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1.
Analyst ; 149(4): 1022-1049, 2024 Feb 12.
Artigo em Inglês | MEDLINE | ID: mdl-38273740

RESUMO

Escherichia coli (E. coli) is a prevalent enteric bacterium and a necessary organism to monitor for food safety and environmental purposes. Developing efficient and specific methods is critical for detecting and monitoring viable E. coli due to its high prevalence. Conventional culture methods are often laborious and time-consuming, and they offer limited capability in detecting potentially harmful viable but non-culturable E. coli in the tested sample, which highlights the need for improved approaches. Hence, there is a growing demand for accurate and sensitive methods to determine the presence of viable E. coli. This paper scrutinizes various methods for detecting viable E. coli, including culture-based methods, molecular methods that target DNAs and RNAs, bacteriophage-based methods, biosensors, and other emerging technologies. The review serves as a guide for researchers seeking additional methodological options and aiding in the development of rapid and precise assays. Moving forward, it is anticipated that methods for detecting E. coli will become more stable and robust, ultimately contributing significantly to the improvement of food safety and public health.


Assuntos
Bacteriófagos , Técnicas Biossensoriais , Escherichia coli/genética , Inocuidade dos Alimentos , Microbiologia de Alimentos
2.
Biochem Biophys Res Commun ; 640: 80-87, 2023 01 15.
Artigo em Inglês | MEDLINE | ID: mdl-36502635

RESUMO

Deficiency in human coagulation factor VIII (FVIII) causes hemophilia A (HA). Patients with HA may suffer from spontaneous bleeding, which can be life-threatening. Recombinant FVIII (rFVIII) is an established treatment and prevention agent for bleeding in patients with HA. Human plasma-derived FVIII (pdFVIII), commonly used in clinical practice, is relatively difficult to prepare. In this study, we developed a novel B-domain-deleted rFVIII, produced and formulated without the use of animal or human serum-derived components. rFVIII promoted the generation of activated factor X and downstream thrombin, and, similar to that of other available FVIII preparations, its activity was inhibited by FVIII inhibitors. In addition, rFVIII has ideal binding affinity to human von Willebrand factor. Activated FVIII (FVIIIa) could be degraded by activated protein C and lose its procoagulant activity. In vitro, commercially available recombinant FVIII (Xyntha) and pdFVIII were used as controls, and there were no statistical differences between rFVIII and commercial FVIII preparations, which demonstrates the satisfactory efficacy and potency of rFVIII. In vivo, HA mice showed that infusion of rFVIII rapidly corrected activated partial thromboplastin time, similar to Xyntha. Moreover, different batches of rFVIII were comparable. Overall, our results demonstrate the potential of rFVIII as an effective strategy for the treatment of FVIII deficiency.


Assuntos
Fator VIII , Proteínas Recombinantes , Animais , Humanos , Camundongos , Fator VIII/farmacologia , Fator VIII/uso terapêutico , Hemofilia A/tratamento farmacológico , Hemorragia , Modelos Animais , Proteínas Recombinantes/farmacologia , Proteínas Recombinantes/uso terapêutico
3.
Microb Ecol ; 85(4): 1253-1264, 2023 May.
Artigo em Inglês | MEDLINE | ID: mdl-35581504

RESUMO

Located between the South and the East China Sea, the Taiwan Straits (TWS) are a marine shelf-channel area, with unique hydrological and geomorphological features affected by rivers inflow and with recent algal blooms with red tide events. This study aimed at assessing microbial distribution and function and their modulation in response to environmental gradients. Surface (0.5 m) water samples from 16 stations along five north to south transects were collected; total prokaryotic abundance by epifluorescence microscope and carbon substrate utilization patterns by Biolog Ecoplates were estimated. Spatially, a patchy microbial distribution was found, with the highest microbial metabolic levels and prokaryotic abundance in the TWS area between Minjiang River estuary and Pingtan Island, and progressive decreases towards offshore stations. Complex carbon sources and carbohydrates were preferentially metabolized. This study provides a snapshot of the microbial abundance and activity in TWS as a model site of aquatic ecosystems impacted from land inputs; obtained data highlights that microbial metabolism is more sensitive than abundance to environmental changes.


