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The ubiquitination function in diabetic nephropathy (DN) has attracted much attention, but there is a lack of information on its ubiquitylome profile. To examine the differences in protein content and ubiquitination in the kidney between db/db mice and db/m mice, we deployed liquid chromatography-mass spectrometry (LC-MS/MS) to conduct analysis. We determined 145 sites in 86 upregulated modified proteins and 66 sites in 49 downregulated modified proteins at the ubiquitinated level. Moreover, 347 sites among the 319 modified proteins were present only in the db/db mouse kidneys, while 213 sites among the 199 modified proteins were present only in the db/m mouse kidneys. The subcellular localization study indicated that the cytoplasm had the highest proportion of ubiquitinated proteins (31.87%), followed by the nucleus (30.24%) and the plasma membrane (20.33%). The enrichment analysis revealed that the ubiquitinated proteins are mostly linked to tight junctions, oxidative phosphorylation, and thermogenesis. Podocin, as a typical protein of slit diaphragm, whose loss is a crucial cause of proteinuria in DN. Consistent with the results of ubiquitination omics, the K261R mutant of podocin induced the weakest ubiquitination compared with the K301R and K370R mutants. As an E3 ligase, c-Cbl binds to podocin, and the regulation of c-Cbl can impact the ubiquitination of podocin. In conclusion, in DN, podocin ubiquitination contributes to podocyte injury, and K261R is the most significant site. c-Cbl participates in podocin ubiquitination and may be a direct target for preserving the integrity of the slit diaphragm structure, hence reducing proteinuria in DN.
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Nefropatías Diabéticas , Péptidos y Proteínas de Señalización Intracelular , Proteínas de la Membrana , Podocitos , Proteínas Proto-Oncogénicas c-cbl , Ubiquitinación , Animales , Nefropatías Diabéticas/metabolismo , Nefropatías Diabéticas/patología , Podocitos/metabolismo , Podocitos/patología , Ratones , Proteínas Proto-Oncogénicas c-cbl/metabolismo , Proteínas Proto-Oncogénicas c-cbl/genética , Péptidos y Proteínas de Señalización Intracelular/metabolismo , Péptidos y Proteínas de Señalización Intracelular/genética , Proteínas de la Membrana/metabolismo , Proteínas de la Membrana/genética , Masculino , Ratones Endogámicos C57BLRESUMEN
Oleanolic acid (OA) is a natural triterpenoid with therapeutic potential for a multitude of diseases. However, the precise mechanism by which OA influences stress-induced apoptosis of intestinal epithelial cells remains elusive. Therefore, the effect of OA on intestinal diseases under stressful conditions and its possible mechanisms have been investigated. In a hydrogen peroxide (H2 O2 )-induced oxidative stress model, OA attenuated H2 O2 -induced apoptosis in a concentration-dependent manner. To investigate the underlying mechanisms, the gene expression profile of OA on IPEC-J2 cells was analyzed using an RNA sequencing system. Results from gene ontology and Kyoto encyclopedia of genes and genomes analysis confirmed that OA may mitigate the cytotoxic effects of H2 O2 by downregulating gene expression through the MAPK signaling pathway. Furthermore, Quantitative real-time polymerase chain reaction results validated the differentially expressed genes data. Western blot analysis further demonstrated that OA effectively suppressed the expression level of c-Jun protein induced by H2 O2 in IPEC-J2 cells. Collectively, our results indicate that OA pretreatment significantly attenuated H2 O2 -induced apoptosis in intestinal epithelial cells through suppressing c-Jun and MAPK pathway.
