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Tumour metabolic reprogramming is pivotal for tumour survival and proliferation. Investigating potential molecular mechanisms within the heterogeneous and clinically aggressive triple-negative breast cancer (TNBC) subtype is essential to identifying novel therapeutic targets. Accordingly, we investigated the role of branched-chain α-keto acid dehydrogenase kinase (BCKDK) in promoting tumorigenesis in TNBC. We analysed The Cancer Genome Atlas dataset and immunohistochemically stained surgical specimens to investigate BCKDK expression and its prognostic implications in TNBC. The effects of BCKDK on tumorigenesis were assessed using cell viability, colony formation, apoptosis, and cell cycle assays, and subsequently validated in vivo. Metabolomic screening was performed via isotope tracer studies. The downstream target was confirmed using mass spectrometry and a co-immunoprecipitation experiment coupled with immunofluorescence analysis. Upstream transcription factors were also examined using chromatin immunoprecipitation and luciferase assays. BCKDK was upregulated in TNBC tumour tissues and associated with poor prognosis. BCKDK depletion led to reduced cell proliferation both in vitro and vivo. MYC-associated zinc finger protein (MAZ) was confirmed as the major transcription factor directly regulating BCKDK expression in TNBC. Mechanistically, BCKDK interacted with glucose-6-phosphate dehydrogenase (G6PD), leading to increased flux in the pentose phosphate pathway for macromolecule synthesis and detoxification of reactive oxygen species. Forced expression of G6PD rescued the growth defect in BCKDK-deficient cells. Notably, the small-molecule inhibitor of BCKDK, 3,6-dichlorobenzo(b)thiophene-2-carboxylic acid, exhibited anti-tumour effects in a patient-derived tumour xenograft model. Our findings hold significant promise for developing targeted therapies aimed at disrupting the MAZ/BCKDK/G6PD signalling pathway, offering potential advancements in treating TNBC through metabolic reprogramming.
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Proliferación Celular , Glucosa , Glucosafosfato Deshidrogenasa , Neoplasias de la Mama Triple Negativas , Regulación hacia Arriba , Neoplasias de la Mama Triple Negativas/metabolismo , Neoplasias de la Mama Triple Negativas/patología , Neoplasias de la Mama Triple Negativas/genética , Humanos , Femenino , Glucosafosfato Deshidrogenasa/metabolismo , Glucosafosfato Deshidrogenasa/genética , Animales , Línea Celular Tumoral , Ratones , Glucosa/metabolismo , Factores de Transcripción/metabolismo , Regulación Neoplásica de la Expresión Génica , Ratones DesnudosRESUMEN
Heart failure (HF) constitutes a major determinant of outcome in chronic kidney disease (CKD) patients. The main pattern of HF in CKD patients is preserved ejection fraction (HFpEF), and left ventricular diastolic dysfunction (LVDD) is a frequent pathophysiological mechanism and specific preclinical manifestation of HFpEF. Therefore, exploring and intervention of the factors associated with risk for LVDD is of great importance in reducing the morbidity and mortality of cardiovascular disease (CVD) complications in CKD patients. We designed this retrospective cross-sectional study to collect clinical and echocardiographic data from 339 nondialysis CKD patients without obvious symptoms of HF to analyze the proportion of asymptomatic left ventricular diastolic dysfunction (ALVDD) and its related factors associated with risk by multivariate logistic regression analysis. Among the 339 nondialysis CKD patients, 92.04% had ALVDD. With the progression of CKD stage, the proportion of ALVDD gradually increased. The multivariate logistic regression analysis revealed that increased age (OR 1.237; 95% confidence interval (CI) 1.108-1.381, per year), diabetic nephropathy (DN) and hypertensive nephropathy (HTN) (OR 25.000; 95% CI 1.355-48.645, DN and HTN vs chronic interstitial nephritis), progression of CKD stage (OR 2.785; 95% CI 1.228-6.315, per stage), increased mean arterial pressure (OR 1.154; 95% CI 1.051-1.268, per mmHg), increased urinary protein (OR 2.825; 95% CI 1.484-5.405, per g/24 h), and low blood calcium (OR 0.072; 95% CI 0.006-0.859, per mmol/L) were factors associated with risk for ALVDD in nondialysis CKD patients after adjusting for other confounding factors. Therefore, dynamic monitoring of these factors associated with risk, timely diagnosis and treatment of ALVDD can delay the progression to symptomatic HF, which is of great importance for reducing CVD mortality, and improving the prognosis and quality of life in CKD patients.
