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Rust fungi are the largest group of obligate plant pathogens and cause severe damage to global forests and agricultural security. Meliosma myriantha, a tree species native to East Asia (China, Japan, and Korea), is vulnerable to three rust species: Neophysopella meliosmae, N. meliosmae-myrianthae, and N. vitis. The early symptoms of infection are indistinguishable between these species, making an accurate and rapid diagnosis challenging. The urediniospores of N. meliosmae-myrianthae and N. vitis are also known to infect economically relevant grapevines (Vitis spp.) and ivies (Parthenocissus spp.), respectively, rendering early detection and identification even more important. To address this issue, we developed a multiplex quantitative polymerase chain reaction assay equipped with TaqMan probes targeting the internal transcribed spacer rDNA sequences specific to the three rust pathogens. This assay successfully detected minute quantities (5 fg for N. meliosmae-myrianthae and 50 fg for N. meliosmae and N. vitis) of DNA from the three Neophysopella species and demonstrated consistent reliability when applied to fresh and herbarium samples collected from M. myriantha, grapevines, and ivies. In conclusion, this novel assay is a rapid and robust diagnostic tool for the three rust pathogens, N. meliosmae, N. meliosmae-myrianthae, and N. vitis, and offers the potential to identify and detect their global movement and spread to grapevines, ivies, and trees.
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Basidiomycota , DNA Fúngico , Reação em Cadeia da Polimerase Multiplex , Doenças das Plantas , Basidiomycota/genética , Basidiomycota/isolamento & purificação , Basidiomycota/classificação , Doenças das Plantas/microbiologia , DNA Fúngico/genética , Reação em Cadeia da Polimerase Multiplex/métodos , DNA Espaçador Ribossômico/genética , Reação em Cadeia da Polimerase em Tempo Real/métodos , Reprodutibilidade dos Testes , Vitis/microbiologiaRESUMO
Despite the well-known advantages of lipid vesicles for drug and gene delivery, structural instability limits their practical applications and requires strictly regulated conditions for transport and storage. Chemical crosslinking and in situ polymerization have been suggested to increase the membrane rigidity and dispersion stability of lipid vesicles. However, such chemically modified lipids sacrifice the dynamic nature of lipid vesicles and obfuscate their in vivo metabolic fates. Here, we present highly robust multilamellar lipid vesicles through the self-assembly of preformed, cationic large unilamellar vesicles (LUVs) with hydrolyzed collagen peptides (HCPs). The cationic LUVs undergo vesicle-to-vesicle attachment and structural reorganization through polyionic complexation with HCPs, resulting in the formation of multilamellar collagen-lipid vesicles (MCLVs). The resulting MCLVs exhibit excellent structural stability against variations in pH and ionic strength and the addition of surfactants. Particularly, the MCLVs maintain their structural stability against repeated freeze-thaw stresses, proving the unprecedented stabilization effect of biological macromolecules on lipid lamellar structures. This work provides a practically attractive technique for the simple and quick fabrication of structurally robust lipid nanovesicles without covalent crosslinkers, organic solvents, and specialized instruments.
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Tensoativos , Lipossomas Unilamelares , Lipossomas Unilamelares/química , Tensoativos/química , Cátions , Lipídeos/química , Peptídeos , Lipossomos/químicaRESUMO
The highly selective detection of trace gases using transparent sensors at room temperature remains challenging. Herein, transparent nanopatterned chemiresistors composed of aligned 1D Au-SnO2 nanofibers, which can detect toxic NO2 gas at room temperature under visible light illumination is reported. Ten straight Au-SnO2 nanofibers are patterned on a glass substrate with transparent electrodes assisted by direct-write, near-field electrospinning, whose extremely low coverage of sensing materials (≈0.3%) lead to the high transparency (≈93%) of the sensor. The sensor exhibits a highly selective, sensitive, and reproducible response to sub-ppm levels of NO2 , and its detection limit is as low as 6 ppb. The unique room-temperature NO2 sensing under visible light emanates from the localized surface plasmonic resonance effect of Au nanoparticles, thereby enabling the design of new transparent oxide-based gas sensors without external heaters or light sources. The patterning of nanofibers with extremely low coverage provides a general strategy to design diverse compositions of gas sensors, which can facilitate the development of a wide range of new applications in transparent electronics and smart windows wirelessly connected to the Internet of Things.