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Peróxido de Hidrógeno , Ácido Oleanólico , Peróxido de Hidrógeno/farmacología , Peróxido de Hidrógeno/metabolismo , Ácido Oleanólico/farmacología , Línea Celular , Apoptosis , Estrés Oxidativo , Células Epiteliales/metabolismoRESUMEN
BACKGROUND: Nme2ABE8e has been constructed and characterized as a compact, accurate adenine base editor with a less restrictive dinucleotide protospacer-adjacent motif (PAM: N4CC) but low editing efficiency at challenging loci in human cells. Here, we engineered a subset of domain-inlaid Nme2Cas9 base editors to bring the deaminase domain closer to the nontarget strand to improve editing efficiency. RESULTS: Our results demonstrated that Nme2ABE8e-797 with adenine deaminase inserted between amino acids 797 and 798 has a significantly increased editing efficiency with a wide editing window ranging from 4 to 18 bases in mammalian cells, especially at the sites that were difficult to edit by Nme2ABE8e. In addition, by swapping the PAM-interacting domain of Nme2ABE8e-797 with that of SmuCas9 or introducing point mutations of eNme2-C in Nme2ABE8e-797, we created Nme2ABE8e-797Smu and Nme2ABE8e-797-C, respectively, which exhibited robust activities at a wide range of sites with N4CN PAMs in human cells. Moreover, the modified domain-inlaid Nme2ABE8e can efficiently restore or install disease-related loci in Neuro-2a cells and mice. CONCLUSIONS: These novel Nme2ABE8es with increased on-target DNA editing and expanded PAM compatibility will expand the base editing toolset for efficient gene modification and therapeutic applications.
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Proteína 9 Asociada a CRISPR , Sistemas CRISPR-Cas , Animales , Humanos , Ratones , Proteína 9 Asociada a CRISPR/genética , Adenina/química , Edición Génica/métodos , ADN/genética , Mamíferos/genéticaRESUMEN
Polyploids are essential in plant evolution and species formation, providing a rich genetic reservoir and increasing species diversity. Complex polyploids with higher ploidy levels often have a dosage effect on the phenotype, which can be highly detrimental to gametes, making them rare. In this study, offspring plants resulting from an autoallotetraploid (RRRC) derived from the interspecific hybridization between allotetraploid Raphanobrassica (RRCC, 2n = 36) and diploid radish (RR, 2n = 18) were obtained. Fluorescence in situ hybridization (FISH) using C-genome-specific repeats as probes revealed two main genome configurations in these offspring plants: RRRCC (2n = 43, 44, 45) and RRRRCC (2n = 54, 55), showing more complex genome configurations and higher ploidy levels compared to the parental plants. These offspring plants exhibited extensive variation in phenotypic characteristics, including leaf type and flower type and color, as well as seed and pollen fertility. Analysis of chromosome behavior showed that homoeologous chromosome pairing events are widely observed at the diakinesis stage in the pollen mother cells (PMCs) of these allopolyploids, with a range of 58.73% to 78.33%. Moreover, the unreduced C subgenome at meiosis anaphase II in PMCs was observed, which provides compelling evidence for the formation of complex allopolyploid offspring. These complex allopolyploids serve as valuable genetic resources for further analysis and contribute to our understanding of the mechanisms underlying the formation of complex allopolyploids.
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Aneuploidia , Cromosomas de las Plantas , Poliploidía , Raphanus , Raphanus/genética , Cromosomas de las Plantas/genética , Hibridación Fluorescente in Situ , Brassica/genética , Hibridación Genética , Meiosis/genética , Genoma de Planta , Polen/genética , FenotipoRESUMEN
Although large-scale and accurate identification of cis-regulatory elements on pig protein-coding and long non-coding genes has been reported, similar study on pig miRNAs is still lacking. Here, we systematically characterized the cis-regulatory elements of pig miRNAs in muscle and fat by adopting miRNAomes, ChIP-seq, ATAC-seq, RNA-seq and Hi-C data. In total, the cis-regulatory elements of 257 (85.95%) expressed miRNAs including 226 known and 31 novel miRNAs were identified. Especially, the miRNAs associated with super-enhancers, active promoters, and "A" compartment were significantly higher than those associated by typical enhancers, prompters without H3K27ac, and "B" compartment, respectively. The tissue specific transcription factors were the primary determination of core miRNA expression pattern in muscle and fat. Moreover, the miRNA promoters are more evolutionarily conserved than miRNA enhancers, like other type genes. Our study adds additional important information to existing pig epigenetic data and provides essential resource for future in-depth investigation of pig epigenetics.
