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Severe erosion wear is found on valve spools, which threatens the safety and reliability of these units. The use of the plasma beam spraying surfacing method can significantly improve the corrosion resistance and sealing performance of hydraulic valve spools, reduce material waste, and reduce maintenance costs. The effects of the co-addition of CeO2 and SiC particles on the morphology, surface cracks, microstructure, precipitated phases, and wear property of plasma-beam-sprayed Fe55-based coatings on 1025 steel were investigated using OM, EDS, ultra-deep field microscopy, and a wet sand rubber wheel friction tester, respectively. The dendrite exhibited a directional growth pattern perpendicular to the substrate and the transitional states of the microstructure with the co-addition of CeO2 and SiC particles. CeO2 or SiC reduced the liquid phase diffusion coefficient DL of Cr and C and resulted in a decrease in the G/R ratio. The dendrites changed into equiaxed grains. The main phase composition of the Fe55 welding layer was Cr7C3, γ-Fe. The martensite in the surfacing layer and the carbides formed Cr7C3, which can improve the hardness of the surfacing layer. The grain boundaries consisted mainly of a reticular eutectic structure. The uniform distribution of the Cr7C3 hard phase in the Fe55+1.5 wt% SiC+0.01 wt% CeO2 resulted in a uniformly worn surface. The sub-wear mechanisms during the friction process were micro-ploughing and micro-cutting. The hardness and toughness of Fe55+1.5 wt% SiC+0.01 wt% CeO2 were well-matched, avoiding excessive micro-cutting and microplastic deformation. A low content of CeO2 could lead to the formation of equiaxed grain and effectively improve the uniformity of the microstructure. The wear-resistant layer of Fe55+1.5 wt% SiC+0.01 wt% CeO2 can effectively improve the service life and long-term sealing performance of the valve spools.
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Carbonate stress has profound impacts on both agricultural and industrial production. Although a number of salinity-tolerant genes have been reported and applied in plants, there is a lack of research on the role of cell wall-related genes in resistance to carbonate. Likewise, in industry, current strategies have not been able to more effectively address the conflict between stress-induced microalgal biofuel accumulation and microalgal growth inhibition. It is of great significance to study the adaptation mechanism of carbonate-tolerant organisms and to explore related genes for future genetic modification. In this study, the role of the cell wall in the NaHCO3-tolerant chlorella JB17 was investigated. We found that JB17 possesses a relatively thick cell wall with a thickness of 300-600 nm, which is much higher than that of the control chlorella with a thickness of about 100 nm. Determination of the cell wall polysaccharide fractions showed that the cellulose content in the JB17 cell wall increased by 10.48% after NaHCO3 treatment, and the decrease in cellulose levels by cellulase digestion inhibited its resistance to NaHCO3. Moreover, the saccharide metabolome revealed that glucose, rhamnose, and trehalose levels were higher in JB17, especially rhamnose and trehalose, which were almost 40 times higher than in control chlorella. Gene expression detection identified an up-regulated expressed gene after NaHCO3 treatment, JbKOBITO1, overexpression of which could improve the NaHCO3 tolerance of Chlamydomonas reinhardtii. As it encodes a glycosyltransferase-like protein that is involved in cellulose synthesis, the strong tolerance of JB17 to NaHCO3 may be partly due to the up-regulated expression of JbKOBITO 1 and JbKOBITO 1-mediated cellulose accumulation. The above results revealed a critical role of cellulose in the NaHCO3 resistance of JB17, and the identified NaHCO3-tolerance gene will provide genetic resources for crop breeding in saline-alkali soils and for genetic modification of microalgae for biofuel production.
