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Accurate in vivo imaging of G-quadruplexes (G4) is critical for understanding the emergence and progression of G4-associated diseases like cancer. However, existing in vivo G4 fluorescent probes primarily operate within the near-infrared region (NIR-I), which limits their application accuracy due to the short emission wavelength. The transition to second near-infrared (NIR-II) fluorescent imaging has been of significant interest, as it offers reduced autofluorescence and deeper tissue penetration, thereby facilitating more accurate in vivo imaging. Nonetheless, the advancement of NIR-II G4 probes has been impeded by the absence of effective probe design strategies. Herein, through a "step-by-step" rational design approach, we have successfully developed NIRG-2, the first small-molecule fluorescent probe with NIR-II emission tailored for in vivo G4 detection. Molecular docking calculations reveal that NIRG-2 forms stable hydrogen bonds and strong π-π interactions with G4 structures, which effectively inhibit twisted intramolecular charge transfer (TICT) and, thereby, selectively illuminate G4 structures. Due to its NIR-II emission (940 nm), large Stokes shift (90 nm), and high selectivity, NIRG-2 offers up to 47-fold fluorescence enhancement and a tissue imaging depth of 5 mm for in vivo G4 detection, significantly outperforming existing G4 probes. Utilizing NIRG-2, we have, for the first time, achieved high-contrast visualization of tumor metastasis through lymph nodes and precise tumor resection. Furthermore, NIRG-2 proves to be highly effective and reliable in evaluating surgical and drug treatment efficacy in cancer lymphatic metastasis models. We are optimistic that this study not only provides a crucial molecular tool for an in-depth understanding of G4-related diseases in vivo but also marks a promising strategy for the development of clinical NIR-II G4-activated probes.
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Colorantes Fluorescentes , G-Cuádruplex , Imagen Óptica , Colorantes Fluorescentes/química , Colorantes Fluorescentes/síntesis química , Humanos , Animales , Metástasis de la Neoplasia , Ratones , Simulación del Acoplamiento Molecular , Diseño de Fármacos , Rayos Infrarrojos , Línea Celular Tumoral , Estructura MolecularRESUMEN
Polysulfide shuttle and sluggish sulfur redox kinetics remain key challenges in lithium-sulfur batteries. Previous researches have shown that introducing oxygen into transition metal sulfides helps to capture polysulfides and enhance their conversion kinetics. Based on this, further investigations are conducted to explore the impact of oxygen doping levels on the physical-chemical properties and electrocatalytic performance of MoS2. The findings reveal that MoS2 doped with high-content oxygen exhibits enhanced conductivity and polysulfides conversion kinetics compared to MoS2 with low-content oxygen doping, which can be attributed to the alteration of crystal structure from 2H-phase to the 1T-phase, the introduction of increased Li-O interactions, and the effect of defects resulting from high-oxygen doping. Consequently, the lithium-sulfur batteries using high-oxygen doped MoS2 as a catalyst deliver a high discharge capacity of 1015 mAh g-1 at 0.25C and maintain 78.5% capacity after 300 more cycles. Specifically, lithium-sulfur batteries employing paper-based electrodedemonstrate an areal capacity of 3.91 mAh cm-2 at 0.15C, even with sulfur loading of 4.1 mg cm-2 and electrolyte of 6.7 µL mg-1. These results indicate that oxygen doping levels can modify the properties of MoS2, and high-oxygen doped MoS2 shows promise as an efficient catalyst for lithium-sulfur batteries.
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BACKGROUND: Dihydromyricetin (DHM), a flavonoid compound of natural origin, has been identified in high concentrations in ampelopsis grossedentata and has a broad spectrum of biological and pharmacological functions, particularly in regulating glucose and lipid metabolism. The objective of this research was to examine how DHM affected nonalcoholic fatty liver disease (NAFLD) and its underlying mechanisms involved in the progression of NAFLD in a rat model subjected to a high-fat diet (HFD). Additionally, the study examines the underlying mechanisms in a cellular model of steatohepatitis using palmitic acid (PA)-treated HepG2 cells, with a focus on the potential correlation between autophagy and hepatic insulin resistance (IR) in the progress of NAFLD. METHODS: SD rats were exposed to a HFD for a period of eight weeks, followed by a treatment with DHM (at doses of 50, 100, and 200 mg·kg-1·d-1) for additional six weeks. The HepG2 cells received a 0.5 mM PA treatment for 24 h, either alone or in conjunction with DHM (10 µM). The histopathological alterations were assessed by the use of Hematoxylin-eosin (H&E) staining. The quantification of glycogen content and lipid buildup in the liver was conducted by the use of PAS and Oil Red O staining techniques. Serum lipid and liver enzyme levels were also measured. Autophagic vesicle and autolysosome morphology was studied using electron microscopy. RT-qPCR and/or western blotting techniques were used to measure IR- and autophagy-related factors levels. RESULTS: The administration of DHM demonstrated efficacy in ameliorating hepatic steatosis, as seen in both in vivo and in vitro experimental models. Moreover, DHM administration significantly increased GLUT2 expression, decreased G6Pase and PEPCK expression, and improved IR in the hepatic tissue of rats fed a HFD and in cells exhibiting steatosis. DHM treatment elevated Beclin 1, ATG 5, and LC3-II levels in hepatic steatosis models, correlating with autolysosome formation. The expression of AMPK levels and its downstream target PGC-1α, and PPARα were decreased in HFD-fed rats and PA-treated hepatocytes, which were reversed through DHM treatment. AMPK/ PGC-1α and PPARα knockdown reduced the impact of DHM on hepatic autophagy, IR and accumulation of hepatic lipid. CONCLUSIONS: Our findings revealed that AMPK/ PGC-1α, PPARα-dependent autophagy pathways in the pathophysiology of IR and hepatic steatosis has been shown, suggesting that DHM might potentially serve as a promising treatment option for addressing this disease.