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Epigenómica , MicroARNs , Animales , Secuenciación de Inmunoprecipitación de Cromatina , MicroARNs/genética , Músculos , Secuencias Reguladoras de Ácidos Nucleicos , Porcinos/genéticaRESUMEN
BACKGROUND: Tumor cells adapt to hypoxia by regulating transcription factors that involved in regulation of metabolism, angiogenesis, cell proliferation, and apoptosis. Under hypoxic conditions, hypoxia-inducible factor-1 (HIF-1), consisting of HIF-1α and HIF-1ß subunits, acts as a key transcription factor mediating the adaptive cellular responses. Caspase-3 is a key apoptosis-related protease that plays a role in tumor growth and development. Studies have shown that caspase-3 could be regulated by HIF-1α under pathological conditions. Therefore, HIF-1α and caspase-3 expression may be related to the poor prognosis of tumors. In this study, we analyzed the possible relationships between these two signaling factors in correlation with the clinical behavior of PTC. METHODS: We detected the expression levels of HIF-1α and caspase-3 in 70 samples of PTC and para-cancerous tissues (control group) by immunohistochemistry (IHC). Furthermore, various clinicopathological parameters were assessed to determine their correlations with HIF-1α and caspase-3 expressions. RESULTS: First, HIF-1α and caspase-3 expressions (60% and 37.1%, respectively) increased significantly in the PTC samples as compared to normal tissues (2.9% for both HIF-1α and caspase-3) (p < 0.05) as determined by IHC. Second, although there was no significant difference between the expression of HIF-1α and caspase-3 in regard to gender, age distribution, tumor size, lymph node metastasis, and BRAFV600E mutation (all p > 0.05), HIF-1α and caspase-3 expressions were associated with capsule invasion and cell subtypes of PTC (p < 0.05). The percent positivity of caspase-3 expression in tall-cell variant (TCV) was the highest (63.6%). Third, HIF-1α expression was positively correlated with that of caspase-3 (rs = 0.326; p < 0.05). CONCLUSIONS: Overexpression of HIF-1α and caspase-3 is associated with carcinogenesis. These factors might serve as promising predictors of aggressive PTC. These findings also suggest their potential as therapeutic targets.
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Subunidad alfa del Factor 1 Inducible por Hipoxia , Neoplasias de la Tiroides , Humanos , Cáncer Papilar Tiroideo , Caspasa 3 , Inmunohistoquímica , Neoplasias de la Tiroides/patología , HipoxiaRESUMEN
The stubborn and complex structure of lignocellulose hinders the valorization of each component of cellulose, hemicellulose, and lignin in the biorefinery industries. Therefore, efficient pretreatment is an essential and prerequisite step for lignocellulose biorefinery. Recently, a considerable number of studies have focused on peroxyacetic acid (PAA) pretreatment in lignocellulose fractionation and some breakthroughs have been achieved in recent decades. In this article, we aim to highlight the challenges of PAA pretreatment and propose a roadmap towards lignocellulose fractionation by PAA for future research. As a novel promising pretreatment method towards lignocellulosic fractionation, PAA is a strong oxidizing agent that can selectively remove lignin and hemicellulose from lignocellulose, retaining intact cellulose for downstream upgrading. PAA in lignocellulose pretreatment can be divided into commercial PAA, chemical activation PAA, and enzymatic in-situ generation of PAA. Each PAA for lignocellulose fractionation shows its own advantages and disadvantages. To meet the theme of green chemistry, enzymatic in-situ generation of PAA has aroused a great deal of enthusiasm in lignocellulose fractionation. Furthermore, mass balance and techno-economic analyses are discussed in order to evaluate the feasibility of PAA pretreatment in lignocellulose fractionation. Ultimately, some perspectives and opportunities are proposed to address the existing limitations in PAA pretreatment towards biomass biorefinery valorization. In summary, from the views of green chemistry, enzymatic in-situ generation of PAA will become a cutting-edge topic research in the lignocellulose fractionation in future.
