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33 pages, 7990 KiB  
Article
Phenotypic, Physiological, and Gene Expression Analysis for Nitrogen and Phosphorus Use Efficienies in Three Popular Genotypes of Rice (Oryza sativa Indica)
by Bhumika Madan and Nandula Raghuram
Plants 2024, 13(18), 2567; https://doi.org/10.3390/plants13182567 - 13 Sep 2024
Viewed by 393
Abstract
Crop nitrogen (N) and phosphorus (P) use efficiencies (NUE/PUE) are important to minimize wastage and nutrient pollution, but no improved crop for both is currently available. We addressed them together in rice, in the view of its high consumption of NPK fertilizers. We [...] Read more.
Crop nitrogen (N) and phosphorus (P) use efficiencies (NUE/PUE) are important to minimize wastage and nutrient pollution, but no improved crop for both is currently available. We addressed them together in rice, in the view of its high consumption of NPK fertilizers. We analyzed 46 morphophysiological parameters for the N/P response in three popular indica genotypes, namely, BPT 5204, Panvel 1, and CR Dhan 301 at low, medium, and normal N/P doses. They include 18 vegetative, 15 physiological, and 13 reproductive parameters. The segregation of significantly N/P-responsive parameters correlating with NUE/PUE revealed 21 NUE, 22 PUE, and 12 common parameters. Feature selection analyses revealed the common high-ranking parameters including the photosynthetic rate at the reproductive stage, tiller number, root–shoot ratio, culm thickness, and flag leaf width. The venn selection using the reported NUE/PUE-related candidate genes in rice revealed five genes in common for both, namely OsIAA3, OsEXPA10, OsCYP75B4, OsSultr3;4, and OsFER2, which were associated with three of the common traits for NUE/PUE. Their expression studies using qRT-PCR revealed the opposite regulation in contrasting genotypes for OsSultr3;4 and OsEXPA10 in N-response and for OsFER2 in P-response, indicating their role in contrasting N/P use efficiencies. Overall, CR Dhan 301 has the highest NUE and PUE followed by Panvel 1 and BPT5204 among the studied genotypes. Full article
(This article belongs to the Section Plant Nutrition)
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Figure 1

Figure 1
<p>Scatter plots showing significant correlations of yield with N-responsive parameters. Correlations of the mean values of each of the parameters, (<b>i</b>–<b>xxii</b>) from three genotypes at normal, medium, and low doses of N with corresponding yield values at <span class="html-italic">p</span>-values of &lt;0.01, 0.02, 0.05, and 0.001 showing significant correlations; <span class="html-italic">p</span>-value of &lt;0.1 means less significant correlation, ns means non-significant correlation.</p>
Full article ">Figure 2
<p>Scatter plots showing significant correlations of yield with P-responsive parameters. Correlations of the mean values of each of the parameters, (<b>i</b>–<b>xvii</b>) from three genotypes at normal, medium, and low doses of P with corresponding yield values at <span class="html-italic">p</span>-values of &lt;0.01, 0.02, 0.05, and 0.001 showing significant correlations; <span class="html-italic">p</span>-value of &lt;0.1 means less significant correlation, ns means non-significant correlation.</p>
Full article ">Figure 3
<p>Scatter plots showing significant correlations of PFP-N with N-responsive parameters. Correlations of the mean values of each of the parameters, (<b>i</b>–<b>xxi</b>) from three genotypes at normal, medium, and low doses of N with corresponding PFP values at <span class="html-italic">p</span>-values of &lt;0.01, 0.02, 0.05, and 0.001 showing significant correlations; <span class="html-italic">p</span>-value of &lt;0.1 means less significant correlation, ns means non-significant correlation.</p>
Full article ">Figure 4
<p>Scatter plots showing significant correlations of PFP-P with P-responsive parameters. Correlations of the mean values of each of the parameters, (<b>i</b>–<b>xx</b>) from three genotypes at normal, medium, and low doses of P with corresponding PFP values at <span class="html-italic">p</span>-values of &lt;0.01, 0.02, 0.05, and 0.001 showing significant correlations; <span class="html-italic">p</span>-value of &lt;0.1 means less significant correlation, ns means non-significant correlation.</p>
Full article ">Figure 5
<p>Scatter plots showing significant correlations of HI with N-responsive parameters. Correlations of the mean values of each of the parameters, (<b>i</b>–<b>xx</b>) from three genotypes at normal, medium, and low doses of N with corresponding HI values at <span class="html-italic">p</span>-values of &lt;0.01, 0.02, 0.05, and 0.001 showing significant correlations; <span class="html-italic">p</span>-value of &lt;0.1 means less significant correlation, ns means non-significant correlation.</p>
Full article ">Figure 6
<p>Scatter plots showing significant correlations of HI with P-responsive parameters. Correlations of the mean values of each of the parameters, (<b>i</b>–<b>xx</b>) from three genotypes at normal, medium, and low doses of P with corresponding HI values at <span class="html-italic">p</span>-values of &lt;0.01, 0.02, 0.05, and 0.001 showing significant correlations; <span class="html-italic">p</span>-value of &lt;0.1 means less significant correlation, ns means non-significant correlation.</p>
Full article ">Figure 7
<p>Loading plots of first two principal components of N-response (<b>i</b>) and P-response (<b>ii</b>). PCA was performed using the data of all parameters at all doses of N and P combined, plotted using Minitab v21.4.2 statistical software.</p>
Full article ">Figure 8
<p>PLS-DA 2D score plots of the N (<b>i</b>) and P (<b>ii</b>) response of three genotypes (BPT for BPT 5204, Pnvl for Panvel 1, and CRDh for CR Dhan 301). PLS-DA was performed for all 46 measured parameters at 3 doses using MetaboAnalyst 6.0 software.</p>
Full article ">Figure 9
<p>Variable Importance plots showing ranking of parameters. Plots for NUE (<b>i</b>) and PUE (<b>ii</b>) parameters as analyzed by RandomForest feature selection tool using MetaboAnalyst 6.0 software. The common parameters for NUE and PUE were plotted separately (<b>iii</b>,<b>iv</b>), in view of the different average values for N/P-response in the respective parameters.</p>
Full article ">Figure 10
<p>N dose-wise response of common parameters (<b>i</b>–<b>xii</b>) associated with NUE and PUE. These are average values of 20 replicates, plotted using GraphPad Prism. Two-way ANOVA was performed to check the significance of the effect of dose, genotypes, and their interaction on the average value of the parameters. <span class="html-italic">p</span>-values are summarized with asterisks and calculated with respect to LN (10%N) as control. <span class="html-italic">p</span>-values &lt; 0.01–0.05 indicated as *, <span class="html-italic">p</span>-values &lt; 0.001–0.01 indicated as **, <span class="html-italic">p</span>-values &lt; 0.0001–0.001 indicated as ***, and <span class="html-italic">p</span>-values &lt; 0.0001 indicated as **** and ns means non-significant.</p>
Full article ">Figure 11
<p>P dose-wise response of common parameters (<b>i–xii</b>) associated with NUE and PUE. These are average values of 20 replicates, plotted using GraphPad Prism. Two-way ANOVA was performed to check the significance of the effect of dose, genotypes, and their interaction on the average value of the parameters. <span class="html-italic">P</span>-values are summarized with asterisks and calculated with respect to LP (5%P) as control. <span class="html-italic">p</span>-values &lt; 0.01–0.05 indicated as *, <span class="html-italic">p</span>-values &lt; 0.001–0.01 indicated as **, <span class="html-italic">p</span>-values &lt; 0.0001–0.001 indicated as ***, and <span class="html-italic">p</span>-values &lt; 0.0001 indicated as **** and ns means non-significant.</p>
Full article ">Figure 12
<p>Yield, PFP, and HI of all three genotypes under 3 N and P doses (<b>i</b>). Number of filled seeds per panicle under three N doses (<b>ii</b>). PFP under three N doses (<b>iii</b>). Percentage harvest index under three N doses (<b>iv</b>). Number of filled seeds per panicle under three P doses (<b>v</b>). PFP under three P doses (<b>vi</b>). Percentage harvest index under three P doses. These are average values of 20 replicates, plotted using GraphPad Prism v6.01. Two-way ANOVA was performed to check the significance of the effect of dose, genotypes, and their interaction on the average value of the parameters.</p>
Full article ">Figure 13
<p>RT-qPCR graphs showing log<sub>2</sub> fold change values in five NUE/PUE common genes. These are represented as mean ± SE from two biological replicates in all three genotypes grown under (<b>i</b>–<b>v</b>) 50% N (7.5 mM) and 100% N (15 mM) with 10% N (1.5 mM) as control (<b>vi</b>–<b>x</b>) 10% P (0.1 mM) and 100% P (1 mM) with 5% P (0.05 mM) as control, <span class="html-italic">OsFER2</span> (Os12g0106000), <span class="html-italic">OsEXPA10</span> (Os04g0583500), <span class="html-italic">OsSultr3;4</span> (Os06g0143700), <span class="html-italic">OsCYP75B4</span> (Os10g0317900), and <span class="html-italic">OsIAA3</span> (Os01g0231000). Two-way ANOVA was performed to check the significance of the effect of dose, genotypes, and their interaction on the Log<sub>2</sub> fold change values of the individual genes. <span class="html-italic">p</span>-values are summarized with asterisks and calculated with respect to Log<sub>2</sub> fold change values at medium doses for individual genes. <span class="html-italic">p</span>-values &lt; 0.01–0.05 indicated as *, <span class="html-italic">p</span>-values &lt; 0.001–0.01 indicated as **, <span class="html-italic">p</span>-values &lt; 0.0001–0.001 indicated as ***, and <span class="html-italic">p</span>-values &lt; 0.0001 indicated as **** and ns means non-significant.</p>
Full article ">
12 pages, 4063 KiB  
Article
Selection and Validation of Reference Genes for Quantitative Real-Time PCR Analysis in Cockroach Parasitoid Tetrastichus hagenowii (Ratzeburg)
by Renke Dong, Fengming Cao, Jincong Yu, Yuan Yuan, Jiahui Wang, Zining Li, Chunxue Zhu, Sheng Li and Na Li
Insects 2024, 15(9), 668; https://doi.org/10.3390/insects15090668 - 3 Sep 2024
Viewed by 570
Abstract
Parasitoid wasps play a crucial role in the efficient control of pests, a substantial menace to human health and well-being. Tetrastichus hagenowii (Ratzeburg) stands out as the most effective egg parasitoid wasp for controlling American cockroaches, but accurate and stable reference genes for [...] Read more.