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Flavonoles , Resistencia a la Insulina , Enfermedad del Hígado Graso no Alcohólico , Ratas , Animales , Ratones , Enfermedad del Hígado Graso no Alcohólico/patología , PPAR alfa/metabolismo , Proteínas Quinasas Activadas por AMP/metabolismo , Resistencia a la Insulina/fisiología , Ratas Sprague-Dawley , Hígado/patología , Metabolismo de los Lípidos , Ácido Palmítico/metabolismo , Ácido Palmítico/farmacología , Ácido Palmítico/uso terapéutico , Autofagia , Dieta Alta en Grasa , Ratones Endogámicos C57BLRESUMEN
BACKGROUND: Gastric cancer is characterized by high invasiveness, heterogeneity, and late diagnosis, leading to high incidence and mortality rates. It is a significant public health concern globally. Early prevention is crucial in reducing the occurrence of gastric cancer, and dietary prevention, particularly focusing on carotenoids, has been considered a convenient and effective approach. However, the association between carotenoid intake and gastric cancer incidence remains controversial. METHODS: A systematic search was conducted in PubMed, Ovid Embase, Web of Science, and Cochrane databases from inception to January 5, 2023. Two reviewers independently screened search results, extracted relevant data, and evaluated study quality. Statistical analysis was performed using the "metan" command in STATA 16 software. Random-effects or fixed-effects models were chosen based on the magnitude of heterogeneity among studies. RESULTS: This study included a total of 35 publications, consisting of 23 case-control studies and 12 cohort studies. Meta-analysis of case-control studies showed that alpha-carotene (OR = 0.71, 95% CI: 0.55-0.92), beta-carotene (OR = 0.62, 95% CI: 0.53-0.72), and lutein (OR = 0.82, 95% CI: 0.69-0.97) significantly reduced the risk of gastric cancer, while beta-cryptoxanthin (OR = 0.88, 95% CI: 0.75-1.04) and lycopene (OR = 0.86, 95% CI: 0.73-1.00) showed no significant correlation. Meta-analysis of cohort studies indicated no significant associations between any of the five carotenoids and gastric cancer incidence (alpha-carotene: RR = 0.81, 95% CI: 0.54-1.23; beta-carotene: RR = 0.86, 95% CI: 0.64-1.16; beta-cryptoxanthin: RR = 0.86, 95% CI: 0.64-1.16; lutein: RR = 0.94, 95% CI: 0.69-1.29; lycopene: RR = 0.89, 95% CI: 0.69-1.14). CONCLUSIONS: The relationship between carotenoids and gastric cancer incidence may vary depending on the type of study conducted. Considering that evidence from cohort studies is generally considered stronger than evidence from case-control studies, and high-quality randomized controlled trials show no significant association between carotenoids and gastric cancer incidence, current evidence does not support the supplementation of carotenoids for gastric cancer prevention. Further targeted research is needed to explore the association between the two.
