The molecular epidemiology of a dengue virus outbreak in Taiwan: population wide versus infrapopulation mutation analysis.

Dengue virus (DENV) causes approximately 390 million dengue infections worldwide every year. There were 22,777 reported DENV infections in Tainan, Taiwan in 2015. In this study, we sequenced the C-prM-E genes from 45 DENV 2015 strains, and phylogenetic analysis based on C-prM-E genes revealed that all strains were classified as DENV serotype 2 Cosmopolitan genotype. Sequence analysis comparing different DENV-2 genotypes and Cosmopolitan DENV-2 sequences prior to 2015 showed a clade replacement event in the DENV-2 Cosmopolitan genotype. Additionally, a major substitution C-A314G (K73R) was found in the capsid region which may have contributed to the clade replacement event. Reverse genetics virus rgC-A314G (K73R) showed slower replication in BHK-21 and C6/36 cells compared to wildtype virus, as well as a decrease in NS1 production in BHK-21-infected cells. After a series of passaging, the C-A314G (K73R) mutation reverted to wildtype and was thus considered to be unstable. Next generation sequencing (NGS) of three sera collected from a single DENV2-infected patient at 1-, 2-, and 5-days post-admission was employed to examine the genetic diversity over-time and mutations that may work in conjunction with C-A314G (K73R). Results showed that the number of haplotypes decreased with time in the DENV-infected patient. On the fifth day after admission, two new haplotypes emerged, and a single non-synonymous NS4A-L115I mutation was identified. Therefore, we have identified a persistent mutation C-A314G (K73R) in all of the DENV-2 isolates, and during the course of an infection, a single new non-synonymous mutation in the NS4A region appears in the virus population within a single host. The C-A314G (K73R) thus may have played a role in the DENV-2 2015 outbreak while the NS4A-L115I may be advantageous during DENV infection within the host.

Advances in the localization of pulmonary nodules: a comprehensive review.

In recent years, with the widespread use of chest CT, the detection rate of pulmonary nodules has significantly increased (Abtin and Brown, J Clin Oncol 31:1002-8, 2013). Video-assisted thoracoscopic surgery (VATS) is the most commonly used method for suspected malignant nodules. However, for nodules with a diameter less than 1 cm, or located more than 1.5 cm from the pleural edge, especially ground-glass nodules, it is challenging to achieve precise intraoperative localization by manual palpation (Ciriaco et al., Eur J Cardiothorac Surg 25:429-33, 2004). Therefore, preoperative accurate localization of such nodules becomes a necessary condition for precise resection. This article provides a comprehensive review and analysis of the research progress in pulmonary nodule localization, focusing on four major localization techniques: Percutaneous puncture-assisted localization, Bronchoscopic preoperative pulmonary nodule localization, 3D Printing-Assisted Localization, and intraoperative ultrasound-guided pulmonary nodule localization.

Multiplex Asymmetric PCR by Combining the Amplification Refractory Mutation System with the Homo-Tag-Assisted Nondimer System.

Asymmetric PCR is widely used to produce single-stranded amplicons (ss-amplicons) for various downstream applications. However, conventional asymmetric PCR schemes are susceptible to events that affect primer availability, which can be exacerbated by multiplex amplification. In this study, a new multiplex asymmetric PCR approach that combines the amplification refractory mutation system (ARMS) with the homo-Tag-assisted nondimer system (HANDS) is described. ARMS-HANDS (A-H) PCR utilizes equimolar-tailed forward and reverse primers and an excess Tag primer. The tailed primer pairs initiate exponential symmetric amplification, whereas the Tag primer drives linear asymmetric amplification along fully matched strands but not one-nucleotide mismatched strands, thereby generating excess ss-amplicons. The production of ss-amplicons is validated using agarose gel electrophoresis, sequencing, and melting curve analysis. Primer dimer alleviation is confirmed by both the reduced Loss function value and a 20-fold higher sensitivity in an 11-plex A-H PCR assay than in an 11-plex conventional asymmetric PCR assay. Moreover, A-H PCR demonstrates unbiased amplification by its allele quantitative ability in correct identification of all 31 trisomy 21 samples among 342 clinical samples. A-H PCR is a new generation of multiplex asymmetric amplification approach with various applications, especially when sensitive and quantitative detection is required.

Enhanced environmental stress resistance and functional properties of the curcumin-shellac nano-delivery system: Anti-flocculation of poly-γ-glutamic acid.