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Understanding the infiltration and solidification processes of liquid 5083Al alloy into Al2O3 three-dimensional reticulated porous ceramic (Al2O3(3D) RPC) is essential for optimizing the microstructure and properties of Al2O3(3D)/5083Al interpenetrating phase composites (IPCs) prepared by low-pressure infiltration process (LPIP). This study employs ProCAST software to simulate the infiltration and solidification processes of liquid 5083Al with pouring velocities (PV) of 0.4 m/s infiltrating into Al2O3(3D) RPC preforms with varying porosities at different pouring temperatures (PT) to prepare Al2O3(3D)/5083Al IPCs using LPIP. The results demonstrate that pore diameter of Al2O3(3D) RPC preforms and PT of liquid 5083Al significantly influence the of the infiltration. Solidification process analysis reveals that the Al2O3(3D) RPC preform with smaller pore diameters allows the lower pouring velocity of 5083Al to solidify faster compared to the preform with larger pore diameters. Al2O3(3D)/5083Al IPCs were prepared successfully from Al2O3(3D) RPC porosity of 15 PPI with liquid 5083Al at PV 0.4 m/s and PT 800 °C using LPIP, resulting in nearly fully dense composites, where both Al2O3(3D) RPCs and 5083Al interpenetrate throughout the microstructure. The infiltration and solidification defects were reduced under air pressure of 0.3 MPa (corresponding to PV of 0.4 m/s) during LPIP. Finite volume method simulations are in good agreement with experimental data, validating the suitability of the simplified model for Al2O3(3D) RPCs in the infiltration simulation.
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Protein aggregation is one of the hallmarks of aging and aging-related diseases, especially for the neurodegenerative diseases (NDs) such as Alzheimer's disease (AD), Parkinson's disease (PD), Huntington's disease (HD), Amyotrophic lateral sclerosis (ALS), and others. In these diseases, many pathogenic proteins, such as amyloid-ß, tau, α-Syn, Htt, and FUS, form aggregates that disrupt the normal physiological function of cells and lead to associated neuronal lesions. Protein aggregates in NDs are widely recognized as one of the important targets for the treatment of these diseases. Natural products, with their diverse biological activities and rich medical history, represent a great treasure trove for the development of therapeutic strategies to combat disease. A number of in vitro and in vivo studies have shown that natural products, by virtue of their complex molecular scaffolds that specifically bind to pathogenic proteins and their aggregates, can inhibit the formation of aggregates, disrupt the structure of aggregates and destabilize them, thereby alleviating conditions associated with NDs. Here, we systematically reviewed studies using natural products to improve disease-related symptoms by reducing or inhibiting the formation of five pathogenic protein aggregates associated with NDs. This information should provide valuable insights into new directions and ideas for the treatment of neurodegenerative diseases.
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Enfermedad de Alzheimer , Productos Biológicos , Enfermedades Neurodegenerativas , Enfermedad de Parkinson , Humanos , Enfermedades Neurodegenerativas/metabolismo , Agregado de Proteínas , Productos Biológicos/farmacología , Productos Biológicos/uso terapéuticoRESUMEN
Metabolism plays an important role in regulating aging at several levels, and metabolic reprogramming is the main driving force of aging. Due to the different metabolic needs of different tissues, the change trend of metabolites during aging in different organs and the influence of different levels of metabolites on organ function are also different, which makes the relationship between the change of metabolite level and aging more complex. However, not all of these changes lead to aging. The development of metabonomics research has opened a door for people to understand the overall changes in the metabolic level in the aging process of organisms. The omics-based "aging clock" of organisms has been established at the level of gene, protein and epigenetic modifications, but there is still no systematic summary at the level of metabolism. Here, we reviewed the relevant research published in the last decade on aging and organ metabolomic changes, discussed several metabolites with high repetition rate, and explained their role in vivo, hoping to find a group of metabolites that can be used as metabolic markers of aging. This information should provide valuable information for future diagnosis or clinical intervention of aging and age-related diseases.