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Background Multicenter studies are required to validate the added benefit of using deep convolutional neural network (DCNN) software for detecting malignant pulmonary nodules on chest radiographs. Purpose To compare the performance of radiologists in detecting malignant pulmonary nodules on chest radiographs when assisted by deep learning-based DCNN software with that of radiologists or DCNN software alone in a multicenter setting. Materials and Methods Investigators at four medical centers retrospectively identified 600 lung cancer-containing chest radiographs and 200 normal chest radiographs. Each radiograph with a lung cancer had at least one malignant nodule confirmed by CT and pathologic examination. Twelve radiologists from the four centers independently analyzed the chest radiographs and marked regions of interest. Commercially available deep learning-based computer-aided detection software separately trained, tested, and validated with 19 330 radiographs was used to find suspicious nodules. The radiologists then reviewed the images with the assistance of DCNN software. The sensitivity and number of false-positive findings per image of DCNN software, radiologists alone, and radiologists with the use of DCNN software were analyzed by using logistic regression and Poisson regression. Results The average sensitivity of radiologists improved (from 65.1% [1375 of 2112; 95% confidence interval {CI}: 62.0%, 68.1%] to 70.3% [1484 of 2112; 95% CI: 67.2%, 73.1%], P < .001) and the number of false-positive findings per radiograph declined (from 0.2 [488 of 2400; 95% CI: 0.18, 0.22] to 0.18 [422 of 2400; 95% CI: 0.16, 0.2], P < .001) when the radiologists re-reviewed radiographs with the DCNN software. For the 12 radiologists in this study, 104 of 2400 radiographs were positively changed (from false-negative to true-positive or from false-positive to true-negative) using the DCNN, while 56 of 2400 radiographs were changed negatively. Conclusion Radiologists had better performance with deep convolutional network software for the detection of malignant pulmonary nodules on chest radiographs than without. © RSNA, 2019 Online supplemental material is available for this article. See also the editorial by Jacobson in this issue.
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Neoplasias Pulmonares/diagnóstico por imagem , Nódulos Pulmonares Múltiplos/diagnóstico por imagem , Redes Neurais de Computação , Interpretação de Imagem Radiográfica Assistida por Computador/métodos , Radiografia Torácica/métodos , Nódulo Pulmonar Solitário/diagnóstico por imagem , Adulto , Idoso , Feminino , Humanos , Pulmão/diagnóstico por imagem , Masculino , Pessoa de Meia-Idade , Reprodutibilidade dos Testes , Estudos Retrospectivos , Sensibilidade e Especificidade , Adulto JovemRESUMO
Biopanning refers to the processes of screening peptides with a high affinity to a target material. Microfluidic biopanning has advantages compared to conventional biopanning which requires large amounts of the target material and involves inefficient multiple pipetting steps to remove nonspecific or low-affinity peptides. Here, we fabricate a microfluidic biopanning system to identify a new gold-binding peptide (GBP). A polydimethylsiloxane microfluidic device is fabricated and bonded to a glass slide with a gold pattern that is deposited by electron-beam evaporation. The microfluidic biopanning system can provide high adjustability in the washing step during the biopanning process because the liquid flow rate and the resulting shear stress can be precisely controlled. The surface plasmon resonance analysis shows that the binding affinity of the identified GBP is comparable to previously reported GBPs. Moreover, molecular dynamics simulations are performed to understand its binding affinity against the gold surface in detail. Theoretical calculations suggest that the association and dissociation rates of the GBPs depend on their sequence-dependent conformations and interactions with the gold surface. These findings provide insight into designing efficient biopanning tools and peptides with a high affinity for various target materials.