Assuntos
Ecossistema , Estuários , Taiwan , Rios , China , Monitoramento Ambiental , Carbono
4.
Environ Sci Technol ; 57(1): 297-309, 2023 01 10.
Artigo em Inglês | MEDLINE | ID: mdl-36576880

RESUMO

To explore the effect of different types of dissolved organic matter (DOM) decomposition on nutrient cycling pathways and the microbial loop, four lakes with different DOM sources were investigated monthly. In Lake Tangxun, Dolichospermum decay released highly labile dissolved organic nitrogen into the water column. This induced bacterial organic nitrogen decomposition, as indicated by the increased abundance of gltB, gltD, gdh, and glnA as well as aminopeptidase activity. Genes associated with dissimilatory nitrate reduction to ammonium further fueled ammonium accumulation, driving Microcystis blooms in the summer. In Lake Zhiyin, fish bait deposits (high nitrogen, similar to Dolichospermum detritus) also caused Microcystis blooms. Detritus from Microcystis decomposition then produced high levels of labile dissolved organic phosphorus, inducing phosphatase activity and increasing soluble reactive phosphorus concentrations from September to April in Lakes Tangxun and Zhiyin. The high refractory DOM from macrophytes in Lake Houguan led to insufficient nutrient availability, leading to nutrient mutualism between algae and bacteria. The high levels of labile dissolved organic carbon from terrestrial detritus in Lake Yandong increased bacterial biomass and production, resulting in low chlorophyll content due to the competitive relationship between algal and bacterial nutrient requirements. Therefore, different DOM compositions induce unique connections among available nutrients, algae, and bacteria in the microbial loop.


Assuntos
Compostos de Amônio , Cianobactérias , Lagos/química , Lagos/microbiologia , Matéria Orgânica Dissolvida , Nitrogênio/análise , China
5.
Environ Sci Technol ; 57(7): 2992-3001, 2023 02 21.
Artigo em Inglês | MEDLINE | ID: mdl-36753734

RESUMO

Regime shifts between nitrogen (N) and phosphorus (P) limitation, which trigger cyanobacterial succession, occur in shallow eutrophic lakes seasonally. However, the underlying mechanism is not yet fully illustrated. We provide a novel insight to address this from interactions between sediment P and nitrification through monthly field investigations including 204 samples and microcosm experiments in Lake Chaohu. Total N to P mass ratios (TN/TP) varied significantly across seasons especially during algal bloom in summer, with the average value being 26.1 in June and descending to 7.8 in September gradually, triggering dominant cyanobacterial succession from Microcystis to Dolichospermum. The regulation effect of sediment N/P on water column TN/TP was stronger in summer than in other seasons. Iron-bound P and alkaline phosphatase activity in sediment, rather than ammonium, contributed to the higher part of nitrification. Furthermore, our microcosm experiments confirmed that soluble active P and enzymatic hydrolysis of organic P, accumulating during algal bloom, fueled nitrifiers and nitrification in sediments. These processes promoted lake N removal and led to relative N deficiency in turn. Our results highlight that N and P cycles do not exist independently but rather interact with each other during lake eutrophication, supporting the dual N and P reduction program to mitigate eutrophication in shallow eutrophic lakes.


Assuntos
Cianobactérias , Lagos , Nitrificação , Fósforo/análise , Nitrogênio/análise , Eutrofização , China
6.
Environ Res ; 217: 114941, 2023 01 15.
Artigo em Inglês | MEDLINE | ID: mdl-36435493

RESUMO

Effective screening feed substitutes for improving water quality in aquaculture systems has become a trending research topic now. In this study, three typical organic agricultural wastes, including sugar cane bagasse (SC), coconut shell powder (CS), and corn cob powder (CC), were selected to evaluate their potential roles on the optimization of water quality and natural bait compared to aquafeeds. Fish feed resulted in the highest growth rate of fish but the worst water quality. Organic detritus addition markedly improved the water quality, especially soluble reactive phosphorus (SRP, decrease of 56-61%) and ammonium (decrease of 16% in SC, 47% in CC). Specially, SC induced core microbes to mediate nutrients transformation and recycling (N2-fixation, ammonification, nitrification, dissimilatory nitrate reduction to ammonia and organic nutrients decomposition), which facilitated the primary productivity based on their positive relationships. This further reduced the available nutrients (especially SRP) in the water and built a mutually beneficial microbial loop. In addition, SC addition increased the abundance of genes involved in amino acids biosynthesis pathways, photosynthesis, and carbon fixation. These results led to energy transfer to higher trophic levels. The addition of CC had a better effect than SC in terms of lower nitrogen levels and a higher fish growth rate (19% in CC, 5% in SC). However, low temperatures and carbon accumulation jointly drive the anaerobic decomposition, resulting in unhealthy microbial loops and low fish growth rates. In contrast to the direct consumption of fish feed, organic detritus can induce more natural bait to provide food for fish by regulating the microbial loop, as showed by the microbial community composition in the water and fish gut. To comprehensively assess water quality, natural bait, and fish growth and quality, certain organic detritus should be considered as an auxiliary material to partially replace feed for healthy and sustainable aquaculture systems.