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Insuficiencia Renal Crónica , Disfunción Ventricular Izquierda , Humanos , Femenino , Masculino , Persona de Mediana Edad , Estudios Retrospectivos , Estudios Transversales , Insuficiencia Renal Crónica/complicaciones , Insuficiencia Renal Crónica/fisiopatología , Disfunción Ventricular Izquierda/etiología , Disfunción Ventricular Izquierda/fisiopatología , Anciano , Medición de Riesgo , Progresión de la Enfermedad , Factores de Riesgo , Ecocardiografía , Hipertensión/complicaciones , Modelos Logísticos , Nefropatías Diabéticas/complicaciones , Nefropatías Diabéticas/fisiopatología , Diástole , Volumen Sistólico , Enfermedades Asintomáticas , Hipertensión Renal , NefritisRESUMEN
Rationale: In the physiological states, the act of scratching protects the person from harmful substances, while in certain pathological conditions, the patient suffers from chronic itch, both physically and mentally. Chronic itch sufferers are more sensitive to mechanical stimuli, and mechanical hyperknesis relief is essential for chronic itch treatment. While neuropeptide Y-Y1 receptor (NPY-Y1R) system is known to play a crucial role in modulating mechanical itch in physiological conditions, it is elusive how they are altered during chronic itch. We hypothesize that the negative regulatory effect of Y1Rs on Tac2 neurons, the key neurons that transmit mechanical itch, declines during chronic itch. Methods: We combined transgenic mice, chemogenetic manipulation, immunofluorescence, rabies virus circuit tracing, and electrophysiology to investigate the plasticity of Y1Rs on Tac2 neurons during chronic itch. Results: We found that Tac2 neurons receive direct input from Npy neurons and that inhibition of Npy neurons induces activation of Tac2 neurons. Moreover, the expression of Y1Rs on Tac2 neurons is reduced, and the regulatory effect is also reduced during chronic itch. Conclusion: Our study clarifies the plasticity of Y1Rs on Tac2 neurons during chronic itch and further elucidates the mechanism by which NPY-Y1R system is responsible for modulating mechanical itch. We highlight Y1Rs as a promising therapeutic target for mechanical hyperknesis during chronic itch.
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Neuropéptido Y , Receptores de Neuropéptido Y , Humanos , Ratones , Animales , Neuropéptido Y/metabolismo , Neuropéptido Y/farmacología , Receptores de Neuropéptido Y/genética , Receptores de Neuropéptido Y/metabolismo , Neuronas/metabolismo , Prurito/metabolismoRESUMEN
Feeding effects are crucial for evaluating the capacity of zooplankton to regulate phytoplankton populations within freshwater ecosystems. To examine the impact of the bloom-forming cyanobacteria Raphidiopsis raciborskii, which occurs in tropical and subtropical freshwaters, on the growth of zooplankton Daphnia in relation to toxins, filament length and fatty acid content, we fed D. magna with R. raciborskii only (cylindrospermopsin (CYN)-producing and non-CYN-producing, as the negative controls), Chlorella pyrenoidosa only (as the positive control) and a mixed diet containing R. raciborskii (CYN-producing and non-CYN-producing) and C. pyrenoidosa. Consequently, our findings revealed that the toxic effect of CYN-producing R. raciborskii strains on Daphnia was mitigated by the coexistence of C. pyrenoidosa containing stearidonic acid (SDA, C18:4 ω3) in mixed diets. This was evident in the elevated survival rate compared that from diets containing only R. raciborskii and a significantly higher reproduction and population intrinsic increase rate compared to diets consisting of only R. raciborskii or C. pyrenoidos. Additionally, a strong positive correlation was observed between arachidonic acid (ARA, 20:4ω6) and the population intrinsic increase rate of Daphnia; notably, R. raciborskii strains were found to be rich in the ω6 polyunsaturated fatty acid ARA. These outcomes reinforce the crucial role of polyunsaturated fatty acids in predicting the population increase of crustacean zooplankton, which has long been neglected. Furthermore, our results underscore the potential effectiveness of zooplankton, particularly in temperate lakes, in controlling CYN-producing R. raciborskii populations.