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Ouro/metabolismo , Peptídeos/metabolismo , Dimetilpolisiloxanos/química , Dispositivos Lab-On-A-Chip , Microfluídica/instrumentação , Microfluídica/métodos , Simulação de Dinâmica Molecular , Peptídeos/química , Ligação Proteica , Conformação ProteicaRESUMO
BACKGROUND: Stafne mandibular bone cavity (SMBC) is an asymptomatic radiolucent lesion observed in the mandible on X-ray images, with well-defined borders. This lesion does not require treatment, and size changes are very rare. The purpose of this article is to summarize the radiological and clinical features of SMBC, aiming to prevent misinterpretation of this benign lesion as other pathologies and aid in differential diagnosis within the field of dental practice. METHODS: A total of 32 patients who visited our hospital and were radiologically diagnosed with SMBC based on cone-beam computed tomography (CBCT) and panoramic images between 2005 and 2021 were included in this study. Among them, surgical intervention was performed in one case. Gender and age distribution of the patients, radiographic location and size of the lesion, relationship with the mandibular canal in CBCT, presence of cortical bone erosion in the mandibular lingual area in panoramic images, and the amount of residual cortical bone on the affected side in CBCT were investigated. RESULT: Patients were 26 men (81.3%) and 6 women, with a mean age of 54.3 years. The average horizontal and vertical length was 16.6 mm and 10.6 mm. Out of a total of 32 cases, 29 cases were found in the posterior region of the mandibular body, while 3 cases were in the angle of the mandible. Lesions located below the mandibular canal were observed in 29 cases, while lesions involving the mandibular canal were present in 3 cases. Erosion of the mandibular lingual cortical bone was observed in 11 cases (34.4%), while 21 cases (65.6%) showed no erosion on panoramic images. Among the total of 14 cases (43.8%) where the cortical bone on the affected side was invaded, the average residual cortical bone thickness was 1.1 mm. CONCLUSION: SMBC is a benign lesion primarily found in the mandibular angle and posterior body of the mandible. In most cases, treatment is not necessary, and differentiation from other lesions can be achieved by understanding its clinical characteristics and features on panoramic radiographs and CBCT.
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The two electron cyclotron emission imaging (ECEI) systems installed at adjacent ports (G and H) on the KSTAR tokamak incorporate large-aperture mm-wave optics, broadband electronics, and high speed digitization (up to 1 MSa/s) for 2D and quasi-3D visualization of MHD-scale fluid dynamics. Recently, the ECEI systems have been proved to be capable of visualization of smaller scale fluctuations albeit with a limited spatiotemporal resolution and even capable of measurement of ion cyclotron harmonic waves by direct high-speed sampling of the ECE IF signals. A four-channel prototype subsystem with a higher sampling rate up to 16 GS/s has been integrated into the G-port ECEI system, enabling the measurement of plasma waves in the GHz range in the form of modulated ECE signals and characterization of high-frequency turbulence during the evolution of pedestal. To achieve higher toroidal resolution in the turbulence measurement, the H-port ECEI system is now being upgraded to have a toroidally dual detector array of 2(toroidal) × 12(vertical) × 8(radial) channel configuration and a high-speed subsystem of 2(toroidal) × 4 channel configuration. The new mm-wave optics has been designed via beam propagation simulation, and the measured performance of the fabricated lens indicates a toroidal resolution of 8-10 cm depending on the focus position and zoom factor, allowing for the measurement of parallel wavenumber up to kâ â¼ 0.8 cm-1.
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Lipid vesicles are widely used for drug and gene delivery, but their structural instability reduces in vivo efficacy and requires specialized handling. To address these limitations, strategies like lipid cross-linking and polymer-lipid conjugation are suggested to enhance stability and biological efficacy. However, the in vivo metabolism of these altered lipids remains unclear, necessitating further studies. A new stabilization technique without chemical modification is urgently needed. Here, a bio-mimetic approach for fabricating robust multilamellar lipid vesicles to enhance in vivo delivery and stabilization of protein antigens is presented. This method leverages 1-O-acylceramide, a natural skin lipid, to facilitate the self-assembly of lipid nanovesicles. Incorporating 1-O-acylceramide, anchoring lipid bilayers akin to its role in the stratum corneum, provides excellent stability under environmental stresses, including freeze-thaw cycles. Encapsulating ovalbumin as a model antigen and the adjuvant monophosphoryl lipid A demonstrates the vesicle's potential as a nanovaccine platform. In vitro studies show enhanced immune responses with both unilamellar and multilamellar vesicles, but in vivo analyses highlight the superior efficiency of multilamellar vesicles in inducing higher antibody and cytokine levels. This work suggests ceramide-induced multilamellar lipid vesicles as an effective nanovaccine platform for enhanced antigen delivery and stability.