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Steel fiber-reinforced ultra-high-performance concrete (UHPC) is becoming an important type of concrete reinforcement. After mixing with the reinforced steel fibers, the UHPC has perfect flex resistance, shear strength, crack resistance, shock resistance, and anti-seepage. In this study, the influence of straight, corrugated, and hooked brass-coated steel fibers (BCSFs) on the microstructure, mechanical properties, and crack expansion mechanism of ultra-high-performance concrete (UHPC) with varying content of 1-6 wt.% under different curing times were investigated. Field emission scanning electron microscopy and energy dispersive X-ray spectrometry were employed to characterize the microstructure of the BCSF-reinforced UHPC mix specimens. X-ray computed tomography was employed to determine the porosity of the BCSF-reinforced UHPC mix specimens. The obtained results indicate the flexural strength and compressive strength of BCSF-reinforced UHPC mix specimens are enhanced, along with increasing the content of BCSFs reinforcement with different shapes (straight, corrugated, and hooked). The embedded BCSFs play a major role in the adhesive property and stress transfer of the BCSFs-UHPC matrix interface. Different from many studies, the flexural strength of mix UHPC with straight BCSFs is higher than those with corrugated and hooked BCSFs. However, the compressive strength of UHPC with corrugated BCSFs is higher than those with straight and hooked BCSFs. The flexural strength of mix UHPC with 6 wt.% straight BCSFs at 28 days reaches the maximum value of 26.2 MPa, and the compressive strength of UHPC with 6 wt.% corrugated BCSFs at 28 days reaches the maximum value of 142.3 MPa. With the increase in straight BCSF content from 1 wt.% to 6 wt.%, the porosity in mix UHPC reduces gradually from 18.4% to 8.3%. The length of average crack spacing is dependent on the straight BCSF content. With the increase in straight BCSF content from 1 wt.% to 6 wt.%, the average crack length reduces gradually from 34.2 mm to 12.1 mm, and the average crack width reduces gradually from 0.78 mm to less than 0.1 mm. During crack extension, part of the energy in the UHPC mixture specimen with the 6 wt.% BCSF content flows into the crack tip region converted into the work dissipated during the bridging process. The crack propagation resistance of the UHPC mixture with straight BCSFs was improved compared with those with corrugated and hooked BCSFs. The bond strength between the BCSFs and UHPC matrix was enhanced by using vibrational mixing, and the brass film coated on the BCSFs contributes to increase the flexural and compressive strength of the UHPC mixture.
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Anestesia , Dexmedetomidina , Laparoscopía , Neoplasias del Cuello Uterino , Femenino , HumanosRESUMEN
The anaphase-promoting complex/cyclosome (APC/C) is a complicated cellular component that plays significant roles in regulating the cell cycle process of eukaryotic organisms. The spatiotemporal regulation mechanisms of APC/C in distinct cell cycle transitions are no longer mysterious, and the components of this protein complex are gradually identified and characterized. Given the close relationship between the cell cycle and lifespan, it is urgent to understand the roles of APC/C in lifespan regulation, but this field still seems to have not been systematically summarized. Furthermore, although several reviews have reported the roles of APC/C in cancer, there are still gaps in the summary of its roles in other age-related diseases. In this review, we propose that the APC/C is a novel cellular ageing regulator based on its indispensable role in the regulation of lifespan and its involvement in age-associated diseases. This work provides an extensive review of aspects related to the underlying mechanisms of APC/C in lifespan regulation and how it participates in age-associated diseases. More comprehensive recognition and understanding of the relationship between APC/C and ageing and age-related diseases will increase the development of targeted strategies for human health.
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Proteínas de Ciclo Celular , Senescencia Celular , Humanos , Ciclosoma-Complejo Promotor de la Anafase/metabolismo , Proteínas de Ciclo Celular/metabolismo , Ciclo Celular , Senescencia Celular/genéticaRESUMEN
Mariner-like elements (MLEs) are promising tools for gene cloning, gene expression, and gene tagging. We have characterized two MLE transposons from moso bamboo, Ppmar1 and Ppmar2. Ppmar2, is smaller in size and has higher natural activities, thus making it a more potential genomic tool compared to Ppmar1. Using a two-component system consisting of a transposase expression cassette and a non-autonomous transposon cotransformed in yeast, we investigated the transposition activity of Ppmar2 and created hyperactive transposases. Five out of 19 amino acid mutations in Ppmar2 outperformed the wild-type in terms of catalytic activities, especially with the S347R mutant having 6.7-fold higher transposition activity. Moreover, 36 yeast mutants with single-gene deletion were chosen to screen the effects of the host factors on Ppmar2NA transposition. Compared to the control strain (his3Δ), the mobility of Ppmar2 was greatly increased in 9 mutants and dramatically decreased in 7 mutants. The transposition ability in the efm1Δ mutant was 15-fold higher than in the control, while it was lowered to 1/66 in the rtt10Δ mutant. Transcriptomic analysis exhibited that EFM1 defection led to the significantly impaired DDR2, HSP70 expression and dramatically boosted JEN1 expression, whereas RTT10 defection resulted in significantly suppressed expression of UTP20, RPA190 and RRP5. Protein methylation, chromatin and RNA transcription may affect the Ppmar2NA transposition efficiency in yeast. Overall, the findings provided evidence for transposition regulation and offered an alternative genomic tool for moso bamboo and other plants.
