Genome-Wide Identification and Characterization of RdHSP Genes Related to High Temperature in Rhododendron delavayi.

Heat shock proteins (HSPs) are molecular chaperones that play essential roles in plant development and in response to various environmental stresses. Understanding R. delavayi HSP genes is of great importance since R. delavayi is severely affected by heat stress. In the present study, a total of 76 RdHSP genes were identified in the R. delavayi genome, which were divided into five subfamilies based on molecular weight and domain composition. Analyses of the chromosome distribution, gene structure, and conserved motif of the RdHSP family genes were conducted using bioinformatics analysis methods. Gene duplication analysis showed that 15 and 8 RdHSP genes were obtained and retained from the WGD/segmental duplication and tandem duplication, respectively. Cis-element analysis revealed the importance of RdHSP genes in plant adaptations to the environment. Moreover, the expression patterns of RdHSP family genes were investigated in R. delavayi treated with high temperature based on our RNA-seq data, which were further verified by qRT-PCR. Further analysis revealed that nine candidate genes, including six RdHSP20 subfamily genes (RdHSP20.4, RdHSP20.8, RdHSP20.6, RdHSP20.3, RdHSP20.10, and RdHSP20.15) and three RdHSP70 subfamily genes (RdHSP70.15, RdHSP70.21, and RdHSP70.16), might be involved in enhancing the heat stress tolerance. The subcellular localization of two candidate RdHSP genes (RdHSP20.8 and RdHSP20.6) showed that two candidate RdHSPs were expressed and function in the chloroplast and nucleus, respectively. These results provide a basis for the functional characterization of HSP genes and investigations on the molecular mechanisms of heat stress response in R. delavayi.

Optimizing ultrafast dynamic contrast-enhanced MRI scan duration in the differentiation of benign and malignant breast lesions.

OBJECTIVE: To determine the optimal scan duration for ultrafast DCE-MRI in effectively differentiating benign from malignant breast lesions. METHODS: The study prospectively recruited participants who underwent breast ultrafast DCE-MRI from September 2021 to March 2023. A 30-phase breast ultrafast DCE-MRI on a 3.0-T MRI system was conducted with a 4.5-s temporal resolution. Scan durations ranged from 40.5 s to 135.0 s, during which the analysis is performed at three-phase intervals, forming eight dynamic sets (scan duration [SD]40.5s: 40.5 s, SD54s: 54.0 s, SD67.5s: 67.5 s, SD81s: 81.0 s, SD94.5s: 94.5 s, SD108s: 108.0 s, SD121.5s: 121.5 s, and SD135s: 135.0 s). Two ultrafast DCE-MRI parameters, maximum slope (MS) and initial area under the curve in 60 s (iAUC), were calculated for each dynamic set and compared between benign and malignant lesions. Areas under the receiver operating characteristic curve (AUCs) were used to assess their diagnostic performance. RESULTS: A total of 140 women (mean age, 47 ± 11 years) with 151 lesions were included. MS and iAUC from eight dynamic sets exhibited significant differences between benign and malignant lesions (all p < 0.05), except iAUC at SD40.5s. The AUC of MS (AUC = 0.804) and iAUC (AUC = 0.659) at SD67.5s were significantly higher than their values at SD40.5s (AUC = 0.606 and 0.516; corrected p < 0.05). No significant differences in AUCs for MS and iAUC were observed from SD67.5s to SD135s (all corrected p > 0.05). CONCLUSIONS: Ultrafast DCE-MRI with a 67.5-s scan duration appears optimal for effectively differentiating malignant from benign breast lesions. CRITICAL RELEVANCE STATEMENT: By evaluating scan durations (40.5-135 s) and analyzing two ultrafast DCE-MRI parameters, we found a scan duration of 67.5 s optimal for discriminating between these lesions and offering a balance between acquisition time and diagnostic efficacy. KEY POINTS: Ultrafast DCE-MRI can effectively differentiate malignant from benign breast lesions. A minimum of 67.5-sec ultrafast DCE-MRI scan duration is required to differentiate benign and malignant lesions. Extending the scan duration beyond 67.5 s did not significantly improve diagnostic accuracy.

