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1.
Small Methods ; : e2401160, 2024 Sep 19.
Artigo em Inglês | MEDLINE | ID: mdl-39295467

RESUMO

The on-demand gene regulation is crucial for extensively exploring specific gene functions and developing personalized gene therapeutics, which shows great promise in precision medicines. Although some nucleic acid-based gene regulatory tools (antisense oligonucleotides and small interfering RNAs) are devised for achieving on-demand activation, the introduction of chemical modifications may cause undesired side effects, thereby impairing the gene regulatory efficacy. Herein, a methyl-engineered DNAzyme (MeDz) is developed for the visualization of endogenous alkyltransferase (AGT) and the simultaneous self-sufficiently on-demand gene regulation. The catalytic activity of DNAzyme can be efficiently blocked by O6-methylguanine (O6MeG) modification and specifically restored via the AGT-mediated DNA-repairing pathway. This simply designed MeDz is demonstrated to reveal AGT of varying expression levels in different cells, opening the possibility to explore the AGT-related biological processes. Moreover, the AGT-guided MeDz exhibits cell-selective regulation on the human early growth response-1 (EGR-1) gene, with efficient gene repression in breast cancer cells and low effectiveness in normal cells. The proposed MeDz offers an attractive strategy for on-demand gene regulation, displaying great potential in biomedical applications.

2.
Small ; : e2406545, 2024 Sep 16.
Artigo em Inglês | MEDLINE | ID: mdl-39282814

RESUMO

Artificial DNA circuits represent a versatile yet promising toolbox for in situ monitoring and concomitant regulation of diverse biological events within live cells. Nonetheless, their performance is significantly impeded by the diffusion-dominated slow reaction kinetics and the uncontrollable off-target activation. Herein, a self-localized cascade (SLC) circuit is reported for the robust and efficient microRNA (miRNA) analysis in living cells. The SLC circuit consists of the cell-specific localization module and the analyte-specific signal amplification module. By integrating the reaction probes of these two modules, the complexity of the system is reduced to realize the responsive co-localization of circuitry probes and the simultaneous cascade signal amplification. Taking advantage of the specifically activated, self-localized, and cascade design, the SLC circuit successfully achieves the robust miRNA-21 (miR-21) imaging and the accurate cells differentiation. Moreover, the reverse regulation mechanism is successfully explored between messenger RNA (mRNA) and miRNA through the engineered SLC circuit and further elucidates the underlying signaling pathways between them. Therefore, the SLC circuit provides a powerful tool for the sensitive detection of intracellular biomolecules and the study of the corresponding cell regulatory mechanisms.

3.
Food Res Int ; 195: 114984, 2024 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-39277245

RESUMO

This study investigated the impact of processing temperatures (190 °C, 210 °C, and 230 °C) and durations (7 min, 10 min, and 14 min) on the formation of Maillard reaction products (MRPs) and antioxidant activities in pan baked buns. Key Maillard reaction indicators, including glyoxal (GO), methylglyoxal (MGO), 5-hydroxymethylfurfural (5-HMF), melanoidins, and fluorescent advanced glycation end products (AGEs) were quantified. The results demonstrated significant increases in GO, MGO, 5-HMF contents (p < 0.05), and antioxidant activities (p < 0.05) when the buns were baked at 210 °C for 14 min, 230 °C for 10 min and 14 min. However, the interior MRPs of baked buns were minimally affected by the baking temperature and duration. Prolonged heating temperatures and durations exacerbated MRPs production (43.8 %-1038 %) in the bottom crust. Nonetheless, this process promoted the release of bound phenolic compounds and enhanced the antioxidant activity. Heating induces the thermal degradation of macromolecules in food, such as proteins and polysaccharides, which releases bound phenolic compounds by disrupting their chemical bonds within the food matrix. Appropriate selections of baking parameters can effectively reduce the formation of MRPs while simultaneously improve sensory quality and health benefit of the pan baked buns. Considering the balance between higher antioxidant properties and lower MRPs, the optimal thermal parameters for pan baked buns were 210 °C for 10 min. Furthermore, a normalized analysis revealed a consistent trend for GO, MGO, 5-HMF, fluorescent AGEs, and melanoidins. Moreover, MRPs were positively correlated with total contents of phenolic compounds, ferric-reducing antioxidant power (FRAP), and color, but negatively correlated with moisture contents and reducing sugars. Additionally, the interaction between baking conditions and Maillard reactions probably contributed to enhanced primary flavors in the final product. This study highlights the importance of optimizing baking parameters to achieve desirable MRPs levels, higher antioxidant activity, and optimal sensory attributes in baked buns.