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In the concept of a microstructured bubble column reactor, microstructuring of the catalyst carrier is realized by introducing a static mesh of thin wires coated with catalyst inside the column. Meanwhile, the wires also serve the purpose of cutting the bubbles, which in turn results in high interfacial area and enhanced interface hydrodynamics. However, there are no models that can predict the fate of bubbles (cut/stuck) passing through these wires, thus making the reactor optimization difficult. In this work, based on several typical bubble-wire interacting configurations, we analyze the outcomes by applying the energy balance of the bubble focusing on buoyancy and surface tension. Two limiting cases of viscosity, corresponding to the ability of the bubble to reconfigure into the lowest energy state, are investigated. Upon analysis, it is observed that a narrow mesh spacing and a smaller bubble Eötvös number generally result in bubbles getting stuck underneath the wire. We have obtained the threshold grid spacing and the critical Eötvös number for bubble passage and bubble cutting, which are verified by the direct numerical simulation results of bubble passing through a single mesh opening. The derived energy balance is generalized to large meshes with multiple openings and different configurations. Finally, a closure model based on the outcomes of energy-balance analysis is proposed for Euler-Lagrange simulations of microstructured bubble columns.
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Submerged macrophytes play crucial roles in maintaining the stability of clear-water states in shallow lakes. Recent stable isotope studies have shown that crustacean zooplankton can utilize submerged macrophyte carbon, but macrophytes alone cannot support the growth and reproduction of such grazers, being deficient in highly unsaturated fatty acids (HUFA). We hypothesized that flagellates feeding on macrophytes can synthesize HUFA and thereby support crustacean zooplankton. To test this hypothesis, we conducted a feeding experiment in which Daphnia magna were provided with a diet of submerged macrophyte Hydrilla verticillata detritus which had been degraded by lake microbes. The chlorophyte Scenedesmus bijuga and undegraded macrophyte detritus were used as controls for comparison of Daphnia's performance. Using biochemical analysis, we examined how the degradation process affected the food quality of the macrophyte. Flagellates were subsequently isolated from the degraded macrophyte and cultured heterotrophically to detect their HUFA synthesis. The 5-day degraded H. verticillata showed significantly higher HUFA concentrations than undegraded macrophyte detritus. They supported better Daphnia performance than undegraded macrophyte, being comparable with S. bijuga. Two isolated flagellates (SL-1 and SL-2), identified as Ochromonas sp. and Poterioochromonas sp., were found to contain HUFA when cultured heterotrophically without dietary sources of fatty acids, suggesting their HUFA synthesis ability. Our results demonstrate that submerged macrophytes may thus indirectly support crustacean zooplankton via flagellate mediation. As crustacean zooplanktons are of key importance for water quality in the grazer control of phytoplankton, this microbial facilitation may contribute to the maintenance of macrophyte clear-water conditions in shallow lakes.
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Carbono , Daphnia , Animales , Lagos , FitoplanctonRESUMEN
In the concept of a microstructured bubble column reactor, meshes coated with catalyst can cut the bubbles, which in turn results in high interfacial area and enhanced interface hydrodynamics. In previous work, we developed a closure model for the fate of bubbles interacting with a wire mesh based on the outcomes of energy balance analysis. In this paper, the model is validated using Euler-Lagrange simulations against two experimental cases of microstructured bubble columns. Before validation of the model, the definition of the deceleration thickness, as used in the calculation of the virtual mass term, is refined to introduce the effects of liquid viscosity and wire thickness. Proceeding with the validation, the inclusion of our cutting closure model results in an excellent match of the bubble size reduction by the wire mesh with the experimental data. Consequently, the simulations produce a more accurate prediction of the reactor performance for the gaseous reaction in view of the pH and gas holdup profiles. The effect of liquid viscosity on the bubble size reduction by the wire mesh is replicated accurately as well. Noticeably, the significance of bubble coalescence and breakup in bubble dynamics overperforms the role of bubble cutting at high superficial gas velocities; thus, further improvement is needed there. Finally, based on the validated cutting model, we share some perspectives on the design of wire meshes to increase the bubble interfacial area.