Modulation of Gut Microbial Community and Metabolism by Bacillus licheniformis HD173 Promotes the Growth of Nursery Piglets Model.

Maintaining the balance and stability of the gut microbiota is crucial for the gut health and growth development of humans and animals. Bacillus licheniformis (B. licheniformis) has been reported to be beneficial to the gut health of humans and animals, whereas the probiotic effects of a new strain, B. licheniformis HD173, remain uncertain. In this study, nursery piglets were utilized as animal models to investigate the extensive impact of B. licheniformis HD173 on gut microbiota, metabolites, and host health. The major findings were that this probiotic enhanced the growth performance and improved the health status of the nursery piglets. Specifically, it reduced the level of pro-inflammatory cytokines IL-1ß and TNF-α in the serum while increasing the level of IL-10 and SOD. In the gut, B. licheniformis HD173 reduced the abundance of pathogenic bacteria such as Mycoplasma, Vibrio, and Vibrio metschnikovii, while it increased the abundance of butyrate-producing bacteria, including Oscillospira, Coprococcus, and Roseburia faecis, leading to an enhanced production of butyric acid. Furthermore, B. licheniformis HD173 effectively improved the gut metabolic status, enabling the gut microbiota to provide the host with stronger metabolic abilities for nutrients. In summary, these findings provide scientific evidence for the utilization of B. licheniformis HD173 in the development and production of probiotic products for maintaining gut health in humans and animals.

Preparation and property studies of ferric sulfoaluminate cement based on Bayer red mud and phosphogypsum.

The Bayer red mud (RM) and phosphogypsum (PG) accumulation have caused significant environmental contamination. However, practical and effective resource utilization technologies are still lacking currently. This work aims to develop ferric sulfoaluminate cement (FSAC) employing low-cost materials including Bayer red mud, phosphogypsum, and other materials. This method effectively improves the utilization rate of Bayer red mud and phosphogypsum. Under the premise of ensuring the performance of FSAC, the utilization rate of solid waste can reach up to 48.56%. The effects of different red mud dosages on cement mineral formation, workability, and mechanical properties are investigated. Then, untreated phosphogypsum is adopted as a retarder for FSAC, and the hydration process, working properties, mechanical properties, types of hydration products, and morphology of FSAC are explored. The results suggest that the crystal transformation of Ye'elemite C 4 A 3 S ¯  is promoted with the increase of Bayer red mud content. Cubic crystal system Ye'elemite C 4 A 3 S ¯ - c  with higher hydration activity is generated, which increases the early strength of cement but greatly reduces the setting time, hindering the later strength growth. Untreated phosphogypsum can effectively delay the early hydration process of FSAC, prolong the setting time of cement, and increase the strength of FSAC in the later stage. When the dosage of Bayer red mud and phosphogypsum is 17.64% and 9.21%, respectively, with phosphogypsum dosage of 20%, the prepared FSAC has satisfactory mechanical properties, and the 3-day and 90-day compressive strengths are 34.6 MPa and 57.1 MPa, respectively. In addition, the study of heavy metal leaching indicates that the FSAC prepared by Bayer red mud, phosphogypsum, and other raw materials will generate no environment pollution, and the solidification of heavy metal elements in the cement slurry is superior.

Adipose exosomal noncoding RNAs: Roles and mechanisms in metabolic diseases.

Exosomes are extracellular vesicles, measuring 40-160 nm in diameter, that are released by many cell types and tissues, including adipose tissue. Exosomes are critical mediators of intercellular communication and their contents are complex and diverse. In recent years, accumulating evidence has proved that multiple adipose tissue-derived exosomal noncoding RNAs (ncRNAs), including microRNAs (miRNAs), long noncoding RNAs (lncRNAs) and circular RNAs (circRNAs), play pivotal roles in the pathogenesis of diverse metabolic diseases, such as obesity. In this narrative review, we focus on the adipose tissue-derived exosomal ncRNAs, especially exosomal miRNAs, and their dysregulation in multiple types of metabolic diseases. A deeper understanding of the role of adipose tissue-derived exosomal ncRNAs may help provide new diagnostic and treatment methods for metabolic diseases.

