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Search Results (3,665)

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Keywords = genetic programming

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30 pages, 3948 KiB  
Article
A Two-Stage Genetic Algorithm for Beam–Slab Structure Optimization
by Zhexi Yang and Wei-Zhen Lu
Buildings 2024, 14(9), 2932; https://doi.org/10.3390/buildings14092932 - 16 Sep 2024
Abstract
Beam–slab structures account for 50–65% of a building’s total dead load and contribute to 20% of the overall cost and CO2 emissions. Despite their importance, conventional beam–slab structural optimization methods often lack search efficiency and accuracy, making them less effective for practical [...] Read more.
Beam–slab structures account for 50–65% of a building’s total dead load and contribute to 20% of the overall cost and CO2 emissions. Despite their importance, conventional beam–slab structural optimization methods often lack search efficiency and accuracy, making them less effective for practical engineering applications. Such limitations arise from the optimization problem involving a complex solution space, particularly when considering components’ arrangement, dimensions, and load transfer paths simultaneously. To address the research gap, this study proposes a novel two-stage genetic algorithm, optimizing beam–slab layout in the first stage and component topological relationships and dimensions in the second stage. Numerical experiments on the prototype case indicate that the algorithm can generate results that meet engineering accuracy requirements within 100 iterations, outperforming comparable algorithms in both efficiency and accuracy. Additionally, this heuristic approach stands out for its independence from prior dataset training and its minimal parameter adjustment requirement, making it highly accessible to engineers without programming expertise. Statistical analysis of the algorithm’s optimization process and case studies demonstrate its robustness and adaptability to various beam–slab structural optimization problems, revealing its significant potential for practical engineering scenarios. Full article
24 pages, 706 KiB  
Review
Cross-Disciplinary Rapid Scoping Review of Structural Racial and Caste Discrimination Associated with Population Health Disparities in the 21st Century
by Drona P. Rasali, Brendan M. Woodruff, Fatima A. Alzyoud, Daniel Kiel, Katharine T. Schaffzin, William D. Osei, Chandra L. Ford and Shanthi Johnson
Societies 2024, 14(9), 186; https://doi.org/10.3390/soc14090186 - 16 Sep 2024
Abstract
A cross-disciplinary rapid scoping review was carried out, generally following the PRISMA-SCR protocol to examine historical racial and caste-based discrimination as structural determinants of health disparities in the 21st century. We selected 48 peer-reviewed full-text articles available from the University of Memphis Libraries [...] Read more.
A cross-disciplinary rapid scoping review was carried out, generally following the PRISMA-SCR protocol to examine historical racial and caste-based discrimination as structural determinants of health disparities in the 21st century. We selected 48 peer-reviewed full-text articles available from the University of Memphis Libraries database search, focusing on three selected case-study countries: the United States (US), Canada, and Nepal. The authors read each article, extracted highlights, and tabulated the thematic contents on structural health disparities attributed to racism or casteism. The results link historical racism/casteism to health disparities occurring in Black and African American, Native American, and other ethnic groups in the US; in Indigenous peoples and other visible minorities in Canada; and in the Dalits of Nepal, a population racialized by caste, grounded on at least four foundational theories explaining structural determinants of health disparities. The evidence from the literature indicates that genetic variations and biological differences (e.g., disease prevalence) occur within and between races/castes for various reasons (e.g., random gene mutations, geographic isolation, and endogamy). However, historical races/castes as socio-cultural constructs have no inherently exclusive basis of biological differences. Disregarding genetic discrimination based on pseudo-scientific theories, genetic testing is a valuable scientific means to achieve the better health of the populations. Epigenetic changes (e.g., weathering—the early aging of racialized women) due to the DNA methylation of genes among racialized populations are markers of intergenerational trauma due to racial/caste discrimination. Likewise, chronic stresses resulting from intergenerational racial/caste discrimination cause an “allostatic load”, characterized by an imbalance of neuronal and hormonal dysfunction, leading to occurrences of chronic diseases (e.g., hypertension, diabetes, and mental health) at disproportionate rates among racialized populations. Major areas identified for reparative policy changes and interventions for eliminating the health impacts of racism/casteism include areas of issues on health disparity research, organizational structures, programs and processes, racial justice in population health, cultural trauma, equitable healthcare system, and genetic discrimination. Full article
(This article belongs to the Topic Diversity Competence and Social Inequalities)
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<p>Flowchart of the literature search strategy.</p>
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9 pages, 620 KiB  
Brief Report
Utilization of Assisted Reproductive Technologies in Breeding Auliekol Cattle: A Comparative Study
by Altyn Kulpiisova, Kairly Yessengaliyev, Gulsara Kassimova, Ainat Kozhakhmetova, Bakytkanym Kadraliyeva, Abeldinov Rustem, Alma Temirzhanova, Nadezhda Burambayeva, Salbak Chylbak-ool, Elena Pakhomova, Nurzhan Abekeshev, Gulnara Baikadamova, Zhomart Kemeshev, Alexandra Tegza, Arman Issimov and Peter White
Life 2024, 14(9), 1167; https://doi.org/10.3390/life14091167 - 15 Sep 2024
Viewed by 205
Abstract
This study evaluates the utilization of in vitro embryo production (IVEP) technology for the conservation and breeding of the Auliekol cattle breed, a primary beef breed in Kazakhstan facing population decline due to the cessation of breeding programs and the incursion of transboundary [...] Read more.
This study evaluates the utilization of in vitro embryo production (IVEP) technology for the conservation and breeding of the Auliekol cattle breed, a primary beef breed in Kazakhstan facing population decline due to the cessation of breeding programs and the incursion of transboundary diseases. We assessed the effect of consecutive ovum pick-up (OPU) procedures on oocyte yield and embryo production in Auliekol and Aberdeen Angus cows. A total of 2232 and 3659 oocytes were aspirated from Auliekol and Aberdeen Angus donors, respectively, with significantly higher yields and embryo production observed in Aberdeen Angus cows. The application of a meiotic block using Butyrolactone I (BLI) and subsequent in vitro fertilization (IVF) protocols was employed, with embryo development monitored up to the morula/blastocyst stage. Results indicated that Auliekol cows exhibited lower oocyte recovery, cleavage, and blastocyst rates compared to Aberdeen Angus cows, likely due to genetic characteristics. Despite the challenges, IVEP presents a valuable tool for the preservation and future propagation of the Auliekol breed, highlighting the need for further research to enhance reproductive outcomes and conservation strategies. Full article
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<p>Inverted microscopic picture of Blastocysts produced in vitro on the seventh day of development. DIC magnification ×100.</p>
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20 pages, 4626 KiB  
Article
Genetic Diversity of Common Bean (Phaseolus vulgaris L.) Landraces Based on Morphological Traits and Molecular Markers
by Evaldo de Paula, Rafael Nunes de Almeida, Talles de Oliveira Santos, José Dias de Souza Neto, Elaine Manelli Riva-Souza, Sheila Cristina Prucoli Posse, Maurício Novaes Souza, Aparecida de Fátima Madella de Oliveira, Alexandre Cristiano Santos Júnior, Jardel Oliveira Santos, Samy Pimenta, Cintia dos Santos Bento and Monique Moreira Moulin
Plants 2024, 13(18), 2584; https://doi.org/10.3390/plants13182584 - 15 Sep 2024
Viewed by 211
Abstract
The objective of this study was to evaluate the genetic diversity among traditional common bean accessions through morphological descriptors and molecular markers. Sixty-seven common bean accessions from the Germplasm bank of the Instituto Federal of Espírito Santo—Campus de Alegre were evaluated. For this, [...] Read more.