Assuntos
Antioxidantes , Culinária , Furaldeído , Produtos Finais de Glicação Avançada , Temperatura Alta , Reação de Maillard , Aldeído Pirúvico , Antioxidantes/análise , Antioxidantes/química , Furaldeído/análogos & derivados , Furaldeído/análise , Furaldeído/química , Aldeído Pirúvico/química , Culinária/métodos , Humanos , Glioxal/química , Paladar , Polímeros/química , Pão/análise
4.
Foods ; 13(16)2024 Aug 20.
Artigo em Inglês | MEDLINE | ID: mdl-39200532

RESUMO

The current study investigated the impact of germination duration on the functional components (vitamin C, γ-aminobutyric acid (GABA), polyphenols, flavonoids) and antioxidant activity of germs and cotyledons of the germinated Chinese chestnut (Castanea mollissima). We utilized seeds of the "Zaofeng" Chinese chestnut to germinate, and sowed the seeds in wet sand at 22 °C and 85% relative humidity. The germination rate, length, diameter, and fresh weight of the sprouts were investigated at 0, 2, 4, 6, 8, and 10 days after sowing, and the kinetic changes of amylose, amylopectin, sugar components, soluble protein, vitamin C, GABA, total phenols, flavonoids, and the DPPH and ABTS free radical scavenging activity in the germs and cotyledons were monitored, respectively. The findings revealed that the germination rate and germ biomass increased continuously during germination. The germination rate reached 90% on the 8th day after sowing. Germination reduced amylose in cotyledons from 42.3% to 34.2%, amylopectin from 42.9% to 25.8%, total sugar from 12.6% to 11.4%, and vitamin C from 1.45 mg/g to 0.77 mg/g. Meanwhile, soluble protein in the embryos rose from 0.31% to 0.60%, vitamin C from 21.1 to 29.4 mg/g, GABA from 0.49 to 1.68 mg/g, total flavonoids from 53.6 to 129.7 mg/g, and ABTS antioxidant activity from 1.52 to 3.27 µmol TE/g. The average contents of D-fructose, inositol, vitamin C, GABA, polyphenols, and flavonoids and the DPPH and ABTS antioxidant activity in germs were as high as 22.5, 6, 35, 7.5, 10, 20, and 10 and 20-fold those of cotyledons, respectively. Especially, the average content of glucose in germ was as high as 80-fold that of cotyledon. D-xylulose, D-galacturonic acid, and D-ribose were only found in germs, but not in cotyledons. Considering the germ biomass and functional components content, germs of Chinese chestnuts germinated at 22 °C for 8 days are considered the most suitable raw material for functional food products. In conclusion, controlled germination not only enhances the physicochemical and functional properties of Chinese chestnut germs but also reduces the caloric content and improves the nutritional composition of the cotyledons appropriately. Moreover, the comprehensive evaluation of compositional changes and functionality in the embryo and cotyledon of Chinese chestnuts will provide a solid foundation for subsequent functional food processing utilizing germinated Chinese chestnuts.