Assuntos
Agricultura , Qualidade da Água , Animais , Estudos de Viabilidade , Pós , Aquicultura/métodos , Nitrogênio/análise , Peixes/metabolismo
7.
Electrophoresis ; 43(21-22): 2141-2155, 2022 11.
Artigo em Inglês | MEDLINE | ID: mdl-35661383

RESUMO

The utilization of an alternating current electric field provides a good means to achieve controlled coalescence between paired inner cores encapsulated in water-in-oil-in-water double-emulsion (DE) droplets. Although previous studies have experimentally determined the conditions under which inter-core electrokinetic fusion occurs, the transient interfacial dielectrophoretic (DEP) dynamics key to understand the underlying fluid mechanics is still unclear from a physical point of view. By coupling DEP motion of two-phase flow to phase-field formulation, bulk-coupled numerical simulations are conducted to characterize the spatial-temporal evolution of the surface charge wave and the resulting nonlinear electrical force induced at both the core/shell and medium/shell oil/water interfaces. The effect of interfacial charge relaxation and droplet geometry on inter-core attractive dipolar interaction is investigated within a wide parametric space, and four distinct device operation modes, including normal inter-core fusion, shell elongation, partial core leakage, and complete core release, are well distinguished from one another by flow regime argumentation. Our results herein reveal for the first time the hitherto unknown transient electrohydrodynamic fluid motion of DE droplet driven by Maxwell-Wagner structural polarization. The dynamic simulation method proposed in present study points out an effective outlet to predict the nonlinear electrokinetic behavior of multicore DE droplets for realizing a more controlled triggering of microscale reactions for a wide range of applications in drug discovery, skin care, and food industry.


Assuntos
Eletricidade , Emulsões/química
8.
Electrophoresis ; 43(21-22): 2074-2092, 2022 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-36030405

RESUMO

We introduce herein an effective way for continuous delivery and position-switchable trapping of nanoparticles via field-effect control on hybrid electrokinetics (HEK). Flow field-effect transistor exploiting HEK delicately combines horizontal linear electroosmosis and transversal nonlinear electroosmosis of a shiftable flow stagnation line (FSL) on gate terminals under DC-biased AC forcing. The microfluidic nanoparticle concentrator proposed herein makes use of a simple device geometry, in which an individual or a series of planar metal strips serving as gate electrode (GE) are subjected to a hybrid gate voltage signal and arranged in parallel between a pair of 3D driving electrodes. On the application of a DC-biased AC electric field across channel length direction, all the GE are electrochemically polarized, and the action of imposed hybrid electric field on the multiple-frequency bipolar counterions within the composite-induced double layer generates two counter-rotating induced-charge electroosmotic (ICEO) micro-vortices on top of each GE. Symmetry breaking in ICEO flow profile occurs once the gate voltage deviates from natural floating potential of corresponding GE. The gate voltage offset not only results in an additional pump motion of working fluid for enhanced electroosmotic transport but also directly changes the location of FSL where nanoparticles are preferentially collected by field-effect HEK. Our results of field-effect control on HEK are supposed to guide an elaborate design of flexible electrokinetic frameworks embedding coplanar metal strips for a high degree of freedom analyte manipulation in modern micro-total-analytical systems.