Parasitoid wasps play a crucial role in the efficient control of pests, a substantial menace to human health and well-being. Tetrastichus hagenowii (Ratzeburg) stands out as the most effective egg parasitoid wasp for controlling American cockroaches, but accurate and stable reference genes for quantitative real-time polymerase chain reaction of T. hagenowii genes are still lacking. In this study, we assessed seven candidate nuclear genes, including α-tubulin (α-TUB), elongation factor-1-alpha (EF-1α), β-actin (Actin), ribosomal protein 49 (RP49), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), nicotinamide adenine dinucleotide (NADH), and elongation factor 2 (EF2) of T. hagenowii. By analyzing expression stability with four algorithms (Delta Ct, geNorm, NormFinder, and BestKeeper), as well as comprehensive ranking with RefFinder, we identified α-TUB as the most stable reference gene for the larval, pupal, female adult, and male adult stages. Subsequently, we estimated the transcript levels of vitellogenin (Vg) and cuticle protein (CP) after normalization with α-TUB across various developmental stages. Significantly higher expression levels of CP and Vg were observed in pupae and female adults, respectively, consistent with previous findings in other insects. This study offers a reliable reference gene for normalizing transcription levels of T. hagenowii genes. Full article
(This article belongs to the Section Insect Molecular Biology and Genomics)
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Figure 1

Figure 1
<p>Melting curve to verify the reference gene amplification efficiency and uniformity. A single peak indicates a single RT-qPCR product. <span class="html-italic">n</span> = 6 for each developmental stage.</p>
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<p>Agarose gel electrophoresis to test the amplification of RT-qPCR primers for candidate reference genes. The RT-qPCR product size for each gene was shown in <a href="#insects-15-00668-t001" class="html-table">Table 1</a>. Three biological replicates of mixed samples from the larval, pupal, female adult, and male adult stages were prepared for each gene. The figure shows DNA bands for cDNA samples and the absence of bands for water negative controls.</p>
Full article ">Figure 3
<p>Ct values of the candidate genes evaluated in larvae, pupae, and adults of <span class="html-italic">T. hagenowii</span><b>.</b> The upper and lower whiskers indicate the maximum and minimum values, respectively. The upper and lower edges of the box represent the upper and lower quartiles, while the middle black line signifies the median. The length of each graphic (whiskers) corresponds to its variation. <span class="html-italic">n</span> = 4 for each developmental stage.</p>
Full article ">Figure 4
<p>Stability rankings of the seven candidate reference genes in <span class="html-italic">T. hagenowii</span> calculated separately by Delta Ct, BestKeeper, NormFinder, geNorm, and RefFinder. The stability values and data are presented from the least stable (left) to the most stable gene (right). (<b>A</b>). Delta Ct; (<b>B</b>). BestKeeper; (<b>C</b>). NormFinder; (<b>D</b>). geNorm; (<b>E</b>). RefFinder.</p>
Full article ">Figure 5
<p>Relative expression levels of <span class="html-italic">Vg</span> (<b>A</b>) and <span class="html-italic">CP</span> (<b>B</b>) across developmental stages in <span class="html-italic">T. hagenowii</span>. Four biological replicates were prepared for each developmental stage analyzed using RT-qPCR. Transcript levels were normalized with <span class="html-italic">α-TUB</span> or <span class="html-italic">Actin</span> using the 2<sup>−ΔΔCT</sup> method and are presented as ratios relative to that of larvae (mean = 1). Columns represent mean values, with error bars indicating standard error (SE). One-way ANOVA was used to assess differences among developmental stages. Different letters above bars mean significant difference between different groups.</p>
Full article ">
29 pages, 10764 KiB  
Article
In Silico Drug Screening for Hepatitis C Virus Using QSAR-ML and Molecular Docking with Rho-Associated Protein Kinase 1 (ROCK1) Inhibitors
by Joshua R. De Borja and Heherson S. Cabrera
Computation 2024, 12(9), 175; https://doi.org/10.3390/computation12090175 - 31 Aug 2024
Viewed by 884
Abstract
The enzyme ROCK1 plays a pivotal role in the disruption of the tight junction protein CLDN1, a downstream effector influencing various cellular functions such as cell migration, adhesion, and polarity. Elevated levels of ROCK1 pose challenges in HCV, where CLDN1 serves as a [...] Read more.
The enzyme ROCK1 plays a pivotal role in the disruption of the tight junction protein CLDN1, a downstream effector influencing various cellular functions such as cell migration, adhesion, and polarity. Elevated levels of ROCK1 pose challenges in HCV, where CLDN1 serves as a crucial entry factor for viral infections. This study integrates a drug screening protocol, employing a combination of quantitative structure–activity relationship machine learning (QSAR-ML) techniques; absorption, distribution, metabolism, and excretion (ADME) predictions; and molecular docking. This integrated approach allows for the effective screening of specific compounds, using their calculated features and properties as guidelines for selecting drug-like candidates targeting ROCK1 inhibition in HCV treatment. The QSAR-ML model, validated with scores of 0.54 (R2), 0.15 (RMSE), and 0.71 (CCC), demonstrates its predictive capabilities. The ADME-Docking study’s final results highlight notable compounds from ZINC15, specifically ZINC000071318464, ZINC000073170040, ZINC000058568630, ZINC000058591055, and ZINC000058574949. These compounds exhibit the best ranking Vina scores for protein–ligand binding with the crystal structure of ROCK1 at the C2 pocket site. The generated features and calculated pIC50 bioactivity of these compounds provide valuable insights, facilitating the identification of structurally similar candidates in the ongoing exploration of drugs for ROCK1 inhibition. Full article
(This article belongs to the Special Issue 10th Anniversary of Computation—Computational Biology)
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Figure 1

Figure 1
<p>Outline of the entire methods section as well as the programs and web servers used in the following study. Data mining (EBI-ChEMBL, PubChem and ZINC15 databases) and pre-processing (R language in RStudio IDE). Feature engineering, statistical analysis and QSAR-ML performed in all in Python scripting using Jupyter notebook and Google Colab alongside modules from Mordred, RDKit, scikit-learn and Joblib. ADME prediction for generating the bioavailability radar and BOILED-EGG representation of the compounds. Molecular docking carried out using CB-Dock2 for cavity detection and protein–ligand docking scoring calculation.</p>
Full article ">Figure 2
<p>Bioactivity distributions. (<b>a</b>) HCV (NS5B polymerase, NS3, and NS3/4A proteases) set; (<b>b</b>) ROCK1 set. The frequency distribution shows the number of each inhibitor compound considered in three classes as either “active” (pIC<sub>50</sub> &gt; 5.4), “intermediate” (5.00 &lt; pIC<sub>50</sub> &gt; 5.39), or “inactive” (pIC<sub>50</sub> &lt; 4.99). The screening process for bioactivity distribution is detailed in the <a href="#app1-computation-12-00175" class="html-app">Supplementary Material (Supplement S1: Code Repository and Dataset Repository)</a>.</p>
Full article ">Figure 3
<p>Histogram plots of the logarithmic distribution of the <span class="html-italic">y</span> (pIC<sub>50</sub>) values visualized at 100 bins. Where frequency represents the count values and how each y datasets are logarithmically distributed after activity sampling is employed. (<b>a</b>) Train set; (<b>b</b>) test set; (<b>c</b>) validation set.</p>
Full article ">Figure 3 Cont.
<p>Histogram plots of the logarithmic distribution of the <span class="html-italic">y</span> (pIC<sub>50</sub>) values visualized at 100 bins. Where frequency represents the count values and how each y datasets are logarithmically distributed after activity sampling is employed. (<b>a</b>) Train set; (<b>b</b>) test set; (<b>c</b>) validation set.</p>
Full article ">Figure 4
<p>Scatter and prediction error plots of NuSVR between the actual (true values of <span class="html-italic">y</span>) vs. predicted values of <span class="html-italic">y</span> in each dataset, not accounting for the training. (<b>a</b>) Test set; (<b>b</b>) validation set.</p>
Full article ">Figure 4 Cont.
<p>Scatter and prediction error plots of NuSVR between the actual (true values of <span class="html-italic">y</span>) vs. predicted values of <span class="html-italic">y</span> in each dataset, not accounting for the training. (<b>a</b>) Test set; (<b>b</b>) validation set.</p>
Full article ">Figure 5
<p>The PKA dataset as an external validation and its metrics. (<b>a</b>) Logarithmic distribution of <span class="html-italic">y</span>; (<b>b</b>) regression and prediction error plots between true <span class="html-italic">y</span> and predicted <span class="html-italic">y</span> values in the set. The external validation dataset is computed separately but followed the same process as the other datasets. <a href="#app1-computation-12-00175" class="html-app">Supplementary Material (Supplement S1: Code and Dataset Repository)</a>.</p>
Full article ">Figure 6
<p>The top 25 features in the <span class="html-italic">y</span>-axis that NuSVR mostly relies upon for it to have a good R<sup>2</sup> represented in the <span class="html-italic">x</span>-axis after calculations were carried out using the permutation_importance function from scikit-learn.</p>
Full article ">Figure 7
<p>BOILED-Egg representation in SwissADME of the screened compounds from QSAR-ML represented by their last three PubChem ID numbers for the evaluation of their gastrointestinal absorption and blood–brain barrier penetration.</p>
Full article ">Figure 8
<p>Three-dimensional structure of ROCK1. (<b>a</b>) The retrieved X-ray crystal structure of ROCK1 (PDB ID: 1S1C) at 2.60 Å resolution from RCSB Protein Data Bank. Its cavity pockets were detected using CurPocket with their IDs from CB-Dock2: (<b>b</b>) C1; (<b>c</b>) C2; (<b>d</b>) C3; (<b>e</b>) C4; (<b>f</b>) C5.</p>
Full article ">Figure 8 Cont.
<p>Three-dimensional structure of ROCK1. (<b>a</b>) The retrieved X-ray crystal structure of ROCK1 (PDB ID: 1S1C) at 2.60 Å resolution from RCSB Protein Data Bank. Its cavity pockets were detected using CurPocket with their IDs from CB-Dock2: (<b>b</b>) C1; (<b>c</b>) C2; (<b>d</b>) C3; (<b>e</b>) C4; (<b>f</b>) C5.</p>
Full article ">Figure 9
<p>Protein–ligand complexes are visualized in PlayMolecule. The top CurPockets ranked based on Autodock Vina’s scoring function in each of the protein–ligand complexes: (<b>a</b>) (Velpatasvir)1S1C-ZINC000203686879 at C2; (<b>b</b>) 1S1C-ZINC000071318464 at C2; (<b>c</b>) 1S1C-ZINC000071296700 at C1; (<b>d</b>) 1S1C-ZINC000071315829 at C1; (<b>e</b>) 1S1C-ZINC000073170040 at C2; (<b>f</b>) 1S1C-ZINC000058568630 at C2; (<b>g</b>) 1S1C-ZINC000073196364 at C1; (<b>h</b>) 1S1C-ZINC000058591055 at C2; (<b>i</b>) 1S1C-ZINC000058568675 at C1; (<b>j</b>) 1S1C-ZINC000058574949 at C2.</p>
Full article ">Figure 9 Cont.