5.
Vet Parasitol ; 331: 110284, 2024 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-39126893

RESUMO

Neospora caninum (N. caninum) is an obligate intracellular Apicomplexa parasite that causes abortions in dairy cows and incurs substantial to significant economic losses in the global dairy farming industry. Cordycepin, a nucleoside antibiotic derived from Chinese medicine Cordyceps militaries, exhibits diverse biological activities. However, it remains unclear whether cordycepin possesses inhibitory effects against N. caninum infection. Therefore, this study aimed to establish both in vivo and in vitro models of N. caninum to investigate the potential impact of cordycepin against N. caninum infection. We successfully established an in vitro model of N. caninum infection in RAW264.7 cells, followed by qRT- PCR analysis to detect the content of N. caninum DNA within the cells. The effects of cordycepin on N. caninum was observed using the Giemsa method on RAW264.7, and the rate of cell infection was calculated. Cordycepin exhibited inhibitory effects on N. caninum tachyzoites in vitro, preserving cellular integrity and reducing the rate of cell infection. In mice, we established an in vivo model of N. caninum infection and detected N. caninum presence in tissues using. Real-time fluorescence quantitative PCR. Histopathological changes were observed through Hematoxylin-eosin staining. Liver function was assessed by using glutamic acid aminotransferase (ALT) and aspartic acid aminotransferase (AST) kits. Oxidative stress status was measured using catalase (CAT), malondialdehyde (MDA), and glutathione (GSH) kits. Compared with the model group, mice treated with cordycepin showed reduced clinical symptoms, increased food intake, and their body weight (P=0.0143, P=0.0068) was significantly higher than those in the model group. Furthermore, cordycepin treatment significantly alleviated hepatic cord disorders, hepatocellular swelling, detachment, and vacuolization; duodenal epithelial detachment and shortening of villi caused by N. caninum infection. Cordycepin administration reduced the increase in ALT (P=0.01, P=0.008) and AST (P<0.001) levels caused by N. caninum infection, while ameliorating hepatocyte swelling, necrosis, and detachment as well as inflammatory cell infiltration within mice liver; it also led to shortened or even disappeared duodenal villi along with and oedema of the submucosa. Analysis of oxidative stress showed that cordycepin ameliorated the damage caused by N. caninum by reducing MDA (P=0.03, P=0.02, P=0.005) and increasing CAT (P=0.004, P<0.001) and GSH (P=0.004, P<0.001) levels. In conclusion, this study reports for the first time on cordycepin's efficacy against N. caninum infection providing a potential candidate drug for neosporosis treatment.


Assuntos
Coccidiose , Desoxiadenosinas , Neospora , Animais , Neospora/efeitos dos fármacos , Coccidiose/tratamento farmacológico , Coccidiose/veterinária , Coccidiose/parasitologia , Camundongos , Desoxiadenosinas/farmacologia , Desoxiadenosinas/uso terapêutico , Feminino , Células RAW 264.7 , Fígado/parasitologia , Fígado/efeitos dos fármacos , Camundongos Endogâmicos BALB C , Coccidiostáticos/farmacologia , Coccidiostáticos/uso terapêutico
6.
Anal Chem ; 96(31): 12854-12861, 2024 08 06.
Artigo em Inglês | MEDLINE | ID: mdl-39042763

RESUMO

Sensitive and reliable microRNA imaging in living cells has significant implications for clinical diagnosis and monitoring. Catalytic DNA circuits have emerged as potent tools for tracking these intracellular biomarkers and probing the corresponding biochemical processes. However, their utility is hindered by the low resistance to external interference, leading to undesired off-site activation and consequent signal leakage. Therefore, achieving the endogenous control of the DNA circuit's activation is preferable to the reliable target analysis in living cells. In this study, we attempted to address this challenge by engineering a simple yet effective endogenous glutathione (GSH)-regulated hybridization chain reaction (HCR) circuit for acquiring high-contrast miRNA imaging. Initially, the HCR hairpin reactants were blocked by the engineered disulfide-integrated DNA duplex, thus effectively passivating their sensing function. And the precaged HCR hairpin was liberated by the cell-specific GSH molecule, thus initiating the HCR system for selectively amplified detection of microRNA-21 (miR-21). This approach prevented unwanted signal leakage before exposure into target cells, thus ensuring robust miR-21 imaging with high accuracy and reliability in specific tumor cells. Moreover, the endogenously responsive HCR circuit established a link between the small regulatory factors and miRNA, thereby enhancing the signal gain. In summary, the endogenously activatable DNA circuit represents a versatile toolbox for robust bioanalysis and exploration of potential signaling pathways in living cells.


Assuntos
Glutationa , MicroRNAs , MicroRNAs/análise , MicroRNAs/metabolismo , Glutationa/metabolismo , Glutationa/análise , Humanos , Hibridização de Ácido Nucleico
7.
Small ; : e2401954, 2024 May 11.
Artigo em Inglês | MEDLINE | ID: mdl-38733233

RESUMO

Achiral solvents are commonly utilized to induce the self-assembly of chiral molecules. This study demonstrates that achiral solvents can trigger helicity inversion in the assemblies of dansyl amphiphiles and control the excited-state "majority rule" in assemblies composed of pure enantiomers, through variation of the cosolvent ratio. Specifically, enantiomers of dansyl amphiphiles self-assemble into helical structures with opposite handedness in methanol (MeOH) and acetonitrile (MeCN), together with inversed circular dichroism and circularly polarized luminescence (CPL) signals. When a mixture of MeOH and MeCN is employed, the achiral cosolvents collectively affect the CPL of the assemblies in a way similar to that of "mixed enantiomers". The dominant cosolvent governs the CPL signal. As the cosolvent composition shifts from pure MeCN to MeOH, the CPL signals undergo a significant inversion and amplification, with two maxima observed at ≈20% MeOH and 20% MeCN. This study deepens the comprehension of how achiral solvents modulate helical nanostructures and their excited-state chiroptical properties.