9.
Soft Matter ; 18(3): 609-616, 2022 Jan 19.
Artigo em Inglês | MEDLINE | ID: mdl-34929022

RESUMO

The heart beating phenomenon of room temperature liquid metal (LM) mercury has attracted much attention in the past years, but its research and application are limited because of the low vapor pressure and high toxicity. Here, a fundamental scientific finding is reported that the non-toxic eutectic gallium indium (EGaIn) alloy droplets beat periodically at a certain frequency based on a floating electrode under the stimulation of the direct current (DC) field. The essential characteristics of heart beating are the displacement and the projected area change of the LM droplet. The mechanism of this phenomenon is the self-regulation of interfacial tension caused by chemical oxidation, chemical corrosion, and continuous electrowetting. In this article, a series of experiments are also carried out to examine the effects of different factors on the heartbeat, such as voltage, the volume of the droplet, the droplet immersion depth, the electrolyte solution concentration, the distance of electrodes, and the type of floating electrode. Finally, the heartbeat state and application boundary of the LM droplet under different conditions are summarized by imitating the human life process. The periodic changes of the LM droplet under an external DC electric field provide a new method to simulate the beating of the heart artificially, and can be applied to the research of organ chip fluid pumping in the future.


Assuntos
Eletroumectação , Gálio , Eletricidade , Eletrodos , Frequência Cardíaca , Humanos
10.
Electrophoresis ; 42(7-8): 939-949, 2021 04.
Artigo em Inglês | MEDLINE | ID: mdl-32705697

RESUMO

We introduce herein an efficient microfluidic approach for continuous transport and localized collection of nanoparticles via hybrid electrokinetics, which delicately combines linear and nonlinear electrokinetics driven by a composite DC-biased AC voltage signal. The proposed technique utilizes a simple geometrical structure, in which one or a series of metal strips serving as floating electrode (FE) are attached to the substrate surface and arranged in parallel between a pair of coplanar driving electrodes (DE) in a straight microchannel. On application of a DC-biased AC electric field across the channel, nanoparticles can be transported continuously by DC bulk electroosmotic flow, and then trapped selectively onto the metal strips due to AC-field induced-charge electrokinetic (ICEK) phenomenon, which behaves as counter-rotating micro-vortices around the ideally polarizable surfaces of FE. Finite-element simulation is carried out by coupling the dual-frequency electric field, flow field and sample mass transfer in sequence, for guiding a practical design of the microfluidic nanoparticle concentrator. With the optimal device geometry, the actual performance of the technique is investigated with respect to DC bias, AC voltage amplitude, and field frequency by using both latex nanospheres (∼500 nm) and BSA molecules (∼10 nm). Our experimental observation indicates nanoparticles are always enriched into a narrow bright band on the surface of each FE, and a horizontal concentration gradient even emerges in the presence of multiple metal strips, which therefore permits localized analyte enrichment. The proposed trapping method is supposed to guide an elaborate design of flexible electrokinetic frameworks embedding FE for continuous-flow analyte manipulation in modern microfluidic systems.


Assuntos
Microfluídica , Nanopartículas , Eletricidade , Eletro-Osmose
11.
J Soils Sediments ; 21(10): 3427-3437, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34075310

RESUMO

Purpose: Nitrogen (N) and phosphorus (P) are the key elements leading to eutrophication, and it is important to jointly control N and P release from sediments into the water column. Methods: Different mixed materials including P sorbent, natural organic carbon (C), and an oxidizing agent were applied in a 1-year pilot-scale experiment. Results: The addition of iron-rich (IR) clay and Phoslock agent promoted the formation of iron bound P (Fe(OOH)~P) and calcium bound P (CaCO3~P) in sediments, respectively. IR clay offered more advantages in immobilization of phosphorus as refractory P, and the Phoslock agent more effectively reduced the risk of P release into water, which was expressed as a low equilibrium P concentration (EPC0). Mixtures of sugarcane (SU) detritus and IR clay exhibited high carbohydrate (CHO) contents, which further fuelled both denitrification and dissimilatory nitrate reduction to ammonium (DNRA). This indicated that the SU dosage should be controlled to avoid DNRA over denitrification. Attention should be given to the fact that SU introduction significantly promoted the generation of an anaerobic state, leading to the desorption and release of Fe(OOH)~P, which could be alleviated by using Oxone. Multienzyme activity analysis showed that P and N transformation shifted from P desorption to organic P hydrolysis and from ammonification to denitrification and DNRA, respectively. Conclusion: We recommend the use of P sorbent and organic C combined with oxidizing agents as effective mixed materials for sediment remediation, which could enhance P adsorption and provide electron donors for denitrification, while also avoiding the generation of anoxia.