<p>Protein–ligand complexes are visualized in PlayMolecule. The top CurPockets ranked based on Autodock Vina’s scoring function in each of the protein–ligand complexes: (<b>a</b>) (Velpatasvir)1S1C-ZINC000203686879 at C2; (<b>b</b>) 1S1C-ZINC000071318464 at C2; (<b>c</b>) 1S1C-ZINC000071296700 at C1; (<b>d</b>) 1S1C-ZINC000071315829 at C1; (<b>e</b>) 1S1C-ZINC000073170040 at C2; (<b>f</b>) 1S1C-ZINC000058568630 at C2; (<b>g</b>) 1S1C-ZINC000073196364 at C1; (<b>h</b>) 1S1C-ZINC000058591055 at C2; (<b>i</b>) 1S1C-ZINC000058568675 at C1; (<b>j</b>) 1S1C-ZINC000058574949 at C2.</p>
Full article ">Figure 10
<p>Protein–ligand contact residues and bonds are in PlayMolecule’s Plexview. Only the docked complexes at the C2 site are represented, (<b>a</b>) (Velpatasvir)1S1C-ZINC000203686879; (<b>b</b>) 1S1C-ZINC000071318464; (<b>c</b>) 1S1C-ZINC000073170040; (<b>d</b>) 1S1C-ZINC000058568630; (<b>e</b>) 1S1C-ZINC000058591055; (<b>f</b>) 1S1C-ZINC000058574949.</p>
Full article ">Figure 10 Cont.
<p>Protein–ligand contact residues and bonds are in PlayMolecule’s Plexview. Only the docked complexes at the C2 site are represented, (<b>a</b>) (Velpatasvir)1S1C-ZINC000203686879; (<b>b</b>) 1S1C-ZINC000071318464; (<b>c</b>) 1S1C-ZINC000073170040; (<b>d</b>) 1S1C-ZINC000058568630; (<b>e</b>) 1S1C-ZINC000058591055; (<b>f</b>) 1S1C-ZINC000058574949.</p>
Full article ">
14 pages, 3609 KiB  
Article
Applying Molecular Modeling to the Design of Innovative, Non-Symmetrical CXCR4 Inhibitors with Potent Anticancer Activity
by Miquel Martínez-Asensio, Lluís Sàrrias, Gema Gorjón-de-Pablo, Miranda Fernández-Serrano, Judith Camaló-Vila, Albert Gibert, Raimon Puig de la Bellacasa, Jordi Teixidó, Gaël Roué, José I. Borrell and Roger Estrada-Tejedor
Int. J. Mol. Sci. 2024, 25(17), 9446; https://doi.org/10.3390/ijms25179446 - 30 Aug 2024
Viewed by 546
Abstract
The identification of new compounds with potential activity against CXC chemokine receptor type 4 (CXCR4) has been broadly studied, implying several chemical families, particularly AMD3100 derivatives. Molecular modeling has played a pivotal role in the identification of new active compounds. But, has its [...] Read more.
The identification of new compounds with potential activity against CXC chemokine receptor type 4 (CXCR4) has been broadly studied, implying several chemical families, particularly AMD3100 derivatives. Molecular modeling has played a pivotal role in the identification of new active compounds. But, has its golden age ended? A virtual library of 450,000 tetraamines of general structure 8 was constructed by using five spacers and 300 diamines, which were obtained from the corresponding commercially available cyclic amines. Diversity selection was performed to guide the virtual screening of the former database and to select the most representative set of compounds. Molecular docking on the CXCR4 crystal structure allowed us to rank the selection and identify those candidate molecules with potential antitumor activity against diffuse large B-cell lymphoma (DLBCL). Among them, compound A{17,18} stood out for being a non-symmetrical structure, synthetically feasible, and with promising activity against DLBCL in in vitro experiments. The focused study of symmetrical-related compounds allowed us to identify potential pre-hits (IC50~20 µM), evidencing that molecular design is still relevant in the development of new CXCR4 inhibitor candidates. Full article
(This article belongs to the Section Biochemistry)
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Figure 1

Figure 1
<p>(<b>a</b>) Structure of bicyclam AMD3100 (<b>1</b>) and monocyclam AMD3465 (<b>2</b>); (<b>b</b>) example of the general structure of non-cyclam CXCR4 inhibitors reported by Zhan et al. [<a href="#B11-ijms-25-09446" class="html-bibr">11</a>] and Fang et al. [<a href="#B12-ijms-25-09446" class="html-bibr">12</a>].</p>
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<p>General structure of tetramines <b>3</b> and <b>4</b> and hit compound <b>5</b> (chiral centers are labeled with asterisks).</p>
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<p>General procedure for the discovery of novel CXCR4 inhibitors. The structure of tetraamines <b>8</b> was considered for the creation of the combinatorial library used in the structure-based virtual screening of potential CXCR4 inhibitors. Diversity selection and molecular modeling were used for the identification of those compounds to be synthesized and biologically tested. Finally, compound <b>A</b>{<span class="html-italic">17</span>,<span class="html-italic">18</span>} was identified as a pre-hit.</p>
Full article ">Figure 4
<p>(<b>a</b>) Projection of the assessed combinatorial library on the first two principal components. Selected compounds are colored in blue and the space described by the full combinatorial library is in orange. (<b>b</b>) Molecular weight distribution for both sets. (<b>c</b>) Density distribution of docking scores obtained for the selected molecules.</p>
Full article ">Figure 5
<p>Non-symmetrical compound <b>A</b>{<span class="html-italic">17</span>,<span class="html-italic">18</span>} resulting from the virtual screening protocol and its corresponding two symmetrical compounds <b>A</b>{<span class="html-italic">17</span>,<span class="html-italic">17</span>} and <b>A</b>{<span class="html-italic">18</span>,<span class="html-italic">18</span>}.</p>
Full article ">Figure 6
<p>Interaction mechanism predicted by molecular docking and molecular dynamics simulations for compounds <b>A</b>{<span class="html-italic">18</span>,<span class="html-italic">18</span>}, <b>A</b>{<span class="html-italic">17</span>,<span class="html-italic">18</span>}, and <b>A</b>{<span class="html-italic">17</span>,<span class="html-italic">17</span>}.</p>
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<p>MTT cell viability and proliferation assay results in PBMCs from different healthy donors (<span class="html-italic">n</span> = 3) and in Karpas 422 (GCB-DLBCL) and HBL-1 (ABC-DLBCL) cell lines (<span class="html-italic">n</span> = 3) at 24 h for all compounds under study. Data are presented as means ± SD. Statistics: One-Way ANOVA statistical 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.</p>
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<p>Molecular structure of spacers (<b>A</b>–<b>E</b>) chosen for the creation of the virtual combinatorial library.</p>
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<p>Synthesis of diamines <b>9</b>{<span class="html-italic">17</span>} and <b>9</b>{<span class="html-italic">18</span>} from <span class="html-italic">N</span>-ethylpiperazine <b>10</b>{<span class="html-italic">17</span>} and <span class="html-italic">N</span>-methylcyclohexanamine <b>10</b>{<span class="html-italic">18</span>}.</p>
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<p>(<b>a</b>) Synthesis of symmetrical tetraamines <b>A</b>{<span class="html-italic">17</span>,<span class="html-italic">17</span>} and <b>A</b>{<span class="html-italic">18</span>,<span class="html-italic">18</span>}. (<b>b</b>) Synthesis of tetraamine <b>A</b>{<span class="html-italic">17</span>,<span class="html-italic">18</span>}.</p>
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26 pages, 3120 KiB  
Article
Multi-Omics Analysis Revealed the rSNPs Potentially Involved in T2DM Pathogenic Mechanism and Metformin Response
by Igor S. Damarov, Elena E. Korbolina, Elena Y. Rykova and Tatiana I. Merkulova
Int. J. Mol. Sci. 2024, 25(17), 9297; https://doi.org/10.3390/ijms25179297 - 27 Aug 2024
Viewed by 455
Abstract
The goal of our study was to identify and assess the functionally significant SNPs with potentially important roles in the development of type 2 diabetes mellitus (T2DM) and/or their effect on individual response to antihyperglycemic medication with metformin. We applied a bioinformatics approach [...] Read more.
The goal of our study was to identify and assess the functionally significant SNPs with potentially important roles in the development of type 2 diabetes mellitus (T2DM) and/or their effect on individual response to antihyperglycemic medication with metformin. We applied a bioinformatics approach to identify the regulatory SNPs (rSNPs) associated with allele-asymmetric binding and expression events in our paired ChIP-seq and RNA-seq data for peripheral blood mononuclear cells (PBMCs) of nine healthy individuals. The rSNP outcomes were analyzed using public data from the GWAS (Genome-Wide Association Studies) and Genotype-Tissue Expression (GTEx). The differentially expressed genes (DEGs) between healthy and T2DM individuals (GSE221521), including metformin responders and non-responders (GSE153315), were searched for in GEO RNA-seq data. The DEGs harboring rSNPs were analyzed using the Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG). We identified 14,796 rSNPs in the promoters of 5132 genes of human PBMCs. We found 4280 rSNPs to associate with both phenotypic traits (GWAS) and expression quantitative trait loci (eQTLs) from GTEx. Between T2DM patients and controls, 3810 rSNPs were detected in the promoters of 1284 DEGs. Based on the protein-protein interaction (PPI) network, we identified 31 upregulated hub genes, including the genes involved in inflammation, obesity, and insulin resistance. The top-ranked 10 enriched KEGG pathways for these hubs included insulin, AMPK, and FoxO signaling pathways. Between metformin responders and non-responders, 367 rSNPs were found in the promoters of 131 DEGs. Genes encoding transcription factors and transcription regulators were the most widely represented group and many were shown to be involved in the T2DM pathogenesis. We have formed a list of human rSNPs that add functional interpretation to the T2DM-association signals identified in GWAS. The results suggest candidate causal regulatory variants for T2DM, with strong enrichment in the pathways related to glucose metabolism, inflammation, and the effects of metformin. Full article
(This article belongs to the Special Issue Advances in Molecular Research of Diabetes Mellitus)
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<p>Scheme of the main stages in the search for rSNPs and their further analysis. The solid gray frame shows the stages of the search for rSNPs; the dotted gray frame, further analysis of the rSNP panel; and the cyan italic, data sources.</p>
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<p>Venn diagram showing the number of the rSNPs localized to allele-specific transcription factor binding sites (ANANASTRA), rSNPs associated phenotypic traits (GWAS data), and eQTLs effects (GTEx data).</p>
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<p>Volcano plot of DEGs. The horizontal axis stands for log2 fold change and the vertical axis, for –log<sub>10</sub> (adjusted <span class="html-italic">p</span>-value). Statistically significant DEGs harboring rSNPs in their promoters are marked red.</p>
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<p>Network chart illustrating the link of top ten significant KEGG pathways according to the enrichment analysis with upregulated hub genes.</p>
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<p>PPI network in the significant modules analyzed for KEGG and GO enrichment. (<b>A</b>) First module. (<b>B</b>) Second module. (<b>C</b>) Third module. In the PPI network, nodes show proteins and edges, their interaction. Hub proteins are denoted with larger symbols and the KEGG- and GO-annotated proteins, with the corresponding color (see the legend).</p>
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<p>Graphical visualization of DEG representation in (<b>A</b>) insulin and (<b>B</b>) PI3K/Akt signaling pathways (KEGG data). Colors of nodes show the direction and value (log2FC) of expression alteration.</p>
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<p>Graphical visualization of DEG representation in (<b>A</b>) insulin and (<b>B</b>) PI3K/Akt signaling pathways (KEGG data). Colors of nodes show the direction and value (log2FC) of expression alteration.</p>
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<p>Volcano plot of DEGs. The horizontal axis shows the log<sub>2</sub> fold change and the vertical axis, –log<sub>10</sub> (adjusted <span class="html-italic">p</span>-value). Significant DEGs with detected rSNPs are colored red.</p>
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13 pages, 3092 KiB  
Article
Screening Wheat Genotypes for Specific Genes Linked to Drought Tolerance
by Ahmed Sallam, Mohamed M. H. El-Defrawy, Mona F. A. Dawood and Mostafa Hashem
Genes 2024, 15(9), 1119; https://doi.org/10.3390/genes15091119 - 24 Aug 2024
Viewed by 778
Abstract
Drought stress, which significantly affects growth and reduces grain yield, is one of the main problems for wheat crops. Producing promising drought-tolerant wheat cultivars with high yields is one of the main targets for wheat breeders. In this study, a total of seven [...] Read more.