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Lignina , Ácido Peracético , Biomasa , Celulosa , Lignina/química , Oxidantes , Ácido Peracético/farmacologíaRESUMEN
Hsp90 has long been recognized as an attractive and crucial molecular target for cancer therapy. Gambogic acid (GA), the main active compound of Gamboge hanburyi, has been reported as a natural inhibitor of Hsp90. Here, we present the structure-activity relationship of Garcinia xanthones analogues as Hsp90 inhibitors and identify that compound 25, with a simplified skeleton, had an improved inhibitory effect toward Hsp90. Compound 25 inhibited the ATPase activity of Hsp90 with an IC50 value of 3.68±0.18µM. It also exhibited potent antiproliferative activities in some solid tumor cells. In SK-BR-3 cells with high Hsp90 expression, compound 25 induced the degradation of Hsp90 client proteins including Akt and Erk1/2 without causing the heat shock response. Additionally, compound 25 inhibited angiogenesis in HUVEC cells through Hsp90 regulation of the HIF-1α pathway. These results demonstrate that compound 25 as an Hsp90 inhibitor with a new structure could be further studied for the development of tumor therapy.
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Inhibidores de la Angiogénesis/química , Inhibidores de la Angiogénesis/farmacología , Garcinia/química , Proteínas HSP90 de Choque Térmico/antagonistas & inhibidores , Xantonas/química , Xantonas/farmacología , Antineoplásicos/química , Antineoplásicos/farmacología , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Proteínas HSP90 de Choque Térmico/metabolismo , Células Endoteliales de la Vena Umbilical Humana , Humanos , Subunidad alfa del Factor 1 Inducible por Hipoxia/metabolismo , Neoplasias/tratamiento farmacológico , Neoplasias/metabolismo , Relación Estructura-ActividadRESUMEN
DDO-6101, a natural-product-like caged xanthone discovered previously in our laboratory based on the pharmacophoric scaffold of Garcinia natural product gambogic acid (GA), shows potent cytotoxicity in vitro but poor efficacy in vivo due to its poor druglike properties. In order to improve the druglike properties and in vivo cytotoxic potency, a novel series of 19 prenyl group-modified derivatives of DDO-6101 was synthesized and evaluated for their in vitro antitumor activity and druglike properties. The SAR and SPR information of these compounds was also obtained. In the light of the in vitro antitumor activity and druglike properties such as aqueous solubility and permeability, compound 6f (named as DDO-6306) was advanced into in vivo efficacy experiment. The results showed that DDO-6306 is more potent than DDO-6101 in vivo and is a promising antitumor candidate for further evaluation.
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Antineoplásicos/química , Antineoplásicos/farmacología , Productos Biológicos/química , Xantonas/química , Xantonas/farmacología , Animales , Antineoplásicos/síntesis química , Apoptosis/efectos de los fármacos , Peso Corporal/efectos de los fármacos , Línea Celular Tumoral , Permeabilidad de la Membrana Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Ensayos de Selección de Medicamentos Antitumorales , Garcinia/química , Garcinia/metabolismo , Humanos , Ratones , Relación Estructura-Actividad , Trasplante Heterólogo , Xantonas/síntesis químicaRESUMEN
Due to the presence of various harmful compounds in polluted water, toxicological tests should be combined to evaluate whether a body of water is polluted. The water quality of four rivers flowing into Dianchi Lake, which is well known for its heavy and lasting pollution, was investigated in this study using a combination of GC-MS analysis and cytotoxicity and mutagenicity tests. GC-MS analysis showed that the rivers investigated contained a variety of chemicals and suggested that severe water pollution existed. In addition, the water obtained from the four rivers induced acute cytotoxicity in CHO cells and dose-dependent mutagenicity in four Salmonella typhimurium strains (TA97, TA98, TA100 and TA102). The integration of GC-MS analysis and short-term in vitro toxicological tests might be a valuable approach that can be used to monitor the health hazards that may result from water pollution.