8.
Plants (Basel) ; 13(9)2024 May 05.
Artigo em Inglês | MEDLINE | ID: mdl-38732488

RESUMO

Dioscorea alata, commonly known as "greater yam", is a vital crop in tropical and subtropical regions of the world, yet it faces significant threats from anthracnose disease, mainly caused by Colletotrichum gloeosporioides. However, exploring disease resistance genes in this species has been challenging due to the difficulty of genetic mapping resulting from the loss of the flowering trait in many varieties. The receptor-like kinase (RLK) gene family represents essential immune receptors in plants. In this study, genomic analysis revealed 467 RLK genes in D. alata. The identified RLKs were distributed unevenly across chromosomes, likely due to tandem duplication events. However, a considerable number of ancient whole-genome or segmental duplications dating back over 100 million years contributed to the diversity of RLK genes. Phylogenetic analysis unveiled at least 356 ancient RLK lineages in the common ancestor of Dioscoreaceae, which differentially inherited and expanded to form the current RLK profiles of D. alata and its relatives. The analysis of cis-regulatory elements indicated the involvement of RLK genes in diverse stress responses. Transcriptome analysis identified RLKs that were up-regulated in response to C. gloeosporioides infection, suggesting their potential role in resisting anthracnose disease. These findings provide novel insights into the evolution of RLK genes in D. alata and their potential contribution to disease resistance.

9.
Poult Sci ; 103(6): 103717, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38643746

RESUMO

Gliotoxin (GT) belongs to the epipolythiodioxopiperazine (ETP) family, which is considered a crucial virulence determinant among the secondary metabolites produced by Aspergillus fumigatus. The metabolites are commonly found in food and feed, contributing to the invasion and immune escape of Aspergillus fumigatus, thereby posing a significant threat to the health of livestock, poultry, and humans. Heterophil extracellular traps (HETs), a novel form of innate immune defense, have been documented in the chicken's innate immune systems for capturing and eliminating invading microbes. However, the effects and mechanisms of GT on the production of duck HETs in vitro remain unknown. In this study, we first confirmed the presence of HETs in duck innate immune systems and further investigated the molecular mechanism underlying GT-induced HETs release. Our results demonstrate that GT can trigger typical release of HETs in duck. The structures of GT-induced HETs structures were characterized by DNA decoration, citrullinated histones 3, and elastase. Furthermore, NADPH oxidase, glycolysis, ERK1/2 and p38 signaling pathway were found to regulate GT-induced HETs. In summary, our findings reveal that gliotoxin activates HETs release in the early innate immune system of duck while providing new insights into the immunotoxicity of GT towards ducks.


Assuntos
Patos , Gliotoxina , Imunidade Inata , Animais , Imunidade Inata/efeitos dos fármacos , Armadilhas Extracelulares/efeitos dos fármacos , Imunotoxinas/toxicidade
10.
Int J Lab Hematol ; 46(4): 678-686, 2024 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-38600718

RESUMO

INTRODUCTION: To identify the differentially expressed genes of acute myeloid leukaemia (AML) and construct and verify a survival prognosis model combined with patient survival information. METHODS: The TARGET database was searched to identify differentially expressed peripheral blood genes in children with AML and healthy children. A gene set functional analysis and pathway analysis were performed using gene ontology and the KEGG pathway. A prognostic model for children with AML was constructed using univariate Cox, LASSO Cox regression and multivariate Cox regression analyses. Time-dependent receiver operating characteristic (ROC) curves were adopted to assess the predictive capacity of the prognostic models. RESULTS: In total, 1640 differentially expressed genes were screened (1119 upregulated and 521 downregulated genes). The differentially expressed genes were mainly involved in nutrient metabolism and cytochrome P450 metabolism. Six key genes related to the prognosis of AML, FAM157A, GPR78, IRX5, RP4-800G7.1, RP11-179H18.5 and RP11-61N20.3, were identified. Kaplan-Meier curves indicated that 3-year and 5-year overall survival was significantly higher in the low-risk group than in the high-risk group. The area under the ROC curve was 0.722. At different stages of AML, FAM157A and RP4-800G7.1 exhibited significant differences in expression. The expression levels of FAM157A were significantly decreased in AML, whereas the expression levels of GPR78, IRX5, RP4-800G7.1, RP11-179H18.5 and RP11-61N20.3 were significantly increased in AML. CONCLUSION: A prognosis-related gene model of AML was successfully constructed, and the expression levels of the model genes varied with AML stage.