Drought stress, which significantly affects growth and reduces grain yield, is one of the main problems for wheat crops. Producing promising drought-tolerant wheat cultivars with high yields is one of the main targets for wheat breeders. In this study, a total of seven drought-tolerant wheat genotypes were screened for the presence of 19 specific drought tolerance genes. The genotypes were tested under normal and drought conditions for two growing seasons. Four spike traits, namely, spike length (SPL), grain number per spike (GNPS), number of spikelets per spike (NSPS), and grain yield per spike (GYPS), were scored. The results revealed that drought stress decreased the SPL, GNPS, NSPS, and GYPS, with ranges ranging from 2.14 (NSPS) to 13.92% (GNPS) and from 2.40 (NSPS) to 11.09% (GYPS) in the first and second seasons, respectively. ANOVA revealed high genetic variation among the genotypes for each trait under each treatment. According to the drought tolerance indices, Omara 007 presented the highest level of drought tolerance (average of sum ranks = 3), whereas both Giza-36 genotypes presented the lowest level of drought tolerance (average of sum ranks = 4.8) among the genotypes tested. Among the 19 genes tested, 11 were polymorphic among the selected genotypes. Omara 007 and Omara 002 presented the greatest number of specific drought tolerance genes (nine) tested in this study, whereas Sohag-5, Giza-36, and PI469072 presented the lowest number of drought tolerance genes (four). The number of different genes between each pair of genotypes was calculated. Seven different genes were found between Omara 007 and Giza-36, Omara 007 and Sohag-5, and Omara 002 and PI469072. The results of this study may help to identify the best genotypes for crossing candidate genotypes, and not merely to genetically improve drought tolerance in wheat. Full article
(This article belongs to the Collection Feature Papers: 'Plant Genetics and Genomics' Section)
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<p>The reduction due to drought of studied traits in both seasons (<b>a</b>), and average of sum ranks (ASR) of all stress indices for each genotype in two seasons (<b>b</b>). SPL: spike length, NSPS: number of spikes per spike, GNPS: grain number per spike, GYPS: grain yield per spike.</p>
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<p>Agarose gel electrophoresis of DREB genes used in this study. Names of genotypes are supplied in <a href="#app1-genes-15-01119" class="html-app">Supplementary Table S1</a>.</p>
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<p>Agarose gel electrophoresis of DREB genes used in this study. Names of genotypes are supplied in <a href="#app1-genes-15-01119" class="html-app">Supplementary Table S1</a>.</p>
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<p>The position of the detected genes among the seven genotypes by PCR on wheat chromosomes.</p>
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<p>Number of genes and the average of ASR (two growing seasons) for each genotype (<b>a</b>) and the numbers of different genes found between genotypes (<b>b</b>).</p>
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19 pages, 3692 KiB  
Article
Screening and Evaluation of Biomechanical Properties and Morphological Characteristics of Peduncles in Foxtail Millet
by Lili Zhang, Guofang Xing, Zhenyu Liu, Yanqing Zhang, Hongbo Li, Yuanmeng Wang, Jiaxin Lu, Nan An, Zhihong Zhao, Zeyu Wang, Yuanhuai Han and Qingliang Cui
Agriculture 2024, 14(9), 1437; https://doi.org/10.3390/agriculture14091437 - 23 Aug 2024
Viewed by 380
Abstract
Mechanized harvesting is a crucial step in the agricultural production of foxtail millet (Setaria italica), as its peduncles are susceptible to bending and breaking during the harvesting process, leading to yield losses and deterioration in grain quality. To evaluate the suitability [...] Read more.
Mechanized harvesting is a crucial step in the agricultural production of foxtail millet (Setaria italica), as its peduncles are susceptible to bending and breaking during the harvesting process, leading to yield losses and deterioration in grain quality. To evaluate the suitability of foxtail millet for mechanical harvesting, this study comprehensively analyzed the biomechanical properties of the peduncles and related biological morphological characteristics of 116 foxtail millet accessions, establishing a system for indicator screening and comprehensive evaluation. Using partial correlation analysis and R-type cluster analysis, four biomechanical and seven related morphological indices of the peduncle were screened from 22 candidate indicators, with their coefficient of variation ranging from 6% to 80%. The entropy method was used to assign weights to the selected indices, with biomechanical factors contributing 47.4%, peduncle morphology 20.2%, spike morphology 27.6%, and plant height 4.8%. The Technique for Order Preference by Similarity to Ideal Solution (TOPSIS) and Rank-Sum Ratio (RSR) methods were applied to rank and grade the classification of the 116 foxtail millet varieties into four performance groups: Excellent (8 varieties), Good (50 varieties), Moderate (51 varieties), and Poor (7 varieties). This study provides a scientific basis for the selection and evaluation of foxtail millet varieties. Full article
(This article belongs to the Section Agricultural Technology)
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<p>The field sampling site and test samples: (<b>a</b>) The field sampling site; (<b>b</b>) Samples of foxtail millet plants used for measuring plant height; (<b>c</b>) Samples of stem and peduncle in foxtail millet; (<b>d</b>) Samples of spike in foxtail millet.</p>
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<p>Tests for the biomechanical properties of the peduncle in foxtail millet: (<b>a</b>) Physical diagram of the bending test; (<b>b</b>) Schematic diagram of the bending test; (<b>c</b>) Physical diagram of the shear test; (<b>d</b>) Schematic diagram of the shear test. Component 1 is a sensor, Component 2 is the clamp for the bending test, Component 3 is the peduncle, Component 4 is the base, and Component 5 is the clamp for the shear test.</p>
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<p>Load-displacement curves for the bending and shear tests of the peduncle in foxtail millet: (<b>a</b>) Load-displacement curves for the bending test; (<b>b</b>) Load-displacement curves for the shear test.</p>
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<p>Heatmap and hierarchical clustering diagram of the partial correlation coefficients for the 15 morphological characteristics in 116 foxtail millet accessions. The variables on the axes are abbreviations of each indicator. (<b>a</b>) Partial correlation and matrix heatmap; (<b>b</b>) Partial correlation plot highlighting significant correlations with rectangles and indicating the hierarchical clustering order.</p>
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<p>Heatmap and hierarchical clustering diagram of the partial correlation coefficients for the seven biomechanical properties in 116 foxtail millet accessions. The variables on the axes are abbreviations of each indicator. (<b>a</b>) Partial correlation and matrix heatmap; (<b>b</b>) Partial correlation plot highlighting significant correlations with rectangles and indicating the hierarchical clustering order.</p>
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<p>Radar chart for seven morphological characteristics of the plant and four biomechanical properties of the peduncle in 116 foxtail millet accessions. (<b>a</b>) Radar chart for seven morphological characteristics; (<b>b</b>) Radar chart for four biomechanical properties.</p>
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27 pages, 8277 KiB  
Article
High-Resolution Identification of Sound Sources Based on Sparse Bayesian Learning with Grid Adaptive Split Refinement
by Wei Pan, Daofang Feng, Youtai Shi, Yan Chen and Min Li
Appl. Sci. 2024, 14(16), 7374; https://doi.org/10.3390/app14167374 - 21 Aug 2024
Viewed by 369
Abstract
Sound source identification technology based on a microphone array has many application scenarios. The compressive beamforming method has attracted much attention due to its high accuracy and high-resolution performance. However, for the far-field measurement problem of large microphone arrays, existing methods based on [...] Read more.