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Neoplasias Gástricas , beta Caroteno , Humanos , beta Caroteno/uso terapéutico , Licopeno , Luteína/uso terapéutico , Neoplasias Gástricas/epidemiología , Neoplasias Gástricas/prevención & control , beta-Criptoxantina , Factores de Riesgo , Carotenoides/uso terapéuticoRESUMEN
CONTEXT AND OBJECTIVES: Persistent organic pollutants (POPs) are a group of organic chemical compounds potentially toxic to human health. The objectives of this study were 1) to describe the levels of POPs biomarkers in blood samples from French women collected during the 1990s and to compare them with levels measured in two more recent French studies, 2) to identify POPs exposure profiles, and 3) to explore their main determinants. METHODS: 73 POPs biomarkers were measured in the blood of 468 women from the French E3N cohort (aged 45-73 years), collected between 1994 and 1999: 28 per- and polyfluoroalkyl substances, 27 organochlorine pesticides, 14 polychlorinated biphenyls and 4 polybrominated diphenyl ethers. POPs biomarker levels were described and compared with levels measured in two more recent French studies conducted by the French National Public Health Agency, the ENNS and Esteban studies. Principal component analysis was performed on POPs quantified in at least 75% of samples to identify the main exposure profiles. Linear regression models were used to estimate the associations between anthropometric, socio-demographic and lifestyle characteristics and exposure to these profiles. RESULTS: Among the 73 biomarkers measured, 41 were quantified in more than 75% of samples. Levels of most pollutants that were also measured in the Esteban of ENNS studies have decreased over time. Six POPs exposure profiles were revealed, explaining 62.1% of the total variance. Most of the characteristics studied were associated with adherence to at least one of these profiles. CONCLUSION: This study highlighted that most of the pollutants for which a comparison was possible decreased over the 10 or 20 years following the E3N blood collection, and identified those which, on the contrary, tended to increase. The health effects of the profiles identified could be assessed in future studies. The determinants identified should be confirmed in larger populations.
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Biomarcadores , Contaminantes Orgánicos Persistentes , Humanos , Femenino , Francia , Persona de Mediana Edad , Anciano , Contaminantes Orgánicos Persistentes/sangre , Biomarcadores/sangre , Exposición a Riesgos Ambientales/análisis , Estudios de Cohortes , Hidrocarburos Clorados/sangre , Contaminantes Ambientales/sangre , Monitoreo del AmbienteRESUMEN
BACKGROUND: Semen cryopreservation is a critical tool for breed improvement and preservation of biodiversity. However, instability of sperm freezability affects its application. The Mediterranean buffalo is one of the river-type buffaloes with the capacity for high milk production. Until now, there is no specific cryopreservation system for Mediterranean buffalo, which influences the promotion of excellent cultivars. To improve the semen freezing extender used in cryopreservation of Mediterranean buffalo, different protein datasets relating to freezability sperm were analyzed by iTRAQ-based proteomics. This study will be beneficial for further understanding the sperm freezability mechanism and developing new cryopreservation strategy for buffalo semen. RESULTS: 2652 quantified proteins were identified, including 248 significantly differentially expressed proteins (DEP). Gene Ontology (GO) analysis indicated that many these were mitochondrial proteins, enriched in the molecular function of phospholipase A2 activity and enzyme binding, and biological processes of regulation of protein kinase A signaling and motile cilium assembly. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis identified 17 significant pathways, including oxidative phosphorylation (OXPHOS). Furthermore, 7 DEPs were verified using parallel reaction monitoring or western blot, which confirmed the accuracy of the iTRAQ data. Peroxiredoxin 6 (PRDX6), which expressed 1.72-fold higher in good freezability ejaculate (GFE) compared to poor freezability ejaculate (PFE) sperms, was selected to explore the function in sperm freezability by adding recombinant PRDX6 protein into the semen freezing extender. The results showed that the motility, mitochondrial function and in vitro fertilization capacity of frozen-thawed sperm were significantly increased, while the oxidation level was significantly decreased when 0.1 mg/L PRDX6 was added compared with blank control. CONCLUSIONS: Above results revealed the metabolic pattern of freezability of Mediterranean buffalo sperms was negatively associated with OXPHOS, and PRDX6 had protective effect on cryo-damage of frozen-thawed sperms.
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Búfalos , Preservación de Semen , Animales , Masculino , Peroxiredoxina VI/genética , Peroxiredoxina VI/análisis , Proteómica , Preservación de Semen/veterinaria , Preservación de Semen/métodos , Espermatozoides/fisiología , Criopreservación/veterinaria , Criopreservación/métodos , Proteínas Recombinantes , Motilidad EspermáticaRESUMEN
Semen cryopreservation is used for the propagation of variety among species and domestic breeding. Mitochondria are implicated in sperm freezability, and their proteins are prone to succinylation, but the relationship between sperm freezability and mitochondrial protein succinylation is unclear. In this study, six bulls were classified as having good or poor freezability ejaculates (GFE or PFE, each 3 bulls). The fresh sperm mitochondrial membrane potential (MMP) and pan succinylation level of the two groups were first detected. Then the lysine succinylome and fatty acid content of the two groups were analyzed using label-free LC-MS/MS and GC-MS/MS in multiple reaction monitoring (MRM) modes, respectively. The results indicated that the GFE sperm had significantly higher MMPs than the PFE group (p < 0.05). A total of 1393 succinylation sites corresponding to 426 proteins were assessed and 5 succinylated peptides of the GFE group were markedly upregulated, while 3 were significantly downregulated (FC > 2.0 - < 0.5 and p-value < 0.05) when compared to the PFE group. Forty-six succinylated proteins were identified to have consistent presence/absence expression. The upregulated succinylated proteins in the GFE sperm were enriched in lipid metabolic processes. A total of 31 fatty acids were further subjected to quantitative analysis of which 23 including arachidic (C20:0), linolenic (C18:3n3), and docosahexaenoic acids (C22:6n3) were decreased in GFE sperm when compared with PFE (p < 0.05). These results suggest that lysine succinylation can potentially influence the sperm freezability of Mediterranean buffaloes through mitochondrial lipid metabolism. This novel study provides our understanding of sperm succinylation and the molecular basis for the mechanism of sperm freezability.