12.
J Environ Sci (China) ; 103: 255-267, 2021 May.
Artigo em Inglês | MEDLINE | ID: mdl-33743907

RESUMO

In order to better understand the contribution of nutrients regeneration pathway, release potential and transformation pattern to cyanobacterial growth and succession, 7 sampling sites in Lake Chaohu with different bloom degree were studied every two months from February to November 2018. The carbon, nitrogen (N) and phosphorus (P) forms or fractions in surface, interstitial water and sediments as well as extracellular enzymatic activities, P sorption, specific microbial abundance and community composition in sediments were analyzed. P regeneration pathway was dominated by iron-bound P desorption and phosphorus-solubilizing bacteria solubilization in severe-bloom and slight-bloom area respectively, which both resulted in high soluble reactive phosphorus (SRP) accumulation in interstitial water. However, in severe-bloom area, higher P release potential caused the strong P release and algal growth, compared to slight-bloom area. In spring, P limitation and N selective assimilation of Dolichospermum facilitated nitrate accumulation in surface water, which provided enough N source for the initiation of Microcystis bloom. In summer, the accumulated organic N in Dolichospermum cells during its bloom was re-mineralized as ammonium to replenish N source for the sustainable development of Microcystis bloom. Furthermore, SRP continuous release led to the replacement of Dolichospermum by Microcystis with the advantage of P quick utilization, transport and storage. Taken together, the succession from Dolichospermum to Microcystis was due to both the different forms of N and P in water column mediated by different regeneration and transformation pathways as well as release potential, and algal N and P utilization strategies.


Assuntos
Cianobactérias , Microcystis , China , Eutrofização , Lagos , Nutrientes , Fósforo/análise
13.
Electrophoresis ; 41(10-11): 778-792, 2020 06.
Artigo em Inglês | MEDLINE | ID: mdl-31943244

RESUMO

We study herein numerically the use of induced-charge electrokinetic phenomena to enable a flexible control of ion transport of dilute electrolyte in a straight ion concentration polarization system. The effect of three convection modes of induced-charge electrokinetic phenomena, including induced-charge electroosmosis, flow-field effect transistor, and alternating-current electroosmosis (ACEO), on convective arrestment of diffusive wave-front propagation is investigated by developing a cross-scale and fully coupled transient numerical simulation model, wherein multiple frequency electrochemical polarization and nonlinear diffuse charge dynamics in spatiotemporally varying solution conductivity are taken into account. We demonstrate by detailed comparative simulation studies that ACEO vortex flow field above a metal strip array arranged along the anodic chamber's bottom surface serves as the most efficient way for adjusting the salt density distribution at micrometer and even millimeter dimension, due to its high flexibility in controlling the stirring flow state with the introduction of two extra electrical parameters. The specific operating status is determined by whether the electrode array is floating in potential (induced-charge electroosmosis) or biased to ground (flow-field effect transistor) or forced to oscillate at another Fourier mode (ACEO). These results prove useful for on-chip electric current control with electroconvective stirring.


Assuntos
Eletro-Osmose/métodos , Íons/química , Microfluídica/métodos , Simulação por Computador , Condutividade Elétrica , Eletrólitos/química , Dinâmica não Linear
14.
Electrophoresis ; 40(6): 979-992, 2019 03.
Artigo em Inglês | MEDLINE | ID: mdl-30256428

RESUMO

Induced-charge electroosmosis (ICEO) has attracted tremendous popularity for driving fluid motion from the microfluidic community since the last decade, while less attention has been paid to ICEO-based nanoparticle manipulation. We propose herein a unique concept of hybrid electroosmotic kinetics (HEK) in terms of bi-phase ICEO (BICEO) actuated in a four-terminal spiral electrode array, for effective electrokinetic enrichment of fluorescent polystyrene nanoparticles on ideally polarizable metal strips. First, by alternating the applied AC voltage waves between consecutive discrete terminals, the flow stagnation lines where the sample nanoparticles aggregate can be switched in time between two different distribution modes. Second, we innovatively introduce the idea of AC field-effect flow control on BICEO; by altering the combination of gating voltage sequence, not only the number of circulative particle trapping lines is doubled, but the collecting locations can be flexibly reconfigured as well. Third, hydrodynamic streaming of DC-biased BICEO is tested in our device design, wherein the global linear electroosmosis dominates BICEO contributed from both AC and DC components, resulting in a reduction of particle enrichment area, while with a sharp increase in sample transport speed inside the bulk phase. The flow field associated with HEK is predicted using a linear asymptotic analysis under Debye-Huckel limit, with the simulation results in qualitative agreement with in-lab observations of nanoparticle trapping by exploiting a series of improved ICEO techniques. This work provides an affordable and field-deployable platform for real-time nanoparticle trapping in the context of dilute electrolyte.