Sound source identification technology based on a microphone array has many application scenarios. The compressive beamforming method has attracted much attention due to its high accuracy and high-resolution performance. However, for the far-field measurement problem of large microphone arrays, existing methods based on fixed grids have the defect of basis mismatch. Due to the large number of grid points representing potential sound source locations, the identification accuracy of traditional grid adjustment methods also needs to be improved. To solve this problem, this paper proposes a sound source identification method based on adaptive grid splitting and refinement. First, the initial source locations are obtained through a sparse Bayesian learning framework. Then, higher-weight candidate grids are retained, and local regions near them are split and updated. During the iteration process, Green’s function and the source strength obtained in the previous iteration are multiplied to get the sound pressure matrix. The robust principal component analysis model of the Gaussian mixture separates and replaces the sound pressure matrix with a low-rank matrix. The actual sound source locations are gradually approximated through the dynamically adjusted sound pressure low-rank matrix and optimized grid transfer matrix. The performance of the method is verified through numerical simulations. In addition, experiments on a standard aircraft model are conducted in a wind tunnel and speakers are installed on the model, proving that the proposed method can achieve fast, high-precision imaging of low-frequency sound sources in an extensive dynamic range at long distances. Full article
(This article belongs to the Special Issue Noise Measurement, Acoustic Signal Processing and Noise Control)
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<p>Microphone array measurement model. The red circles represent the actual positions of sound sources, the grey circles indicate the positions of discrete grids, the red arrows illustrate the radiation process from the sound source to the microphone array, and the grey arrows depict the sound source identification process.</p>
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<p>Schematic diagram of the method. In the figure, the red stars represent the locations of the real sound sources, and the arrow represents the process guidance of the method.</p>
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<p>Simulation experiment setup. (<b>a</b>) Distribution of microphones; (<b>b</b>) Measurement schematic. (<b>c</b>) The locations of the three sound sources.</p>
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<p>Analysis of the impact of decay factor <math display="inline"><semantics> <mi>δ</mi> </semantics></math> on sound source identification results. (<b>a</b>) Impact on RMSEL and RMSEM. (<b>b</b>) Impact on the number of iterations and calculation efficiency.</p>
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<p>Identification error statistics of the methods at different frequencies for the SNR of 20 dB. (<b>a</b>) RMSEL. (<b>b</b>) RMSEM.</p>
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<p>Identification error statistics of the methods at different SNRs for the frequency of 500 Hz. (<b>a</b>) RMSEL. (<b>b</b>) RMSEM.</p>
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<p>Comparison of arrays: (<b>a</b>) 4 m, 193 microphones; (<b>b</b>) 3 m, 150 microphones; (<b>c</b>) 2.2 m, 107 microphones; (<b>d</b>) 1.6 m, 64 microphones; (<b>e</b>) 1.2 m, 45 microphones. The red dots represent the locations of the microphones.</p>
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<p>Error statistics of the method identification for <span class="html-italic">f</span> = 500 Hz, <span class="html-italic">SNR</span> = 20 dB. (<b>a</b>) RMSEL. (<b>b</b>) RMSEM.</p>
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<p>Sound source identification results of <span class="html-italic">f</span> = 500 Hz, SNR = 20 dB. (<b>a</b>) SBL; (<b>b</b>) OMP; (<b>c</b>) FISTA; (<b>d</b>) off-grid-G<span class="html-italic">L</span><sub>1</sub>; (<b>e</b>) RPCA-SBL; (<b>f</b>) VG-SBL. The different colors in the figure correspond to the colorbar on the right side of the figure, representing different amplitudes.</p>
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<p>Sound source identification results of <span class="html-italic">f</span> = 500 Hz, SNR = 20 dB. (<b>a</b>) SBL; (<b>b</b>) OMP; (<b>c</b>) FISTA; (<b>d</b>) off-grid-G<span class="html-italic">L</span><sub>1</sub>; (<b>e</b>) RPCA-SBL; (<b>f</b>) VG-SBL. The different colors in the figure correspond to the colorbar on the right side of the figure, representing different amplitudes.</p>
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<p>The experimental setup. (<b>a</b>) Relative locations between the wind tunnel, the aircraft standard model, and the microphone array. (<b>b</b>) Localized view of aircraft standard model and the microphone array. (<b>c</b>) Arrangement of the three Bluetooth speakers on the aircraft standard model.</p>
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<p>Time–frequency spectrum of a single channel signal under the working condition of 40 m/s@300 Hz. (<b>a</b>) Time domain spectrum. (<b>b</b>) Power spectrum. 300 Hz is marked with a red circle.</p>
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<p>Sound source identification results of 40 m/s@300 Hz: (<b>a</b>) SBL; (<b>b</b>) OMP; (<b>c</b>) FISTA; (<b>d</b>) off-grid-G<span class="html-italic">L</span><sub>1</sub>; (<b>e</b>) RPCA-SBL; (<b>f</b>) VG-SBL. The different colors in the figure correspond to the colorbar on the right side of the figure, representing different amplitudes.</p>
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<p>Sound source identification results of 40 m/s@300 Hz: (<b>a</b>) SBL; (<b>b</b>) OMP; (<b>c</b>) FISTA; (<b>d</b>) off-grid-G<span class="html-italic">L</span><sub>1</sub>; (<b>e</b>) RPCA-SBL; (<b>f</b>) VG-SBL. The different colors in the figure correspond to the colorbar on the right side of the figure, representing different amplitudes.</p>
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<p>Dynamic range of sound source identification.</p>
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<p>The schematic distribution of the identified locations of the three sound sources for the different methods.</p>
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<p>The amplitude identification errors of three sound sources by different methods: (<b>a</b>) Source A; (<b>b</b>) Source B; (<b>c</b>) Source C.</p>
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<p>The error box plots. (<b>a</b>) RMSEL. (<b>b</b>) RMSEM. To facilitate distinction, different colors are used to represent different methods.</p>
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31 pages, 3790 KiB  
Article
MISAO: Ultra-Short-Term Photovoltaic Power Forecasting with Multi-Strategy Improved Snow Ablation Optimizer
by Xu Zhang, Jun Ye, Shenbing Ma, Lintao Gao, Hui Huang and Qiman Xie
Appl. Sci. 2024, 14(16), 7297; https://doi.org/10.3390/app14167297 - 19 Aug 2024
Viewed by 475
Abstract
The increase in installed PV capacity worldwide and the intermittent nature of solar resources highlight the importance of power prediction for grid integration of this technology. Therefore, there is an urgent need for an effective prediction model, but the choice of model hyperparameters [...] Read more.
The increase in installed PV capacity worldwide and the intermittent nature of solar resources highlight the importance of power prediction for grid integration of this technology. Therefore, there is an urgent need for an effective prediction model, but the choice of model hyperparameters greatly affects the prediction performance. In this paper, a multi-strategy improved snowmelt algorithm (MISAO) is proposed for optimizing intrinsic computing-expressive empirical mode decomposition with adaptive noise (ICEEMDAN) and weighted least squares support vector machine for PV power forecasting. Firstly, a cyclic chaotic mapping initialization strategy is used to generate a uniformly distributed high-quality population, which facilitates the algorithm to enter the appropriate search domain quickly. Secondly, the Gaussian diffusion strategy enhances the local exploration ability of the intelligences and extends their search in the solution space, effectively preventing them from falling into local optima. Finally, a stochastic follower search strategy is employed to reserve better candidate solutions for the next iteration, thus achieving a robust exploration–exploitation balance. With these strategies, the optimization performance of MISAO is comprehensively improved. In order to comprehensively evaluate the optimization performance of MISAO, a series of numerical optimization experiments were conducted using IEEE CEC2017 and test sets, and the effectiveness of each improvement strategy was verified. In terms of solution accuracy, convergence speed, robustness, and scalability, MISAO was compared with the basic SAO, various state-of-the-art optimizers, and some recently developed improved algorithms. The results showed that the overall optimization performance of MISAO is excellent, with Friedman average rankings of 1.80 and 1.82 in the two comparison experiments. In most of the test cases, MISAO delivered more accurate and reliable solutions than its competitors. In addition, the altered algorithm was applied to the selection of hyperparameters for the ICEEMDAN-WLSSVM PV prediction model, and seven neural network models, including WLSSVM, ICEEMDAN-WLSSVM, and MISAO-ICEEMDAN-WLSSVM, were used to predict the PV power under three different weather types. The results showed that the models have high prediction accuracy and stability. The MAPE, MAE and RMSE of the proposed model were reduced by at least 25.3%, 17.8% and 13.3%, respectively. This method is useful for predicting the output power, which is conducive to the economic dispatch of the grid and the stable operation of the power system. Full article
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<p>Inspiration source of SAO.</p>
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<p>Trend curve of DDF over iterations.</p>
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<p>Flow chart of the proposed MISAO.</p>
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<p>The convergence curves of some functions.</p>
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<p>Friedman test results of MISAO and peer algorithms in CEC2017.</p>
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<p>Flow chart of combined prediction.</p>
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<p>Sunny power decomposition results.</p>
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<p>Comparison of forecast results of sunny weather.</p>
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<p>Comparison of forecast results of cloudy weather.</p>
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<p>Comparison of forecast results of rainy weather.</p>
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<p>Comparison of forecast results without weather classification.</p>
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21 pages, 9431 KiB  
Article
Anti-Skin Aging Potential, Antibacterial Activity, Inhibition of Single-Stranded DNA-Binding Protein, and Cytotoxic Effects of Acetone-Extracted Passiflora edulis (Tainung No. 1) Rind Extract on Oral Carcinoma Cells
by Yen-Hua Huang and Cheng-Yang Huang
Plants 2024, 13(16), 2194; https://doi.org/10.3390/plants13162194 - 8 Aug 2024
Viewed by 558
Abstract
The passion fruit, Passiflora edulis, recognized for its rich nutritional properties, has long been used for its varied ethnobotanical applications. This study investigates the therapeutic potential of P. edulis var. Tainung No. 1 rind extracts by examining their polyphenolic content (TPC), total [...] Read more.