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BACKGROUND: Ammonium is an important raw material for biomolecules and life activities, and the toxicity of ammonium is also an important ecological and agricultural issue. Ammonium toxicity in yeast has only recently been discovered, and information on its mechanism is limited. In recent years, environmental pollution caused by nitrogen-containing wastewater has been increasing. In addition, the use of yeast in bioreactors to produce nitrogen-containing compounds has been developed. Therefore, research on resistance mechanisms that allow yeast to grow under conditions of high concentrations of ammonium has become more and more important. RESULTS: To further understand the resistance mechanism of yeast to grow under high concentration of ammonium, we used NH4Cl to screen a yeast non-essential gene-deletion library. We identified 61 NH4Cl-sensitive deletion mutants from approximately 4200 mutants in the library, then 34 of them were confirmed by drop test analysis. Enrichment analysis of these 34 genes showed that biosynthesis metabolism, mitophagy, MAPK signaling, and other pathways may play important roles in NH4Cl resistance. Transcriptome analysis under NH4Cl stress revealed 451 significantly upregulated genes and 835 significantly downregulated genes. The genes are mainly enriched in: nitrogen compound metabolic process, cell wall, MAPK signaling pathway, mitophagy, and glycine, serine and threonine metabolism. CONCLUSIONS: Our results present a broad view of biological pathways involved in the response to NH4Cl stress, and thereby advance our understanding of the resistance genes and cellular transcriptional regulation under high concentration of ammonium.
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Compuestos de Amonio , Saccharomyces cerevisiae , Compuestos de Amonio/toxicidad , Genoma Fúngico , Nitrógeno/metabolismo , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismo , TranscriptomaRESUMEN
Upon stress challenges, proteins/RNAs undergo liquid-liquid phase separation (LLPS) to fine-tune cell physiology and metabolism to help cells adapt to adverse environments. The formation of LLPS has been recently linked with intracellular pH, and maintaining proper intracellular pH homeostasis is known to be essential for the survival of organisms. However, organisms are constantly exposed to diverse stresses, which are accompanied by alterations in the intracellular pH. Aging processes and human diseases are also intimately linked with intracellular pH alterations. In this review, we summarize stress-, aging-, and cancer-associated pH changes together with the mechanisms by which cells regulate cytosolic pH homeostasis. How critical cell components undergo LLPS in response to pH alterations is also discussed, along with the functional roles of intracellular pH fluctuation in the regulation of LLPS. Further studies investigating the interplay of pH with other stressors in LLPS regulation and identifying protein responses to different pH levels will provide an in-depth understanding of the mechanisms underlying pH-driven LLPS in cell adaptation. Moreover, deciphering aging and disease-associated pH changes that influence LLPS condensate formation could lead to a deeper understanding of the functional roles of biomolecular condensates in aging and aging-related diseases.