Assuntos
Eletro-Osmose/instrumentação , Técnicas Analíticas Microfluídicas/instrumentação , Nanopartículas/química , Eletricidade , Eletro-Osmose/métodos , Desenho de Equipamento , Cinética , Microeletrodos
15.
Environ Sci Technol ; 52(10): 5653-5661, 2018 05 15.
Artigo em Inglês | MEDLINE | ID: mdl-29688011

RESUMO

Dolichospermum flos-aquae (formerly Anabaena flos-aquae) is a diazotrophic cyanobacterium causing harmful blooms worldwide, which is partly attributed to its capacity to compete for nitrogen (N) and phosphorus (P). Preventing the blooms by reducing P alone or both N and P has caused debate. To test the effects alone and together on the growth of cyanobacteria, we performed culture experiments in different eutrophication scenarios. N2 fixation in terms of heterocyst density, nitrogenase activity and nifH expression increased significantly in P-replete cultures, suggesting that P enrichment facilitates N2 fixation. Correspondingly, the expression of genes involved in P uptake, e.g., those involved in P-transport ( pstS) and the hydrolysis of phosphomonoesters ( phoD), was upregulated in P-deficient cultures. Interestingly, N addition enhanced not only the expression of these genes but also polyphosphate formation and alkaline phosphatase activity in P-deficient cultures relative to the P-replete cultures, as evidenced by qualitative (enzyme-labeled fluorescence) and quantitative (fluorogenic spectrophotometry) measurements. Furthermore, after N addition, cell activity and growth increased in the P-deficient cultures, underscoring the risk of N enrichment in P-limited systems. The eco-physiological responses shown here help further our understanding of the mechanism of N and P colimitation and underscore the importance of dual N and P reduction in controlling cyanobacterial blooms.


Assuntos
Dolichospermum flosaquae , Fósforo , Eutrofização , Nitrogênio , Nutrientes
16.
Biochem Biophys Res Commun ; 479(2): 224-230, 2016 10 14.
Artigo em Inglês | MEDLINE | ID: mdl-27638309

RESUMO

To elucidate the key miRNAs and the signalling pathways that are involved in porcine oocyte maturation, we performed a deep sequencing analysis of the miRNAs of pig germinal vesicle (GV) oocytes and metaphase II (MII) oocytes. Seven differentially expressed (DE) miRNAs were identified and the expression levels of miR-21 and miR-27b-3p were further confirmed by QPCR analysis. The target genes of 7 DE miRNAs were predicted and subjected to pathway analysis. Interestingly, fatty acid metabolism and fatty acid biosynthesis were the top two significantly enriched molecular functions during oocyte maturation. Heat map, which was built with 7 DE miRNAs and the enriched the molecular functions, revealed that miR-21, miR-27b-3p, miR-10a-5p and miR-10b-5p were involved in fatty acid metabolism. In particular, the regulatory role of miR-27b-3p on peroxisome proliferator-activated receptor-γ (PPARγ) was confirmed by their inversed expression patterns in GV and MII oocytes and luciferase report assays. In addition, we observed that PPARγ agonist (rosiglitazone) treatment significantly enhanced porcine oocyte maturation rate and early embryo developmental competent. Taken together, our results demonstrated that miR-27b and its target, PPARγ, play the vital roles in pig oocyte maturation through regulating the fatty acid metabolism. These data increased our understanding of the regulatory gene networks in porcine oocyte maturation and development.