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Ovalbumina , Animais , Camundongos , Ovalbumina/química , Ovalbumina/imunologia , Materiais Biomiméticos/química , Materiais Biomiméticos/farmacologia , Vacinação/métodos , Lipídeo A/química , Lipídeo A/análogos & derivados , Vacinas/química , Vacinas/imunologia , Ceramidas/química , Lipídeos/química , Nanopartículas/química , Feminino , Camundongos Endogâmicos C57BLRESUMO
We aimed to evaluate the association between inflammation-based prognostic markers and mortality after hip replacement. From March 2010 to June 2020, we identified 5,369 consecutive adult patients undergoing hip replacement with C-reactive protein (CRP), albumin, and complete blood count measured within six months before surgery. Receiver operating characteristic (ROC) curves were generated to evaluate predictabilities and estimate thresholds of CRP-to-albumin ratio (CAR), neutrophil-to-lymphocyte ratio (NLR), and platelet-to-lymphocyte ratio (PLR). Patients were divided according to threshold, and mortality risk was compared. The primary outcome was one-year mortality, and overall mortality was also analyzed. One-year mortality was 2.9%. Receiver operating characteristics analysis revealed areas under the curve of 0.838, 0.832, 0.701, and 0.732 for CAR, NLR, PLR, and modified Glasgow Prognostic Score, respectively. The estimated thresholds were 2.10, 3.16, and 11.77 for CAR, NLR, and PLR, respectively. According to the estimated threshold, high CAR and NLR were associated with higher one-year mortality after adjustment (1.0% vs. 11.7%; HR = 2.16; 95% CI 1.32-3.52; p = 0.002 for CAR and 0.8% vs. 9.6%; HR = 2.05; 95% CI 1.24-3.39; p = 0.01 for NLR), but PLR did not show a significant mortality increase (1.4% vs. 7.4%; HR = 1.12; 95% CI 0.77-1.63; p = 0.57). Our study demonstrated associations of preoperative levels of CAR and NLR with postoperative mortality in patients undergoing hip replacement. Our findings may be helpful in predicting mortality in patients undergoing hip replacement.
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Artroplastia de Quadril , Biomarcadores , Proteína C-Reativa , Inflamação , Humanos , Artroplastia de Quadril/mortalidade , Feminino , Masculino , Idoso , Prognóstico , Inflamação/sangue , Biomarcadores/sangue , Proteína C-Reativa/metabolismo , Proteína C-Reativa/análise , Pessoa de Meia-Idade , Neutrófilos , Curva ROC , Linfócitos , Idoso de 80 Anos ou mais , Estudos Retrospectivos , Albumina Sérica/análise , Plaquetas/patologiaRESUMO
Introduction: The large-scale artificial intelligence (AI) language model chatbot, Chat Generative Pre-Trained Transformer (ChatGPT), is renowned for its ability to provide data quickly and efficiently. This study aimed to assess the medical responses of ChatGPT regarding anesthetic procedures. Methods: Two anesthesiologist authors selected 30 questions representing inquiries patients might have about surgery and anesthesia. These questions were inputted into two versions of ChatGPT in English. A total of 31 anesthesiologists then evaluated each response for quality, quantity, and overall assessment, using 5-point Likert scales. Descriptive statistics summarized the scores, and a paired sample t-test compared ChatGPT 3.5 and 4.0. Results: Regarding quality, "appropriate" was the most common rating for both ChatGPT 3.5 and 4.0 (40 and 48%, respectively). For quantity, responses were deemed "insufficient" in 59% of cases for 3.5, and "adequate" in 69% for 4.0. In overall assessment, 3 points were most common for 3.5 (36%), while 4 points were predominant for 4.0 (42%). Mean quality scores were 3.40 and 3.73, and mean quantity scores were - 0.31 (between insufficient and adequate) and 0.03 (between adequate and excessive), respectively. The mean overall score was 3.21 for 3.5 and 3.67 for 4.0. Responses from 4.0 showed statistically significant improvement in three areas. Conclusion: ChatGPT generated responses mostly ranging from appropriate to slightly insufficient, providing an overall average amount of information. Version 4.0 outperformed 3.5, and further research is warranted to investigate the potential utility of AI chatbots in assisting patients with medical information.