The objective of this study was to evaluate the genetic diversity among traditional common bean accessions through morphological descriptors and molecular markers. Sixty-seven common bean accessions from the Germplasm bank of the Instituto Federal of Espírito Santo—Campus de Alegre were evaluated. For this, 25 specific morphological descriptors were used, namely 12 quantitative and 13 qualitative ones. A diversity analysis based on morphological descriptors was carried out using the Gower algorithm. For molecular characterization, 23 ISSR primers were used to estimate dissimilarity using the Jaccard Index. Based on the dendrograms obtained by the UPGMA method, for morphological and molecular characterization, high genetic variability was observed between the common bean genotypes studied, evidenced by cophenetic correlation values in the order of 0.99, indicating an accurate representation of the dissimilarity matrix by the UPGMA clustering. In the morphological characterization, high phenotypic diversity was observed between the accessions, with grains of different shapes, colors, and sizes, and the accessions were grouped into nine distinct groups. Molecular characterization was efficient in separating the genotypes in the Andean and Mesoamerican groups, with the 23 ISSR primers studied generating an average of 6.35 polymorphic bands. The work identified divergent accessions that can serve different market niches, which can be indicated as parents to form breeding programs in order to obtain progenies with high genetic variability. Full article
(This article belongs to the Special Issue Characterization and Conservation of Vegetable Genetic Resources)
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<p>Phenotypic diversity of common bean (<span class="html-italic">P. vulgaris</span> L.) accessions belonging to Ifes—Campus de Alegre germplasm bank.</p>
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<p>Phenotypic diversity of common bean (<span class="html-italic">P. vulgaris</span> L.) accessions belonging to Ifes—Campus de Alegre germplasm bank.</p>
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<p>Phenotypic diversity of common bean (<span class="html-italic">P. vulgaris</span> L.) accessions belonging to Ifes—Campus de Alegre germplasm bank.</p>
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<p>A dendrogram of genetic dissimilarity created using the Gower distance, based on quantitative and qualitative descriptors, for the 67 common bean accessions (cophenetic correlation = 0.86). The numbers I, II, III, IV, V, VI, VII, VIII, and IX refer to groups that include the genetically closest accessions.</p>
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<p>The dendrogram obtained with UPGMA from the Jaccard dissimilarity matrix of 67 accessions of the common bean based on 146 polymorphic ISSR markers (cophenetic correlation = 0.99). Numbers I and II refer to the groups that include the genetically closest accessions, separated according to gene pools: I—accessions of Andean origin; II—accessions of Mesoamerican origin.</p>
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22 pages, 1024 KiB  
Article
Coordinated Planning of Soft Open Points and Energy Storage Systems to Enhance Flexibility of Distribution Networks
by Jingyu Li, Yifan Zhang, Chao Lv, Guangchen Liu, Zhongtian Ruan and Feiyang Zhang
Appl. Sci. 2024, 14(18), 8309; https://doi.org/10.3390/app14188309 (registering DOI) - 14 Sep 2024
Viewed by 270
Abstract
With the large-scale penetration of distributed generation (DG), the volatility problems of active distribution networks (ADNs) have become more prominent, which can no longer be met by traditional regulation means and need to be regulated by introducing flexible resources. Soft open points (SOP) [...] Read more.
With the large-scale penetration of distributed generation (DG), the volatility problems of active distribution networks (ADNs) have become more prominent, which can no longer be met by traditional regulation means and need to be regulated by introducing flexible resources. Soft open points (SOP) and energy storage systems (ESS) can regulate the tidal currents on spatial and temporal scales, respectively, to improve the flexibility of ADN. To this end, in-depth consideration of DG admission is given to establish flexibility assessment indicators from the power side of ADN. The conditional deep convolution generative adversarial network (C-DCGAN) is used to generate the output scenario of DG. On this basis, the SOP and ESS two-layer planning models, which take account of the potential for improvement in the flexibility of ADN, are constructed. Among them, the upper layer is the site selection and volume determination layer, which considers the economy of the system with the optimization objective of minimizing the annual integrated cost; the lower layer is the operation optimization layer, which considers the flexibility of the system and takes the highest average daily flexibility level as the optimization objective. The planning model is solved using genetic algorithm-particle swarm optimization (GA-PSO) and second-order cone programming (SOCP). The case analysis verifies the rationality and effectiveness of the planning model. Full article
(This article belongs to the Special Issue New Insights into Power Systems)
17 pages, 2374 KiB  
Article
Advancing Cancer Care in Colombia: Results of the First In Situ Implementation of Comprehensive Genomic Profiling
by Juan Javier López Rivera, Paula Rueda-Gaitán, Laura Camila Rios Pinto, Diego Alejandro Rodríguez Gutiérrez, Natalia Gomez-Lopera, Julian Lamilla, Fabio Andrés Rojas Aguirre, Laura Bernal Vaca and Mario Arturo Isaza-Ruget
J. Pers. Med. 2024, 14(9), 975; https://doi.org/10.3390/jpm14090975 (registering DOI) - 14 Sep 2024
Viewed by 196
Abstract
Background: Comprehensive genomic profiling (CGP) identifies genetic alterations and patterns that are crucial for therapy selection and precise treatment development. In Colombia, limited access to CGP tests underscores the necessity of documenting the prevalence of treatable genetic alterations. This study aimed to describe [...] Read more.
Background: Comprehensive genomic profiling (CGP) identifies genetic alterations and patterns that are crucial for therapy selection and precise treatment development. In Colombia, limited access to CGP tests underscores the necessity of documenting the prevalence of treatable genetic alterations. This study aimed to describe the somatic genetic profile of specific cancer types in Colombian patients and assess its impact on treatment selection. Methods: A retrospective cohort study was conducted at Clínica Colsanitas S.A. from March 2023 to June 2024. Sequencing was performed on the NextSeq2000 platform with the TruSight Oncology 500 (TSO500) assay, which simultaneously evaluates 523 genes for DNA analysis and 55 for RNA; additionally, analyses were performed with the SOPHiA DDM software. The tumor mutational burden (TMB), microsatellite instability (MSI), and programmed cell death ligand 1 (PDL1) were assessed. Results: Among 111 patients, 103 were evaluated, with gastrointestinal (27.93%), respiratory (13.51%), and central nervous system cancers (10.81%) being the most prevalent. TP53 (37%), KMT2C (28%), and KRAS (21%) were frequent mutations. Actionable findings were detected in 76.7% of cases, notably in digestive (20 patients) and lung cancers (8 patients). MSI was stable at 82.52% and high at 2.91%, whilst TMB was predominantly low (91.26%). Conclusions: The test has facilitated access to targeted therapies, improving clinical outcomes in Colombian patients. This profiling test is expected to increase opportunities for personalized medicine in Colombia. Full article
(This article belongs to the Section Omics/Informatics)
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<p>Radial chart demonstrating the distribution of cancer types across various body systems, digit indicates the number of patients affected by system. Note that the digestive system, for example, includes GIST, liver cancer, colorectal cancer, pancreatic cancer, stomach cancer, and bile duct cancer [<a href="#B21-jpm-14-00975" class="html-bibr">21</a>].</p>
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<p>(<b>a</b>) The number of genetic variants identified in various cancer types, with each color representing a specific cancer type such as thyroid gland (blue), sarcoma (orange), and melanoma (pink). The horizontal bars represent the total number of variants found for each gene across these cancers, with TP53 and KMT2C having the highest number of variants (41 and 31, respectively). (<b>b</b>) A breakdown of the types of genetic variants found, including amplifications, deletions, and missense mutations. Each variant type is color-coded, as described in the legend on the right.</p>
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<p>Distribution of clinically relevant and potentially clinically relevant results. Out of 103 patients with solid tumors who underwent genomic profiling, 79 received results with clinical relevance. Among these, 64 showed potential treatment benefits, 11 had resistance-related findings, and 2 had diagnostic significance. In addition, 24 patients received results with potential clinical relevance, while 8 samples were insufficient for analysis.</p>
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<p>TMB and MSI across different cancer types. (<b>A</b>) Bar chart displaying the frequency of cancer types with high (TMB &gt; 10) and low (TMB ≤ 10) tumor mutational burden. (<b>B</b>) Bar chart illustrating the frequency of MSI status across different cancer types. The chart categorizes MSI status into high, equivocal, stable, and rejected MSI.</p>
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<p>TMB and MSI across different cancer types. (<b>A</b>) Bar chart displaying the frequency of cancer types with high (TMB &gt; 10) and low (TMB ≤ 10) tumor mutational burden. (<b>B</b>) Bar chart illustrating the frequency of MSI status across different cancer types. The chart categorizes MSI status into high, equivocal, stable, and rejected MSI.</p>
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<p>PD-L1 expression across different tumor types. Bar chart visualizing the PD-L1 expression by tumor type. The chart contrasts the number of tumors with PD-L1 expression (lilac bars) versus those that are PD-L1 negative (purple bars).</p>
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15 pages, 275 KiB  
Review
Ethical Issues in Living Donor Kidney Transplantation: An Update from a Psychosocial Perspective
by Valentina Martinelli, Estella L. L. Lumer, Matteo Chiappedi, Pierluigi Politi, Marilena Gregorini, Teresa Rampino, Andrea Peri, Andrea Pietrabissa and Laura Fusar-Poli
Healthcare 2024, 12(18), 1832; https://doi.org/10.3390/healthcare12181832 - 13 Sep 2024
Viewed by 194
Abstract
Living donor kidney transplantation (LDKT) currently represents the treatment of choice for patients with end-stage renal failure. LDKT is a serious event with profound psychological, interpersonal, familial, and social implications. Over the last few years, there has been an exponential growth in living [...] Read more.