Assuntos
Regulação da Expressão Gênica , MicroRNAs/genética , Oócitos/metabolismo , PPAR gama/genética , Regiões 3' não Traduzidas/genética , Animais , Sequência de Bases , Western Blotting , Análise por Conglomerados , Ácidos Graxos/metabolismo , Feminino , Perfilação da Expressão Gênica/métodos , Técnicas de Maturação in Vitro de Oócitos , MicroRNAs/classificação , Oócitos/citologia , Oócitos/efeitos dos fármacos , PPAR gama/agonistas , PPAR gama/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Rosiglitazona , Homologia de Sequência do Ácido Nucleico , Transdução de Sinais/genética , Suínos , Tiazolidinedionas/farmacologia
17.
Genes Cells ; 18(1): 42-55, 2013 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-23231677

RESUMO

Induced pluripotent stem cells (iPSCs) are flourishing in the investigation of cell reprogramming. However, we still know little about the sequential molecular mechanism during somatic cell reprogramming (SCR). Here, we first observed rapid generation of colonies whereas mouse embryonic fibroblasts (MEFs) were induced by OCT4, SOX2, KLF4 (OSK), and vitamin C for 7 days. The colony's global transcriptional profiles were analyzed using Affymetrix microarray. Microarray data confirmed that SCR was a process in which transcriptome got reversed and pluripotent genes expressed de novo. There were many changes, especially substantial growth expression of epigenetic factors, on transcriptome during the transition from Day 7 to iPSCs indicating that this period may provide 'flexibility' genome structure, chromatin remodeling, and epigenetic modifications to rebind to the transcriptional factors. Several biological processes such as viral immune response, apoptosis, cell fate specification, and cell communication were mainly involved before Day 7 whereas cell cycle, DNA methylation, and histone modification were mainly involved after Day 7. Furthermore, it was suggested that p53 signaling contributed to the transition 'hyperdynamic plastic' cell state and assembled cell niche for SCR, and small molecular compounds useful for chromatin remodeling can enhance iPSCs by exciting epigenetic modification rather than the exogenous expression of more TFs vectors.


Assuntos
Reprogramação Celular , Fibroblastos/citologia , Células-Tronco Pluripotentes Induzidas/citologia , Transcriptoma , Animais , Apoptose , Ácido Ascórbico/metabolismo , Comunicação Celular , Ciclo Celular , Montagem e Desmontagem da Cromatina , Metilação de DNA , Epigênese Genética , Fibroblastos/metabolismo , Perfilação da Expressão Gênica , Regulação da Expressão Gênica no Desenvolvimento , Genoma , Células-Tronco Pluripotentes Induzidas/metabolismo , Fator 4 Semelhante a Kruppel , Fatores de Transcrição Kruppel-Like/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Fator 3 de Transcrição de Octâmero/metabolismo , Análise de Sequência com Séries de Oligonucleotídeos , Fatores de Transcrição SOXB1/metabolismo , Proteína Supressora de Tumor p53/metabolismo
18.
Neuro Endocrinol Lett ; 35(6): 523-30, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25433842

RESUMO

OBJECTIVES: Post-translational modifications of lysine residues of histones can result in a series of functional changes. Lysine 79 of histone H3 (H3K79) can be methylated specifically by the Dot1 family of histone lysine methyltransferases. Although multiple developmental abnormalities in Dot1L-deficient mouse embryos have been studied, the biological function of H3K79 methylation in mammal oocytes remains unclear. Here, the distribution of Dot1L, methyltransferase of residue lys79 of histone H3 (H3K79) in mouse, and its effect on mouse oocytes meiosis were investigated to examine whether there are changes in the pattern of distribution and effect of Dot1L on mouse oocytes meiosis. METHODS: The mRNA level of Dot1l in mouse oocytes was examined using real-time qPCR (RT-qPCR) technique. The distribution and level of Dot1L protein and H3K79 methylation were examined using immunofluorescence and western-blot techniques, respectively. The down regulation of Dot1l in mouse oocytes was conducted using siRNA injection technique. RESULTS: Dot1L was detected diffuse staining in the nuclear of mouse GV (Germinal Vesicle) stage oocytes. The Dot1l expression and H3K79 methylation level were suppressed effectively with anti-Dot1l siRNA injection. In Dot1L deficient, accompanying with BubR1 (MAD3/Bub1b) remains on the chromosome, the mouse oocytes was blocked in metaphase of meiosis I. The histone deacetylation was also incomplete in Dot1L-deficient mouse oocytes. CONCLUSION: Dot1L protein is well distributed in mouse GV stage oocytes. Dot1L and H3K79 methylation play important roles in meiosis progression and are supposed to be associated with chromosome deacetylation of mouse oocytes.