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Preservación de Semen , Animales , Masculino , Preservación de Semen/métodos , Búfalos/metabolismo , Lisina/metabolismo , Metabolismo de los Lípidos , Cromatografía Liquida , Espectrometría de Masas en Tándem , Espermatozoides/metabolismo , Criopreservación/métodosRESUMEN
The cryotolerance of semen obtained from Mediterranean buffalo bulls usually is more likely to deteriorate during the summer. To obtain the optimal sperm for fertility, the physiological status and reproductive performance of Mediterranean buffalo bulls in the summer and spring were first analysed by assessing blood serum and seminal plasma samples; then, the lipid profiles of seminal plasma were investigated by LC-MS/MS. The T, T3 and SOD levels of serum and seminal plasma in the spring were significantly higher than in the summer (p < .05). The results suggest that T3 level is positively correlated with semen cryotolerance; sphingolipids are potential markers for semen cryotolerance of Mediterranean buffalo. To our knowledge, this is the first report of targeted lipidomics in semen cryotolerance.
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Bison , Preservación de Semen , Masculino , Animales , Semen/fisiología , Búfalos/fisiología , Cromatografía Liquida/veterinaria , Espectrometría de Masas en Tándem/veterinaria , Espermatozoides/fisiología , Análisis de Semen/veterinaria , Análisis de Semen/métodos , Preservación de Semen/veterinaria , Preservación de Semen/métodos , LípidosRESUMEN
The aim of this study was to investigate the effect of circCUL2 on the proliferation, invasion and migration of retinoblastoma cells by regulating the miR-214-5p/E2F2 axis. qRT-PCR and western blot were performed to detect the expressions of circCUL2, miR-214-5p and E2F2 in tumor tissues and adjacent normal tissues from retinoblastoma patients, and in normal human retinal epithelial cells ARPE-19 and human retinoblastoma cells Y79 and SO-Rb50. qRT-PCR and western blot were performed for the detection of RNA levels of circCUL2 and miR-214-5p and the mRNA and protein levels of E2F2, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay for cell proliferation ability, Transwell assay for cell invasion ability, and scratch assay for cell migration ability. Luciferase dual reporter assay was used to detect the targeting relationship between circCUL2 and miR-214-5p, and between miR-214-5p and E2F2. CircCUL2 and E2F2 were lowly expressed, while miR-214-5p was highly expressed in retinoblastoma tumor tissues and cells. Transfection with pcDNA3.1-CUL2 or miR-214-5p inhibitor inhibited the proliferation, invasion and migration of Y79 and SO-Rb50 cells compared with the negative control; while transfection with sh-CUL2 or miR-214-5p mimics promoted the proliferation, invasion and migration of Y79 and SO-Rb50 cells. CircCUL2 negatively regulated miR-214-5p, while miR-214-5p negatively regulated E2F2. Overexpression of miR-214-5p or silencing of E2F2 in SO-Rb50 cells partially reversed the inhibitory effect of circCUL2 on the proliferation, invasion and migration of retinoblastoma cells. CircCUL2 inhibited the proliferation, invasion and migration of retinoblastoma cells by regulating the miR-214-5p/E2F2 axis.