Treatment of metastatic TFE3 microphthalmia transcription factor translocation renal cell carcinoma: a case report.

Background: Microphthalmia-associated transcription factor/transcription factor E (MiTF/TFE) translocation renal cell carcinoma (RCC) is a rare type of non-clear cell RCC (nccRCC), which is more common in females. Currently, there is no standardized treatment for advanced metastatic microphthalmia translocation RCC (MiT-RCC). The main treatment modalities include surgery, chemotherapy, immunotherapy, anti-vascular endothelial growth factor or vascular endothelial growth factor receptor (VEGFR) inhibitors, mammalian target of rapamycin (mTOR) inhibitors, and targeted therapy against the mesenchymal-epithelial transition (MET) factor signaling pathway. Case Description: We present the case of an 8-year-old male patient with hematuria and paroxysmal urinary pain. Based on tumor genetic testing results and targeted drug matching analysis, the patient underwent tumor biopsy, tumor radical surgery with vascular osteotomy, and cervicothoracic lymph node dissection. The patient was then treated with a combination of immunotherapy [sintilimab, a drug directed against programmed cell death receptor-1 (PD-1)] and VEGFR tyrosine kinase inhibitor (TKI) (from pazopanib to sunitinib). Throughout the 10 cycles of conventional chemotherapy (seven courses of sintilimab since the start of the third chemotherapy treatment), the patient's condition remained stable, with no tumor recurrence at the primary site. However, in the later stages, the patient developed a large amount of ascites, and the family requested discontinuation of treatment, ultimately leading to the patient's death. Conclusions: In this case report, we summarize the therapeutic strategy of a young patient with metastatic transcription factor E3 (TFE3) MiT-RCC. For this disease, early immunotherapy and the use of precision-targeted drugs may have a favorable impact on the survival prognosis of the patient but may still be of less benefit in children with advanced multiple metastases. Therefore, further research on tumor driver genes, among other treatment components, is urgently needed to improve precision therapy.

Effect of Chitosan and Its Water-Soluble Derivatives on Antioxidant Activity.

The antioxidant activity of chitosan (CS) and three water-soluble derivatives was analyzed comparatively by in vitro and in vivo experiments, including hydroxypropyl chitosan (HPCS), quaternary ammonium salt of chitosan (HACC), and carboxymethyl chitosan (CMCS). The results show that chitosan and its water-soluble derivatives have a scavenging ability on DPPH radicals, superoxide radicals, and hydroxyl radicals, and a reducing ability. A remarkable difference (p < 0.05) was found for HACC and HPCS compared with CS on DPPH radicals, hydroxyl radicals, and reducing ability. The antioxidant ability of the four chitosan samples was in the order of HPCS > HACC > CMCS > CS. Furthermore, antioxidant activity of all samples increased gradually in a concentration-dependent manner. The in vivo result indicates that oral CS and its derivatives samples result in a decrease in lipid peroxides (LPO) and free fatty acids (FFA) levels in serum with an increase in superoxide dismutase (SOD) activity. Especially for the HPCS and HACC groups, the LPO, FFA, and SOD activity in serum was different significantly in comparison with the high-fat controlgroup (HF) (p < 0.05). These results indicate that chitosan and its derivatives can be used as good antioxidants, and the antioxidant activity might be related to the molecular structure of chitosan derivatives.

Molecular Regulation of Porcine Skeletal Muscle Development: Insights from Research on CDC23 Expression and Function.