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Mutágenos/análisis , Ríos/química , Contaminantes Químicos del Agua/análisis , Animales , Cromatografía de Gases y Espectrometría de Masas , Lagos/química , Pruebas de Mutagenicidad , Mutágenos/toxicidad , Contaminantes Químicos del Agua/toxicidadRESUMEN
The growing interest in utilizing lignin for dye removal has gained momentum, but there is limited information on the intricate relationship between lignin structural characteristics and adsorption efficacy, especially for its biochar derivatives. This study focused on three types of lignin and their corresponding biochar derivatives. Among them, ZnCl2-activated acidic/alkali densified lignin preparation of lignin-derived active carbon exhibited superior adsorption performance, achieving 526.32 mg/g for methylene blue and 2156.77 mg/g for congo red. Its exceptional adsorption capacity was attributed to its unique structural properties, including low alkyl and O-alkyl group content and high aromatic carbon levels. Furthermore, the adsorption mechanisms adhered to pseudo-second-order kinetics and the Langmuir model, signifying a spontaneous process. Intriguingly, lignin-derived active carbon also demonstrated remarkable recovery capabilities. These findings provide valuable insights into the impact of structural attributes on lignin and its biochar's adsorption performance.
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Carbón Orgánico , Lignina , Lignina/química , Adsorción , Carbón Orgánico/química , Cinética , Azul de Metileno/química , Rojo Congo/químicaRESUMEN
The antimicrobial peptide (AMP) LI is a fusion product of antimicrobial peptide LL37 produced by human neutrophils and Indolicidin secreted by bovine neutrophils. LI retained the antimicrobial activity of the parental peptides and showed high cell selectivity. In this study, the flexible linker Gly-Ser-Gly (G-S-G) was used to ligate LI into dimeric LIG, and constructed the Pichia pastoris (P. pastoris) expression vector pPIC9K-6×His-3×FLAG-LIG. The total protein expression of P. pastoris GS115 reached the highest level (189.6â¯mg/L) after 96â¯h induction with 3 % methanol at the initial pH value of 7.0. Finally, 5.9â¯mg/L of recombinant LIG (rLIG) was obtained after enterokinase digestion and purification. The rLIG had high antimicrobial activity and low hemolytic activity. Compared with monomer LI, GSG linked dimeric LIG, which had no significant change in antimicrobial activity and had good salt ions stability. In this study, the dimeric antimicrobial peptide LIG was successfully expressed, which provided a new idea for the expression of AMPs in the P. pastoris expression system, and had important significance for the application of AMPs.
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Antiinfecciosos , Saccharomycetales , Animales , Bovinos , Humanos , Péptidos Antimicrobianos , Pichia/metabolismo , Antiinfecciosos/farmacología , Proteínas Recombinantes/genética , Proteínas Recombinantes/farmacologíaRESUMEN
The prevalence of struvite and other phosphate minerals in eutrophic environments has a significant effect on the transport and transformation of environmental heavy metals, but their competitive immobilization characteristics and mechanisms for heavy metals remain unclear. Three different sources of struvite (BS, CSHS, and CSS) were obtained respectively by biosynthesis and chemical synthesis with or without humic acid to investigate their competitive immobilization characteristics and mechanism of heavy metals in the Pb(II)-Cd(II)-Zn(II) composite system. The results showed that the immobilization of heavy metals by struvite is physico-chemical adsorption and the affinity (in descending order) is Pb(II) >> Cd(II)/Zn(II). Cd(II) promotes the immobilization of Pb(II)/Zn(II) by BS. The order of the selective strength by struvite for Pb(II) is BS >> CSS ≈ CSHS. The study indicates that the difference between struvite holding heavy metal ions is related to the material composition and heavy metal types, and BS shows best selective immobilization for Pb(II) in the Pb(II)-Cd(II)-Zn(II) composite system. This study provides a theoretical basis for understanding the environmental geochemical role and eco-environmental effects of struvite.