Curcumin was widely designed as nanoparticles to remove application restrictions. The occurrence of flocculation is a primary factor limiting the application of the curcumin nano-delivery system. To enhance the environmental stress resistance and functional properties of shellac-curcumin nanoparticles (S-Cur-NPs), γ-polyglutamic acid (γ-PGA) was utilized as an anti-flocculant. The encapsulation efficiency and loading capacity of S-Cur-NPs were also improved with γ-PGA incorporation. FTIR and XRD analysis confirmed the presence of amorphous characteristics in S-Cur-NPs and the combination of γ-PGA and shellac was driven by hydrogen bonding. The hydrophilic, thermodynamic, and surface potential of S-Cur-NPs was improved by the incorporation of γ-PGA. This contribution of γ-PGA on S-Cur-NPs effectively mitigated the flocculation occurrence during heating, storage, and in-vitro digestive treatment. Furthermore, it was revealed that γ-PGA enhanced the antibacterial and antioxidant properties of S-Cur-NPs and effectively protected the functional activity against heating, storage, and in-vitro digestion. Release studies conducted in simulated gastrointestinal fluids revealed that S-Cur-NPs have targeted intestinal release properties. Overall, the design of shellac with γ-PGA was a promising strategy to relieve the application stress of shellac and curcumin in the food industry.

Self-healing, environmentally stable and adhesive hydrogel sensor with conductive cellulose nanocrystals for motion monitoring and character recognition.

Conductive hydrogel-based sensors offer diverse applications in artificial intelligence, wearable electronic devices and character recognition management. However, it remains a significant challenge to maintain their satisfactory performances under extreme climatic conditions. Herein, a stretchable, self-adhesive, self-healing and environmentally stable conductive hydrogel was developed through free radical polymerization of hydroxyethyl acrylate (HEA) and poly(ethylene glycol) methacrylate (PEG) as the skeleton, followed by the incorporation of polyaniline-coated cellulose nanocrystal (CNC@PANI) as the conductive and reinforced nanofiller. Encouragingly, the as-prepared hydrogel (CHP) exhibited decent mechanical strength, satisfactory self-adhesion, prominent self-healing property (95.04 % after 60 s), excellent anti-freezing performance (below -60 °C) and outstanding moisture retention. The assembled sensor derived from CHP hydrogel possessed a low detection limit (0.5 % strain), high strain sensitivity (GF = 1.68) and fast response time (96 ms). Remarkably, even in harsh environmental temperatures from -60 °C to 80 °C, it reliably detected subtle and large-scale human motion for a long-term process (>10,000 cycles), manifesting its exceptional environmental tolerance. More interestingly, this hydrogel-based sensor could be assembled into a "writing board" for accurate handwritten numeral recognition. Therefore, the as-obtained multifunctional hydrogel could be a promising material applied in human motion detection and character recognition platforms even in harsh surroundings.

Overexpression of the alfalfa (Medicago sativa) gene, MsKMS1, negatively regulates seed germination in transgenic tobacco (Nicotiana tabacum).

Soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE)-associated proteins are a class of transmembrane proteins involved in intracellular trafficking pathways. However, the functions of many SNARE domain-containing proteins remain unclear. We have previously identified a SNARE-associated gene in alfalfa (Medicago sativa ) KILLING ME SLOWLY1 (MsKMS1 ), which is involved in various abiotic stresses. In this study, we investigated the function of MsKMS1 in the seed germination of transgenic tobacco (Nicotiana tabacum ). Phylogenetic analysis showed that MsKMS1 was homologous to the SNARE-associated or MAPR component-related proteins of other plants. Germination assays revealed that MsKMS1 negatively regulated seed germination under normal, D-mannitol and abscisic acid-induced stress conditions, yet MsKMS1 -overexpression could confer enhanced heat tolerance in transgenic tobacco. The suppressive effect on germination in MsKMS1 -overexpression lines was associated with higher abscisic acid and salicylic acid contents in seeds. This was accompanied by the upregulation of abscisic acid biosynthetic genes (ZEP and NCED ) and the downregulation of gibberellin biosynthetic genes (GA20ox2 and GA20ox3 ). Taken together, these results suggested that MsKMS1 negatively regulated seed germination by increasing abscisic acid and salicylic acid contents through the expression of genes related to abscisic acid and gibberellin biosynthesis. In addition, MsKMS1 could improve heat tolerance during the germination of transgenic tobacco seeds.