The passion fruit, Passiflora edulis, recognized for its rich nutritional properties, has long been used for its varied ethnobotanical applications. This study investigates the therapeutic potential of P. edulis var. Tainung No. 1 rind extracts by examining their polyphenolic content (TPC), total flavonoid content (TFC), anti-skin aging activities against key enzymes such as elastase, tyrosinase, and hyaluronidase, and their ability to inhibit bacterial growth, single-stranded DNA-binding protein (SSB), and their cytotoxic effects on oral carcinoma cells. The acetone extract from the rind exhibited the highest levels of TPC, TFC, anti-SSB, and antibacterial activities. The antibacterial effectiveness of the acetone-extracted rind was ranked as follows: Escherichia coli > Pseudomonas aeruginosa > Staphylococcus aureus. A titration curve for SSB inhibition showed an IC50 value of 313.2 μg/mL, indicating the potency of the acetone extract in inhibiting SSB. It also significantly reduced the activity of enzymes associated with skin aging, particularly tyrosinase, with a 54.5% inhibition at a concentration of 100 μg/mL. Gas chromatography–mass spectrometry (GC–MS) analysis tentatively identified several major bioactive compounds in the acetone extract, including stigmast-5-en-3-ol, vitamin E, palmitic acid, stigmasterol, linoleic acid, campesterol, and octadecanoic acid. Molecular docking studies suggested some of these compounds as potential inhibitors of tyrosinase and SSB. Furthermore, the extract demonstrated anticancer potential against Ca9-22 oral carcinoma cells by inhibiting cell survival, migration, and proliferation and inducing apoptosis. These results underscore the potential of P. edulis (Tainung No. 1) rind as a promising candidate for anti-skin aging, antibacterial, and anticancer applications, meriting further therapeutic investigation. Full article
(This article belongs to the Special Issue Biological Activities of Plant Extracts 2023)
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<p>Molecular docking analysis of tyrosinase. (<b>A</b>) The crystal structure of tyrosinase (PDB ID 2Y9X) is shown, with the complexed molecule 2-hydroxycyclohepta-2,4,6-trien-1-one removed for clarity. Active site copper ions are highlighted in brown. (<b>B</b>) Docking analysis demonstrating that four compounds from the extract exhibit higher binding affinities than kojic acid: campesterol (cyan), stigmasterol (orange), stigmast-5-en-3-ol (deepsalmon), and vitamin E (light magenta). These compounds have the capacity to dock into the tyrosinase active site, potentially obstructing substrate access and inhibiting enzyme activity through various binding poses. (<b>C</b>–<b>F</b>) Binding modes of stigmasterol, stigmast-5-en-3-ol, campesterol, and vitamin E with tyrosinase, illustrating their interactions within the enzyme’s active site. The blue dashed lines indicate hydrogen bonds, and the black dashed lines indicate hydrophobic interactions. The numbers labeled on the dashed lines represent the distances in angstroms.</p>
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<p>Inhibition of SSB by <span class="html-italic">P. edulis</span> rind extracts. (<b>A</b>) SSB binding to ssDNA. SSB from <span class="html-italic">K. pneumoniae</span> across a range of concentrations (0, 18, 37, 77, 155, 310, 625, 1250, 2500, and 5000 nM) was incubated with a biotinylated dT35 oligonucleotide. A streptavidin–horseradish peroxidase conjugate was used to detect the ssDNA and the resulting complexes. C indicates the formed complex. (<b>B</b>–<b>D</b>) Inhibition of ssDNA-binding activity of SSB by <span class="html-italic">P. edulis</span> rind extracts obtained using acetone (<b>B</b>), methanol (<b>C</b>), and ethanol (<b>D</b>). SSB (310 nM) was treated with varying concentrations of each extract (0, 31, 63, 125, 250, 500, 1000, 2000, and 3000 μg/mL) to assess inhibition of binding activity.</p>
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<p>Molecular docking analysis of SSB. (<b>A</b>) Crystal structure of <span class="html-italic">K. pneumoniae</span> SSB. <span class="html-italic">K. pneumoniae</span> SSB is a homotetramer. The charge distribution pattern is shown for clarity to indicate possible binding sites. (<b>B</b>) Docking analysis depicting the four most prevalent compounds from the extract, each individually docked into SSB: stigmasterol (orange), stigmast-5-en-3-ol (deepsalmon), campesterol (cyan), and vitamin E (lightmagenta). (<b>C</b>) The structure of <span class="html-italic">P. aeruginosa</span> SSB bound by ssDNA dT20. The ssDNA within the complex crystal structure of the <span class="html-italic">P. aeruginosa</span> SSB tetramer is highlighted in yellow. Given the unavailability of an ssDNA-complexed structure for <span class="html-italic">K. pneumoniae</span> SSB, the complexed structure of the <span class="html-italic">P. aeruginosa</span> SSB is utilized for comparative analysis of the ssDNA-binding mode in <span class="html-italic">K. pneumoniae</span> SSB. (<b>D</b>) Superimposed structures of ssDNA bound by <span class="html-italic">P. aeruginosa</span> SSB alongside the docked compounds bound by <span class="html-italic">K. pneumoniae</span> SSB, suggesting potential ssDNA-binding sites in <span class="html-italic">K. pneumoniae</span> SSB. (<b>E</b>–<b>G</b>) Binding modes of stigmasterol (<b>E</b>), stigmast-5-en-3-ol (<b>F</b>), and campesterol (<b>G</b>) to <span class="html-italic">K. pneumoniae</span> SSB are illustrated, highlighting their interactions within the binding sites.</p>
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<p>Anticancer potential of acetone-extracted <span class="html-italic">P. edulis</span> rind on Ca9-22 gingival carcinoma cells. (<b>A</b>) Overview of the rind extract’s influence on Ca9-22 cell viability, migration, proliferation, and nuclear condensation. (<b>B</b>) Results from the Trypan blue exclusion assay, illustrating cell viability post-treatment with varying concentrations of the rind extract. (<b>C</b>) Hoechst staining analysis depicting the extent of apoptosis and DNA fragmentation across different concentrations of the rind extract. (<b>D</b>) Wound healing assay images capturing Ca9-22 cell migration before and 24 h post-extraction treatment at various concentrations. (<b>E</b>) Clonogenic assay results evaluating the ability of Ca9-22 cells to form colonies under different concentrations of the rind extract, reflecting their survival and proliferative capabilities. Statistical significance relative to the control is indicated by * for <span class="html-italic">p</span> &lt; 0.05, ** for <span class="html-italic">p</span> &lt; 0.01, and *** for <span class="html-italic">p</span> &lt; 0.001. A control medium containing 1% DMSO served as the negative control and caused deaths at the rate of 0%, reduced migration by 0%, suppressed proliferation and colony formation by 0%, and did not induce apoptosis in Ca9-22 cells.</p>
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11 pages, 1245 KiB  
Article
Quantitative Trait Loci Mapping and Candidate Gene Analysis for Fiber Quality Traits in Upland Cotton
by Xiaoyun Jia, Hongxia Zhao, Jijie Zhu, Shijie Wang, Miao Li and Guoyin Wang
Agronomy 2024, 14(8), 1719; https://doi.org/10.3390/agronomy14081719 - 5 Aug 2024
Viewed by 538
Abstract
Superior fiber quality is one of the most important objectives in cotton breeding. To detect the genetic basis underlying fiber quality, an F2 population containing 413 plants was constructed by crossing Jifeng 914 and Jifeng 173, both of which have superior fiber quality, [...] Read more.
Superior fiber quality is one of the most important objectives in cotton breeding. To detect the genetic basis underlying fiber quality, an F2 population containing 413 plants was constructed by crossing Jifeng 914 and Jifeng 173, both of which have superior fiber quality, with Jifeng 173 being better. Five fiber quality traits were investigated in the F2, F2:3, F2:4, and F2:5 populations. Quantitative trait loci (QTL) mapping was conducted based on a high-density genetic map containing 11,488 single nucleotide polymorphisms (SNPs) and spanning 4202.12 cM in length. Transgressive segregation patterns and complex correlations in the five tested traits were observed. A total of 108 QTLs were found, including 13 major effect QTLs that contributed more than 10% toward phenotypic variation (PV) and 9 stable QTLs that could be repeatedly mapped in different generations. Chromosome A7 contained 12 QTL, ranking the first. No QTL was found on chromosomes D1 and D11. Two QTLs could be repeatedly detected in three populations, including qFL-D3-2 in F2, F2:4, and F2:5 with 9.18–21.45% of PV and qFS-A11-1 in F2:3, F2:4, and F2:5 with 6.05–10.41% of PV. Another seven stable QTLs could be detected in two populations, including four major effect QTLs: qFL-A12-3, qFS-D10-2, qMC-D6-2, and qMC-D8-1. Fourteen QTL-overlapping regions were found, which might explain the complex correlations among the five phenotypic traits. Four regions on chromosome A11, D3, D6, and D10 covered by both stable and major effect QTLs are promising for further fine mapping. The genomic regions of the two QTLs detected in three populations and the four major effect QTLs contain 810 genes. Gene functional analysis revealed that the annotated genes are mainly involved in protein binding and metabolic pathways. Fifteen candidate genes in the qFL-D3-2 region are highly expressed in fiber or ovules during fiber initiation, elongation, secondary cell wall thickening, or maturation stages. qRT-PCR revealed that Ghir_D03G005440.1 and Ghir_D03G011310.1 may play a role in promoting fiber initiation, while Ghir_D03G006470.1 may be beneficial for promoting fiber elongation. This study provides more information for revealing the molecular genetic basis underlying cotton fiber quality. Full article
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<p>Distribution of stable QTL, major QTL, and QTL-overlapping regions on the genetic map. Note: FL, fiber length; FS, fiber strength; MC, micronaire; FU, fiber uniformity; FE, fiber elongation rate.</p>
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<p>Gene expression patterns in the ovules. Note: JF914, Jifeng 914; JF173, Jifeng 173; *, <span class="html-italic">p</span> &lt; 0.05; **, <span class="html-italic">p</span> &lt; 0.01.</p>
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<p>Gene expression patterns in the fibers. Note: JF4, Jifeng 4; JF4x, Jifeng 4xuan; *, <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, 677 KiB  
Article
Arabic Lexical Substitution: AraLexSubD Dataset and AraLexSub Pipeline
by Eman Naser-Karajah and Nabil Arman
Data 2024, 9(8), 98; https://doi.org/10.3390/data9080098 - 30 Jul 2024
Viewed by 520
Abstract
Lexical substitution aims to generate a list of equivalent substitutions (i.e., synonyms) to a sentence’s target word or phrase while preserving the sentence’s meaning to improve writing, enhance language understanding, improve natural language processing models, and handle ambiguity. This task has recently attracted [...] Read more.
Lexical substitution aims to generate a list of equivalent substitutions (i.e., synonyms) to a sentence’s target word or phrase while preserving the sentence’s meaning to improve writing, enhance language understanding, improve natural language processing models, and handle ambiguity. This task has recently attracted much attention in many languages. Despite the richness of Arabic vocabulary, limited research has been performed on the lexical substitution task due to the lack of annotated data. To bridge this gap, we present the first Arabic lexical substitution benchmark dataset AraLexSubD for benchmarking lexical substitution pipelines. AraLexSubD is manually built by eight native Arabic speakers and linguists (six linguist annotators, a doctor, and an economist) who annotate the 630 sentences. AraLexSubD covers three domains: general, finance, and medical. It encompasses 2476 substitution candidates ranked according to their semantic relatedness. We also present the first Arabic lexical substitution pipeline, AraLexSub, which uses the AraBERT pre-trained language model. The pipeline consists of several modules: substitute generation, substitute filtering, and candidate ranking. The filtering step shows its effectiveness by achieving an increase of 1.6 in the F1 score on the entire AraLexSubD dataset. Additionally, an error analysis of the experiment is reported. To our knowledge, this is the first study on Arabic lexical substitution. Full article
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<p>The fuzzy scoring scale–synonymy strength.</p>
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<p>The substitution generation of AraLexSub for the target word prediction. The sentence is [<span style="lang:ar">لا يظهر تحيزاً لأحد بعينه</span>/ Do not show bias towards anyone in particular] with the target word [<span style="lang:ar">عين</span>/ particular]. [MASK], [CLS], and [SEP] are Bert special symbols, where [MASK] is used to mask the word, [CLS] is added before each input instance, and [SEP] is a unique separator token.</p>
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24 pages, 6425 KiB  
Article
Bioaffinity Ultrafiltration Combined with HPLC-ESI-qTOF-MS/MS for Screening Potential Bioactive Components from the Stems of Dendrobium fimbriatum and In Silico Analysis
by Yu-Hui Hsieh, Wu-Chang Chuang, Ming-Chung Lee, Yu-Hsin Fan, Nai-Kuei Huang and Jih-Jung Chen
Antioxidants 2024, 13(8), 918; https://doi.org/10.3390/antiox13080918 - 29 Jul 2024
Viewed by 680
Abstract
Dendrobium fimbriatum is a perennial herb, and its stems are high-grade tea and nourishing medicinal materials. Various solvent extracts of D. fimbriatum were evaluated for their anti-inflammatory, anti-acetylcholinesterase (AChE), antioxidant, and anti-α-glucosidase properties. Acetone and EtOAc extracts showed significant antioxidant effects. Acetone, n [...] Read more.
