<span class="html-italic">n</span> = 3). **: <span class="html-italic">p</span> &lt; 0.01 ***: <span class="html-italic">p</span> &lt; 0.001. MT_10: MT-treated group (10 mg/mL).</p>
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<p>Volcano plot and KEGG pathway of differentially expressed genes of fifth-instar <span class="html-italic">Bombyx mori</span> larvae following MT treatment. (<b>a</b>) Volcano plot of differentially expressed genes of fifth-instar <span class="html-italic">Bombyx mori</span> larvae following MT treatment. The Y-axis represents the -log10 significance. The X-axis represents the log<sub>2</sub> (fold change). The red dots represent upregulated genes, the green dots represent downregulated genes, and the blue dots represent unchanged genes. (<b>b</b>) The 15 most enriched KEGG pathways based on upregulated genes in the <span class="html-italic">Bombyx mori</span> transcriptome induced by MT. (<b>c</b>) The 15 most enriched KEGG pathways based on downregulated genes in the <span class="html-italic">Bombyx mori</span> transcriptome induced by MT.</p>
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<p>Heatmaps of antioxidation, detoxification, and lysosome gene expression in the present study. (<b>a</b>) The expression level of the peroxisome pathway. (<b>b</b>) The expression level of the cytochrome P450 pathway. (<b>c</b>) The expression level of the lysosome pathway. (<b>d</b>) The expression levels of genes related to the amino acid biosynthesis pathway. (<b>e</b>) The expression levels of genes involved in protein export. The color scale is shown at the upper left, ranging from the lowest (blue) to the highest (red) log10 (expression) value. (<b>f</b>) Quantitative reverse transcriptase–polymerase chain reaction validation of differentially expressed genes related to the peroxisome pathway in silkworms induced by MT. (<b>g</b>) Reverse transcription quantitative transcriptase–polymerase chain reaction (RT-qPCR) validation of differentially expressed genes related to the peroxisome pathway in silkworms induced by MT. The data are presented as the means ± SDs (<span class="html-italic">n</span> = 3). *: <span class="html-italic">p</span> &lt; 0.05 **: <span class="html-italic">p</span> &lt; 0.01. The heatmaps represent the log<sub>10</sub> (averaged values of 3 replicates of FPKM).</p>
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<p>Motif analysis of three CYPs and homologous sequences. (<b>a</b>) Motif analysis of BmorCYP4C1 (KWMTBOMO15838) and homologous sequences. (<b>b</b>) Motif analysis of BmorCYP6B1 (KWMTBOMO12722) and homologous sequences. (<b>c</b>) Motif analysis of BmorCYP4C3 (KWMTBOMO01330) and homologous sequences. Black lines indicate heme-binding domain motif pattern (FXXGXRXCXG). Bmor: <span class="html-italic">Bombyx mori</span>, Atra: <span class="html-italic">Amyelois transitella</span>, Bany: <span class="html-italic">Bicyclus anynana</span>, Bman: <span class="html-italic">Bombyx mandarina</span>, Ccro: <span class="html-italic">Colias croceus</span>, Ceur: <span class="html-italic">Colias eurytheme</span>, Eedi: <span class="html-italic">Euphydryas editha</span>, <span class="html-italic">Harm: Helicoverpa armigera</span>, Lnin: <span class="html-italic">Leptosia nina</span>, Lsti: <span class="html-italic">Loxostege sticticalis</span>, Msex: <span class="html-italic">Manduca sexta</span>, Mhyp: <span class="html-italic">Maniola hyperantus</span>, Mjur: <span class="html-italic">Maniola jurtina</span>, Mlor: <span class="html-italic">Mythimna loreyi</span>, Nio: <span class="html-italic">Nymphalis io</span>, Ofur: <span class="html-italic">Ostrinia furnacalis</span>, Onub: <span class="html-italic">Ostrinia nubilalis</span>, Paeg: <span class="html-italic">Pararge aegeria</span>, Papo: <span class="html-italic">Parnassius apollo</span>, Pmne: <span class="html-italic">Parnassius mnemosyne</span>, Pgos: <span class="html-italic">Pectinophora gossypiella</span>, Pbra: <span class="html-italic">Pieris brassicae</span>, Pnap: <span class="html-italic">Pieris napi</span>, Pint: Plodia interpunctella, Sexi: <span class="html-italic">Spodoptera exigua</span>, Sins: <span class="html-italic">Streltzoviella insularis</span>, Tin: <span class="html-italic">Trichoplusia ni</span>, Vata: Vanessa atalanta, Vcar: <span class="html-italic">Vanessa cardui</span>, Vtam: <span class="html-italic">Vanessa