Genome assembly and multi-omic analyses reveal the mechanisms underlying flower color formation in Torenia fournieri.

Torenia fournieri Lind. is an ornamental plant that is popular for its numerous flowers and variety of colors. However, its genomic evolutionary history and the genetic and metabolic bases of flower color formation remain poorly understood. Here, we report the first T. fournieri reference genome, which was resolved to the chromosome scale and was 164.4 Mb in size. Phylogenetic analyses clarified relationships with other plant species, and a comparative genomic analysis indicated that the shared ancestor of T. fournieri and Antirrhinum majus underwent a whole genome duplication event. Joint transcriptomic and metabolomic analyses identified many metabolites related to pelargonidin, peonidin, and naringenin production in rose (TfR)-colored flowers. Samples with blue (TfB) and deep blue (TfD) colors contained numerous derivatives of petunidin, cyanidin, quercetin, and malvidin; differences in the abundances of these metabolites and expression levels of the associated genes were hypothesized to be responsible for variety-specific differences in flower color. Furthermore, the genes encoding flavonoid 3-hydroxylase, anthocyanin synthase, and anthocyanin reductase were differentially expressed between flowers of different colors. Overall, we successfully identified key genes and metabolites involved in T. fournieri flower color formation. The data provided by the chromosome-scale genome assembly establish a basis for understanding the differentiation of this species and will facilitate future genetic studies and genomic-assisted breeding.

Integrated physiological, metabolomic, and transcriptomic analyses elucidate the regulation mechanisms of lignin synthesis under osmotic stress in alfalfa leaf (Medicago sativa L.).

Alfalfa, an essential forage crop known for its high yield, nutritional value, and strong adaptability, has been widely cultivated worldwide. The yield and quality of alfalfa are frequently jeopardized due to environmental degradation. Lignin, a constituent of the cell wall, enhances plant resistance to abiotic stress, which often causes osmotic stress in plant cells. However, how lignin responds to osmotic stress in leaves remains unclear. This study explored the effects of osmotic stress on lignin accumulation and the contents of intermediate metabolites involved in lignin synthesis in alfalfa leaves. Osmotic stress caused an increase in lignin accumulation and the alteration of core enzyme activities and gene expression in the phenylpropanoid pathway. We identified five hub genes (CSE, CCR, CADa, CADb, and POD) and thirty edge genes (including WRKYs, MYBs, and UBPs) by integrating transcriptome and metabolome analyses. In addition, ABA and ethylene signaling induced by osmotic stress regulated lignin biosynthesis in a contradictory way. These findings contribute to a new theoretical foundation for the breeding of high-quality and resistant alfalfa varieties.

Ionic Conductive Cellulose-Based Hydrogels with Superior Long-Lasting Moisture and Antifreezing Features for Flexible Strain Sensor Applications.

Nowadays, wearable devices derived from flexible conductive hydrogels have attracted enormous attention. Nevertheless, the utilization of conductive hydrogels in practical applications under extreme conditions remains a significant challenge. Herein, a series of inorganic salt-ion-enhanced conductive hydrogels (HPE-LiCl) consisting of hydroxyethyl cellulose, hydroxyethyl acrylate, lithium chloride, and ethylene glycol/water binary solvent were fabricated via a facile one-pot method. Apart from outstanding self-adhesion, high stretchability, and remarkable fatigue resistance, the HPE-LiCl hydrogels possessed especially excellent antifreezing and long-lasting moisture performances, which could maintain satisfactory flexibility and electric conductivity over extended periods of time, even in challenging conditions such as extremely low temperatures (as low as -40 °C) and high temperatures (as high as 80 °C). Consequently, the HPE-LiCl-based sensor could timely and accurately monitor various human motion signals even in adverse environments and after long-term storage. Hence, this work presents a facile strategy for the design of long-term reliable hydrogels as smart strain sensors, especially used in extreme environments.

VP1 codon deoptimization and high-fidelity substitutions in 3D polymerase as potential vaccine strategies for eliciting immune responses against enterovirus A71.