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Cadmio , Metales Pesados , Estruvita , Plomo , Adsorción , Metales Pesados/química , Zinc/químicaRESUMEN
Bioactive peptides (BPs) are protein fragments with beneficial effects on metabolism, physiology, and diseases. This review focuses on proteolytic BPs, which are produced by the action of gut microbiota on proteins in food and have demonstrated to influence the composition of gut microbes. And gut microbiota are candidate targets of BPs to alleviate oxidative stress, enhance immunity, and control diseases, including diabetes, hypertension, obesity, cancer, and immune and neurodegenerative diseases. Despite promising results, further research is needed to understand the mechanisms underlying the interactions between BPs and gut microbes, and to identify and screen more BPs for industrial applications. Overall, BPs offer potential as therapeutic agents for various diseases through their interactions with gut microbes, highlighting the importance of continued research in this area.
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Currently, the Cas9 and Cas12a systems are widely used for genome editing, but their ability to precisely generate large chromosome fragment deletions is limited. Type I-E CRISPR mediates broad and unidirectional DNA degradation, but controlling the size of Cas3-mediated DNA deletions has proven elusive thus far. Here, we demonstrate that the endonuclease deactivation of Cas9 (dCas9) can precisely control Cas3-mediated large-fragment deletions in mammalian cells. In addition, we report the elimination of the Y chromosome and precise retention of the Sry gene in mice using CRISPR/Cas3 and dCas9-controlled CRISPR/Cas3, respectively. In conclusion, dCas9-controlled CRISPR/Cas3-mediated precise large-fragment deletion provides an approach for establishing animal models by chromosome elimination. This method also holds promise as a potential therapeutic strategy for treating fragment mutations or human aneuploidy diseases that involve additional chromosomes.
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Proteínas Asociadas a CRISPR , Sistemas CRISPR-Cas , Ratones , Humanos , Animales , Edición Génica , Cromosoma Y , Proteínas Asociadas a CRISPR/genética , ADN/genética , Mamíferos/genéticaRESUMEN
Cold stress in low-temperature environments can trigger changes in gene expression, but epigenomics regulation of temperature stability in vital tissues, including the fat and diencephalon, is still unclear. Here, we explore the cold-induced changes in epigenomic features in the diencephalon and fat tissues of two cold-resistant Chinese pig breeds, Min and Enshi black (ES) pigs, utilizing H3K27ac CUT&Tag, RNA-seq, and selective signature analysis. Our results show significant alterations in H3K27ac modifications in the diencephalon of Min pigs and the fat of ES pigs after cold exposure. Dramatic changes in H3K27ac modifications in the diencephalon of Min pig are primarily associated with genes involved in energy metabolism and hormone regulation, whereas those in the fat of ES pig are primarily associated with immunity-related genes. Moreover, transcription factors PRDM1 and HSF1, which show evidence of selection, are enriched in genomic regions presenting cold-responsive alterations in H3K27ac modification in the Min pig diencephalon and ES pig fat, respectively. Our results indicate the diversity of epigenomic response mechanisms to cold exposure between Min and ES pigs, providing unique epigenetic resources for studies of low-temperature adaptation in large mammals.