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Lipid-based nanocarriers have been extensively utilized for the solubilization and cutaneous delivery of water-insoluble active ingredients in skincare formulations. However, their practical application is often limited by structural instability, leading to premature release and degradation of actives. Here we present highly robust multilamellar nanovesicles, prepared by the polyionic self-assembly of unilamellar vesicles with hydrolyzed collagen peptides, to stabilize all-trans-retinol and enhance its cutaneous delivery. Our results reveal that the reinforced multilayer structure substantially enhances dispersion stability under extremely harsh conditions, like freeze-thaw cycles, and stabilizes the encapsulated retinol. Interestingly, these multilamellar vesicles exhibit significantly lower cytotoxicity to human dermal fibroblasts than their unilamellar counterparts, likely due to their smaller particle number per weight, minimizing potential disruptions to cellular membranes. In artificial skin models, retinol-loaded multilamellar vesicles effectively upregulate collagen-related gene expression while suppressing the synthesis of metalloproteinases. These findings suggest that the robust multilamellar vesicles can serve as effective nanocarriers for the efficient delivery and stabilization of bioactive compounds in cutaneous applications.
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Administração Cutânea , Colágeno , Fibroblastos , Lipídeos , Nanopartículas , Vitamina A , Vitamina A/administração & dosagem , Vitamina A/química , Humanos , Colágeno/química , Lipídeos/química , Nanopartículas/química , Fibroblastos/efeitos dos fármacos , Estabilidade de Medicamentos , Pele/metabolismo , Portadores de Fármacos/química , Sobrevivência Celular/efeitos dos fármacos , Absorção Cutânea , Pele ArtificialRESUMO
Background: We sought to investigate the prognostic value of preoperative C-reactive protein (CRP)-to-albumin ratio (CAR) for the prediction of mortality in patients undergoing off-pump coronary artery bypass grafting (OPCAB). Methods: From January 2010 to August 2016, adult patients undergoing OPCAB were analyzed retrospectively. In a total of 2,082 patients, preoperative inflammatory markers including CAR, CRP, neutrophil-to-lymphocyte ratio, and platelet-to-lymphocyte ratio were recorded. Receiver operating characteristic (ROC) curves were used to determine the optimal threshold and compare the predictive values of the markers. The patients were divided into two groups according to the cut-off value of CAR, and then the outcomes were compared. The primary end point was 1-year mortality. Results: During the 1-year follow-up period, 25 patients (1.2%) died after OPCAB. The area under the curve of CAR for 1-year mortality was 0.767, which was significantly higher than other inflammatory markers. According to the calculated cut-off value of 1.326, the patients were divided into two groups: 1,580 (75.9%) patients were placed in the low CAR group vs. 502 (24.1%) patients in the high CAR group. After adjustment with inverse probability weighting, high CAR was significantly associated with increased risk of 1-year mortality after OPCAB (Hazard ratio, 5.01; 95% Confidence interval, 2.01-12.50; p < 0.001). Conclusions: In this study, we demonstrated that preoperative CAR was associated with 1-year mortality following OPCAB. Compared to previous inflammatory markers, CAR may offer superior predictive power for mortality in patients undergoing OPCAB. For validation of our findings, further prospective studies are needed.
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Colorectal cancer is one of the most common cancers in the world. While colonoscopy is an effective screening technique, navigating an endoscope through the colon to detect polyps is challenging. A 3D map of the observed surfaces could enhance the identification of unscreened colon tissue and serve as a training platform. However, reconstructing the colon from video footage remains difficult. Learning-based approaches hold promise as robust alternatives, but necessitate extensive datasets. Establishing a benchmark dataset, the 2022 EndoVis sub-challenge SimCol3D aimed to facilitate data-driven depth and pose prediction during colonoscopy. The challenge was hosted as part of MICCAI 2022 in Singapore. Six teams from around the world and representatives from academia and industry participated in the three sub-challenges: synthetic depth prediction, synthetic pose prediction, and real pose prediction. This paper describes the challenge, the submitted methods, and their results. We show that depth prediction from synthetic colonoscopy images is robustly solvable, while pose estimation remains an open research question.