Enterovirus A71 (EV-A71) can induce severe neurological complications and even fatal encephalitis in children, and it has caused several large outbreaks in Taiwan since 1998. We previously generated VP1 codon-deoptimized (VP1-CD) reverse genetics (rg) EV-A71 viruses (rgEV-A71s) that harbor a high-fidelity (HF) 3D polymerase. These VP1-CD-HF rgEV-A71s showed lower replication kinetics in vitro and decreased virulence in an Institute of Cancer Research (ICR) mouse model of EV-A71 infection, while still retaining their antigenicity in comparison to the wild-type virus. In this study, we aimed to further investigate the humoral and cellular immune responses elicited by VP1-CD-HF rgEV-A71s to assess the potential efficacy of these EV-A71 vaccine candidates. Following intraperitoneal (i.p.) injection of VP1-CD-HF rgEV-A71s in mice, we observed a robust induction of EV-A71-specific neutralizing IgG antibodies in the antisera after 21 days. Splenocytes isolated from VP1-CD-HF rgEV-A71s-immunized mice exhibited enhanced proliferative activities and cytokine production (IL-2, IFN-γ, IL-4, IL-6, and TNF-α) upon re-stimulation with VP1-CD-HF rgEV-A71, as compared to control mice treated with adjuvant only. Importantly, administration of antisera from VP1-CD-HF rgEV-A71s-immunized mice protected against lethal EV-A71 challenge in neonatal mice. These findings highlight that our generated VP1-CD-HF rgEV-A71 viruses are capable of inducing both cellular and humoral immune responses, supporting their potential as next-generation EV-A71 vaccines for combating EV-A71 infection.IMPORTANCEEV-A71 can cause severe neurological diseases and cause death in young children. Here, we report the development of synthetic rgEV-A71s with the combination of codon deoptimization and high-fidelity (HF) substitutions that generate genetically stable reverse genetics (rg) viruses as potential attenuated vaccine candidates. Our work provides insight into the development of low-virulence candidate vaccines through a series of viral genetic editing for maintaining antigenicity and genome stability and suggests a strategy for the development of an innovative next-generation vaccine against EV-A71.

3D edible scaffolds with yeast protein: A novel alternative protein scaffold for the production of high-quality cell-cultured meat.

The purpose of this study was to develop a novel edible scaffold by utilizing yeast proteins, which could partially replace collagen and produce hypoallergenic, odorless, and highly nutritious cell-cultured meat that meets the demands of a more significant number of consumers. The scaffold comprised proanthocyanidins, dialdehyde chitosan, collagen, and different proportions of yeast proteins (YP). The results indicated that the scaffold possessed excellent mechanical properties and biocompatibility, and supported cell proliferation and myogenic differentiation. Additionally, we evaluated the texture characteristics of the cultured meat models and traditional beef and discovered that the YP30 cultured meat model had similar springiness and chewiness as beef. Subsequently, further analyzed the similarity between the cultured meat models and traditional beef in appearance, taste, and nutrition. Further results illustrated that the yeast protein cultured meat model exhibited a complete model structure and comparable color and taste to beef after frying. Moreover, it was concluded that the protein content of the YP30 cultured meat model was closer to that of beef. These findings suggested that the edible scaffold using yeast proteins has enormous potential to facilitate the sustainable development of the cell-cultured meat industry.

Production of green-natural and "authentic" cultured meat based on proanthocyanidins-dialdehyde chitosan-collagen ternary hybrid edible scaffolds.

Cultured meat has the potential to fulfill the meat demand for the growing human population, but cultured meat development will be required to simplify the production process and produce naturally cultured meat, such as no longer stripping off scaffolders and adding artificial dyes. In this study, proanthocyanidins (PC) and dialdehyde chitosan (DAC) were employed as dual crosslinkers with collagen to prepare a hybrid 3D edible scaffold for the production of high-quality cell-cultured meat. The results revealed that the scaffold was biocompatible and could offer robust mechanical support and adhesion sites for bovine myoblasts, enabling long-term cell culture. Meanwhile, the Col-PC-DAC scaffold promoted the myogenic differentiation of bovine myoblasts and extracellular matrix protein secretion, further affecting the texture of cultured meat. After cooking the cultured meat and beef, it was shown that the cultured meat had some similarities to beef in color and flavor. Importantly, our findings demonstrate that cultured meat can acquire a color remarkably similar to that of conventional beef without the need for artificial dyeing. This breakthrough not only simplifies the production process but also ensures a more natural and appealing appearance of cultured meat. In conclusion, the proanthocyanidins-dialdehyde chitosan-collagen hybrid 3D edible scaffolds provide a new option for producing cultured meat that satisfies consumer expectations.