Dendrobium fimbriatum is a perennial herb, and its stems are high-grade tea and nourishing medicinal materials. Various solvent extracts of D. fimbriatum were evaluated for their anti-inflammatory, anti-acetylcholinesterase (AChE), antioxidant, and anti-α-glucosidase properties. Acetone and EtOAc extracts showed significant antioxidant effects. Acetone, n-hexane, and EtOAc extracts revealed potent inhibition against α-glucosidase. EtOAc, n-hexane, and dichloromethane extracts displayed significant anti-AChE activity. Among the isolated constituents, gigantol, moscatin, and dendrophenol showed potent antioxidant activities in FRAP, DPPH, and ABTS radical scavenging tests. Moscatin (IC50 = 161.86 ± 16.45 μM) and dendrophenol (IC50 = 165.19 ± 13.25 μM) displayed more potent anti-AChE activity than chlorogenic acid (IC50 = 236.24 ± 15.85 μM, positive control). Dendrophenol (IC50 = 14.31 ± 3.17 μM) revealed more efficient anti-NO activity than quercetin (positive control, IC50 = 23.09 ± 1.43 μM). Analysis of AChE and iNOS inhibitory components was performed using molecular docking and/or the bioaffinity ultrafiltration method. In bioaffinity ultrafiltration, the binding affinity of compounds to the enzyme (acetylcholinesterase and inducible nitric oxide synthase) was determined using the enrichment factor (EF). Among the main components of the EtOAc extract from D. fimbriatum stem, moscatin, dendrophenol, gigantol, and batatasin III with acetylcholinesterase exhibited the highest binding affinities, with affinity values of 66.31%, 59.48%, 54.60%, and 31.87%, respectively. Moreover, the affinity capacity of the identified compounds with inducible nitric oxide synthase can be ranked as moscatin (88.99%) > dendrophenol (65.11%) > gigantol (44.84%) > batatasin III (27.18%). This research suggests that the bioactive extracts and components of D. fimbriatum stem could be studied further as hopeful candidates for the prevention or treatment of hyperglycemia, oxidative stress-related diseases, and nervous disorders. Full article
(This article belongs to the Special Issue Antioxidant Capacity of Natural Products)
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<p>Dried stems of <span class="html-italic">Dendrobium fimbriatum</span> were used in this study and collected from Mingjian Township, Nantou County, Taiwan.</p>
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<p>Chemical structures of gigantol (<b>1</b>), moscatin (<b>2</b>), batatasin III (<b>3</b>), and dendrophenol (<b>4</b>) from <span class="html-italic">D. fimbriatum</span>.</p>
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<p>Inhibitory activities of moscatin and dendrophenol against LPS-induced iNOS expression in RAW 264.7 murine macrophages are assessed by Western blot. (<b>A</b>) The inhibitory effect of moscatin against LPS-induced iNOS in RAW 264.7 macrophage cell line. (<b>B</b>) The inhibitory activity of dendrophenol against LPS-induced iNOS in RAW 264.7 macrophage cell line. (<b>C</b>) The inhibition rate line chart of moscatin against LPS-induced iNOS in RAW 264.7 macrophage cell line. (<b>D</b>) The inhibition rate line chart of dendrophenol against LPS-induced iNOS in RAW 264.7 macrophage cell line. Quantification data of iNOS/β-actin are shown as mean ± SD (n = 3). Quercetin is applied as a positive control. * <span class="html-italic">p</span> &lt; 0.05 compared with the control group, <sup>#</sup> <span class="html-italic">p</span> &lt; 0.05 compared with the LPS group.</p>
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<p>HPLC chromatograms of the potential AChE inhibitors in the EtOAc extract of <span class="html-italic">D. fimbriatum</span> stems obtained by bioaffinity ultrafiltration. (<b>A</b>) Schematic diagram of bioaffinity ultrafiltration assay. (<b>B</b>) HPLC chromatogram (280 nm) of the chemical components in the EtOAc extract of <span class="html-italic">D. fimbriatum</span> stem obtained by bioaffinity ultrafiltration. The black line represents <span class="html-italic">D. fimbriatum</span> stem extract without ultrafiltration, while the red and blue lines represent <span class="html-italic">D. fimbriatum</span> stem extract with active and inactive AChE and iNOS, respectively.</p>
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<p>(<b>A</b>) Interaction of gigantol with the active sites of <span class="html-italic">E. electric</span> AChE. (<b>B</b>) Interaction of dendrophenol with the active sites of <span class="html-italic">E. electric</span> AChE. (<b>C</b>) Interaction of batatasin III with the active sites of <span class="html-italic">E. electric</span> AChE. (<b>D</b>) Interaction of moscatin with the active sites of <span class="html-italic">E. electric</span> AChE. (<b>E</b>) Interaction of gigantol with the active sites of <span class="html-italic">M. musculus</span> iNOS. (<b>F</b>) Interaction of dendrophenol with the active sites of <span class="html-italic">M. musculus</span> iNOS. (<b>G</b>) Interaction of batatasin III with the active sites of <span class="html-italic">M. musculus</span> iNOS. (<b>H</b>) Interaction of moscatin with the active sites of <span class="html-italic">M. musculus</span> iNOS.</p>
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<p>(<b>A</b>) Interaction of gigantol with the active sites of <span class="html-italic">E. electric</span> AChE. (<b>B</b>) Interaction of dendrophenol with the active sites of <span class="html-italic">E. electric</span> AChE. (<b>C</b>) Interaction of batatasin III with the active sites of <span class="html-italic">E. electric</span> AChE. (<b>D</b>) Interaction of moscatin with the active sites of <span class="html-italic">E. electric</span> AChE. (<b>E</b>) Interaction of gigantol with the active sites of <span class="html-italic">M. musculus</span> iNOS. (<b>F</b>) Interaction of dendrophenol with the active sites of <span class="html-italic">M. musculus</span> iNOS. (<b>G</b>) Interaction of batatasin III with the active sites of <span class="html-italic">M. musculus</span> iNOS. (<b>H</b>) Interaction of moscatin with the active sites of <span class="html-italic">M. musculus</span> iNOS.</p>
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<p>(<b>A</b>) Interaction of gigantol with the active sites of <span class="html-italic">E. electric</span> AChE. (<b>B</b>) Interaction of dendrophenol with the active sites of <span class="html-italic">E. electric</span> AChE. (<b>C</b>) Interaction of batatasin III with the active sites of <span class="html-italic">E. electric</span> AChE. (<b>D</b>) Interaction of moscatin with the active sites of <span class="html-italic">E. electric</span> AChE. (<b>E</b>) Interaction of gigantol with the active sites of <span class="html-italic">M. musculus</span> iNOS. (<b>F</b>) Interaction of dendrophenol with the active sites of <span class="html-italic">M. musculus</span> iNOS. (<b>G</b>) Interaction of batatasin III with the active sites of <span class="html-italic">M. musculus</span> iNOS. (<b>H</b>) Interaction of moscatin with the active sites of <span class="html-italic">M. musculus</span> iNOS.</p>
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<p>(<b>A</b>) Interaction of gigantol with the active sites of <span class="html-italic">E. electric</span> AChE. (<b>B</b>) Interaction of dendrophenol with the active sites of <span class="html-italic">E. electric</span> AChE. (<b>C</b>) Interaction of batatasin III with the active sites of <span class="html-italic">E. electric</span> AChE. (<b>D</b>) Interaction of moscatin with the active sites of <span class="html-italic">E. electric</span> AChE. (<b>E</b>) Interaction of gigantol with the active sites of <span class="html-italic">M. musculus</span> iNOS. (<b>F</b>) Interaction of dendrophenol with the active sites of <span class="html-italic">M. musculus</span> iNOS. (<b>G</b>) Interaction of batatasin III with the active sites of <span class="html-italic">M. musculus</span> iNOS. (<b>H</b>) Interaction of moscatin with the active sites of <span class="html-italic">M. musculus</span> iNOS.</p>
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<p>(<b>A</b>) Interaction of gigantol with the active sites of <span class="html-italic">E. electric</span> AChE. (<b>B</b>) Interaction of dendrophenol with the active sites of <span class="html-italic">E. electric</span> AChE. (<b>C</b>) Interaction of batatasin III with the active sites of <span class="html-italic">E. electric</span> AChE. (<b>D</b>) Interaction of moscatin with the active sites of <span class="html-italic">E. electric</span> AChE. (<b>E</b>) Interaction of gigantol with the active sites of <span class="html-italic">M. musculus</span> iNOS. (<b>F</b>) Interaction of dendrophenol with the active sites of <span class="html-italic">M. musculus</span> iNOS. (<b>G</b>) Interaction of batatasin III with the active sites of <span class="html-italic">M. musculus</span> iNOS. (<b>H</b>) Interaction of moscatin with the active sites of <span class="html-italic">M. musculus</span> iNOS.</p>
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20 pages, 3102 KiB  
Article
A Transformative Technology Linking Patient’s mRNA Expression Profile to Anticancer Drug Efficacy
by Chen Yeh, Shu-Ti Lin and Hung-Chih Lai
Onco 2024, 4(3), 143-162; https://doi.org/10.3390/onco4030012 - 14 Jul 2024
Viewed by 1073
Abstract
As precision medicine such as targeted therapy and immunotherapy often have limited accessibility, low response rate, and evolved resistance, it is urgent to develop simple, low-cost, and quick-turnaround personalized diagnostic technologies for drug response prediction with high sensitivity, speed, and accuracy. The major [...] Read more.