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As the most common cardiovascular disease, atherosclerosis (AS), is a leading cause of high mortality in patients with chronic renal failure. Rab27a has been reported to regulate the progression of cardiovascular and renal diseases. Nevertheless, little studies investigated the role and mechanism of Rab27a in uremic-accelerated AS (UAAS). An animal model of UAAS was established in apolipoprotein E knockout (apoE-/-) mice using 5/6 nephrectomy (NX). We conducted in vitro and in vivo functional experiments to explore the role of Rab27a in UAAS, including the presence of oxidized low-density lipoprotein (ox-LDL). Rab27a expression was upregulated in the plaque tissues of NX apoE-/- mice. The knockout of Rab27a (Rab27a-/-) reduced AS-induced artery injury, as manifested by the reductions of plaque area, collagen deposition, inflammation and lipid droplet. Besides, cholesterol efflux was increased, while the expression of lipid metabolism-related proteins and the secretions of pro-inflammatory factors were decreased in ox-LDL-induced NX Rab27a-/- apoE-/- mice group. Further, Rab27a deletion inhibited the activation of nuclear factor κB (NF-κB) pathway. In conclusion, our study indicated that Rab27a deficiency attenuated foam cell formation and macrophage inflammation, depending on the NF-κB pathway activation, to inhibit AS progression in uremic apoE-/- mice. This finding may provide a new targeting strategy for UAAS therapy.
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Aterosclerosis , Células Espumosas , Animales , Ratones , Ratones Noqueados , Células Espumosas/metabolismo , FN-kappa B/metabolismo , Aterosclerosis/genética , Aterosclerosis/metabolismo , Macrófagos/metabolismo , Inflamación/metabolismo , Apolipoproteínas E/genética , Apolipoproteínas E/metabolismo , Apolipoproteínas/metabolismo , Proteínas rab27 de Unión a GTP/metabolismoRESUMEN
Organic carbon derived from terrestrial plants contributes to aquatic consumers, e.g., zooplankton in lakes. The degree of the contribution depends on the availability of terrestrial organic carbon in lake organic pool and the transfer efficiency of the carbon. Terrestrial organic carbon is poor-quality food for zooplankton with a mismatch of nutrition content and was incorporated to zooplankton with much lower efficiency than phytoplankton. Contributions of terrestrial carbon to zooplankton generally decrease with an increase in phytoplankton production, indicating a preferential incorporation of phytoplankton in previous investigations. However, in eutrophic lakes, the dominating cyanobacteria were of poor quality and incorporated to consumers inefficiently too. In that case, zooplankton in eutrophic wetlands, where cyanobacteria dominate the phytoplankton production and massive terrestrial plants are inundated, may not preferentially incorporate poor food-quality phytoplankton resource to their biomass. Therefore, we hypothesize that carbon contributions of terrestrial vegetation to zooplankton and to lake particulate organic pool should be similar in such aquatic ecosystems. We tested this hypothesis by sampling zooplankton and carbon sources in Ming Lake (Jinan University Campus, southern China) which was overgrown by terrestrial plants after drying and re-flooded. After 60 days of observations at weekly (or biweekly) intervals, applying stable carbon (13C), nitrogen (15 N), and hydrogen (2H) isotopic analysis and a stable isotope mixing model, we estimated the occurrence of extensive carbon contribution (≥ 50%) of flooded terrestrial plants to cladocerans and copepods. Contribution of inundated terrestrial plants to cladocerans was similar to that to lake particulate organic pool. Thus, our study quantified the role of terrestrial carbon in eutrophic wetlands, enhancing our understanding of cross-ecosystem interactions in food webs with an emphasis on the resource quality.