Targeting ST8SIA6-AS1 counteracts KRASG12C inhibitor resistance through abolishing the reciprocal activation of PLK1/c-Myc signaling.

BACKGROUND: KRASG12C inhibitors (KRASG12Ci) AMG510 and MRTX849 have shown promising efficacy in clinical trials and been approved for the treatment of KRASG12C-mutant cancers. However, the emergence of therapy-related drug resistance limits their long-term potential. This study aimed to identify the critical mediators and develop overcoming strategies. METHODS: By using RNA sequencing, RT-qPCR and immunoblotting, we identified and validated the upregulation of c-Myc activity and the amplification of the long noncoding RNA ST8SIA6-AS1 in KRASG12Ci-resistant cells. The regulatory axis ST8SIA6-AS1/Polo-like kinase 1 (PLK1)/c-Myc was investigated by bioinformatics, RNA fluorescence in situ hybridization, RNA immunoprecipitation, RNA pull-down and chromatin immunoprecipitation. Gain/loss-of-function assays, cell viability assay, xenograft models, and IHC staining were conducted to evaluate the anti-cancer effects of co-inhibition of ST8SIA6-AS1/PLK1 pathway and KRAS both in vitro and in vivo. RESULTS: KRASG12Ci sustainably decreased c-Myc levels in responsive cell lines but not in cell lines with intrinsic or acquired resistance to KRASG12Ci. PLK1 activation contributed to this ERK-independent c-Myc stability, which in turn directly induced PLK1 transcription, forming a positive feedback loop and conferring resistance to KRASG12Ci. ST8SIA6-AS1 was found significantly upregulated in resistant cells and facilitated the proliferation of KRASG12C-mutant cancers. ST8SIA6-AS1 bound to Aurora kinase A (Aurora A)/PLK1 and promoted Aurora A-mediated PLK1 phosphorylation. Concurrent targeting of KRAS and ST8SIA6-AS1/PLK1 signaling suppressed both ERK-dependent and -independent c-Myc expression, synergistically led to cell death and tumor regression and overcame KRASG12Ci resistance. CONCLUSIONS: Our study deciphers that the axis of ST8SIA6-AS1/PLK1/c-Myc confers both intrinsic and acquired resistance to KRASG12Ci and represents a promising therapeutic target for combination strategies with KRASG12Ci in the treatment of KRASG12C-mutant cancers.

Identification of Cuproptosis-Related circRNA-miRNA-mRNA Network in Laser-Induced Choroidal Neovascularization Models and in Peripheral Blood Mononuclear Cells of Patients with Neovascular Age-Related Macular Degeneration.

INTRODUCTION: The aims of this study were to investigate the molecular alterations of cuproptosis-related genes and to construct the cuproptosis-related circular RNA (circRNA)-microRNA (miRNA)-mRNA networks in neovascular age-related macular degeneration (nAMD). METHODS: The transcriptional profiles of laser-induced choroid neovascularization (CNV) mouse models and nAMD patient samples were obtained from sequencing and from the GEO database (GSE146887), respectively. The expression levels of ten cuproptosis-related genes (FDX1, DLAT, LIAS, DLD, PDHB, MTF1, CDKN2A, GLS, LIPT1, and PDHA1) were extracted and verified in both mouse CNV models and patient peripheral blood mononuclear cells (PBMCs) samples. The cuproptosis-related circRNA-miRNA-mRNA network was further constructed based on miRNet database, the dataset GSE131646 of small RNA expression profile, and the dataset GSE140178 of circRNA expression profile in mouse CNV models. RESULTS: The significant upregulation of Cdkn2a and Mtf1 and the downregulation of other 5 cuproptosis-related genes were verified in the mouse CNV model, but only CDKN2A significantly upregulated in PBMCs of patients with nAMD. Four miRNAs were detected in the intersection between miRNet prediction and sequencing data: miR-129-5p, miR-129-2-3p, miR-182-5p, and miR-615-3p. There were 9 circRNAs at the intersection of hsa-miR-182-5p and hsa-miR-615-3p predictions, one circRNA predicted by hsa-miR-129-5p and GSE140178 (hsa-circASH1L), and one circRNA predicted by hsa-miR-182-5p and hsa-miR-615-3p (hsa-circNPEPPS). CONCLUSION: This study suggested the repression of cuproptosis in nAMD pathologies and constructed a cuproptosis-related network of 8 cuproptosis-related genes, 4 miRNAs, and 11 circRNAs.