Cell division cycle 23 (CDC23) is a component of the tetratricopeptide repeat (TPR) subunit in the anaphase-promoting complex or cyclosome (APC/C) complex, which participates in the regulation of mitosis in eukaryotes. However, the regulatory model and mechanism by which the CDC23 gene regulates muscle production in pigs are largely unknown. In this study, we investigated the expression of CDC23 in pigs, and the results indicated that CDC23 is widely expressed in various tissues and organs. In vitro cell experiments have demonstrated that CDC23 promotes the proliferation of myoblasts, as well as significantly positively regulating the differentiation of skeletal muscle satellite cells. In addition, Gene Set Enrichment Analysis (GSEA) revealed a significant downregulation of the cell cycle pathway during the differentiation process of skeletal muscle satellite cells. The protein-protein interaction (PPI) network showed a high degree of interaction between genes related to the cell cycle pathway and CDC23. Subsequently, in differentiated myocytes induced after overexpression of CDC23, the level of CDC23 exhibited a significant negative correlation with the expression of key factors in the cell cycle pathway, suggesting that CDC23 may be involved in the inhibition of the cell cycle signaling pathway in order to promote the differentiation process. In summary, we preliminarily determined the function of CDC23 with the aim of providing new insights into molecular regulation during porcine skeletal muscle development.

Genome-Wide Identification and Co-Expression Networks of WOX Gene Family in Nelumbo nucifera.

WUSCHEL-related homeobox (WOX) genes are a class of plant-specific transcription factors, regulating the development of multiple tissues. However, the genomic characterizations and expression patterns of WOX genes have not been analyzed in lotus. In this study, 15 NnWOX genes were identified based on the well-annotated reference genome of lotus. According to the phylogenetic analysis, the NnWOX genes were clustered into three clades, i.e., ancient clade, intermediate clade, and WUS clade. Except for the conserved homeobox motif, we further found specific motifs of NnWOX genes in different clades and divergence gene structures, suggesting their distinct functions. In addition, two NnWOX genes in the ancient clade have conserved expression patterns and other NnWOX genes exhibit different expression patterns in lotus tissues, suggesting a low level of functional redundancy in lotus WOX genes. Furthermore, we constructed the gene co-expression networks for each NnWOX gene. Based on weighted gene co-expression network analysis (WGCNA), ten NnWOX genes and their co-expressed genes were assigned to the modules that were significantly related to the cotyledon and seed coat. We further performed RT-qPCR experiments, validating the expression levels of ten NnWOX genes in the co-expression networks. Our study reveals comprehensive genomic features of NnWOX genes in lotus, providing a solid basis for further function studies.

Deep learning radiomics on shear wave elastography and b-mode ultrasound videos of diaphragm for weaning outcome prediction.

PURPOSE: We proposed an automatic method based on deep learning radiomics (DLR) on shear wave elastography (SWE) and B-mode ultrasound videos of diaphragm for two classification tasks, one for differentiation between the control and patient groups, and the other for weaning outcome prediction. MATERIALS AND METHODS: We included a total of 581 SWE and B-mode ultrasound videos, of which 466 were from the control group of 179 normal subjects, and 115 were from the patient group of 35 mechanically ventilated subjects in the intensive care unit (ICU). Among the patient group, 17 subjects successfully weaned and 18 failed. The deep neural network of U-Net was utilized to automatically segment diaphragm regions in dual-modal videos of SWE and B-mode. High-throughput radiomics features were then extracted, the statistical test and least absolute shrinkage and selection operator (LASSO) were applied for feature dimension reduction. The optimal classification models for the two tasks were established using the support vector machine (SVM). RESULTS: The automatic segmentation model achieved Dice score of 87.89 %. A total of 4524 radiomics features were extracted, 10 and 20 important features were left after feature dimension reduction for constructing the two classification models. The best areas under receiver operating characteristic curves of the two models reached 84.01 % and 94.37 %, respectively. CONCLUSIONS: Our proposed DLR methods are innovative for automatic segmentation of diaphragm regions in SWE and B-mode videos and deep mining of high-throughput radiomics features from dual-modal images. The approaches have been proved to be effective for prediction of weaning outcomes.