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Proteínas Cullin/genética , Factor de Transcripción E2F2/genética , MicroARNs/genética , Neoplasias de la Retina/genética , Retinoblastoma/genética , Apoptosis/fisiología , Línea Celular Tumoral , Movimiento Celular/fisiología , Proliferación Celular/fisiología , Regulación Neoplásica de la Expresión Génica , Humanos , ARN Circular/genética , ARN Mensajero/genética , Retinoblastoma/patologíaRESUMEN
The Switch/sucrose nonfermentable (SWI/SNF) chromatin remodelling complex is closely related to chromatin openness and gene transcriptional activity. To understand if the chromatin openness of donor cells was related to the development efficiency of somatic cell cloning embryos, two buffalo fetal fibroblasts (BFF), BFF1 and BFF3, with significantly different cloned blastocyst development rates (18.4% and 30.9% respectively), were selected in this study. The expression of SWI/SNF complex genes, chromatin openness, and transcript level of these two cell lines were analysed, and the effect of ATP on the expression of the SWI/SNF complex genes was further explored. The results showed that compared with BFF1, the expression of SWI/SNF complex family genes was higher in BFF3 at the G0/G1 phase, where SMARCC1, SMARCC2 and SMARCE1 were significantly different (p < .05). Assay of Transposase Accessible Chromatin sequencing (ATAC-seq) results showed that, at the genome-wide level, BFF3 had more open chromatin, especially which having more open chromatin peaks at SMARCA4, SMARCA2, and RBPMS2 (RNA Binding Protein, mRNA Processing Factor 2) sites. In total, 2,712 differentially expressed genes (DEGs) were identified by the RNA-Seq method, with 1380 up- and 1332 down-regulated genes in BFF3. Interestingly, the ATPase-related genes ATP1B1 and ATP11A were extreme significantly up-regulated in BFF3 (p < .01). The ATP content and the expression of SWI/SNF complex genes in both BFF1 and BFF3 decreased when treated with rotenone. The above results demonstrated that the SWI/SNF complex contributed to chromatin opening, and chromatin opening of donor cells was essential for cloned embryo development.
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Búfalos , Cromatina , Adenosina Trifosfato , Animales , Búfalos/genética , ARN Mensajero , Rotenona , Sacarosa , TransposasasRESUMEN
OBJECTIVE: To investigate the correlation of clinical measurements on normal and abnormal fasting blood glucose (FBG) with benign prostatic hyperplasia (BPH). METHODS: From September 2016 to January 2018, 771 BPH patients were enrolled for further selection. The eligible patients were divided into normal FBG, impaired fasting glucose (IFG), and high risk of type 2 diabetes mellitus (HR-T2DM) groups. Then, relevant parameters were compared among these three groups using Pearson's correlation coefficient. RESULTS: Finally including 443 patients with normal FBG, 113 with IFG and 56 with HR-T2DM. Height, weight, body mass index, smoking status, hemoglobin, serum Na+, serum Cl-, and serum Ca2+ were significantly different between normal and abnormal FBG groups. In IFG/HR-T2DM group, obviously connections were demonstrated for weight with prostate volume (PV), for serum Na+, PV, and serum Cl- with total prostate-specific antigen (t-PSA), for FBG with international prostate symptom score (IPSS). In normal FBG group, significant correlations of age, weight, body mass index, hemoglobin, and serum Ca2+ with PV, of age, systolic blood pressure, PV, and serum Cl- with t-PSA; and of FBG, hemoglobin, and serum Na+ with IPSS were also observed. CONCLUSIONS: Our study suggests that FBG level probably plays an important role in BPH.
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Diabetes Mellitus Tipo 2 , Hiperplasia Prostática , Glucemia , Índice de Masa Corporal , Ayuno , Humanos , MasculinoRESUMEN
Calcium-activated chloride channels (CaCCs)/TMEM16A control diverse fundamental physiological functions, and abnormal function of TMEM16A will lead to various diseases including asthma, hypertension, gastrointestinal hypomotility and cancers. Therefore, TMEM16A as drug targets for related diseases has been increasingly concerned by researchers. In this work, COS were reported as novel natural activators of TMEM16A. It was demonstrated that COS can activate TMEM16A in a concentration dependent manner, with an EC50 of 74.5⯵g/mL. Then, fluorescence experiments and inside-out patch clamp experiments were combined to confirm that COS can directly activate TMEM16A. Further, we compared the activation effects of COS monomers DP2 to DP6, with DP3 the best activator. Molecular simulation was performed to find that the binding sites between DP3 and TMEM16A are E143 and E146 in TMEM16A, and it was speculated that COS and TMEM16A may be combined by electrostatic interaction. Finally, we verified that guinea pig ileum contraction was promoted by COS and the monomers through activating TMEM16A. Collectively, COS are novel efficient natural activators of TMEM16A, with potential to be developed to treatment diseases caused by down-regulation of TMEM16A including gastrointestinal hypomotility.