Living donor kidney transplantation (LDKT) currently represents the treatment of choice for patients with end-stage renal failure. LDKT is a serious event with profound psychological, interpersonal, familial, and social implications. Over the last few years, there has been an exponential growth in living donation programs involving genetically and emotionally related donors, as well as people who donate to an unrelated and unknown subject. The implementation of paired exchange programs, Samaritan donation, and preemptive transplantation raise further ethical issues, which are inextricably linked to the unique psychosocial context of both the donor and the recipient. The present narrative review aims to provide an update on the main ethical challenges related to LDKT. We conducted a comprehensive literature search in PubMed/Medline. The results of the most relevant studies were narratively synthesized from a psychosocial perspective around the four principles of biomedical ethics: autonomy, beneficence, non-maleficence, and justice. Finally, we discussed the potential future directions to provide an effective, patient-centered, and ethical psychosocial assessment and follow-up of living donors and recipients that underwent LDKT. Full article
19 pages, 4026 KiB  
Article
Genome-Wide Analysis of Fruit Color and Carotenoid Content in Capsicum Core Collection
by Nayoung Ro, Hyeonseok Oh, Ho-Cheol Ko, Jungyoon Yi, Young-Wang Na and Mesfin Haile
Plants 2024, 13(18), 2562; https://doi.org/10.3390/plants13182562 - 12 Sep 2024
Viewed by 225
Abstract
This study investigated carotenoid content and fruit color variation in 306 pepper accessions from diverse Capsicum species. Red-fruited accessions were predominant (245 accessions), followed by orange (35) and yellow (20). Carotenoid profiles varied significantly across accessions, with capsanthin showing the highest mean concentration [...] Read more.
This study investigated carotenoid content and fruit color variation in 306 pepper accessions from diverse Capsicum species. Red-fruited accessions were predominant (245 accessions), followed by orange (35) and yellow (20). Carotenoid profiles varied significantly across accessions, with capsanthin showing the highest mean concentration (239.12 μg/g), followed by β-cryptoxanthin (63.70 μg/g) and zeaxanthin (63.25 μg/g). Total carotenoid content ranged from 7.09 to 2566.67 μg/g, emphasizing the diversity within the dataset. Correlation analysis revealed complex relationships between carotenoids, with strong positive correlations observed between total carotenoids and capsanthin (r = 0.94 ***), β-cryptoxanthin (r = 0.87 ***), and zeaxanthin (r = 0.84 ***). Principal component analysis (PCA) identified two distinct carotenoid groups, accounting for 67.6% of the total variance. A genome-wide association study (GWAS) identified 91 significant single nucleotide polymorphisms (SNPs) associated with fruit color (15 SNPs) and carotenoid content (76 SNPs). These SNPs were distributed across all chromosomes, with varying numbers on each. Among individual carotenoids, α-carotene was associated with 28 SNPs, while other carotenoids showed different numbers of associated SNPs. Candidate genes encoding diverse proteins were identified near significant SNPs, potentially contributing to fruit color variation and carotenoid accumulation. These included pentatricopeptide repeat-containing proteins, mitochondrial proton/calcium exchangers, E3 ubiquitin-protein ligase SINAT2, histone–lysine N-methyltransferase, sucrose synthase, and various enzymes involved in metabolic processes. Seven SNPs exhibited pleiotropic effects on multiple carotenoids, particularly β-cryptoxanthin and capsanthin. The findings of this study provide insights into the genetic architecture of carotenoid biosynthesis and fruit color in peppers, offering valuable resources for targeted breeding programs aimed at enhancing the nutritional and sensory attributes of pepper varieties. Full article
(This article belongs to the Special Issue Molecular Marker-Assisted Technologies for Crop Breeding)
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<p>The distribution of 306 pepper accessions based on fruit color at maturity.</p>
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<p>The average carotenoid contents of pepper accessions based on fruit color.</p>
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<p>The average carotenoid contents of pepper accessions based on species. These accessions represent five species: <span class="html-italic">C. annuum</span> (198), <span class="html-italic">C. baccatum</span> (43), <span class="html-italic">C. chinense</span> (43), and <span class="html-italic">C. frutescens</span> (21).</p>
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<p>Heatmap depicting carotenoid correlations in a diverse set of 306 pepper accessions. This visualization presents Pearson correlation coefficients, with a color scale on the right indicating correlation strength and direction. The analysis includes the following carotenoids: violaxanthin, lutein, antheraxanthin, capsorubin, capsanthin, zeaxanthin, β-cryptoxanthin, α-carotene, β-carotene and total carotenoid content. Significance is represented by *, **, and *** for <span class="html-italic">p</span>-values of less than 0.05, 0.01, and 0.001, respectively.</p>
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<p>PCA plot based on the carotenoid data of 306 pepper accessions: (<b>a</b>)—variables and (<b>b</b>)—individuals; each dot represents a single accession. The variables are a—α-carotene, b—antheraxanthin, c—β-carotene, d—β-cryptoxanthin, e—capsanthin, f—capsorubin, g—lutein, h—violaxanthin, i—zeaxanthin, and j—total carotenoid.</p>
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<p>GWAS results for fruit color and carotenoid traits presented as Manhattan plots. The figure comprises plots for fruit color (<b>a</b>) and carotenoids: (<b>b</b>–<b>j</b>) α-carotene, antheraxanthin, β-carotene, β-cryptoxanthin, capsanthin, capsorubin, violaxanthin, zeaxanthin, and total carotenoids. In each plot, SNPs are represented as individual points, with chromosomes distinctly colored and labeled on the <span class="html-italic">x</span>-axis. The <span class="html-italic">y</span>-axis represents association strength as −log10(p). A grey dashed line at −log10(p) = 6.0 denotes the significance threshold (<span class="html-italic">p</span> &lt; 0.05).</p>
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20 pages, 14793 KiB  
Article
Predicted Responses of Genetically Improved Populations to Climate Changes Based on Second-Cycle Douglas-Fir Progeny Tests
by Terrance Z. Ye and Keith J. S. Jayawickrama
Forests 2024, 15(9), 1610; https://doi.org/10.3390/f15091610 - 12 Sep 2024
Viewed by 215
Abstract
The current planting of economically important timber species, such as Douglas-fir, mainly relies on genetically improved seeds from seed orchards. However, published research on the effects of climate change has largely focused on natural populations. To bridge this gap, data from 80 cooperative [...] Read more.