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Colonoscopia , Imageamento Tridimensional , Humanos , Imageamento Tridimensional/métodos , Neoplasias Colorretais/diagnóstico por imagem , Pólipos do Colo/diagnóstico por imagemRESUMO
Phytophthora species, classified under Oomycota, cause significant damage to various crops and trees. The present study introduced Phytophthora species, P. nagaii and P. tentaculata, new to Korea, which pose notable risks to their respective host plants. Our research provided a comprehensive description of these species taking into account their cultural features, morphological characteristics, and molecular phylogenetic analysis using the internal transcribed spacer rDNA region and cytochrome c oxidase subunit mtDNA genes (cox1 and cox2) sequences. In addition, this study first evaluated the sensitivity of P. nagaii and P. tentaculata to five anti-oomycete fungicides, finding both species most responsive to picarbutrazox and P. tentaculata resistant to fluazinam. The data can guide targeted treatment strategies and offer insights into effective control methods. The findings expand our understanding of the diversity, distribution, and management of Phytophthora species in Korea.
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This study aimed to identify causal variants associated with important carcass traits such as weight and meat quality in Hanwoo cattle. We analyzed missense mutations extracted from imputed sequence data (ARS-UCD1.2) and performed an exon-specific association test on the carcass traits of 16,970 commercial Hanwoo. We found 33, 2, 1, and 3 significant SNPs associated with carcass weight (CW), backfat thickness (BFT), eye muscle area (EMA), and marbling score (MS), respectively. In CW and EMA, the most significant missense SNP was identified at 19,524,263 on BTA14 and involved the PRKDC. A missense SNP in the ZFAND2B, located at 107,160,304 on BTA2 was identified as being involved in BFT. For MS, missense SNP in the ACVR2B gene, located at 11,849,704 in BTA22 was identified as the most significant marker. The contribution of the most significant missense SNPs to genetic variance was confirmed to be 8.47%, 2.08%, 1.73%, and 1.19% in CW, BFT, EMA, and MS, respectively. We generated favorable and unfavorable haplotype combinations based on the significant SNPs for CW. Significant differences in GEBV (Genomic Estimated Breeding Values) were observed between groups with each favorable and unfavorable haplotype combination. In particular, the missense SNPs in PRKDC, MRPL9, and ANKFN1 appear to significantly affect the protein's function and structure, making them strong candidates as causal mutations. These missense SNPs have the potential to serve as valuable markers for improving carcass traits in Hanwoo commercial farms.
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Genoma , Mutação de Sentido Incorreto , Bovinos/genética , Animais , Fenótipo , Carne/análise , GenômicaRESUMO
In Korea, Korea Proven Bulls (KPN) program has been well-developed. Breeding and evaluation of cows are also an essential factor to increase earnings and genetic gain. This study aimed to evaluate the accuracy of cow breeding value by using three methods (pedigree index [PI], pedigree-based best linear unbiased prediction [PBLUP], and genomic-BLUP [GBLUP]). The reference population (n = 16,971) was used to estimate breeding values for 481 females as a test population. The accuracy of GBLUP was 0.63, 0.66, 0.62 and 0.63 for carcass weight (CWT), eye muscle area (EMA), back-fat thickness (BFT), and marbling score (MS), respectively. As for the PBLUP method, accuracy of prediction was 0.43 for CWT, 0.45 for EMA, 0.43 for MS, and 0.44 for BFT. Accuracy of PI method was the lowest (0.28 to 0.29 for carcass traits). The increase by approximate 20% in accuracy of GBLUP method than other methods could be because genomic information may explain Mendelian sampling error that pedigree information cannot detect. Bias can cause reducing accuracy of estimated breeding value (EBV) for selected animals. Regression coefficient between true breeding value (TBV) and GBLUP EBV, PBLUP EBV, and PI EBV were 0.78, 0.625, and 0.35, respectively for CWT. This showed that genomic EBV (GEBV) is less biased than PBLUP and PI EBV in this study. In addition, number of effective chromosome segments (Me) statistic that indicates the independent loci is one of the important factors affecting the accuracy of BLUP. The correlation between Me and the accuracy of GBLUP is related to the genetic relationship between reference and test population. The correlations between Me and accuracy were -0.74 in CWT, -0.75 in EMA, -0.73 in MS, and -0.75 in BF, which were strongly negative. These results proved that the estimation of genetic ability using genomic data is the most effective, and the smaller the Me, the higher the accuracy of EBV.