Assuntos
Regulação Leucêmica da Expressão Gênica , Leucemia Mieloide Aguda , Humanos , Leucemia Mieloide Aguda/genética , Leucemia Mieloide Aguda/mortalidade , Leucemia Mieloide Aguda/diagnóstico , Criança , Prognóstico , Feminino , Masculino , Pré-Escolar , Curva ROC , Perfilação da Expressão Gênica , Estimativa de Kaplan-Meier , Biomarcadores Tumorais/genética , Lactente
11.
Adv Mater ; 36(31): e2403329, 2024 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-38625749

RESUMO

The precise manipulation of supramolecular polymorphs has been widely applied to control the morphologies and functions of self-assemblies, but is rarely utilized for the fabrication of circularly polarized luminescence (CPL) materials with tailored properties. Here, this work reports that an amphiphilic naphthalene-histidine compound (NIHis) readily self-assembled into distinct chiral nanostructures through pathway-dependent supramolecular polymorphism, which shows opposite and multistimuli responsive CPL signals. Specifically, NIHis display assembly-induced CPL from the polymorphic keto tautomer, which become predominant during enol-keto tautomerization shifting controlled by a bulk solvent effect. Interestingly, chiral polymorphs of nanofiber and microbelt with inverted CPL signals can be prepared from the same NIHis monomer in exactly the same solvent compositions and concentrations by only changing the temperature. The tunable CPL performance of the solid microbelts is realized under multi external physical or chemical stimuli including grinding, acid fuming, and heating. In particular, an emission color and CPL on-off switch based on the microbelt polymorph by reversible heating-cooling protocol is developed. This work brings a new approach for developing smart CPL materials via supramolecular polymorphism engineering.

12.
Stem Cell Reports ; 19(4): 579-595, 2024 Apr 09.
Artigo em Inglês | MEDLINE | ID: mdl-38518781

RESUMO

Transcription factors (TFs) are pivotal in guiding stem cell behavior, including their maintenance and differentiation. Using single-cell RNA sequencing, we investigated TFs expressed in endothelial progenitors (EPs) derived from human pluripotent stem cells (hPSCs) and identified upregulated expression of SOXF factors SOX7, SOX17, and SOX18 in the EP population. To test whether overexpression of these factors increases differentiation efficiency, we established inducible hPSC lines for each SOXF factor and found only SOX17 overexpression robustly increased the percentage of cells expressing CD34 and vascular endothelial cadherin (VEC). Conversely, SOX17 knockdown via CRISPR-Cas13d significantly compromised EP differentiation. Intriguingly, we discovered SOX17 overexpression alone was sufficient to generate CD34+VEC+CD31- cells, and, when combined with FGF2 treatment, more than 90% of CD34+VEC+CD31+ EP was produced. These cells are capable of further differentiating into endothelial cells. These findings underscore an undiscovered role of SOX17 in programming hPSCs toward an EP lineage, illuminating pivotal mechanisms in EP differentiation.