The current planting of economically important timber species, such as Douglas-fir, mainly relies on genetically improved seeds from seed orchards. However, published research on the effects of climate change has largely focused on natural populations. To bridge this gap, data from 80 cooperative second-cycle coastal Douglas-fir progeny tests across eight breeding zones in western Washington and Oregon were analyzed. Climate transfer functions for age-12 growth were derived, showing significant results for the US Pacific Northwest. Region-specific transfer functions (Coast, Inland, and Cascade) displayed stronger correlations. Mean annual temperature and mean coldest month temperature were the most important climatic variables explaining growth. The study found that populations from slightly warmer areas tended to grow better but moving populations from colder to warmer areas by 2 °C (analogous to projected global warming) would result in an 8% genetic loss in age-12 height and a 25% genetic loss in age-12 volume. However, substantial diversity in climatic response was found among full-sib families within large breeding zones, suggesting that breeding and selecting suitable families for future climatic conditions within breeding zones is feasible. The study discusses potential strategies to adapt current breeding programs to address the impacts of future climate change while maintaining high population growth rates in Douglas-fir breeding programs. Full article
(This article belongs to the Section Genetics and Molecular Biology)
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<p>Geographic variation in mean annual temperature (MAT) over the decade 2011–2020 (<b>a</b>) and mean annual precipitation (MAP) during the same period (<b>b</b>) across the PNW. Each dot indicates a progeny testing site, color-coded by testing program, included in this study.</p>
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<p>MAT and MAP patterns for parent trees (shown as faded dots) and testing sites (represented by colored symbols): (<b>a</b>) categorized by testing program; (<b>b</b>) grouped by geographic region (Coast, Inland, and Cascade).</p>
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<p>Relative height growth at age 12 as a function of transfer distance for five climate variables: MAT, MCMT, MAP, SHM, and MWMT. The results are derived from pooled transfer functions using data from western WA, western OR, and the PNW regions.</p>
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<p>Relative height growth at age 12 as a function of transfer distance for five climate variables: MAT, MCMT, MAP, SHM, and MWMT. The results are derived from pooled transfer functions using data from western WA, western OR, and the PNW regions.</p>
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<p>Relative height growth at age 12 as a function of transfer distance for five climate variables: MAT, MCMT, MAP, SHM, and MWMT. The results are derived from pooled transfer functions using data from the Coast, Inland, and Cascade regions across both WA and OR.</p>
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<p>Relative height growth at age 12 as a function of transfer distance for five climate variables: MAT, MCMT, MAP, SHM, and MWMT. The results are derived from pooled transfer functions using data from the Coast, Inland, and Cascade regions across both WA and OR.</p>
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<p>Projected increases in MAT from 2015 to (<b>a</b>) 2055 and (<b>b</b>) 2085 in the PNW.</p>
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<p>Projected changes in MAP from 2015 to (<b>a</b>) 2055 and (<b>b</b>) 2085 in the PNW.</p>
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<p>Patterns of family growth response to climate variables (MAT and MAP) across testing sites. Fifty full-sib families from each of the top, middle, and bottom groups, categorized based on predicted genetic gains for height at age 12, are sampled to examine family response patterns to climate change. The color scale represents the predicted genetic gain (%) of age-12 height growth over population means. (1) Pattern (A): families perform best with an MAP of about 2000 mm and prefer a higher MAT. (2) Pattern (B): families reach peak performance either at a low MAT with MAP between 2200 and 3600 mm, or with MAT around 10 °C and MAP around 2300 mm. (3) Pattern (C): similar to (A), families thrive either with MAT above 8.3 °C and MAP around 2000 mm, or with MAT below 9 °C and MAP above 3600 mm. (4) Pattern (D): comparable to (B), families achieve optimal performance with MAT below 7.6 °C and MAP between 2500 and 3500 mm. (5) Pattern (E): families excel with MAT above 9 °C and MAP between 2500 and 2800 mm. (6) Pattern (F): families thrive with MAT between 7.4 and 8.2 °C and MAP between 3200 and 3800 mm.</p>
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23 pages, 4097 KiB  
Review
Current Insights into Weak Seed Dormancy and Pre-Harvest Sprouting in Crop Species
by Angel J. Matilla
Plants 2024, 13(18), 2559; https://doi.org/10.3390/plants13182559 - 12 Sep 2024
Viewed by 265
Abstract
During the domestication of crops, seed dormancy has been reduced or eliminated to encourage faster and more consistent germination. This alteration makes cultivated crops particularly vulnerable to pre-harvest sprouting, which occurs when mature crops are subjected to adverse environmental conditions, such as excessive [...] Read more.
During the domestication of crops, seed dormancy has been reduced or eliminated to encourage faster and more consistent germination. This alteration makes cultivated crops particularly vulnerable to pre-harvest sprouting, which occurs when mature crops are subjected to adverse environmental conditions, such as excessive rainfall or high humidity. Consequently, some seeds may bypass the normal dormancy period and begin to germinate while still attached to the mother plant before harvest. Grains affected by pre-harvest sprouting are characterized by increased levels of α-amylase activity, resulting in poor processing quality and immediate grain downgrading. In the agriculture industry, pre-harvest sprouting causes annual economic losses exceeding USD 1 billion worldwide. This premature germination is influenced by a complex interplay of genetic, biochemical, and molecular factors closely linked to environmental conditions like rainfall. However, the exact mechanism behind this process is still unclear. Unlike pre-harvest sprouting, vivipary refers to the germination process and the activation of α-amylase during the soft dough stage, when the grains are still immature. Mature seeds with reduced levels of ABA or impaired ABA signaling (weak dormancy) are more susceptible to pre-harvest sprouting. While high seed dormancy can enhance resistance to pre-harvest sprouting, it can lead to undesirable outcomes for most crops, such as non-uniform seedling establishment after sowing. Thus, resistance to pre-harvest sprouting is crucial to ensuring productivity and sustainability and is an agronomically important trait affecting yield and grain quality. On the other hand, seed color is linked to sprouting resistance; however, the genetic relationship between both characteristics remains unresolved. The identification of mitogen-activated protein kinase kinase-3 (MKK3) as the gene responsible for pre-harvest sprouting-1 (Phs-1) represents a significant advancement in our understanding of how sprouting in wheat is controlled at the molecular and genetic levels. In seed maturation, Viviparous-1 (Vp-1) plays a crucial role in managing pre-harvest sprouting by regulating seed maturation and inhibiting germination through the suppression of α-amylase and proteases. Vp-1 is a key player in ABA signaling and is essential for the activation of the seed maturation program. Mutants of Vp-1 exhibit an unpigmented aleurone cell layer and exhibit precocious germination due to decreased sensitivity to ABA. Recent research has also revealed that TaSRO-1 interacts with TaVp-1, contributing to the regulation of seed dormancy and resistance to pre-harvest sprouting in wheat. The goal of this review is to emphasize the latest research on pre-harvest sprouting in crops and to suggest possible directions for future studies. Full article
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<p>Wheat seeds exhibiting pre-harvest sprouting (courtesy of J. Barrero-Sánchez; <a href="https://people.csiro.au/B/J/Jose-Barrero" target="_blank">https://people.csiro.au/B/J/Jose-Barrero</a> (accessed on 25 May 2024 )).</p>
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<p>Wheat spike with pre-harvest sprouting (courtesy of Z. Pang and Y. Liang; <a href="mailto:ycliang@zju.edu.cn">ycliang@zju.edu.cn</a>).</p>
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13 pages, 2279 KiB  
Article
Metagenomics Insight into Veterinary and Zoonotic Pathogens Identified in Urban Wetlands of Los Lagos, Chile
by Catherine Opitz-Ríos, Alvaro Burgos-Pacheco, Francisca Paredes-Cárcamo, Javier Campanini-Salinas and Daniel A. Medina
Pathogens 2024, 13(9), 788; https://doi.org/10.3390/pathogens13090788 - 12 Sep 2024
Viewed by 419
Abstract
Wetlands are ecosystems that are essential to ecological balance and biodiversity; nevertheless, human activity is a constant threat to them. Excess nutrients are caused by intensive livestock and agricultural operations, pollution, and population growth, which in turn leads to uncontrolled microbiological development. This [...] Read more.