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Cianobacterias , Zooplancton , Humanos , Animales , Carbono/metabolismo , Lagos , Ecosistema , Biomasa , Cadena Alimentaria , Fitoplancton/metabolismo , Cianobacterias/metabolismoRESUMEN
The Bering-Chukchi shelf is one of the world's most productive areas and characterized by high benthic biomass. Sedimentary microbial communities play a crucial role in the remineralization of organic matter and associated biogeochemical cycles, reflecting both short-term changes in the environment and more consistent long-term environmental characteristics in a given habitat. In order to get a better understanding of the community structure of sediment-associated prokaryotes, surface sediments were collected from 26 stations in the Bering-Chukchi shelf and adjacent northern deep seas in this study. Prokaryote community structures were analyzed by metabarcoding of the 16S rRNA gene, and potential interactions among prokaryotic groups were analyzed by co-occurrence networks. Relationships between the prokaryote community and environmental factors were assessed. Gammaproteobacteria, Alphaproteobacteria, and Flavobacteriia were the dominant bacterial classes, contributing 35.0, 18.9, and 17.3% of the bacterial reads, respectively. The phototrophic cyanobacteria accounted for 2.7% of the DNA reads and occurred more abundantly in the Bering-Chukchi shelf. Prokaryotic community assemblages were different in the northern deep seas compared to the Bering-Chukchi shelf, represented by the lowered diversity and the increased abundant operational Taxonomic Units (OTU), suggesting that the abundant taxa may play more important roles in the northern deep seas. Correlation analysis showed that latitude, water depth, and nutrients were important factors affecting the prokaryote community structure. Abundant OTUs were distributed widely in the study area. The complex association networks indicated a stable microbial community structure in the study area. The high positive interactions (81.8-97.7%) in this study suggested that symbiotic and/or cooperative relationships accounted for a dominant proportion of the microbial networks. However, the dominant taxa were generally located at the edge of the co-occurrence networks rather than in the major modules. Most of the keystone OTUs were intermediately abundant OTUs with relative reads between 0.01 and 1%, suggesting that taxa with moderate biomass might have considerable impacts on the structure and function of the microbial community. This study enriched the understanding of prokaryotic community in surface sediments from the Bering-Chukchi shelf and adjacent sea areas.
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Nitrous oxide (N2O) emissions from lakes exhibit significant spatiotemporal heterogeneity, and quantitative identification of the different N2O production processes is greatly limited, causing the role of nitrification to be undervalued or ignored in models of a lake's N2O emissions. Here, the contributions of nitrification and denitrification to N2O production were quantitatively assessed in the eutrophic Lake Taihu using molecular biology and isotope mapping techniques. The N2O fluxes ranged from -41.48 to 28.84 µmol m-2 d-1 in the lake, with lower N2O concentrations being observed in spring and summer and significantly higher N2O emissions being observed in autumn and winter. The 15N site preference and relevant isotopic evidence demonstrated that denitrification contributed approximately 90% of the lake's gross N2O production during summer and autumn, 27-83% of which was simultaneously eliminated via N2O reduction. Surprisingly, nitrification seemed to act as a key process promoting N2O production and contributing to the lake as a source of N2O emissions. A combination of N2O isotopocule-based approaches and molecular techniques can be used to determine the precise characteristics of microbial N2O production and consumption in eutrophic lakes. The results of this study provide a basis for accurately assessing N2O emissions from lakes at the regional and global scales.
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Lagos , Nitrificación , Desnitrificación , Óxido NitrosoRESUMEN
The southern Chinese coast is one of the most developed regions in China and is an area where harmful algal blooms (HABs) have occurred frequently. In this study, differences in the phytoplankton community between microscopic observations and 18S rDNA metabarcoding were compared in 89 surface water samples collected from the southern Chinese coast and the western South China Sea (SCS). This is the first report investigating the phytoplankton community and HAB species using a combination of morphological and metabarcoding approaches in this sea area. There were substantial differences in phytoplankton community structure detected by the two methods. Microscopic observation revealed diatom predominance in the phytoplankton community, while metabarcoding indicated dinoflagellate dominance. The phytoplankton community structure obtained by microscopic observation better reflects the real situation in the water column. Metabarcoding annotated more species than morphospecies observed by microscopy. Haptophyta and Cryptophyta were the specific phyla detected in metabarcoding but were missed in microscopy due to their small size. Conversely, some taxa were found in microscopic analysis alone, such as species in Dinophysis, Prorocentrum, and Scrippsiella, suggesting some biases during metabarcoding and gaps in sequence databases. Metabarcoding is superior for detecting morphologically cryptic, small-sized and HAB taxa, such as unarmored dinoflagellates, nanosized hatophytes and chlorophytes, as well as multiple species in Alexandrium, Pseudonitzschia, and Chaetoceros in our study. A total of 62 HAB taxa were identified in this study, including blooming and potentially toxic species. Diatom abundances generally decreased southward, while those of dinoflagellates and haptophytes showed the opposite trend. Chlorophytes were mainly distributed in coastal waters, especially in the Pearl River Estuary. Phytoplankton community structures were shaped by nutrients and salinity, and phosphorus was the most limiting factor for phytoplankton growth. The phytoplankton community in the western SCS showed unique characteristics away from those in the coastal sea areas. The results suggest that the combination of morphological and metabarcoding approaches comprehensively reveals the phytoplankton community structure and diversity of HAB species.