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White mold (or Sclerotinia stem rot), caused by Sclerotinia species, is a major air, soil, or seed-transmitted disease affecting numerous crops and wild plants. Microscopic or culture-based methods currently available for their detection and identification are time-consuming, laborious, and often erroneous. Therefore, we developed a multiplex quantitative PCR (qPCR) assay for the discrimination, detection, and quantification of DNA collected from each of the three economically relevant Sclerotinia species, namely, S. sclerotiorum, S. minor, and S. nivalis. TaqMan primer/probe combinations specific for each Sclerotinia species were designed based on the gene sequences encoding aspartyl protease. High specificity and sensitivity of each probe were confirmed for sclerotium and soil samples, as well as pure cultures, using simplex and multiplex qPCRs. This multiplex assay could be helpful in detecting and quantifying specific species of Sclerotinia, and therefore, may be valuable for disease diagnosis, forecasting, and management.
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Global temperatures are steadily increasing, leading to significant changes in microbial diversity and ecology. In the present study, we isolated high-temperature-growing fungi and fungi-like group (Oomycota) strains from freshwater environments of Korea and identified them based on cultural, morphological, and multilocus phylogenetic analyses. As a result, we introduce Saksenaea (Fungi) isolates as a new species, Saksenaea longicolla sp. nov. and record Phytophthora chlamydospora and P. lagoariana (Oomycota) new to Korea. In the growth experiments, they exhibited high-temperature tolerance, which can grow at 35-40 °C but become inactive at 4 °C and below. This study confirms the presence of high-temperature-tolerant fungi and oomycetes in Korea and suggests that the Korean climate conditions are changing in favor of these species. This indicates that climate warming is altering microbial distributions in freshwater environments.
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The genus Pseudoperonospora, an obligate biotrophic group of Oomycota, causes the most destructive foliar downy mildew disease on many economically important crops and wild plants. A previously unreported disease by Pseudoperonospora was found on oriental pickling melon (Cucumis melo var. conomon) in Korea, which is a minor crop cultivated in the temperate climate zone of East Asia, including China, Korea, and Japan. Based on molecular phylogenetic and morphological analyses, the causal agent was identified as Pseudoperonospora cubensis, and its pathogenicity has been proven. Importantly, two phylogenetic clades of P. cubensis, harboring probably two distinct species, were detected within the same plots, suggesting simultaneous coexistence of the two clades. This is the first report of P. cubensis causing downy mildew on oriental pickling melon in Korea, and the confirmation of presence of two phylogenetic clades of this pathogen in Korea. Given the high incidence of P. cubensis and high susceptibility of oriental pickling melon to this disease, phytosanitary measures, including rapid diagnosis and effective control management, are urgently required.
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Various lipid-based nanocarriers have been developed for the co-delivery of protein antigens with immunological adjuvants. However, their in vivo potency in vaccine delivery is limited by structural instability, which causes off-target delivery and low cross-presentation efficacies. Recent works employ covalent cross-linking to stabilize the lipid nanostructures, though the immunogenicity and side effects of chemically modified protein antigens and lipids can cause a long-lasting safety issue. Here robust "conjugation-free" multilamellar protein antigen-lipid hybrid nanovesicles (MPLVs) are introduced through the antigen-mediated self-assembly of unilamellar lipid vesicles for the co-delivery of protein antigens and immunologic adjuvants. The nanocarriers coated with monophosphoryl lipid A and hyaluronic acids elicit highly increase antigen-specific immune responses in vitro and in vivo. The MPLVs increase the generation of immunological surface markers and cytokines in mouse-derived bone-marrow dendritic cells compared to soluble antigens with adjuvants. Besides, the vaccination of mice with the MPLVs significantly increase the production of anti-antigen antibody and interferon-gamma via the activation of CD4+ and CD8+ T cells, respectively. These findings suggest that MPLVs can serve as a promising nanovaccine delivery platform for efficient antigen cross-presentation through the efficient co-delivery of protein antigens with adjuvants.