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Envejecimiento , Proteínas , Fenómenos Fisiológicos Celulares , Humanos , Concentración de Iones de HidrógenoRESUMEN
Sodium bicarbonate (NaHCO3) is an important inorganic salt. It is not only widely used in industrial production and daily life, but is also the main stress in alkaline saline soil. NaHCO3 has a strong ability to inhibit the growth of fungi in both natural environment and daily application. However, the mechanism by which fungi respond to NaHCO3 stress is not fully understood. To further clarify the toxic mechanisms of NaHCO3 stress and identify the specific cellular genes and pathways involved in NaHCO3 resistance, we performed genome-wide screening with NaHCO3 using a Saccharomyces cerevisiae deletion mutant library. A total of 33 deletion mutants with NaHCO3 sensitivity were identified. Compared with wild-type strains, these mutants had significant growth defects in the medium containing NaHCO3. Bioinformatics analysis found that the corresponding genes of these mutants are mainly enriched in the cell cycle, mitophagy, cell wall integrity, and signaling pathways. Further study using transcriptomic analysis showed that 309 upregulated and 233 downregulated genes were only responded to NaHCO3 stress, when compared with yeast transcriptomic data under alkaline and saline stress. Upregulated genes were mainly concentrated in amino acid metabolism, steroid biosynthesis, and cell wall, while downregulated genes were enriched in various cellular metabolisms. In summary, we have identified the cellular pathways and key genes that respond to NaHCO3 stress in the whole genome, providing resource and direction for understanding NaHCO3 toxicity and cellular resistance mechanisms.
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In order to evaluate the application of EEG intelligent detection in gynecological anesthesia depth, the application of ANGEL-6000 EEG depth monitor in laparoscopic gynecological anesthesia was proposed. This method was applied to 60 patients who underwent elective laparoscopic gynecological surgery in our hospital from February to August 2016. Inclusion criteria were ASA i â¼ ii; the average age was (37.8 ± 6.6) years from 20 to 50 years old; the average body weight was (51.53 ± 3.87) kg; conscious and no communication barriers; and patients without instrument ventilation. The patients were divided into observation group and control group according to the random number table method, with 30 patients in each group. The two groups were anesthetized with the same anesthetic drugs, and their consciousness index was monitored. IoC values were recorded before induction of anesthesia (T0), 5 min after intubation (T1), 5 min after operation (T2), intraoperative exploration (T3), at the end of operation (T4), 1 min before extubation (T5), and 5 min after extubation (T6). The dosage of anesthetic drugs, operation time, extubation time, and operation time of the two groups were statistically analyzed. Compared with the operation time of patients in the two groups, the extubation time, awake time, and time out of the operating room of patients in the control group were longer than the observation group. The IoC values of patients in the control group at T0 and T6 time points were lower than those in the observation group at each time point from T1 to T5. Comparison of perioperative dose of remifentanil and atracurium between the two groups was performed. The control group used more propofol dose in perioperative period. The application of neuroelectric signal in laparoscopic gynecological surgery to detect changes in perioperative IoC value can well reflect the level of consciousness of patients and reflect the effect of perioperative stimulation at different time points on the EEG of patients in real time.
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Anestesia Obstétrica , Ginecología , Obstetricia , Propofol , Adulto , Humanos , Persona de Mediana Edad , Remifentanilo , Adulto JovenRESUMEN
In this study, an Al2O33D/5083 Al composite was fabricated by infiltrating a molten 5083 Al alloy into a three-dimensional alumina reticulated porosity ceramics skeleton preform (Al2O33D) using a pressureless infiltration method. The corrosion resistance of 5083 Al alloy and Al2O33D/5083 Al in NaCl solution were compared via electrochemical impedance spectroscopy (EIS), dynamic polarization potential (PDP), and neutral salt spray (NSS) tests. The microstructure of the two materials were investigated by 3D X-ray microscope and scanning electron microscopy aiming at understanding the corrosion mechanisms. Results show that an Al2O33D/5083 Al composite consists of interpenetrating structure of 3D-continuous matrices of continuous networks 5083 Al alloy and Al2O33D phase. A large area of strong interfaces of 5083 Al and Al2O33D exist in the Al2O33D/5083 Al composite. The corrosion development process can be divided into the initial period, the development period, and the stability period. Al2O33D used as reinforcement in Al2O33D/5083 Al composite improves the corrosion resistance of Al2O33D/5083 Al composite via electrochemistry tests. Thus, the corrosion resistance of Al2O33D/5083 Al is higher than that of 5083 Al alloy. The NSS test results indicate that the corrosion resistance of Al2O33D/5083 Al was lower than that of 5083 Al alloy during the initial period, higher than that of 5083 Al alloy during the development period, and there was no obvious difference in corrosion resistance during the stability period. It is considered that the elements in 5083 Al alloy infiltrated into the Al2O33D/5083 Al composite are segregated, and the uniform distribution of the segregated elements leads to galvanic corrosion during the corrosion initial period. The perfect combination of interfaces of Al2O33D and the 5083 Al alloy matrix promotes excellent corrosion resistance during the stability period.