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Diatomeas , Dinoflagelados , ADN Ribosómico , Dinoflagelados/genética , Floraciones de Algas Nocivas , Fósforo , Fitoplancton , AguaRESUMEN
In the past three decades, harmful algal blooms (HAB) have become more frequent and widespread in southeast Chinese sea areas. Resting stages are regarded as the "seed bank" of algal blooms, and play an important role in initiating HABs. The distribution of resting stages in sediments especially those of HAB species can make good predictions about the potential risk of future blooms, however with limited reports. In this study, surface sediment samples were collected in the four sea areas along the southeast Chinese coasts, including Dafeng Port (DF) in the southern Yellow Sea, Xiangshan Bay (XS), Funing Bay (FN), and Dongshan Bay (DS) in the East China Sea. Diversity and community structure of eukaryotic microalgae in surface sediments were assessed by metabarcoding V4 region of the 18S rDNA, focusing on the distribution of HAB species. Biogenic elements including total organic carbon (TOC), total nitrogen (TN), total phosphorus (TP), biogenic silicon (BSi), and moisture content (MC) were analyzed. A total of 454 eukaryotic algal OTUs were detected, which belonged to 31 classes of 9 phyla. Altogether 149 algal species were detected in this study, and 59 taxa have been reported to form resting stages. Eukaryotic algal community was similar in XS, FN and DS of the East China Sea, which were predominated by dinoflagellates. However, algal community was different in DF of the Yellow Sea, and characterized by the dominance of chrysophytes and low OTU richness. The distribution of most abundant HAB species showed positive correlations with TN, BSi, and TOC, suggesting that eutrophication and consequent increase in diatom productivity may have a significant influence on the distribution of HAB species and facilitate the occurrence of HABs. Furthermore, HAB species occurred more abundantly and widely in FN. Our results suggest high potential risks of HABs in the southeast Chinese coast especially in Funing Bay.
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Brain energetics disturbance is a hypothesized cause of depression. Glucose is the predominant fuel of brain energy metabolism; however, the cell-specific change of glucose metabolism and underlying molecular mechanism in depression remains unclear. In this study, we firstly applied 18 F-FDG PET and observed brain glucose hypometabolism in the prefrontal cortex (PFC) of corticosterone-induced depression of rats. Next, astrocytic glucose hypometabolism was identified in PFC slices in both corticosterone-induced depression of rats and cultured primary astrocytes from newborn rat PFC after stress-level corticosterone (100 nM) stimulation. Furthermore, we found the blockage of glucose uptake and the decrease of plasma membrane (PM) translocation of glucose transporter 1 (GLUT1) in astrocytic glucose hypometabolism under depressive condition. Interestingly, thioredoxin interacting protein (TXNIP), a glucose metabolism sensor and controller, was found to be over-expressed in corticosterone-stimulated astrocytes in vivo and in vitro. High TXNIP level could restrict GLUT1-mediated glucose uptake in primary astrocytes in vitro. Adeno-associated virus vector-mediated astrocytic TXNIP over-expression in rat medial PFC suppressed GLUT1 PM translocation, consequently developed depressive-like behavior. Conversely, TXNIP siRNA facilitated GLUT1 PM translocation to recover glucose hypometabolism in corticosterone-exposed cultured astrocytes. Notably, astrocyte-specific knockdown of TXNIP in medial PFC of rats facilitated astrocytic GLUT1 PM translocation, showing obvious antidepressant activity. These findings provide a new astrocytic energetic perspective in the pathogenesis of depression and, more importantly, provide TXNIP as a promising molecular target for novel depression therapy.