A 3-Ketoacyl-CoA Synthase 10 (KCS10) Homologue from Alfalfa Enhances Drought Tolerance by Regulating Cuticular Wax Biosynthesis.

Cuticular wax, forming the first line of defense against adverse environmental stresses, comprises very long-chain fatty acids (VLCFAs) and their derivatives. 3-Ketoacyl-CoA synthase (KCS) is a rate-limiting enzyme for VLCFA biosynthesis. In this study, we isolated KCS10, a KCS gene from alfalfa, and analyzed the effect of gene expression on wax production and drought stress in transgenic plants. MsKCS10 overexpression increased compact platelet-like crystal deposition and promoted primary alcohol biosynthesis through acyl reduction pathways in alfalfa leaves. Overexpression of MsKCS10 induced the formation of coiled-rodlet-like crystals and increased n-alkane content through decarbonylation pathways in tobacco and tomato fruits. Overexpression of MsKCS10 enhanced drought tolerance by limiting nonstomatal water loss, improving photosynthesis, and maintaining osmotic potential under drought stress in transgenic tobacco. In summary, MsKCS10 plays an important role in wax biosynthesis, wax crystal morphology, and drought tolerance, although the mechanisms are different among the plant species. MsKCS10 can be targeted in future breeding programs to improve drought tolerance in plants.

Fe-pillared montmorillonite functionalized chitosan/gelatin foams for efficient removal of organic pollutants by integration of adsorption and Fenton degradation.

A Fe-pillared montmorillonite (Fe-MMT) functionalized bio-based foam (Fe-MMT@CS/G) was developed by using chitosan (CS) and gelatin (G) as the matrix for high-efficiency elimination of organic pollutants through the integration of adsorption and Fenton degradation. The results showed that the mechanical properties of as-obtained foam were strengthened by the addition of certain amounts of Fe-MMT. Interestingly, Fe-MMT@CS/G displayed efficient adsorption ability for charged pollutants under a wide range of pH. The adsorption processes of methyl blue (MB), methylene blue (MEB) and tetracycline hydrochloride (TCH) on Fe-MMT@CS/G were well described by the Freundlich isotherm model and pseudo-second-order kinetic model. The maximum adsorption capacities were 2208.24 mg/g for MB, 1167.52 mg/g for MEB, and 806.31 mg/g for TCH. Electrostatic interactions, hydrogen bonding and van der Waals forces probably involved the adsorption process. As expected, this foam could exhibit better removal properties toward both charged and uncharged organic pollutants through the addition of H2O2 to trigger the Fenton degradation reaction. For non-adsorbable and uncharged bisphenol A (BPA), the removal efficiency was dramatically increased from 1.20 % to 92.77 % after Fenton degradation. Additionally, it presented outstanding recyclability. These results suggest that Fe-MMT@CS/G foam is a sustainable and efficient green material for the alleviation of water pollution.

Macrophage Sult2b1 promotes pathological neovascularization in age-related macular degeneration.

Disordered immune responses and cholesterol metabolism have been implicated in age-related macular degeneration (AMD), the leading cause of blindness in elderly individuals. SULT2B1, the key enzyme of sterol sulfonation, plays important roles in inflammation and cholesterol metabolism. However, the role and underlying mechanism of SULT2B1 in AMD have not been investigated thus far. Here, we report that SULT2B1 is specifically expressed in macrophages in choroidal neovascularization lesions. Sutl2b1 deficiency significantly reduced leakage areas and inhibited pathological angiogenesis by inhibiting M2 macrophage activation in vivo and in vitro. Mechanistically, loss of Sult2b1 activated LXRs and subsequently increased ABCA1 and ABCG1 (ABCA1/G1)-mediated cholesterol efflux from M2 macrophages. LXR inhibition (GSK2033 treatment) in Sult2b1 -/- macrophages reversed M2 polarization and decreased intracellular cholesterol capacity to promote pathological angiogenesis. In contrast to SULT2B1, STS, an enzyme of sterol desulfonation, protected against choroidal neovascularization development by activating LXR-ABCA1/G1 signalling to block M2 polarization. Collectively, these data reveal a cholesterol metabolism axis related to macrophage polarization in neovascular AMD.