Assuntos
Histonas/metabolismo , Meiose/fisiologia , Metiltransferases/metabolismo , Oócitos/metabolismo , Acetilação , Animais , Feminino , Histona-Lisina N-Metiltransferase , Histonas/genética , Metilação , Metiltransferases/genética , Camundongos Endogâmicos ICR , Oócitos/citologia , Processamento de Proteína Pós-Traducional/fisiologia , RNA Mensageiro/genética , RNA Interferente Pequeno/genética
19.
Sci Total Environ ; 925: 171610, 2024 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-38462007

RESUMO

In shallow lakes, there are complex relationships between lake eutrophication and greenhouse gas emissions that deserve to be studied, which are important for solving lake eutrophication, slowing down climate warming, and reducing carbon emissions. In order to explore the relationship and mechanism between eutrophication and greenhouse gases (GHGs), the net GHGs emission flux and transformation of carbon, and nitrogen in 45 shallow freshwater lakes were investigated from May to September 2022. Eutrophication facilitated potential denitrification rate (Dt) without increasing nitrous oxide (N2O) production based on the significantly positive relationship between eutrophication and Dt. This should be attributed to the shift from incomplete (N2O producing process) to complete denitrification (N2 producing process). Compared to NarG mediating nitrate (NO3-) to nitrite (NO2-), fewer eutrophication indicators showed a positive relationship with NosZ mediating N2O to N2, suggesting that more stringent conditions are required for complete denitrification, which was achieved in the lakes we investigated. Optimal reduction in net carbon dioxide (CO2) emissions occurs at high levels of primary productivity, as indicated by the V-shaped relationship between chlorophyll a (Chl a) and CO2 emissions. However, in hyper-eutrophic lakes, there is an upward trend in CO2 production. The possible explanations should include CO2 production and fixation as well as methane (CH4) oxidation. The bell-shaped relationship between the net flux of CH4 emission and Chl a could be explained that CH4 was heavily oxidized due to sufficient oxygen caused by algal bloom. This fact gave evidence for the increase of the net flux of CO2 emission in high primary productivity lakes. Therefore, the relationship and mechanism between net GHGs emission flux and eutrophication remained complex and various.

20.
Water Res ; 249: 120910, 2024 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-38016223

RESUMO

Understanding the long-term variations in basins that undergo large-scale hydroelectric projects is crucial for effective dam operation and watershed management. In this study, comprehensive analyses were conducted on a dataset spanning over 20 years (1998-2018) of hydrological regime and physicochemical parameters from the Yangtze River basin to evaluate the potential impacts of the Three Gorges Dam. Water level significantly increased from 128.75±58.18 m in 2002 to 136.78±55.05 m in 2005, and the mean flow velocity significantly decreased from 2004 to 2010. However, no significant change in the flow was observed in the basin. Meanwhile, remarkable fluctuations in physicochemical parameters, including dissolved oxygen, chemical oxygen demand, conductivity, hardness, and alkalinity, were mainly observed during impoundment (2003-2009). After that, the above parameters tended to stabilize, and some even returned to their original levels. The dam's retention effect significantly reduced the suspended solids (SS) in both up- and downstream, to only one-third of the pre-operation level. And total phosphorus and chemical oxygen demand also significantly decreased with the decline of SS. Particularly, ammonium also showed a significant downward trend, with the up- and downstream of the dam falling by 36.8 % and 26.1 %, respectively. However, the increasing total nitrogen (7.5 % and 20.0 % up- and downstream of the dam, respectively) still threatened the water quality of the basin, especially in the estuaries. Additionally, the significant decline in dissolved oxygen downstream (from 8.53±1.08 mg/L to 8.11±1.36 mg/L) also exacerbated the hypoxia in the Yangtze River estuary. The results demonstrated the long-term impact of the construction of the Three Gorges Dam on the environmental elements of the Yangtze River basin, which provides reference data and guidance for the construction of big dams in major rivers in the future.


Assuntos
Monitoramento Ambiental , Qualidade da Água , Rios , Estuários , Oxigênio , China
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