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Astrocitos , Glucosa , Animales , Astrocitos/metabolismo , Proteínas de Ciclo Celular , Corticosterona/metabolismo , Glucosa/metabolismo , Transportador de Glucosa de Tipo 1/metabolismo , Ratas , Tiorredoxinas/metabolismoRESUMEN
High fructose has been reported to drive glomerular podocyte oxidative stress and then induce podocyte foot process effacement in vivo, which could be partly regarded as podocyte hypermotility in vitro. Atractylodin possesses anti-oxidative effect. The aim of this study was to explore whether atractylodin prevented against fructose-induced podocyte hypermotility via anti-oxidative property. In fructose-exposed conditionally immortalized human podocytes, we found that atractylodin inhibited podocyte hypermotility, and up-regulated slit diaphragm proteins podocin and nephrin, and cytoskeleton protein CD2-associated protein (CD2AP), α-Actinin-4 and synaptopodin expression, which were consistent with its anti-oxidative activity evidenced by up-regulation of catalase (CAT) and superoxide dismutase (SOD) 1 expression, and reduction of reactive oxygen species (ROS) production. Atractylodin also significantly suppressed expression of transient receptor potential channels 6 (TRPC6) and phosphorylated Ca2+/calmodulin-dependent protein kinase IV (CaMK4) in cultured podocytes with fructose exposure. Additionally, in fructose-exposed podocytes, CaMK4 siRNA up-regulated synaptopodin and reduced podocyte hypermotility, whereas, silencing of TRPC6 by siRNA decreased p-CaMK4 expression, inhibited podocyte hypermotility, showing TRPC6/p-CaMK4 signaling activation in podocyte hypermotility under fructose condition. Just like atractylodin, antioxidant N-acetyl-L-cysteine (NAC) could inhibit TRPC6/p-CaMK4 signaling activation to reduce fructose-induced podocytes hypermotility. These results first demonstrated that the anti-oxidative property of atractylodin may contribute to the suppression of podocyte hypermotility via inhibiting TRPC6/p-CaMK4 signaling and restoring synaptopodin expression abnormality.
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Antioxidantes/farmacología , Fructosa/efectos adversos , Furanos/farmacología , Podocitos/efectos de los fármacos , Edulcorantes/efectos adversos , Proteína Quinasa Tipo 4 Dependiente de Calcio Calmodulina/antagonistas & inhibidores , Proteína Quinasa Tipo 4 Dependiente de Calcio Calmodulina/metabolismo , Línea Celular , Movimiento Celular/efectos de los fármacos , Regulación hacia Abajo/efectos de los fármacos , Humanos , Proteínas de Microfilamentos/metabolismo , Estrés Oxidativo/efectos de los fármacos , Fosforilación , Podocitos/fisiología , Proteolisis/efectos de los fármacos , Transducción de Señal/efectos de los fármacos , Canal Catiónico TRPC6/antagonistas & inhibidores , Canal Catiónico TRPC6/metabolismoRESUMEN
Impaired wound healing presents great health risks to patients. While encouraging, the current clinical successes of mesenchymal stromal cell (MSC)-based therapies for tissue repair have been limited. Genetic engineering could endow MSCs with more robust regenerative capacities. Here, we identified that C-C motif chemokine receptor 2 (CCR2) overexpression enhanced the targeted migration and immunoregulatory potential of MSCs in response to C-C motif chemokine ligand 2 (CCL2) in vitro. Intravenously infusion of CCR2-engineered MSCs (MSCsCCR2) exhibited improved homing efficiencies to injured sites and lungs of diabetic mice. Accordingly, MSCCCR2 infusion inhibited monocyte infiltration, reshaped macrophage inflammatory properties, prompted the accumulation of regulatory T cells (Treg cells) in injured sites, and reshaped systemic immune responses via the lung and spleen in mouse diabetic wound models. In summary, CCR2-engineered MSCs restore immunological homeostasis to accelerate diabetic wound healing via their improved homing and immunoregulatory potentials in response to CCL2. Therefore, these findings provide a novel strategy to explore genetically engineered MSCs as tools to facilitate tissue repair in diabetic wounds.