Versatile CO2-responsive Sponges Decorated with ZIF-8 for Bidirectional Separation of Oil/Water and Controllable Removal of Dyes.

The complex wastewater containing water-soluble dyes and water-insoluble oils has given rise to significant environmental concerns that demand urgent remediation. Herein, a novel "smart" multifunctional sponge (ZIF-8@PMS) stepwise decorated with ZIF-8 nanoparticles and CO2-responsive copolymer (poly(2-(diethylamino) ethyl methacrylate-co-3-(trimethoxysilyl)propyl acrylate-co-stearyl methacrylate) was successfully prepared for CO2 controllable oil/water separation and dyes removal. The results revealed that the sponge coated with CO2-responsive copolymer for three cycles (ZIF-8@PMS-3) exhibited optimal comprehensive properties. The ZIF-8@PMS-3 had excellent compressive-resilient characteristics and chemical stability. As expected, it displayed tunable wettability and charged state under the regulation of CO2. Based on these features, ZIF-8@PMS-3 presented highly efficient removal of oil and dyes, even for the dye-containing oil/water emulsions, via a synergistic combination of adsorption and separation methods. The adsorption capacity for oil and various organic solvents ranged from 21.3 to 50 g g-1. The maximum adsorption capacities toward anionic dyes: methyl orange with 1205.89 mg g-1 and methyl blue with 880.00 mg g-1 in the presence of CO2 through electrostatic interaction. In the absence of CO2, it achieved maximum adsorption capacities for cationic dyes, including malachite green with 1246.15 mg g-1 and rhodamine B with 203 mg g-1, primarily driven by π-π interactions. According to distinct adsorption mechanisms, ZIF-8@PMS-3 could selectively adsorb either anionic or cationic dyes by exploiting CO2 as a trigger. Furthermore, the separation efficiencies for both types of oil/water emulsions surpassed 99.9%, with respective fluxes of 1566.99 L m-2 h-1 (water-in-oil emulsion) and 310.37 L m-2 h-1 (oil-in-water emulsion). Additionally, the as-prepared sponges exhibited remarkable antibacterial properties and exceptional recyclability. Therefore, the ZIF-8@PMS-3 holds substantial promise for potential applications in practical industrial wastewater treatment.

Effects of proanthocyanidins and dialdehyde chitosan on the proliferation and differentiation of bovine myoblast for cultured meat production.

Cultured meat technology intends to manufacture meat by cultivating muscle stem cells in vitro, which is an emerging methodology in meat production. However, the insufficient stemness of bovine myoblasts cultivated in vitro declined the ability of cell expansion and myogenic differentiation, which limited the production of cultured meat. Therefore, in this study, we introduced proanthocyanidins (PC, natural polyphenolic compounds) and dialdehyde chitosan (DAC, natural polysaccharides) to explore the effects of proliferation and differentiation of bovine myoblasts in vitro. The experiment results revealed that PC and DAC promoted cell proliferation by improving the transition from G1 to the S phase as well as cell division in G2. Meanwhile, the myogenic differentiation of cells was further boosted by the combined PC and DAC up-regulation of MYH3 expression. Moreover, the study revealed the synergistic effect of PC and DAC on enhancing the structural stability of collagen, and bovine myoblasts demonstrated excellent growth and dispersion ability on collagen scaffolds. It is concluded that both PC and DAC promote the proliferation and differentiation of bovine myoblasts, contributing to the development of cultured meat production systems.

Osmotic stress-induced lignin synthesis is regulated at multiple levels in alfalfa (Medicago sativa L.).

Alfalfa is an important forage crop. Yield and quality are frequently threatened by extreme environments such as drought and salt stress. As a component of the cell wall, lignin plays an important role in the abiotic stress response, the mechanisms of which have not been well clarified. In this study, we combined physiological, transcriptional, and metabolic analyses to reveal the changes in lignin content in alfalfa under mannitol-induced osmotic stress. Osmotic stress enhanced lignin accumulation by increasing G and S units, which was associated with increases in enzyme activities and decreases in 8 intermediate metabolites. Upon combined analysis of the transcriptome and metabolome, we identified five key structural genes and several coexpressed transcription factors, such as MYB and WRKY, which may play a core role in regulating lignin content and composition under osmotic stress. In addition, lignin synthesis was positively regulated by ABA but negatively regulated by ethylene under osmotic stress. These results provide new insight into the regulatory mechanism of lignin synthesis under abiotic stress.