Wetlands are ecosystems that are essential to ecological balance and biodiversity; nevertheless, human activity is a constant threat to them. Excess nutrients are caused by intensive livestock and agricultural operations, pollution, and population growth, which in turn leads to uncontrolled microbiological development. This impairment in water quality can constitute a risk to animal, human, and environmental health. To thoroughly characterize the microbial communities, shotgun metagenomics was used to characterize the taxonomic and functional pattern of microorganisms that inhabit urban wetlands in the Los Lagos Region of Chile. The main objective was to identify microorganisms of veterinary relevance, assess their potential antibiotic resistance, and characterize the main virulence mechanism. As expected, a high diversity of microorganisms was identified, including bacteria described as animal or human pathogens, such as Pasteurella multocida, Pseudomonas aeruginosa, Staphylococcus aureus, and Escherichia coli. Also, a diverse repertory of antimicrobial-resistant genes (ARGs) was detected in metagenomic assembled sequences and inside the sequence of mobile genetic elements, genes that confer mainly resistance to beta-lactams, consistent with the families of antibiotics most used in Chile. In addition, a diverse collection of virulence mechanisms was also identified. Given the significance of the relationship between environmental, animal, and human health—a concept known as One Health—there is a need to establish molecular surveillance programs that monitor the environmental biohazard elements using molecular tools. This work is the first report of the presence of these harmful biological elements in urban wetlands subjected to anthropogenic pressure, located in the south of Chile. Full article
(This article belongs to the Special Issue Spatio-Temporal Analysis of Veterinary Infectious Diseases)
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<p><b>Taxonomy abundance at the species level is represented as a stacked bar plot of each sample.</b> The color pattern of each bar shows the microbial community structure, while the amplitude of each color represents the percentage of abundance of the assigned taxonomy. Remarkable taxa with the most abundance was <span class="html-italic">Pseudomonas</span> sp. SCA2728.1_7 on the Mirasol wetland, represented by the dark blue color, <span class="html-italic">Serratia liquefaciens</span> on Antiñir, denoted by olive color, <span class="html-italic">Pseudomonas fragi</span> on Teodosio2 represented by cyan color, while on Teodosio1 the most abundant specie was <span class="html-italic">Pseudomonas psychrophile</span>, denoted by tomato color.</p>
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<p>Heatmap of bacteria species identified that are included in the list of notifiable diseases. The relative abundance of bacterial species in the cities of Osorno, Llanquihue, Puerto Varas, and Puerto Montt is represented by the colors white (low abundance), yellow (middle abundance), and red (high abundance). The wetland with the highest relative abundance of microorganisms, represented by an intense red color, is the La Paloma wetland, located in the city of Puerto Montt. Conversely, the wetland with the lowest abundance is the Luis Ebel wetland, also in the city of Puerto Montt.</p>
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<p>Heatmap of bacterial species relevant in veterinary and human medicine. The relative abundance of bacterial species in wetlands of the cities of Osorno, Llanquihue, Puerto Varas, and Puerto Montt is presented. The wetland with the highest relative abundance of infectious bacterial species, represented by an intense red color, is the La Paloma wetland in the city of Puerto Montt. Conversely, the wetlands with the lowest abundance are Las Ranas in the city of Llanquihue, Luis Ebel in the city of Puerto Montt, and La Marina in the city of Puerto Varas.</p>
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<p>Identified antimicrobial resistances and classification by antimicrobial families or by antimicrobial compounds in urban wetlands of the Los Lagos region. A wide diversity of ARGs were identified using the NCBI database, with the highest frequency corresponding to the Teodosio Sarao wetland in the city of Llanquihue, followed by the Luis Ebel wetland in the city of Puerto Montt. The number denotes the different genes for the same resistance.</p>
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14 pages, 3399 KiB  
Article
Detection of Quantitative Trait Loci Associated with Alkaline Tolerance Using Recombinant Inbred Line Population Derived from Longdao5 × Zhongyouzao8 at Seedling Stage
by Xijuan Zhang, Kai Liu, Chuanming Yang, Benfu Hou, Xianli Yang, Lizhi Wang, Shize Cui, Yongcai Lai, Zhugang Li and Shukun Jiang
Life 2024, 14(9), 1151; https://doi.org/10.3390/life14091151 - 11 Sep 2024
Viewed by 279
Abstract
Salt–alkaline stress is one of the most stressful occurrences, causing negative effects on plant development and agricultural yield. Identifying and utilizing genes that affect alkaline tolerance is an excellent approach to accelerate breeding processes and meet the needs for remediating saline–alkaline soil. Here, [...] Read more.