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Diabetes Mellitus Experimental , Células Madre Mesenquimatosas , Cicatrización de Heridas , Animales , Diabetes Mellitus Experimental/terapia , Ingeniería Genética , Homeostasis , Humanos , Ratones , Receptores CCR2RESUMEN
OBJECTIVES: The purpose of this study was to analyze the α-SiO2 content, composition, dispersion, morphology, and free radical content of dust between the alveolar and the workplace, to explore the possible changes in the properties (especially the pathogenicity) of dust after it enters the lung. METHODS: We collected the dust in the workplace in HANDAN Coal mine. They were selected by a 400 mesh sieve and was made a suspension of 50 mg/ml, which would be used to perfuse into the trachea of rats. When one week, four weeks, eight weeks, fourteen weeks, twenty weeks after perfusing, we harvested dust in rats alveolar through lung lavage for further processing. RESULTS: In the animal test, typical fibrous nodules appeared 20 weeks after dust exposure. No inflammatory reaction was observed in the saline group. The results of animal experiments showed that there was no significant difference in the content of α-SiO2 between dust in the workplace and the lung lavage (P > 0.05). The content of the Fe element gradually increased with dust exposure time. The 12 elements of Al, Mg, Si, Pb, Mn, Ni, Zn, Cu, Cr, Sb, Cd, and AS were reduced in the experiment group compared with the workplace group. The shape of the dust in the workplace was mostly spherical. The shape of the dust extracted from the lung lavage fluid was mostly blocky and angular, and a few dust edges were sharp, and more than 80% of the particle size was smaller than 5 µm, while less than 1% of the particle size was larger than 10 µm. The amount of hydroxyl radical released by lung lavage dust in phosphate buffer was higher than that of the workplace dust. CONCLUSIONS: After the dust entered the alveoli, the content of α-SiO2 in the dust did not change with dust exposure time, while the content of elements in the dust, the morphology, and dispersion of the dust changed. The ability of dust in alveoli to produce hydroxyl radicals in phosphate buffer was higher than that in the workplace.
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Polvo , Dióxido de Silicio , Animales , Líquido del Lavado Bronquioalveolar , Pulmón , Ratas , Lugar de TrabajoRESUMEN
BACKGROUND: The importance of monoterpenes in grape and wine aroma has compelled researchers to focus on developing methods to increase their abundance. Recent research has revealed that exogenous elicitors can increase the contents of these compounds. This study determined the effects of methyl jasmonate (MeJA) preharvest treatments on the monoterpene profiles of Muscat Hamburg grapes and wine. RESULTS: A total of 27 monoterpenes were identified for Muscat Hamburg grapes and wine. The contents of most of the monoterpenes (free and glycosylated forms) in the grapes and wine increased in response to MeJA. An analysis of the expression of the genes in the terpenoid biosynthesis pathway indicated that the related biosynthetic pathways were activated by MeJA. The transcript levels of some genes were consistent with monoterpene production, including VviCSLinNer, VviGwbOciF, VviPNRLin, VviGT14 and VviUGT85A1L1. The developmental expression patterns of the VviPP2B1 and VviMYB24 transcription factor genes were positively correlated with monoterpene accumulation in ripening grapes. CONCLUSIONS: Our results suggest that MeJA may be useful for improving the aroma quality of grapes and wines.
Asunto(s)
Acetatos/farmacología , Ciclopentanos/farmacología , Frutas/química , Monoterpenos/metabolismo , Oxilipinas/farmacología , Reguladores del Crecimiento de las Plantas/farmacología , Vitis/efectos de los fármacos , Vino/análisis , Vías Biosintéticas/efectos de los fármacos , Frutas/efectos de los fármacos , Frutas/genética , Frutas/metabolismo , Regulación de la Expresión Génica de las Plantas/efectos de los fármacos , Monoterpenos/análisis , Odorantes/análisis , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Vitis/química , Vitis/genética , Vitis/metabolismoRESUMEN
Monoterpenes contribute to the varietal aromas of grapes and wines. We determined the effects of cluster zone leaf removal on the monoterpene profiles of Sauvignon Blanc grape berries and wines, and on the expression of key genes in the terpenoid pathway. Leaf removal at two intensities (half basic, 50%; full basic, 100%) was conducted at two weeks before veraison, veraison, and two weeks after veraison. Half basic leaf removal increased the pH and decreased the tartaric acid content in grapes and wines. The concentrations of most free- and bound-form monoterpenes in grapes were increased by early leaf removal. The total monoterpene contents were increased in wines in the defoliation treatments, but were significantly lower in wines from the full basic leaf removal treatments than in wines from the half basic leaf removal treatments. The defoliation treatments resulted in increased transcript levels of some key genes in terpene biosynthesis (VvPNLinNer1, VvPNLinNer2, VvPNLNGl1, VvPNLNGl2, and VvUGT88A1L1).