Assuntos
Células Endoteliais , Fator 2 de Crescimento de Fibroblastos , Células-Tronco Pluripotentes , Fatores de Transcrição SOXF , Humanos , Antígenos CD34/metabolismo , Diferenciação Celular/genética , Células Endoteliais/metabolismo , Fator 2 de Crescimento de Fibroblastos/genética , Fator 2 de Crescimento de Fibroblastos/metabolismo , Células-Tronco Pluripotentes/metabolismo , Fatores de Transcrição SOXF/genética , Fatores de Transcrição SOXF/metabolismo
13.
Foods ; 13(5)2024 Feb 26.
Artigo em Inglês | MEDLINE | ID: mdl-38472819

RESUMO

The purpose of this study was to clarify the effect of CA (controlled atmosphere, 2-3% O2 + 3% CO2) and NO (nitric oxide, generated by 0.4 nM sodium nitroprusside), alone or combined (CA + NO), on the physio-chemical properties, enzyme activities and antioxidant capacities of chestnuts during storage at 0 °C for 180 d. Compared with control (CT), CA and CA+NO both improved the storage quality of the samples, but only CA resulted in more ethanol production. Moreover, these improvements were further enhanced and ethanol synthesis was inhibited by the addition of NO. A spectrometer was used to assess the production of phenolic content (TPC) and activities of phenylalanine ammonia-lyase (PAL), superoxide dismutas (SOD), peroxidase (POD), catalase (CAT) and polyphenol oxidase (PPO) as influenced by CA or CA+NO treatments. Higher TPC, PAL, SOD, POD, CAT, and lower PPO were observed in CA alone, and more so in the combination with NO group. The increased antioxidant production and enhanced antioxidant activities contributed to scavenging reactive oxygen species (ROS) and reducing malondialdehyde (MDA). This study unveiled the correlations and differences between the effects of CA and CA+NO on storage quality, providing valuable insights into postharvest preservation and suggesting that the combination (CA+NO) was more beneficial for quality maintenance in chestnuts.

14.
Proc Natl Acad Sci U S A ; 121(4): e2317058121, 2024 Jan 23.
Artigo em Inglês | MEDLINE | ID: mdl-38232281

RESUMO

Integration of methanogenic archaea with photocatalysts presents a sustainable solution for solar-driven methanogenesis. However, maximizing CH4 conversion efficiency remains challenging due to the intrinsic energy conservation and strictly restricted substrates of methanogenic archaea. Here, we report a solar-driven biotic-abiotic hybrid (biohybrid) system by incorporating cadmium sulfide (CdS) nanoparticles with a rationally designed methanogenic archaeon Methanosarcina acetivorans C2A, in which the glucose synergist protein and glucose kinase, an energy-efficient route for glucose transport and phosphorylation from Zymomonas mobilis, were implemented to facilitate nonnative substrate glucose for methanogenesis. We demonstrate that the photo-excited electrons facilitate membrane-bound electron transport chain, thereby augmenting the Na+ and H+ ion gradients across membrane to enhance adenosine triphosphate (ATP) synthesis. Additionally, this biohybrid system promotes the metabolism of pyruvate to acetyl coenzyme A (AcCoA) and inhibits the flow of AcCoA to the tricarboxylic acid (TCA) cycle, resulting in a 1.26-fold augmentation in CH4 production from glucose-derived carbon. Our results provide a unique strategy for enhancing methanogenesis through rational biohybrid design and reprogramming, which gives a promising avenue for sustainably manufacturing value-added chemicals.


Assuntos
Trifosfato de Adenosina , Metano , Metano/metabolismo , Transporte de Elétrons , Trifosfato de Adenosina/metabolismo , Metabolismo Energético , Transporte Biológico , Methanosarcina/metabolismo
15.
Dev Comp Immunol ; 153: 105107, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38036049

RESUMO

Artificial insemination has been a predominant technique employed in goat husbandry for breeding purposes. Subsequent to artificial insemination, sperm can elicit inflammation in the reproductive tract, resulting in substantial the accumulation of neutrophils. Recognized as foreign entities, sperm may become entrapped within neutrophil extracellular traps (NETs) released by neutrophils, thereby exploiting their properties of pathogen elimination. Deoxyribonuclease I (DNase I), which is known for disintegrating NETs and causing loss of function, has been utilized to ameliorate liver and brain damage resulting from NETs, as well as to enhance sperm quality. This study investigated the mechanism of sperm-induced NETs and further explored the impact of DNase I on NETs. Sperm quality was evaluated using optical microscopy, while the structure of NETs was observed through immunofluorescence staining. The formation mechanism of NETs was examined using inhibitors and PicoGreen. The findings revealed that sperm induced the formation of NETs, a process regulated by glycolysis, NADPH oxidase, ERK1/2, and p38 signaling pathways. The composition of NETs encompassed DNA, citrullinated histone H3 (citH3), and elastase (NE). DNase I protects sperm by degrading NETs, thereby concurrently preserving the integrity of plasma membrane and motility of sperm. In summary, the release of sperm-induced NETs leads to its damage, but this detrimental effect is counteracted by DNase I through degradation of NETs. These observations provide novel insights into reproductive immunity in goats.