Characterizing structural variants based on graph-genotyping provides insights into pig domestication and local adaption.

Structural variants (SVs), such as deletions (DELs) and insertions (INSs), contribute substantially to pig genetic diversity and phenotypic variation. Using a library of SVs discovered from long-read primary assemblies and short-read sequenced genomes, we map pig genomic SVs with a graph-based method for re-genotyping SVs in 402 genomes. Our results demonstrate that those SVs harboring specific trait-associated genes may greatly shape pig domestication and local adaptation. Further characterization of SVs reveals that some population-stratified SVs may alter the transcription of genes by affecting regulatory elements. We identify that the genotypes of two DELs (296-bp DEL, chr7: 52,172,101-52,172,397; 278-bp DEL, chr18: 23,840,143-23,840,421) located in muscle-specific enhancers are associated with the expression of target genes related to meat quality (FSD2) and muscle fiber hypertrophy (LMOD2 and WASL) in pigs. Our results highlight the role of SVs in domestic porcine evolution, and the identified candidate functional genes and SVs are valuable resources for future genomic research and breeding programs in pigs.

The convenient method and application for monitoring the health of traffic sign poles based on mobile phone.

Traffic sign poles are crucial components of the highway system, and their maintenance mainly relies on the subjective judgment of staff, which is low-efficiency and will lead to mistaken inspections. This paper proposes a convenient and effective method to monitor the health of sign poles by using a mobile phone. It is unknown whether a mobile phone can be used as a conventional acceleration sensor. Therefore, the performance of the mobile phone was initially tested to ensure its suitability for acquiring the acceleration data of the traffic sign pole. The results show that the acceleration sensor of mobile phones is high-performance and can be used as a traditional acceleration sensor under the similar sampling frequency. The mobile phone can measure the 1st, 2nd and 5th natural frequencies of the traffic sign pole. Although the 1st and 5th natural frequencies have a large error with simulation, the 2nd natural frequency is accurate and stable. The looseness of the base connection greatly impacts on the 2nd natural frequency, which can be used as a measure of the loose base connection. The 2nd natural frequency was measured for 21 times and found to conform to a normal distribution. The relationship between the 2nd natural frequency and base connection loosening was investigated, which fitted to the equation. The fitting result is good and can be used to predict the degree of sign pole base connection loosening. Therefore, the mobile phone based convenient health monitoring method for traffic sign poles is feasible.

Feeding mediated web-building plasticity in a cobweb spider.

Behavioral plasticity has been proposed as a means by which animals alter their phenotypes in response to changing conditions. Animals may display behavioral plasticity as a consequence of environmental variation. The detritus-based, bell-shaped cobweb spider Campanicola campanulata is an ideal model to study behavioral plasticity, because its web architecture is easy to be quantified, and the functions of different parts of the web are clear. Though the plasticity of cobweb architecture has been reported in a few species, retreats as important defensive structures have rarely been considered before because retreats in most cobwebs are relatively small compared with the web size. We studied the web-building behaviors of C. campanulata under different feeding regimes. We set up 3 spider treatments with different feeding conditions: marginally well fed, moderately well fed, and extremely well fed, and observed the differences in the web architecture among them. In addition, we measured the mechanical properties of anchor silk, and also calculated the foraging and defense investment of the spiders. The results showed that marginally well-fed spiders build cobwebs with significantly longer length of anchor silk, lower retreat to the ground, more number and longer gumfooted lines, and larger capture area, while extremely well-fed spiders build cobwebs with significantly bigger retreat volume and higher height of retreat to the ground. In addition, marginally well-fed spiders invest significantly less during cobweb construction. However, there was no significant difference between the breaking force and elongation at break in anchor silk among different treatments. These results demonstrated that marginally well-fed spiders invest more in foraging, and extremely well-fed spiders invest more in defense, and the spider made a balance between foraging and predator avoidance in response to changes in physiological state. Our study strengthens the current understanding of web construction in cobweb spiders, especially those facing high costs during retreat construction.