Salt–alkaline stress is one of the most stressful occurrences, causing negative effects on plant development and agricultural yield. Identifying and utilizing genes that affect alkaline tolerance is an excellent approach to accelerate breeding processes and meet the needs for remediating saline–alkaline soil. Here, we employed a mapping population of 176 recombinant inbred lines (RILs) produced from a cross between alkali-tolerant Longdao5 and alkali-sensitive Zhongyouzao8 to identify the quantitative trait loci (QTLs) determining alkali tolerance at the seedling stage. For the evaluation of alkali tolerance, the recovered seedling’s average alkali tolerance index (ATI), root number (RN), root length (RL), seedling dry weight (SW), root dry weight (RW), and seedling height (SH) were assessed, together with their relative alkaline damage rate. Under alkaline stress, the ATI was substantially negative connected with the root number, seedling height, seedling dry weight, and root dry weight; however, it was considerably positive correlated with the relative alkaline damage rate of the root number and root dry weight. A total of 13 QTLs for the root number, root length, seedling height, seedling dry weight, root dry weight, and alkali tolerance index under alkaline stress were identified, which were distributed across chromosomes 1, 2, 3, 4, 5, 7, and 8. All of these QTLs formed two QTL clusters for alkali tolerance on chromosome 5 and chromosome 7, designated AT5 and AT7, respectively. Nine QTLs were identified for the relative alkaline damage rate of the root number, root length, seedling height, seedling dry weight, and root dry weight under alkali stress. These QTLs were located on chromosome 2, 4, 6, 7, 8, 9, and 12. In conclusion, these findings further strengthen our knowledge about rice’s genetic mechanisms for alkaline tolerance. This research offers clues to accelerate breeding programs for new alkaline-tolerance rice varieties. Full article
(This article belongs to the Special Issue Plant Functional Genomics and Breeding)
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<p>The evaluation criteria for the relative degree of leaf alkaline damage of rice seedlings.</p>
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<p>The phenotypic traits associated with alkaline tolerance at the seedling stage between Longdao5 (left) and Zhongyouzao8 (right); (<b>a</b>) 0 days in normal Kimura’s culture solution B; (<b>b</b>) 6 days in normal Kimura’s culture solution B; (<b>c</b>) 9 days in normal Kimura’s culture solution B; (<b>d</b>) 12 days in normal Kimura’s culture solution B; (<b>e</b>) 15 days in normal Kimura’s culture solution B; (<b>f</b>) 18 days in normal Kimura’s culture solution B; (<b>g</b>) 0 days in Kimura’s culture solution B with NaHCO<sub>3</sub>; (<b>h</b>) 6 days in Kimura’s culture solution B with NaHCO<sub>3</sub>; (<b>i</b>) 9 days in Kimura’s culture solution B with NaHCO<sub>3</sub>; (<b>j</b>) 12 days in Kimura’s culture solution B with NaHCO<sub>3</sub>; (<b>k</b>) 3 days of recovery in Kimura’s culture solution B; (<b>l</b>) 6 days of recovery in Kimura’s culture solution B.</p>
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<p>The phenotypic traits associated with alkaline tolerance between Longdao5 and Zhongyouzao8 after 6 days of recovery in Kimura’s culture solution B. CK: cyan; Alkaline stress: carmine pink. (<b>a</b>) The alkali tolerance index between Longdao5 and Zhongyouzao8; (<b>b</b>) the root number between Longdao5 and Zhongyouzao8; (<b>c</b>) the root length between Longdao5 and Zhongyouzao8; (<b>d</b>) the seedling height between Longdao5 and Zhongyouzao8; (<b>e</b>) the seedling dry weight between Longdao5 and Zhongyouzao8; (<b>f</b>) the root dry weight between Longdao5 and Zhongyouzao8.</p>
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<p>Distributions and the LOD value for the root number (RN), root length (RL), seedling height (SH), seedling weight (SW), root weight (RW), and alkali tolerance index (ATI) under the alkaline stress and normal condition at the whole-genome level in RIL populations. (<b>a</b>) The root number distribution of RILs’ population under alkaline stress; (<b>b</b>) the root number distribution of RILs’ population under the normal condition; (<b>c</b>) the root length distribution of RILs’ population under alkaline stress; (<b>d</b>) the root length distribution of RILs’ population under the normal condition; (<b>e</b>) the seedling height distribution of RILs’ population under alkaline stress; (<b>f</b>) the seedling height distribution of RILs’ population under the normal condition; (<b>g</b>) the seedling weight distribution of RILs’ population under alkaline stress; (<b>h</b>) the seedling weight distribution of RILs’ population under the normal condition; (<b>i</b>) the root weight distribution of RILs’ population under alkaline stress; (<b>j</b>) the root weight distribution of RILs’ population under the normal condition; (<b>k</b>) the alkali tolerance index distribution of RILs’ population under alkaline stress; (<b>l</b>) the LOD value of the root number (the upper part) and its relative alkaline damage rate (the lower part) at the whole-genome level in RILs’ populations; (<b>m</b>) the LOD value of root length (the upper part) and its relative alkaline damage rate (the lower part) at the whole-genome level in RILs’ populations; (<b>n</b>) the LOD value of seedling height (the upper part) and its relative alkaline damage rate (the lower part) at the whole-genome level in RILs’ populations; (<b>o</b>) the LOD value of seedling weight (the upper part) and its relative alkaline damage rate (the lower part) at the whole-genome level in RILs’ populations; (<b>p</b>) the LOD value of root weight (the upper part) and its relative alkaline damage rate (the lower part) at the whole-genome level in RILs’ populations; (<b>q</b>) the LOD value of the alkali tolerance index at the whole-genome level in RILs’ populations.</p>
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<p>The correlation coefficients between the ATI and the seedling traits in the RIL population. The values are correlation coefficients. The areas of squares correspond to absolute values of the corresponding r. The blue and orange colors indicate positive and negative correlations, respectively.</p>
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<p>Genomic locations of the QTLs with alkaline tolerance-related traits identified in the RIL population. Green indicates the QTLs for the key trait alkali tolerance index; carmine pink indicates the QTLs for the traits of a seedling and root under alkali stress; blue indicates the QTLs for the relative alkaline damage rate; 1–12 indicates chromosome 1 to chromosome 12.</p>
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13 pages, 1728 KiB  
Article
Whole-Genome Resequencing Identifies SNPs in Sucrose Synthase and Sugar Transporter Genes Associated with Sweetness in Coconut
by Manlika Khongmaluan, Wanchana Aesomnuk, Reajina Dumhai, Mutiara K. Pitaloka, Yong Xiao, Rui Xia, Tippaya Kraithong, Natthaporn Phonsatta, Atikorn Panya, Vinitchan Ruanjaichon, Samart Wanchana and Siwaret Arikit
Plants 2024, 13(18), 2548; https://doi.org/10.3390/plants13182548 - 11 Sep 2024
Viewed by 319
Abstract
Coconut (Cocos nucifera L.) is an important agricultural commodity with substantial economic and nutritional value, widely used for various products, including coconut water. The sweetness is an important quality trait of coconut water, which is influenced by genetic and environmental factors. In [...] Read more.
Coconut (Cocos nucifera L.) is an important agricultural commodity with substantial economic and nutritional value, widely used for various products, including coconut water. The sweetness is an important quality trait of coconut water, which is influenced by genetic and environmental factors. In this study, we utilized next-generation sequencing to identify genetic variations in the coconut genome associated with the sweetness of coconut water. Whole-genome resequencing of 49 coconut accessions, including diverse germplasm and an F2 population of 81 individuals, revealed ~27 M SNPs and ~1.5 M InDels. Sugar content measured by °Bx was highly variable across all accessions tested, with dwarf varieties generally sweeter. A comprehensive analysis of the sugar profiles revealed that sucrose was the major sugar contributing to sweetness. Allele mining of the 148 genes involved in sugar metabolism and transport and genotype–phenotype association tests revealed two significant SNPs in the hexose carrier protein (Cnu01G018720) and sucrose synthase (Cnu09G011120) genes associated with the higher sugar content in both the germplasm and F2 populations. This research provides valuable insights into the genetic basis of coconut sweetness and offers molecular markers for breeding programs aimed at improving coconut water quality. The identified variants can improve the selection process in breeding high-quality sweet coconut varieties and thus support the economic sustainability of coconut cultivation. Full article
(This article belongs to the Section Plant Genetics, Genomics and Biotechnology)
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<p>Population study of the 49 coconut accessions. (<b>A</b>) UPGMA phylogenetic tree. (<b>B</b>) Principal coordinate analysis (PCoA). (<b>C</b>) STRUCTURE analysis. Different genetic groups are highlighted by different colors.</p>
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<p>The sugar content in coconut water from accessions with varying sweetness levels. A stacked bar graph illustrates the quantities of different sugars and sugar alcohols measured in coconut water extracted from 7-month-old fruits.</p>
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<p>Genotype–phenotype association on two genes, Cnu01G018720 and Cnu09G011120. The box plot/violin plots display the three different genotypes at SNP 1_157345938 on the gene Cnu01G018720, analyzed in 49 accessions (<b>A</b>) and 81 F<sub>2</sub> individuals (<b>B</b>). The box plot/violin plots display the three different genotypes at SNP 9_119160271 on the gene Cnu09G011120, analyzed in 49 accessions (<b>D</b>) and 81 F<sub>2</sub> individuals (<b>E</b>). The medians are indicated by solid horizontal lines in the box plots. The structure of Cnu01G018720 (<b>C</b>) and Cnu09G011120 (<b>F</b>) shows UTRs (small blue boxes), exons (large blue boxes) and introns (arrow lines). The grey bar represents the chromosome, with SNP positions in each gene indicated by vertical bars. * <span class="html-italic">p</span> &lt; 0.05; ** <span class="html-italic">p</span> &lt; 0.01; *** <span class="html-italic">p</span> &lt; 0.001; **** <span class="html-italic">p</span> &lt; 0.0001; ns (not significant).</p>
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10 pages, 936 KiB  
Article
Developing a Microsatellite Polymerase Chain Reaction System for Small Yellow Croaker (Larimichthys polyactis) and Its Application in Parentage Assignment
by Eun-Soo Noh, Eun-Ha Shin, Hee-Jeong Kong, Young-Ok Kim and Yong-Woon Ryu
Biology 2024, 13(9), 710; https://doi.org/10.3390/biology13090710 - 11 Sep 2024
Viewed by 263
Abstract
(1) Background: The small yellow croaker, an economically important fish in East Asia, has been subjected to population declines due to overfishing and environmental pressures. The development of effective breeding programs is considered crucial for the species, and accurate parentage assignment is deemed [...] Read more.