Assuntos
Armadilhas Extracelulares , Masculino , Animais , Armadilhas Extracelulares/metabolismo , Cabras , Sêmen , Neutrófilos , Espermatozoides , Desoxirribonuclease I/metabolismo , Desoxirribonuclease I/farmacologia
16.
Ecotoxicol Environ Saf ; 270: 115901, 2024 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-38157799

RESUMO

The toxicity of nanoparticles to freshwater microalgae is of significant importance in maintaining the overall stability of aquatic ecosystems. However, the transport mechanism and toxicity response of microalgae towards nanoplastics (NPs) remain to be further investigated. In this study, we examined the toxicity and internalization mechanisms of polystyrene nanoplastics (PS-NPs) in the microalga Chlorella sorokiniana. The results revealed that the PS-NPs inhibited algal cells' growth and disrupted cell integrity upon contact, leading to cell shrinkage or rupture. Moreover, amino-modified PS-NPs (Nano-PS-NH2) exhibited greater toxicity to C. sorokiniana than carboxyl-modified PS-NPs (Nano-PS-COOH). Furthermore, significant inhibition of PS-NPs internalization was observed when four different endocytosis-related inhibitors were used, indicating that internalized PS-NPs can enter algal cells through endocytic pathways. More importantly, C. sorokiniana exposed to Nano-PS-NH2 responded to the reduction in carbon sources and energy resulting from the suppression of photosynthesis by regulating the metabolism of carbohydrates. These findings elucidate the effects of PS-NPs on C. sorokiniana, including their impact on cell morphology and metabolism, while shedding light on the internalization mechanisms of NPs by C. sorokiniana which deepen our understanding of the toxicity of nanoplastics on algae and provide important theoretical support for solving such aquatic ecological environment problems.


Assuntos
Chlorella , Microalgas , Nanopartículas , Poluentes Químicos da Água , Microplásticos/toxicidade , Poliestirenos/toxicidade , Ecossistema , Poluentes Químicos da Água/toxicidade , Nanopartículas/toxicidade
17.
Angew Chem Int Ed Engl ; 63(6): e202316863, 2024 Feb 05.
Artigo em Inglês | MEDLINE | ID: mdl-38116831

RESUMO

Pathway-dependent self-assembly, in which a single building block forms two or more types of self-assembled nanostructures, is an important topic due to its mimic to the complexity in biology and manipulation of diverse supramolecular materials. Here, we report a pathway-dependent self-assembly using chiral glutamide derivatives (L or D-PAG), which form chiral nanotwist and nanotube through a cooperative slow cooling and an isodesmic fast cooling process, respectively. Furthermore, pathway-dependent self-assembly can be harnessed to control over the supramolecular co-assembly of PAG with a luminophore ß-DCS or a photopolymerizable PCDA. Fast cooling leads to the co-assembled PAG/ß-DCS nanotube exhibiting green circularly polarized luminescence (CPL), while slow cooling to nanofiber with blue CPL. Additionally, fast cooling process promotes the photopolymerization of PCDA into a red chiral polymer, whereas slow cooling inhibits the polymerization. This work not only demonstrates the pathway-dependent control over structural characteristics but also highlights the diverse functions emerged from the different assemblies.