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Anoctamina-1/metabolismo , Quitosano/química , Quitosano/farmacología , Canales de Cloruro/metabolismo , Oligosacáridos/química , Oligosacáridos/farmacología , Animales , Sitios de Unión/efectos de los fármacos , Calcio/metabolismo , Línea Celular , Cobayas , Células HEK293 , Humanos , Íleon/efectos de los fármacos , Íleon/metabolismo , Ratones , Proteínas de Neoplasias/metabolismoRESUMEN
In this paper, the nitrogen atom was inserted into the anthracycline system of the isocryptolepine nucleus to obtain the "Aza"-type structure benzo[4,5]imidazo[1,2-c] quinazoline. A series of "Aza"-type derivatives were designed, synthesized and evaluated for their antifungal activity against six plant fungi in vitro. Among all derivatives, compounds A-0, B-1 and B-2 showed significant antifungal activity against B. cinerea with the EC50 values of 2.72⯵g/mL, 5.90⯵g/mL and 4.00⯵g/mL, respectively. Compound A-2 had the highest activity against M. oryzae with the EC50 values of 8.81⯵g/mL, and compound A-1 demonstrated the most control efficacy against R. solani (EC50, 6.27⯵g/mL). Moreover, compound A-0 was selected to investigate the in vivo tests against B. cinerea and the results indicated that the preventative efficacy of it up to 72.80% at 100⯵g/mL. Preliminary mechanism studies revealed that after treatment with A-0 at 5⯵g/mL, the B. cinerea mycelia appeared curved, collapsed and the cell membrane integrity may be damaged. The reactive oxygen species production, mitochondrial membrane potential and nuclear morphometry of mycelia have been changed, and the membrane function and cell proliferation of mycelia were destroyed. Compounds A-0, A-1, B-1 and B-2 presented weaker toxicities against two cells lines than isocryptolepine. This study lays the foundation for the future development of isocryptolepine derivatives as environmentally friendly and safe agricultural fungicides.
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Antifúngicos/farmacología , Diseño de Fármacos , Hongos/efectos de los fármacos , Fungicidas Industriales/farmacología , Alcaloides Indólicos/farmacología , Quinolinas/farmacología , Antifúngicos/síntesis química , Antifúngicos/química , Relación Dosis-Respuesta a Droga , Fungicidas Industriales/síntesis química , Fungicidas Industriales/química , Alcaloides Indólicos/síntesis química , Alcaloides Indólicos/química , Pruebas de Sensibilidad Microbiana , Estructura Molecular , Plantas/microbiología , Quinolinas/síntesis química , Quinolinas/química , Relación Estructura-ActividadRESUMEN
Autophagy is essential for oocyte maturation and preimplantation embryo development. ATG4C, a member of the ATG4 family, plays a crucial role in the autophagy process. The effect of ATG4C on the early embryonic development in pig has not been studied. In this study, the expression patterns of ATG4C were explored using qRT-PCR and immunofluorescence staining. Different concentrations of serum were added to in vitro maturation (IVM) medium to investigate its effects on oocyte maturation and embryonic development. Finally, the developmental potential of parthenogenetic embryos was detected by downregulating ATG4C in MII stage oocytes under 0 % serum condition. The results revealed that ATG4C was highly expressed in porcine oocytes matured in vitro and in parthenogenetic embryos. Compared with the 10 % serum group, the cumulus cell expansion, first polar body (PB1) extrusion rate, and subsequent developmental competence of embryos were reduced in the 0 % and 5 % serum groups. The mRNA levels of LC3, ATG5, BECLIN1, TFAM, PGC1α, and PINK1 were significantly increased (P < 0.05) in the 0 % serum group. ATG4C was significantly upregulated in the embryos at the 1-cell, 2-cell, 8-cell, and 16-cell stages in the 0 % serum group (P < 0.05). Compared with the negative control group, downregulation of ATG4C significantly decreased the 4-cell, 8-cell, and blastocyst rates (P < 0.05), and the expression of genes related to autophagy, mitochondria, and zygotic genome activation (ZGA) was significantly decreased (P < 0.05). The relative fluorescence intensity of LC3 and mitochondrial content in the ATG4C siRNA group was significantly reduced (P < 0.05). Collectively, the results indicate that ATG4C is highly expressed in porcine oocytes matured in vitro and in early embryos, and inhibition of ATG4C effects embryonic developmental competence by decreasing autophagy, mitochondrial content, and ZGA under serum-free condition.