(1) Background: The small yellow croaker, an economically important fish in East Asia, has been subjected to population declines due to overfishing and environmental pressures. The development of effective breeding programs is considered crucial for the species, and accurate parentage assignment is deemed essential for such programs. (2) Methods: The assembled reference genome of the small yellow croaker was utilized to select highly polymorphic microsatellite markers. A multiplex PCR system was optimized for the simultaneous amplification of these markers. The system’s accuracy was validated using controlled mating pairs and subsequently applied to a group mating scenario. (3) Results: The developed multiplex PCR system demonstrated high accuracy in assigning offspring to their parents in both the controlled and group mating scenarios. (4) Conclusions: The system is presented as a valuable tool for pedigree management, selective breeding, and conservation efforts for the small yellow croaker, facilitating sustainable aquaculture practices and genetic improvement. Full article
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<p>Size range of microsatellite markers for multiplex PCR.</p>
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<p>Analysis of genetic relationships between parents, offspring, and wild groups using PCA.</p>
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19 pages, 6886 KiB  
Article
GSK-3β in Dendritic Cells Exerts Opposite Functions in Regulating Cross-Priming and Memory CD8 T Cell Responses Independent of β-Catenin
by Chunmei Fu, Jie Wang, Tianle Ma, Congcong Yin, Li Zhou, Björn E. Clausen, Qing-Sheng Mi and Aimin Jiang
Vaccines 2024, 12(9), 1037; https://doi.org/10.3390/vaccines12091037 - 10 Sep 2024
Viewed by 532
Abstract
GSK-3β plays a critical role in regulating the Wnt/β-catenin signaling pathway, and manipulating GSK-3β in dendritic cells (DCs) has been shown to improve the antitumor efficacy of DC vaccines. Since the inhibition of GSK-3β leads to the activation of β-catenin, we hypothesize that [...] Read more.
GSK-3β plays a critical role in regulating the Wnt/β-catenin signaling pathway, and manipulating GSK-3β in dendritic cells (DCs) has been shown to improve the antitumor efficacy of DC vaccines. Since the inhibition of GSK-3β leads to the activation of β-catenin, we hypothesize that blocking GSK-3β in DCs negatively regulates DC-mediated CD8 T cell immunity and antitumor immunity. Using CD11c-GSK-3β−/− conditional knockout mice in which GSK-3β is genetically deleted in CD11c-expressing DCs, we surprisingly found that the deletion of GSK-3β in DCs resulted in increased antitumor immunity, which contradicted our initial expectation of reduced antitumor immunity due to the presumed upregulation of β-catenin in DCs. Indeed, we found by both Western blot and flow cytometry that the deletion of GSK-3β in DCs did not lead to augmented expression of β-catenin protein, suggesting that GSK-3β exerts its function independent of β-catenin. Supporting this notion, our single-cell RNA sequencing (scRNA-seq) analysis revealed that GSK-3β-deficient DCs exhibited distinct gene expression patterns with minimally overlapping differentially expressed genes (DEGs) compared to DCs with activated β-catenin. This suggests that the deletion of GSK-3β in DCs is unlikely to lead to upregulation of β-catenin at the transcriptional level. Consistent with enhanced antitumor immunity, we also found that CD11c-GSK-3β−/− mice exhibited significantly augmented cross-priming of antigen-specific CD8 T cells following DC-targeted vaccines. We further found that the deletion of GSK-3β in DCs completely abrogated memory CD8 T cell responses, suggesting that GSK-3β in DCs also plays a negative role in regulating the differentiation and/or maintenance of memory CD8 T cells. scRNA-seq analysis further revealed that although the deletion of GSK-3β in DCs positively regulated transcriptional programs for effector differentiation and function of primed antigen-specific CD8 T cells in CD11c-GSK-3β−/− mice during the priming phase, it resulted in significantly reduced antigen-specific memory CD8 T cells, consistent with diminished memory responses. Taken together, our data demonstrate that GSK-3β in DCs has opposite functions in regulating cross-priming and memory CD8 T cell responses, and GSK-3β exerts its functions independent of its regulation of β-catenin. These novel insights suggest that targeting GSK-3β in cancer immunotherapies must consider its dual role in CD8 T cell responses. Full article
(This article belongs to the Special Issue Vaccines Targeting Dendritic Cells)
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<p>Deletion of GSK-3β in DCs led to augmented antitumor immunity in CD11c-GSK-3β<sup>−/−</sup> mice. WT and CD11c-GSK-3β<sup>−/−</sup> mice (<span class="html-italic">n</span> = 7–9) were inoculated with B16F10 melanoma cells, and tumor sizes were monitored. (<b>A</b>,<b>B</b>) CD11c-GSK-3β<sup>−/−</sup> mice exhibited reduced tumor growth compared to WT mice. Tumor sizes from the day of treatment are shown in (<b>A</b>) and tumor weights at the end of the experiment (day 20) are shown in (<b>B</b>). A linear mixed model (Lme4) was fitted to the data in (<b>A</b>), and ANOVA for the fitted linear mixed model was then performed to determine the difference between groups. Student’s <span class="html-italic">t</span>-tests were used for (<b>B</b>). *** <span class="html-italic">p</span> &lt; 0.001. (<b>C</b>) Photo of the tumors at the day 20 after tumor inoculation. Data are representative of two experiments.</p>
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<p>GSK-3β<sup>−/−</sup> DCs exhibited different expression profiles from β-catenin<sup>active</sup> DCs by scRNA-seq. DCs sorted from spleens of WT (GSK-3β<sup>Flox/Flox</sup>) and CD11c-GSK-3β<sup>−/−</sup> mice, or from WT (β-catenin <sup>Exon3/Exon3</sup>) and CD11c-β-catenin<sup>active</sup> (CD11c-Cre β-catenin<sup>Exon3/Exon3</sup>), were subjected to scRNA-seq as described. (<b>A</b>) Uniform manifold approximation and projection (UMAP) dimensionality reduction mapping analysis of single-cell gene expression of integrated WT (GSK-3β<sup>Flox/Flox</sup>) and GSK-3β<sup>−/−</sup> DCs, and WT (β-catenin<sup>Exon3/Exon3</sup>) and β-catenin<sup>active</sup> DCs. Each dot represents one single cell. A total of 13 clusters were identified and color-coded as indicated. (<b>B</b>) Bubble plots showing the expression of key markers for pDC, cDC1, cDC2, and MoDCs cells among 13 UMAP clusters. The sizes of dots represent the percentages expressed; the color of dot represents the average expression. (<b>C</b>) Bubble