Single-cell transcriptome sequencing reveals tumor heterogeneity in family neuroblastoma.

Neuroblastoma(NB) is the most common extracranial solid tumor in childhood, and it is now believed that some patients with NB have an underlying genetic susceptibility, which may be one of the reasons for the multiplicity of NB patients within a family line. Even within the same family, the samples show great variation and can present as ganglioneuroblastoma or even benign ganglioneuroma. The genomics of NB is still unclear and more in-depth studies are needed to reveal its key components. We first performed single-cell RNA sequencing(sc-RNAseq) analysis on clinical specimens of two family neuroblastoma(FNB) and four sporadic NB cases. A complete transcriptional profile of FNB was constructed from 18,394 cells from FNB, and we found that SDHD may be genetically associated with FNB and identified a prognostic related CAF subtype in FNB: Fib-4. Single-cell flux estimation analysis (scFEA) results showed that malignant cells were associated with arginine spermine, oxaloacetate and hypoxanthine, and that malignant cells metabolize lactate at lower levels than T cells. Our study provides new resources and ideas for the development of the genomics of family NB, and the mechanisms of cell-to-cell interactions and communication and the metabolic landscape will provide new therapeutic targets.

Photoswitchable and Reversible Fluorescent Eutectogels for Conformal Information Encryption.

Smart fluorescent materials that can respond to environmental stimuli are of great importance in the fields of information encryption and anti-counterfeiting. However, traditional fluorescent materials usually face problems such as lack of tunable fluorescence and insufficient surface-adaptive adhesion, hindering their practical applications. Herein, inspired by the glowing sucker octopus, we present a novel strategy to fabricate a reversible fluorescent eutectogel with high transparency, adhesive and self-healing performance for conformal information encryption and anti-counterfeiting. Using anthracene as luminescent unit, the eutectogel exhibits photoswitchable fluorescence and can therefore be reversibly written/erased with patterns by non-contact stimulation. Additionally, different from mechanically irreversible adhesion via glue, the eutectogel can adhere to various irregular substrates over a wide temperature range (-20 to 65 °C) and conformally deform more than 1000 times without peeling off. Furthermore, by exploiting surface-adaptive adhesion, high transparency and good stretchability of the eutectogel, dual encryption can be achieved under UV and stretching conditions to further improve the security level. This study should provide a promising strategy for the future development of advanced intelligent anti-counterfeiting materials.

Searching for new signals for susceptibility to umbilical hernia through genome-wide association analysis in three pig breeds.

Umbilical hernia (UH) is a prevalent congenital disorder in pigs, resulting in considerable economic losses and severe animal welfare issues. In the present study, we conducted a genome-wide association study (GWAS) using the GeneSeek 50K Chip in 2777 pigs (Duroc, n = 1267; Landrace, n = 696; and Yorkshire, n = 814) to explore the candidate genes underlying the risk of umbilical hernia in pigs. After quality control analyses, 2748 animals and 48 524 single nucleotide polymorphisms (SNPs) were retained for subsequent GWAS analysis using the FarmCPU model. The heritability of umbilical hernias was estimated to 0.51 ± 0.04, indicating a reasonable basis for investigating genetic markers associated with this disorder. We identified 54 SNPs and 517 candidate genes that showed significant associations with susceptibility to umbilical hernia across the combined population of the three pig breeds. Gene enrichment analyses highlighted several crucial pathways for platelet degranulation, inflammatory mediator regulation of TRP channels and ion transport. These findings provide further insights into the underlying genetic architecture of umbilical hernias in pigs.

Dual ultrasound-guided totally implantable venous access ports via the right internal jugular vein in pediatric patients with cancer: a preliminary experience in a single institution.