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Desarrollo Embrionario , Regulación del Desarrollo de la Expresión Génica , Técnicas de Maduración In Vitro de los Oocitos , Oocitos , Animales , Porcinos/embriología , Oocitos/metabolismo , Desarrollo Embrionario/fisiología , Técnicas de Maduración In Vitro de los Oocitos/veterinaria , Proteínas Relacionadas con la Autofagia/genética , Proteínas Relacionadas con la Autofagia/metabolismo , Técnicas de Cultivo de Embriones/veterinaria , Femenino , Autofagia , PartenogénesisRESUMEN
The epigenetic modification levels of donor cells directly affect the developmental potential of somatic cell nuclear transfer (SCNT) embryos. BRG1, as an epigenetic modifying enzyme, has not yet been studied in donor cells and SCNT embryos. In this study, BRG1 was overexpressed in porcine fetal fibroblasts (PFFs), its effect on chromatin openness and gene transcription was examined, subsequently, the development potential of porcine SCNT embryos was investigated. The results showed that compared with the control group, the percentage of G1 phase cells was significantly increased (32.3 % ± 0.87 vs 25.7 % ± 0.81, P < 0.05) in the experimental group. The qRT-PCR results showed that the expression of H3K9me3-related genes was significantly decreased (P < 0.05), HAT1 was significantly increased (P < 0.05). Assay of Transposase Accessible Chromatin sequencing (ATAC-seq) results revealed that SMARCA4ãNANOGãSOX2ãMAP2K6 and HIF1A loci had more open chromatin peaks in the experimental group. The RNA-seq results showed that the upregulated genes were mainly enriched in PI3K/AKT and WNT signaling pathways, and the downregulated genes were largely focused on disease development. Interestingly, the developmental rate of porcine SCNT embryos was improved (27.33 % ± 1.40 vs 17.83 % ± 2.02, P < 0.05), the expression of zygotic gene activation-related genes in 4-cell embryos, and embryonic development-related genes in blastocysts was significantly upregulated in the experimental group (P < 0.05). These results suggest that overexpression of BRG1 in donor cells is benefit for the developmental potential of porcine SCNT embryos.
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Técnicas de Transferencia Nuclear , Fosfatidilinositol 3-Quinasas , Animales , Porcinos , Fosfatidilinositol 3-Quinasas/metabolismo , Técnicas de Transferencia Nuclear/veterinaria , Blastocisto/metabolismo , Epigénesis Genética , Desarrollo Embrionario , Cromatina/metabolismo , Embrión de Mamíferos/metabolismo , Clonación de Organismos/veterinariaRESUMEN
Brahma-related gene 1 (BRG1) enhances the pluripotency of embryonic and adult stem cells, however, its effect on induced pluripotent stem cell (iPSC) pluripotency has not been reported. The aim of this study was to investigate the effect of BRG1 on porcine iPSC pluripotency and its mechanisms. The effect of BRG1 on porcine iPSC pluripotency was explored by positive and negative control it. The mechanism was investigated by regulating the WNT/ß-catenin signaling pathway and autophagy flux. The results showed that inhibition of BRG1 decreased pluripotency-related gene expression in porcine iPSCs; while its overexpression had the opposite effect, the expression of WNT/ß-catenin signaling pathway- and autophagy-related genes was significantly up-regulated (P < 0.05) in the BRG1 overexpressed group when compared to the control group. Inhibited pluripotency-related gene or protein expression, decreased autophagy flux, and increased mitochondrial length and mitochondrial membrane potential (MMP) were observed when porcine iPSCs were treated with the WNT/ß-catenin signaling pathway inhibitor IWR-1. Forced BRG1 expression restored porcine iPSC pluripotency, increased autophagy flux, shortened mitochondria, and reduced MMP. Lastly, Compound C was used to activate porcine iPSC autophagy, and it was found that the expression of BRG1 and ß-catenin increased, and pluripotency-related gene and protein expression was up-regulated; these effects were reversed when the BRG1 inhibitor PFI-3 and IWR-1 were added. These results suggested that BRG1 enhanced the pluripotency of porcine iPSCs through WNT/ß-catenin and autophagy pathways.
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Células Madre Pluripotentes Inducidas , beta Catenina , Animales , Porcinos , beta Catenina/genética , Vía de Señalización Wnt/genética , Factores de Transcripción/genética , Factores de Transcripción/metabolismo , AutofagiaRESUMEN
Compared to traditional lithium metal batteries, anode-free lithium metal batteries use bare current collectors as an anode instead of Li metal, making them highly promising for mass production and achieving high-energy density. The current collector, as the sole component of the anode, is crucial in lithium deposition-stripping behavior and greatly impacts the rate of Li depletion from the cathode. In this study, to investigate the lithiophilicity effect of the current collector on the solid electrolyte interface (SEI) film construction and cycling performance of anode-free lithium batteries, various lightweight paper-based current collectors were prepared by electroless plating Cu and lipophilic Ag on low-dust paper (LDP). The areal densities of the as-prepared LDP@Cu, LDP@Cu-Ag, and LDP@Ag were approximately 0.33 mg cm-2. The use of lipophilic Ag-coated collectors with varying loadings allowed for the regulation of lipophilicity. The impacts of these collectors on the distribution of SEI components and Li depletion rate in common electrolytes were investigated. The findings suggest that higher loadings of lipophilic materials, such as Ag, on the current collector increase its lipophilicity but also lead to significant Li depletion during the cycling process in full-cell anode-free Li metal batteries. Thus, moderately lithiophilic current collectors, such as LDP@Cu-Ag, show more potential for Li deposition and striping and stable SEI with a low speed of Li depletion.