tameamea</span>, Zces: <span class="html-italic">Zerene cesonia</span>, Agri: <span class="html-italic">Achroia grisella</span>, Apla: <span class="html-italic">Arctia plantaginis</span>, Csup: <span class="html-italic">Chilo suppressalis</span>, Cinc: <span class="html-italic">Chrysodeixis includens</span>, Cfag: <span class="html-italic">Cydia fagiglandana</span>, Cpom: <span class="html-italic">Cydia pomonella</span>, Cspl: <span class="html-italic">Cydia spendana</span>, Cstr: <span class="html-italic">Cydia strobilella</span>, Dple: <span class="html-italic">Danaus plexippus</span>, Gmel: <span class="html-italic">Galleria mellonella</span>, Hzea: <span class="html-italic">Helicoverpa zea</span>, Lsti: <span class="html-italic">Loxostege sticticalis</span>, Pxut: <span class="html-italic">Papilio xuthus</span>, Pmac: <span class="html-italic">Pieris macdunnoughi</span>, Prap: <span class="html-italic">Pieris rapae</span>, Pxyl: <span class="html-italic">Plutella xylpstella</span>, Sfru: <span class="html-italic">Spodoptera frugiperda</span>, Slit: <span class="html-italic">Spodoptera littoralis</span>, Aage: <span class="html-italic">Aricia, agestis</span>, Asel: <span class="html-italic">Ascotis selenaria</span>, Camp: <span class="html-italic">Cydia amplana</span>, Dsac: <span class="html-italic">Diatraea saccharalis</span>, Lsin: <span class="html-italic">Leptidea sinapis</span>, Mcin: <span class="html-italic">Melitaea cinxia</span>, Pope: <span class="html-italic">Phthorimaea operculella</span>, Tabs: <span class="html-italic">Tuta absoluta</span>, Zfil: <span class="html-italic">Zygaena filipendulae</span>.</p>
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<p>Scheme of CYP responses to MT stress in silkworm midgut.</p>
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18 pages, 693 KiB  
Article
Effects of Stocking Density of the River Shrimp Cryphiops caementarius on Physiological and Performance Responses in a Biofloc System
by Carlos Andres Mendez, María Cristina Morales and Katherina Brokordt
Fishes 2024, 9(10), 377; https://doi.org/10.3390/fishes9100377 - 25 Sep 2024
Viewed by 524
Abstract
This study aimed to evaluate and compare the effects of biofloc technology (BFT) and clear water (CW) on water quality physiological and productive performance of juvenile freshwater Northern River shrimp, Cryphiops caementarius under three stocking densities (100, 200, and 400 shrimp m−2 [...] Read more.
This study aimed to evaluate and compare the effects of biofloc technology (BFT) and clear water (CW) on water quality physiological and productive performance of juvenile freshwater Northern River shrimp, Cryphiops caementarius under three stocking densities (100, 200, and 400 shrimp m−2). Shrimp with an initial body weight of 0.44 ± 0.07 g were stocked in 18 rectangular fiberglass tanks with a water volume 150 L for 290 days. During the experiment, water quality parameters stayed within acceptable ranges for shrimp growth. The highest survival rate was recorded in BFT treatments; however, the growth performance of shrimp in the treatments with the lowest stocking density was higher than that with the highest stocking density, regardless of whether BFT or CW was used. Transcriptional levels of heat shock protein (Hsp70) and superoxide dismutase (SOD) showed significant differences (p < 0.05) between treatments, particularly in BFT. These results indicate that an initial stocking density of 200 shrimp m−2 appears to be appropriate for shrimp juveniles cultured in a BFT system. Thus, this technology emerges as an effective tool for river shrimp farmers looking to increase their stocking densities and improve the efficiency of their production systems in arid zones. Full article
(This article belongs to the Special Issue Biofloc Technology in Aquaculture)
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<p>Relative expression of genes in juveniles of <span class="html-italic">Criphiops caementarius</span> cultured at different initial stocking densities for 290 days. Effect of stocking density on (<b>A</b>) heat shock protein (Hsp70) and (<b>B</b>) superoxide dismutase (SOD). Data are presented as mean ± standard error. Different letters indicated significant differences between experimental groups (<span class="html-italic">p</span> &lt; 0.05, ANOVA followed by Tukey’s test).</p>
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