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Sodium metabisulfite (Na2S2O5) is widely used as a preservative in the food and wine industry. However, it causes varying degrees of cellular damage to organisms. In order to improve our knowledge regarding its cyto-toxicity, a genome-wide screen using the yeast single deletion collection was performed. Additionally, a total of 162 Na2S2O5-sensitive strains and 16 Na2S2O5-tolerant strains were identified. Among the 162 Na2S2O5 tolerance-related genes, the retromer complex was the top enriched cellular component. Further analysis demonstrated that retromer complex deletion leads to increased sensitivity to Na2S2O5, and that Na2S2O5 can induce mislocalization of retromer complex proteins. Notably, phosphatidylinositol 3-monophosphate kinase (PI3K) complex II, which is important for retromer recruitment to the endosome, might be a potential regulator mediating retromer localization and the yeast Na2S2O5 tolerance response. Na2S2O5 can decrease the protein expressions of Vps34, which is the component of PI3K complex. Therefore, Na2S2O5-mediated retromer redistribution might be caused by the effects of decreased Vps34 expression levels. Moreover, both pharmaceutical inhibition of Vps34 functions and deletions of PI3K complex II-related genes affect cell tolerance to Na2S2O5. The results of our study provide a global picture of cellular components required for Na2S2O5 tolerance and advance our understanding concerning Na2S2O5-induced cytotoxicity effects.
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Fosfatidilinositol 3-Quinasas Clase III/genética , Conservantes de Alimentos/efectos adversos , Complejos Multiproteicos/genética , Fosfatidilinositol 3-Quinasas/genética , Sulfitos/efectos adversos , Resistencia a Medicamentos/genética , Endosomas/efectos de los fármacos , Endosomas/genética , Eliminación de Gen , Regulación de la Expresión Génica/efectos de los fármacos , Genoma Fúngico/efectos de los fármacos , Genoma Fúngico/genética , Complejos Multiproteicos/antagonistas & inhibidores , Unión Proteica/efectos de los fármacos , Transporte de Proteínas/efectos de los fármacos , Saccharomyces cerevisiae/efectos de los fármacos , Saccharomyces cerevisiae/genética , Sulfitos/farmacologíaRESUMEN
Interface problems and the destruction of the continuity of the oxide film in the Al matrix usually reduce the corrosion resistance of the material. In this paper, the corrosion resistance of Al matrix composites (AMCs) was improved by introducing the silicon carbide skeletons (SiC3D) obtained with polymer replica technology. SiC3D/6061Al was fabricated by infiltrating molten 6061Al alloy in the oxidized SiC3D using the low-pressure casting method. The corrosion resistance performances of 6061Al and SiC3D/6061Al in NaCl solution were studied by electrochemical, neutral salt spray corrosion (NSS), and salt leaching (SL) tests. Results show corrosion resistance of SiC3D/6061Al is higher than that of 6061Al alloys by open circuit potential (OCP), potentio-dynamic polarization (PDP), and electrochemical impedance spectroscopy (EIS) tests. However, NSS and SL tests show the corrosion resistance of SiC3D/6061Al is lower than that of 6061Al alloy. The reason is a corrosion resistant and anti-oxidation network macrostructure with large interface recombination, few concentrated interfaces, and a small specific area that formed in SiC3D/6061Al. SiC3D cannot damage the continuity of the Al2O3 passivating film, and the network macrostructure greatly improves the corrosion resistance performance.