Objective: To assess the efficacy and safety of dual ultrasound-guided (DUG) totally implantable venous access port (TIVAP) implantation (namely, using ultrasound-guided percutaneous puncture with transesophageal echocardiography-guided catheterization) via the right internal jugular vein (IJV) in pediatric patients with cancer. Methods: Fifty-five children with cancer requiring chemotherapy underwent DUG-TIVAP implantation via the right IJV. Clinical data were recorded, including the procedure success rate, first attempt success rate, and perioperative and postoperative complications. Results: All 55 cases were successfully operated on. The first puncture success rate was 100%. The operation time was 22-41 min, with a mean time of 30.8±5.5 min. The mean TIVAP implantation time was 253±145 days (range 42-520 days). There were no perioperative complications. The postoperative complication rate was 5.4% (3/55), including skin infections around the port in one case, catheter-related infection in one case, and fibrin sheath formation in one case. The ports were all preserved after anti-infection or thrombolytic therapy. No unplanned port withdrawal was recorded in this study. Conclusions: DUG-TIVAP implantation is a technique with a high success rate and a low complication rate; therefore, it provides an alternative for children with cancer. Further randomized controlled studies are needed to confirm the efficacy and safety of DUG-TIVAP via the right IJV in children.

Biobased Intelligent Food-Packaging Materials with Sustained-Release Antibacterial and Real-Time Monitoring Ability.

It has been widely accepted that sustainable polymers derived from renewable resources are able to replace the short-turnover petroleum-based materials and reduce environmental impact in the future. However, their hydrophilic chemical structures rich with oxygen groups could lead to easy growth of bacteria, which greatly limit their applications in packaging materials. Here, we present an intelligent food-packaging material with sustained-release antibacterial and real-time monitoring ability based on totally biobased contents. In detail, sodium alginate with Artemisia argyi emission oil (encapsulated in gelatin-Arabic gum microcapsules) and citric acid-sourced pH-responsive carbon quantum dots (CQDs) are coated on bamboo cellulose papers. The obtained biobased composite material (almost 100% biocarbon content) with antibacterial ability is able to extend the shelf life of fresh shrimps and can be biodegraded. Moreover, owing to the introduction of CQDs, the composite can rapidly (within 1 s) detect slight pH variations (response pH ∼5, 10-9 mol/L of OH-) through an obvious color change (hue value from 305 to 355°). The developed strategy may open up new opportunities in the design of multifunctional biobased composites for intelligent applications.

Development of a Naked Eye CRISPR-Cas12a and -Cas13a Multiplex Point-of-Care Detection of Genetically Modified Swine.

The Rapid Visual CRISPR (RAVI-CRISPR) assay employs Cas12a and Cas13a enzymes for precise gene detection in a sample. However, RAVI-CRISPR is limited in single-tube multiplex detection applications due to the lack of specific single-strand (ss) DNA-fluorescently quenched (ssDNA-FQ) and RNA-fluorescently quenched (ssRNA-FQ) reporter cleavage mechanisms. We report the development of a sensitive and specific dual-gene Cas12a and Cas13a diagnostic system. To optimize the application for field testing, we designed a portable multiplex fluorescence imaging assay that could distinguish test results with the naked eye. Herein, dual gene amplified products from multiplex recombinase polymerase amplification (RPA) were simultaneously detected in a single tube using Cas12a and Cas13a enzymes. The resulting orthogonal DNA and RNA collateral cleavage specifically distinguishes individual and mixed ssDNA-FQ and ssRNA-FQ reporters using the green-red-yellow, fluorescent signal conversion reaction system, detectable with portable blue and ultraviolet (UV) light transilluminators. As a proof-of-concept, reliable multiplex RAVI-CRISPR detection of genome-edited pigs was demonstrated, exhibiting 100% sensitivity and specificity for the analysis of CD163 knockout, lactoferrin (LF) knock-in, and wild-type pig samples. This portable naked-eye multiplex RAVI-CRISPR detection platform can provide accurate point-of-care screening of genetically modified animals and infectious diseases in resource-limited settings.