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Acute kidney injury (AKI) is a major public health problem worldwide that still lacks effective treatments. Recent studies have suggested that ferroptosis is a key mediator of AKI due to its activation of lipid peroxidation. Therefore, we hypothesized that antiferroptosis agents might be a novel potential therapeutic strategy for AKI. Herein, we demonstrated that liquiritigenin (LG), an active ingredient of liquorice, improves renal function by inhibiting vitamin K epoxide reductase complex subunit 1 (VKORC1)-mediated ferroptosis, both in vivo and in vitro. In a folic acid-induced murine AKI model, after a single pre-treatment intravenous injection, LG markedly alleviated the loss of renal function through suppressing ferroptosis induced by iron accumulation. LG prevented mitochondrial morphological changes and upregulated glutathione and glutathione peroxidase 4 levels, while downregulating malonaldehyde and divalent iron levels. An in vitro RNA-sequence analysis suggested that the protective role of LG may involve upregulation of VKORC1. Moreover, knockdown of VKORC1 diminished the renal protective and antiferroptosis roles of LG. Collectively, our findings demonstrated that LG protected against AKI by inhibiting VKORC1-mediated ferroptosis. This suggests that inhibiting ferroptosis might be a novel therapeutic approach in the future.
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The force needed to buckle a thin elastic surface is proportional to its bending rigidity. This fact suggests using a buckling setup to measure the bending modulus of lipid membranes. Extending the work of Noguchi [Phys. Rev. E 83, 061919 (2011)], we systematically derive highly accurate analytical expressions for the forces along and perpendicular to the buckle, and we elucidate some of their counterintuitive properties using the framework of a surface stress tensor. Furthermore, we estimate the corrections to buckling forces due to thermal fluctuations and find them significant only for stresses along the ridges. We then apply this buckling protocol to four different lipid membrane models, which widely differ in their level of resolution and the treatment of solvent, and show that in all cases buckling is a reliable and accurate means for measuring their rigidity. Finally, we show that monitoring both stresses and energies during a simulation offers additional insights into the thermodynamics of curvature elasticity and permits one to predict the bending rigidity for a range of temperatures around the actual simulation temperature.
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Membrana Dobles de Lípidos/química , TermodinámicaRESUMEN
INTRODUCTION: Absent contractility is a kind of esophageal motility disorder. Patients are often diagnosed by endoscopic ultrasonography or high-resolution manometry (HRM). In this paper, we report two cases of absent esophageal contractility first discovered by transabdominal ultrasonography. CASE PRESENTATION: The main symptom of both cases was acid reflux, and the main diagnosis was absent esophageal contractility. They were all discovered by transabdominal ultrasonography. After the treatment of surgery or drugs, the symptoms relieved during follow-up, without any other discomforts. CONCLUSION: Transabdominal ultrasound as a primary screening method for absent contractility is proposed in this paper, which is non-invasive, real-time, and fast. Ultrasound is expected to improve the diagnostic efficiency of peristalsis and patient experience in combination with the above invasive examinations. The use of sonography in the diagnosis of absent contractility has seldom been studied. Further investigation is required to determine whether this technique may better assess absent contractility.
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In this study, an ammonia fiber expansion (AFEX)-assisted deep eutectic solvent (DES) pretreatment method was developed for the rapid separation of wheat straw fractions, which reduced the pretreatment time for DES and improved the pretreatment efficiency. This study describes the feasibility of the AFEX-assisted DES pretreatment in terms of both progressive and parallel relationships and analyzes the subsequent enzymatic effect in generating glucose from cellulose. Ammonia fiber expansion-assisted DES one-pot pretreatment at 120 °C, for 1.5 h resulted in an enzymatic efficiency of 98.0 ± 3.1 %. Moreover, the enzyme efficiency remained greater than 85 % after three recovery cycle experiments. The comparison between regenerated-lignin (d-lignin) and alkaline-lignin showed that regenerated lignin has a lower molecular weight and belongs to para-hydroxy-phenyl-guaiacyl-syringyl (H-G-S) type lignin. This study developed is a green and efficient pretreatment process with great potential in the separation and utilization of biomass fractions.