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Stress granules (SGs) are highly dynamic cytoplasmic foci formed in response to stress. The formation of SGs is reported to be regulated by diverse post-translational protein modifications (PTMs). Among them, ADP-ribosylation is of emerging interest due to its recently identified roles in SG organization. In this review, we summarized the latest advances on the roles of poly(ADP-ribose) (PAR) in the regulation of SG formation and dynamics, including its function in modulating nucleocytoplasmic trafficking and SG recruitment of SG components, as well as its effects on protein phase separation behavior. Moreover, the functional role of PAR chain diversity on dynamic of SG composition is also introduced. Potential future developments on investigating global ADP-ribosylation networks, individual roles of different PARPs, and interactions between ADP-ribosylation and other PTMs in SGs are also discussed.
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The zone refining method is a physical method for effectively purifying metals. Increasing yield and reducing impurity content have always been the focus of its research. This article systematically summarizes the relevant research on the production of high-purity metals by zone refining, including mechanisms, parameter optimization, zone refining types, analysis methods, limitations, and future development directions, and it provides relevant theoretical foundations for the production of high-purity metals as well.
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Lithium hexafluorophosphate (LiPF6) is one of the leading electrolytes in lithium-ion batteries, and its usage has increased tremendously in the past few years. Little is known, however, about its potential environmental and biological impacts. In order to improve our understanding of the cytotoxicity of LiPF6 and the specific cellular response mechanisms to it, we performed a genome-wide screen using a yeast (Saccharomyces cerevisiae) deletion mutant collection and identified 75 gene deletion mutants that showed LiPF6 sensitivity. Among these, genes associated with mitochondria showed the most enrichment. We also found that LiPF6 is more toxic to yeast than lithium chloride (LiCl) or sodium hexafluorophosphate (NaPF6). Physiological analysis showed that a high concentration of LiPF6 caused mitochondrial damage, reactive oxygen species (ROS) accumulation, and ATP content changes. Compared with the results of previous genome-wide screening for LiCl-sensitive mutants, we found that oxidative phosphorylation-related mutants were specifically hypersensitive to LiPF6. In these deletion mutants, LiPF6 treatment resulted in higher ROS production and reduced ATP levels, suggesting that oxidative phosphorylation-related genes were important for counteracting LiPF6-induced toxicity. Taken together, our results identified genes specifically involved in LiPF6-modulated toxicity, and demonstrated that oxidative stress and ATP imbalance maybe the driving factors in governing LiPF6-induced toxicity.
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Fluoruros/toxicidad , Litio/toxicidad , Mitocondrias/efectos de los fármacos , Fosforilación Oxidativa/efectos de los fármacos , Fosfatos/toxicidad , Saccharomyces cerevisiae/efectos de los fármacos , Adaptación Fisiológica/efectos de los fármacos , Adenosina Trifosfato/antagonistas & inhibidores , Adenosina Trifosfato/biosíntesis , Regulación Fúngica de la Expresión Génica/efectos de los fármacos , Ontología de Genes , Estudio de Asociación del Genoma Completo , Mitocondrias/genética , Mitocondrias/metabolismo , Proteínas Mitocondriales/antagonistas & inhibidores , Proteínas Mitocondriales/genética , Proteínas Mitocondriales/metabolismo , Anotación de Secuencia Molecular , Estrés Oxidativo , Especies Reactivas de Oxígeno/agonistas , Especies Reactivas de Oxígeno/metabolismo , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismo , Proteínas de Saccharomyces cerevisiae/antagonistas & inhibidores , Proteínas de Saccharomyces cerevisiae/genética , Proteínas de Saccharomyces cerevisiae/metabolismoRESUMEN
Phenomic studies can provide a systemic overview of the network of interactions between phenotypes, genotypes, and environmental factors. Yeast (Saccharomyces cerevisiae) is one of the most important model organisms for phenomic studies due to the availability of a large variety of genome-wide strain collections. We describe a detailed protocol for performing a yeast phenomic screen for evaluation of protein colocalization via a genome-wide imaging-based screening approach utilizing a GFP-tagged yeast strain collection.