18.
Parasitol Res ; 123(1): 34, 2023 Dec 13.
Artigo em Inglês | MEDLINE | ID: mdl-38087003

RESUMO

Toxoplasma gondii (T. gondii) exhibits a significantly high prevalence of infection in goats, leading to adverse consequences such as abortion and stillbirth in ewes, thereby posing a substantial challenge to the goat farming industry. Neutrophil extracellular traps (NETs) have been shown to capture T. gondii in goats; however, the precise mechanisms underlying NET release in goats remain poorly understood. Therefore, the aim of our research was to elucidate the involved mechanism. We assessed the cytotoxicity of T. gondii on neutrophils using CCK-8 assay, visualized the structure of T. gondii-induced goat NETs through immunofluorescence, quantified ROS release during T. gondii-induced NET formation using fluorescence microplate analysis, and employed inhibitors targeting TLR 2, TLR4, NADPH oxidase, ERK1/2, and P38 MAPK signaling pathways as well as glycolysis to dissect the mechanisms underlying T. gondii-induced NET release. Within 1 h, T. gondii did not exhibit significant cytotoxicity towards neutrophils in our findings. The formation of typical NET structures induced by T. gondii involved DNA, citrullinated histone 3 (citH3), and neutrophil elastase (NE). Additionally, T. gondii significantly stimulated the release of NETs in goats. The process was accompanied by the production of reactive oxygen species (ROS) mediated through NADPH oxidase, p38, and ERK1/2 signaling pathways. Inhibition of these pathways resulted in a decrease in NET release. Moreover, inhibition of TLR 2, TLR4, and glycolysis also led to a reduction in T. gondii-induced NET release. Overall, our study demonstrates that T. gondii can induce characteristic NET structures and elucidates the involvement of various mechanisms including TLR2/TLR4 signaling pathway activation, NADPH oxidase activity modulation via ROS production regulation through p38 MAPK and ERK1/2 signaling pathways, and glycolysis regulation during the innate immune response against T. gondii infection in goats.


Assuntos
Armadilhas Extracelulares , Toxoplasma , Animais , Feminino , Ovinos , Sistema de Sinalização das MAP Quinases , Receptor 2 Toll-Like/genética , Receptor 2 Toll-Like/metabolismo , Receptor 4 Toll-Like/genética , Espécies Reativas de Oxigênio/metabolismo , Cabras , Neutrófilos , Transdução de Sinais , Proteínas Quinases p38 Ativadas por Mitógeno/genética , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo , NADPH Oxidases/metabolismo
19.
Anal Chem ; 95(51): 18731-18738, 2023 12 26.
Artigo em Inglês | MEDLINE | ID: mdl-38096424

RESUMO

The engineering of catalytic hybridization DNA circuits represents versatile ways to orchestrate a complex flux of molecular information at the nanoscale, with potential applications in DNA-encoded biosensing, drug discovery, and therapeutics. However, the diffusive escape of intermediates and unintentional binding interactions remain an unsolved challenge. Herein, we developed a compact, yet efficient, self-regulatory assembly circuit (SAC) for achieving robust microRNA (miRNA) imaging in live cells through DNA-templated guaranteed catalytic hybridization. By integrating the toehold strand with a preblocked palindromic fragment in the stem domain, the proposed miniature SAC system allows the reactant-to-template-controlled proximal hybridization, thus facilitating the bidirectional-sustained assembly and the localization-intensified signal amplification without undesired crosstalk. With condensed components and low reactant complexity, the SAC amplifier realized high-contrast intracellular miRNA imaging. We anticipate that this simple and template-controlled design can enrich the clinical diagnosis and prognosis toolbox.


Assuntos
Técnicas Biossensoriais , DNA Catalítico , MicroRNAs , MicroRNAs/genética , Técnicas Biossensoriais/métodos , Limite de Detecção , DNA/genética , DNA/química , Hibridização de Ácido Nucleico/métodos , DNA Catalítico/química
20.
Adv Mater ; 35(49): e2306504, 2023 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-37694949

RESUMO

Si nanoparticles (NPs) are considered as a promising high-capacity anode material owing to their ability to prevent mechanical failure from drastic volume change during (de)lithiation. However, upon cycling, a quick capacity fading is still observed for Si NPs, and the underlying mechanism remains elusive. In this contribution, it is demonstrated that the quick capacity fading is mainly caused by the generation of dead (electrochemically inert) Si with blocked electron conductivity in a densely composited Si/SEI (solid electrolyte interface) hybrid. This is due to the combined influence of electrolyte-related side reactions and the accompanied agglomeration of Si NPs. A compact, sub-nano scale interfused SiOx /C composite coating onto the Si NPs is constructed, and a highly stabilized electrochemistry is achieved upon long cycling. The SiOx /C coating with electron/ion dual transport paths and robust mechanical flexibility enables a fast and stable lithium ion/electron dual diffusion pathway towards the encapsulated Si. With fast reaction kinetics, stable SEI, and an antiagglomeration feature, the obtained Si@SiOx /C composite demonstrates a stable high capacity. This work unravels new perspectives on the capacity fading of Si NPs and provides an effective encapsulating method to remedy the structure degradation and capacity fading of nano Si.

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