plots depicting expression of top DEGs for UMAP clusters shown in (<b>A</b>). (<b>D</b>) Distribution of cells from WT/GSK-3β<sup>Flox/Flox</sup> and GSK-3β<sup>−/−</sup> (left), or WT/β-catenin<sup>Exon3/Exon3</sup> and β-catenin<sup>active</sup> DCs (right) within each of the 13 clusters as depicted in (<b>A</b>). (<b>E</b>) Venn plot showing the overlap of downregulated DEGs (left) and upregulated DEGs (right) in GSK-3β<sup>−/−</sup> DCs versus WT/GSK-3β<sup>Flox/Flox</sup> DCs (GSK-3β<sup>−/−</sup> vs. WT), and β-catenin<sup>active</sup> and WT/β-catenin<sup>Exon3/Exon3</sup> DCs (β-catenin<sup>active</sup> vs. WT). (<b>F</b>) Volcano plot visualizing expression of DEGs in GSK-3β<sup>−/−</sup> and WT/GSK-3β<sup>Flox/Flox</sup> DCs, and their expression pattern in β-catenin<sup>active</sup> and WT/β-catenin<sup>Exon3/Exon3</sup> DCs. DEGs in GSK-3β<sup>−/−</sup> DCs versus WT/GSK-3β<sup>Flox/Flox</sup> DCs are shown in volcano plot (upper), and expression of downregulated DEGs (lower left) and upregulated DEGs (lower right) in β-catenin<sup>active</sup> and WT/β-catenin<sup>Exon3/Exon3</sup> DCs are analyzed and shown in volcano plots. (<b>G</b>) GO enrichment analysis identifies top regulated biological process pathways in in GSK-3β<sup>−/−</sup> DCs vs. WT/GSK-3β<sup>Flox/Flox</sup> DCs (upper), and β-catenin<sup>active</sup> vs. WT/β-catenin<sup>Exon3/Exon3</sup> DCs (lower).</p>
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<p>Deletion of GSK-3β in DCs does not upregulate β-catenin. (<b>A</b>,<b>B</b>) GSK-3β<sup>−/−</sup> cDCs express similar levels of β-catenin to WT cDCs. WT and GSK-3β<sup>−/−</sup> splenic cDCs were isolated and subjected to Western blot. (<b>A</b>) Expression of GSK-3α/β, β-catenin, and β-actin by Western blotting is shown. One of three experiments is shown. (<b>B</b>) Statistical analysis of β-catenin expression is shown. The relative expression of β-catenin Western blot intensity relative to that of b-actin loading control was calculated, and the ratios for WT cDCs for each experiment were set at 1.0. (<b>C</b>,<b>D</b>) Deletion of GSK-3β in DCs does not upregulate β-catenin. Histogram overlay of β-catenin expression (<b>C</b>) and mean fluorescence intensity (MFI) of β-catenin expression (<b>D</b>) on gated CD11c<sup>+</sup>Bst2<sup>−</sup> cDCs are shown. Student’s <span class="html-italic">t</span>-test, and NS &gt; 0.05. Data shown are representative of at least three experiments.</p>
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<p>Deletion of GSK-3β in DCs abrogated memory CD8 T cell responses despite augmented cross-priming. (<b>A</b>,<b>B</b>) Deletion of GSK-3β in DCs led to significantly augmented cross-priming. WT and DC-GSK-3β<sup>−/−</sup> mice (<span class="html-italic">n</span> = 4) were immunized with anti-DEC-205-OVA with CpG following adoptive transfer of naïve CFSE-labeled Thy1.1<sup>+</sup> OTI cells, and cross-priming was examined at day 4 after immunization. (<b>A</b>) The percentages of Thy1.1<sup>+</sup> OTI cells out of total CD8 T cells, and (<b>B</b>) the percentages of IFN-γ<sup>+</sup> cells out of total Thy1.1<sup>+</sup>CD8<sup>+</sup> OTI cells in both spleen and draining LN are shown. (<b>C</b>) CD8 memory responses were abrogated in CD11c-GSK-3β<sup>−/−</sup> mice upon recall. Immunized WT and CD11c-GSK-3β<sup>−/−</sup> mice (<span class="html-italic">n</span> = 4–5) were recalled at day 21 and examined 5 days later. The percentages of Thy1.1<sup>+</sup> OTI cells out of total CD8 T cells are shown. Student’s <span class="html-italic">t</span>-test. * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, and *** <span class="html-italic">p</span> &lt; 0.001. Data shown are representative of at least two experiments.</p>
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<p>scRNA-seq of OVA-specific CD8 T cells identifies distinct populations and reveals differences between CD8 T cells primed in WT and CD11c-GSK-3β<sup>−/−</sup> mice. WT and CD11c-GSK-3β<sup>−/−</sup> mice adoptively transferred Thy1.1<sup>+</sup> OTI CD8 T cells were immunized with anti-DEC-205-OVA plus CpG. Spleen cells were harvested at day 4 or day 10 after immunization, and FACS-sorted OTI cells were subjected to scRNA-seq as described. (<b>A</b>,<b>B</b>) UMAP-dimensionality reduction mapping analysis of single-cell gene expression data of OTI cells isolated 4 or 10 days following vaccination with ant-DEC-205-OVA. Each dot represents one single cell. A total of 9 clusters were identified and color-coded as indicated. UMAP visualization of single cells from combined OTI cells (<b>A</b>), or OT1 cells from WT or CD11c-GSK-3β<sup>−/−</sup> mice at day 4 and day 10 (<b>B</b>) are shown. (<b>C</b>) Bubble plots depicting expression of top DEGs for UMAP clusters shown in (<b>A</b>). (<b>D</b>) Distribution of OTI cells from either WT or CD11c-GSK-3β<sup>−/−</sup> mice at day 4 or day 10 within each of the 9 clusters as depicted in (<b>A</b>). (<b>E</b>) Bubble plots showing the key signatures for CD8 T cells effector and memory phenotype. (<b>F</b>) Expression of effector markers among the UMAP clusters. Gradient expression levels are color-coded as indicated. (<b>G</b>) Violin plot depicting the module score of gene sets associated with effector on OTI cells from either WT or CD11c-GSK-3β<sup>−/−</sup> mice at day 4 or day 10. *** <span class="html-italic">p</span> &lt; 0.001 and **** <span class="html-italic">p</span> &lt; 0.0001 (<b>H</b>) Signaling pathways that are significantly downregulated or upregulated in OTI cells primed in CD11c-GSK-3β<sup>−/−</sup> mice compared to OTI cells from WT mice at day 4 and day 10.</p>
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<p>Schematic representation of GSK-3β’s dual roles in regulating CD8 T cell responses. Inhibition of GSK-3β is generally believed to upregulate β-catenin, leading to increased IL-10 production, which suppresses cross-priming and reduces memory CD8 T cell responses. However, our studies demonstrate that genetic deletion of GSK-3β in CD11c<sup>+</sup> DCs does not result in β-catenin accumulation (activation). Instead, the deletion of GSK-3β in DCs enhances cross-priming of CD8 T cells, as indicated by an increase in effector cells and a higher effector index, based on scRNA-seq analysis. Despite this enhanced cross-priming, memory CD8 T cells are nearly abrogated in CD11c-GSK-3β<sup>−/−</sup> mice, likely due to a significant loss of both effector and memory CD8 T cell populations. Collectively, these findings reveal novel mechanisms by which GSK-3β exerts opposing effects on CD8 T cell responses.</p>
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