As precision medicine such as targeted therapy and immunotherapy often have limited accessibility, low response rate, and evolved resistance, it is urgent to develop simple, low-cost, and quick-turnaround personalized diagnostic technologies for drug response prediction with high sensitivity, speed, and accuracy. The major challenges of drug response prediction strategies employing digital database modeling are the scarcity of labeled clinical data, applicability only to a few classes of drugs, and losing the resolution at the individual patient level. Although these challenges have been partially addressed by large-scale cancer cell line datasets and more patient-relevant cell-based systems, the integration of different data types and data translation from pre-clinical to clinical utilities are still far-fetched. To overcome the current limitations of precision medicine with a clinically proven drug response prediction assay, we have developed an innovative and proprietary technology based on in vitro patient testing and in silico data analytics. First, a patient-derived gene expression signature was established via the transcriptomic profiling of cell-free mRNA (cfmRNA) from the patient’s blood. Second, a gene-to-drug data fusion and overlaying mechanism to transfer data were performed. Finally, a semi-supervised method was used for the database searching, matching, annotation, and ranking of drug efficacies from a pool of ~700 approved, investigational, or clinical trial drug candidates. A personalized drug response report can be delivered to inform clinical decisions within a week. The PGA (patient-derived gene expression-informed anticancer drug efficacy) test has significantly improved patient outcomes when compared to the treatment plans without PGA support. The implementation of PGA, which combines patient-unique cfmRNA fingerprints with drug mapping power, has the potential to identify treatment options when patients are no longer responding to therapy and when standard-of-care is exhausted. Full article
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<p>Plasma cfmRNA profiling by cancer type, functional cluster, and expression level. (<b>A</b>) The pie chart displayed the distribution of the various functional classes of cfmRNA in lung cancer; (<b>B</b>) representative gene expression heatmaps showing high-, medium-, and low-expressing transcripts involved in different pathways from different cancer types.</p>
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<p>Validation of the selected PGA Lung biomarkers for drug efficacy prediction. (<b>A</b>) The overexpression of PGA Lung biomarkers in most lung tumor tissues from the TCGA database (1145 samples). Significant association of PGA Lung biomarkers with hypoxia (<b>B</b>) and MSI scores (<b>C</b>) in the TCGA PanCancer database (510 LUAD samples).</p>
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<p>Strong correlation of the PGA Lung biomarker expression levels between the plasma and tissue samples. The relative cfmRNA levels were expressed as delta Ct values, whereas the tissue mRNA expression was normalized as fold expression. The data showed a positive correlation between the cfmRNA and tissue mRNA expression (i.e., an inverse relationship between delta Ct and fold expression).</p>
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<p>Single-cell RNA-Seq spatial transcriptomic analysis in lung carcinoma tissues (32,341 cells). The visualization of tumor cells expressing the key lung cancer driver genes EGFR, KRAS, BRAF, MET, HER2, ALK, ROS1, or RET. The expression patterns of EGFR, MET, HER2, and ROS1 were highly overlapped, and these EGFR-/MET-/HER2-/ROS1-coexpressing cells only constituted a small fraction of the entire tumor population. By contrast, the expression profiles of KRAS and BRAF were similar and distributed across the entire section.</p>
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<p>Single-cell RNA-Seq spatial transcriptomic analysis of PGA Lung biomarkers in (<b>A</b>) lung carcinoma tissues (32,341 cells) and (<b>B</b>) dissociated tumor cells from the pleural effusion of lung adenocarcinoma patients (7511 cells). The visualization of tumor cells expressing the representative PGA Lung biomarkers 1–8. The expression patterns of these PGA Lung genes were highly similar and distributed across the entire section, resembling those of KRAS and BRAF. Most significantly, the population of tumor cells expressing PGA Lung biomarkers was found to be PCNA-positive, indicative of high proliferation potential.</p>
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<p>Strong functional genomics similarity between the TCGA lung tumors and lung cancer cell lines. The Spearman correlation and normalized enrichment score (NES) were derived from the expression patterns of overactive genes and the activities of the cancer-related pathways.</p>
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<p>Overview of in silico data fusion, annotation, mapping, and analyses in the PGA Lung test.</p>
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<p>Kaplan–Meier analysis of progression-free survival (PFS) and overall survival (OS) for the treatment of real-world lung cancer patients with or without the support from the PGA Lung test.</p>
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12 pages, 3280 KiB  
Article
Toxicity of Silver–Chitosan Nanocomposites to Aquatic Microcrustaceans Daphnia magna and Thamnocephalus platyurus and Naturally Luminescent Bacteria Vibrio fischeri
by Mariliis Sihtmäe, Jüri Laanoja, Irina Blinova, Anne Kahru and Kaja Kasemets
Nanomaterials 2024, 14(14), 1193; https://doi.org/10.3390/nano14141193 - 12 Jul 2024
Viewed by 635
Abstract
All novel materials should be analyzed for their potential environmental hazard. In this study, the toxicity of different silver–chitosan nanocomposites—potential candidates for wound dressings or antimicrobial surface coatings—was evaluated using environmentally relevant aquatic microcrustaceans Daphnia magna and Thamnocephalus platyurus and naturally luminescent bacteria [...] Read more.
All novel materials should be analyzed for their potential environmental hazard. In this study, the toxicity of different silver–chitosan nanocomposites—potential candidates for wound dressings or antimicrobial surface coatings—was evaluated using environmentally relevant aquatic microcrustaceans Daphnia magna and Thamnocephalus platyurus and naturally luminescent bacteria Vibrio fischeri. Three silver-chitosan nanocomposites (nAgCSs) with different weight ratios of Ag to CS were studied. Citrate-coated silver nanoparticles (nAg-Cit), AgNO3 (ionic control) and low molecular weight chitosan (LMW CS) were evaluated in parallel. The primary size of nAgCSs was ~50 nm. The average hydrodynamic sizes in deionized water were ≤100 nm, and the zeta potential values were positive (16–26 mV). The nAgCSs proved very toxic to aquatic crustaceans: the 48-h EC50 value for D. magna was 0.065–0.232 mg/L, and the 24-h LC50 value for T. platyurus was 0.25–1.04 mg/L. The toxic effect correlated with the shedding of Ag ions (about 1%) from nAgCSs. Upon exposure of V. fischeri to nAgCSs for 30 min, bacterial luminescence was inhibited by 50% at 13–33 mg/L. However, the inhibitory effect (minimum bactericidal concentration, MBC) on bacterial growth upon 1 h exposure was observed at higher concentrations of nAgCSs, 40–65 mg/L. LMW CS inhibited bacterial luminescence upon 30-min exposure at 5.6 mg/L, but bacterial growth was inhibited at a much higher concentration (1 h MBC > 100 mg/L). The multi-trophic test battery, where D. magna was the most sensitive test organism, ranked the silver-chitosan nanocomposites from ‘extremely toxic’ [L(E)C50 ≤ 0.1 mg/L] to ‘very toxic’ [L(E)C50 > 0.1–1 mg/L]. Chitosan was toxic (EC(L)50) to crustaceans at ~12 mg/L, and ranked accordingly as ‘harmful’ [L(E)C50 > 10–100 mg/L]. Thus, silver-chitosan nanocomposites may pose a hazard to aquatic organisms and must be handled accordingly. Full article
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<p>Quantitative (using light scattering/mean count rate; Zetasizer Nano-ZS, Malvern Instruments Ltd., Malvern, UK) and qualitative (visualization of the settling in cuvettes) determination of the stability of studied nanomaterials suspensions during 24 h in DI-water and toxicity testing media (2% NaCl and artificial fresh water, AFW) at 10 mg Ag/L.</p>
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<p>Kinetics of bioluminescence during the first 15 s of exposure of <span class="html-italic">Vibrio fischeri</span> to suspensions of silver-chitosan nanocomposites with silver-chitosan weight ratios of (<b>A</b>) 1:0.3 (nAgCS-0.3), (<b>B</b>) 1:1 (nAgCS-1) and (<b>C</b>) 1:3 (nAgCS-3), (<b>D</b>) citrate-coated silver nanoparticles (nAg-Cit), (<b>E</b>) low molecular-weight chitosan (LMW CS) and (<b>F</b>) silver ions (AgNO<sub>3</sub>). NaCl (2%) served as a control and diluent. Concentrations of test compounds are nominal. Chitosan was tested at an initial pH ranging from 4.3 (100 mg/L) to 6.5 (0.1 mg/L), and at the level of obtained 30-min EC<sub>50</sub> value, the pH of chitosan was 5.2–5.5. RLU—relative light units. 30-min EC<sub>50</sub> values taken from <a href="#nanomaterials-14-01193-t002" class="html-table">Table 2</a> are added to the panels. Please note that the EC<sub>50</sub> values for chitosan are presented as mg compound/L and Ag-containing compounds as mg Ag/L.</p>
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<p>Kinetics of bioluminescence during the first 15 s of exposure of <span class="html-italic">Vibrio fischeri</span> to suspensions of silver-chitosan nanocomposites with silver-chitosan weight ratios of (<b>A</b>) 1:0.3 (nAgCS-0.3), (<b>B</b>) 1:1 (nAgCS-1) and (<b>C</b>) 1:3 (nAgCS-3), (<b>D</b>) citrate-coated silver nanoparticles (nAg-Cit), (<b>E</b>) low molecular-weight chitosan (LMW CS) and (<b>F</b>) silver ions (AgNO<sub>3</sub>). NaCl (2%) served as a control and diluent. Concentrations of test compounds are nominal. Chitosan was tested at an initial pH ranging from 4.3 (100 mg/L) to 6.5 (0.1 mg/L), and at the level of obtained 30-min EC<sub>50</sub> value, the pH of chitosan was 5.2–5.5. RLU—relative light units. 30-min EC<sub>50</sub> values taken from <a href="#nanomaterials-14-01193-t002" class="html-table">Table 2</a> are added to the panels. Please note that the EC<sub>50</sub> values for chitosan are presented as mg compound/L and Ag-containing compounds as mg Ag/L.</p>
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<p>Colony forming ability (viability) of bacteria <span class="html-italic">Vibrio fischeri</span> on agar plates after 1 and 24 h exposure to the different concentrations of silver-chitosan nanocomposites with silver-chitosan weight ratios of 1:0.3 (nAgCS-0.3), 1:1 (nAgCS-1) and 1:3 (nAgCS-3), citrate-coated silver nanoparticles (nAg-Cit), AgNO<sub>3</sub> and low molecular weight chitosan (LMW CS) in 2% NaCl solution. Please note that the EC<sub>50</sub> values for chitosan are presented as mg compound/L and Ag-containing compounds as mg Ag/L. The photos were taken in the dark.</p>
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<p>Accumulation of the silver-chitosan nanocomposites (nAgCSs) in the gut of alive particle-ingesting microcrustaceans: <span class="html-italic">Thamnocephalus platyurus</span> (<b>A</b>) and <span class="html-italic">Daphnia magna</span> (<b>B</b>). See also <a href="#nanomaterials-14-01193-t002" class="html-table">Table 2</a>. A Nikon stereo microscope (SMZ1270) was used for imaging. Please note the different sizes of scale bars.</p>
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