RESUMEN
Ultrasound Localization Microscopy (ULM) surpasses the constraints imposed by acoustic diffraction, achieving sub-wavelength resolution visualization of microvasculature through the precise localization of minute microbubbles (MBs). Nonetheless, the analysis of densely populated regions with overlapping MB point spread responses introduces significant localization errors, limiting the use of technique to low-concentration conditions. This raises a trade-off issue between localization efficiency and MB density. In this work, we present a new deep learning framework that combines Transformer and U-Net architectures, termed ULM-TransUNet. As a non-linear model, it is able to learn the complex data patterns of overlapping MBs in dense conditions for accurate localization. To evaluate the performance of ULM-TransUNet, a series of numerical simulations and in vivo experiments are carried out. Numerical simulation results indicate that ULM-TransUNet achieves high-quality ULM imaging, with improvements of 21.93 % in detection rate, 17.36 % in detection precision, and 20.53 % in detection sensitivity, compared to previous state-of-the-art deep learning (DL) method (e.g., ULM-UNet). For the in vivo experiments, ULM-TransUNet achieves the highest spatial resolution (9.4 µm) and rapid inference speed (26.04 ms/frame). Furthermore, it consistently detects more small vessels and resolves closely spaced vessels more effectively. The outcomes of this work imply that ULM-TransUNet can potentially enhance the microvascular imaging performance on high-density MB conditions.
Asunto(s)
Microburbujas , Ultrasonografía , Animales , Ultrasonografía/métodos , Microscopía/métodos , Aprendizaje Profundo , Microvasos/diagnóstico por imagen , Ratones , Simulación por ComputadorRESUMEN
Lithium-sulfur (Li-S) batteries are expected to be one of the next generations of high-energy-density battery systems due to their high theoretical energy density of 2600 Wh kg-1. Embracing the trends toward flexibility, lightweight design, and cost-effectiveness, paper-based electrodes offer a promising alternative to traditional coated cathodes in Li-S batteries. Within paper-based electrodes, conductive fibers such as carbon nanotubes (CNTs) play a crucial role. They help to form a three-dimensional network within the paper matrix to ensure structural integrity over extended cycling while mitigating the shuttle effect by confining sulfur within the cathode. Herein, we explore how variously functionalized CNTs, serving as conductive fibers, impact the physical and electrochemical characteristics of paper-based sulfur cathodes in Li-S batteries. Specifically, graphitized hydroxylated carbon nanotubes (G-CNTs) exhibit remarkable capacity at low currents owing to their excellent conductivity and interaction with lithium polysulfide (LiPS), achieving the highest initial specific capacity of 1033 mAh g-1 at 0.25 C (1.1 mA cm-2). Aminated multi-walled carbon nanotubes (NH2-CNTs) demonstrate an enhanced affinity for LiPS due to the -NH2 groups. However, the uneven distribution of these fibers may induce electrode surface passivation during charge-discharge cycles. Notably, hydroxylated multi-walled carbon nanotubes (OH-CNTs) can establish a uniform and stable 3D network with plant fibers, showcasing superior mechanical properties and helping to mitigate Li2S agglomeration while preserving the electrode porosity. The paper-based electrode integrated with OH-CNTs even retains a specific capacity of approximately 800 mAh g-1 at about 1.25 C (5 mA cm-2), demonstrating good sulfur utilization and rate capacity compared to other CNT variants.
RESUMEN
2-Phosphonobutane-1,2,4,-tricarboxylic acid (PBTC) is an orthophosphate compound widely used as an antiscalant chemical and corrosion inhibitor in manufacturing. However, PBTC poses persistent environmental concerns due to its stability and resistance to conventional water treatment. In addressing the issues of PBTC in aquatic systems, Al-based metal-organic frameworks (MOFs) have been developed and applied as sustainable adsorbents. The materials are synthesized from terephthalic acid (TPA) linkers derived from upcycling products of post-consumer polyethylene terephthalate (PET) bottles. The PET-derived linker was prepared using alkaline hydrolysis followed by acidification and employed in forming MIL-53 (Al), with a comparative assessment against the corresponding MOFs made from commercial-grade TPA. The structures and properties of the materials were characterized with microscopic and spectroscopic methods. The synthesized adsorbents achieved a phosphate adsorption capacity of 826 mg/g at pH 5, with kinetics fitting a pseudo-second-order model and isotherm patterns aligning with Langmuir, Freundlich, and Sips models, indicative of diverse adsorption on heterogeneous surfaces. The results highlight the role of electrostatic interactions and hydrogen bonding mechanisms in PBTC adsorption. The eco-friendly materials with high adsorption performance offer an innovative route for sustainable waste management and water purification.