RESUMEN
RNAi and Polycomb repression play evolutionarily conserved and often coordinated roles in transcriptional silencing. Here, we show that, in the protozoan Tetrahymena thermophila, germline-specific internally eliminated sequences (IESs)-many related to transposable elements (TEs)-become transcriptionally activated in mutants deficient in the RNAi-dependent Polycomb repression pathway. Germline TE mobilization also dramatically increases in these mutants. The transition from noncoding RNA (ncRNA) to mRNA production accompanies transcriptional activation of TE-related sequences and vice versa for transcriptional silencing. The balance between ncRNA and mRNA production is potentially affected by cotranscriptional processing as well as RNAi and Polycomb repression. We posit that interplay between RNAi and Polycomb repression is a widely conserved phenomenon, whose ancestral role is epigenetic silencing of TEs.
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Elementos Transponibles de ADN/genética , Proteínas del Grupo Polycomb/genética , Proteínas Protozoarias/genética , Interferencia de ARN , Tetrahymena thermophila/genética , Activación Transcripcional/genética , Epigénesis Genética , Silenciador del Gen , Mutación , ARN Mensajero/genética , ARN no Traducido/genéticaRESUMEN
Due to the inherent tendency to be blocked by obstacles, reliability is a major challenge for visible light communication (VLC). The intelligent reflective surface (IRS) is an effective way to reduce or eliminate the influence of blockage in the VLC system. However, the complexity increases correspondingly due to complex shadow analysis when access points (APs), IRSs, and obstacles coexist. We proposed a robust layout optimization scheme for the IRS-VLC system to resist blockages. First, we model the random obstructions based on spatial geometry methods. Second, we build the optimization problem model considering consistent illumination, achievable data rate (ADR), the positions of the APs, and the directions of the IRS array. In order to achieve this objective, we develop an anti-occlusion method based on the non-dominated sorting genetic algorithm-II (NSGA-II) to get the Pareto front and use enhanced measurement functions (ASFs) to extract the best solution. Simulation results show that the proposed scheme works well in the IRS-VLC system. It is noteworthy that the rectangle layout has always demonstrated superior performance in the IRS-VLC system compared to other traditional layouts.
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Biofilms formed by spoilage and pathogenic bacteria increase microbial persistence, causing an adverse influence on the quality of seafood. The mono-species biofilms are widely reported, however, the contamination of multi-species biofilms and their matrix in food environments are still not fully understood. Here, we assessed the contamination of multi-species biofilms in three seafood processing environments with different hygiene levels by detecting bacterial number and three biofilm matrix components (carbohydrates, extracellular DNA (eDNA), and proteins). Samples comprising seven food matrix surfaces and eight food processing equipment surfaces were collected from two seafood processing plants (XY and XC) and one seafood market (CC). The results showed that the bacterial counts ranged from 1.89 to 4.91 CFU/cm2 and 5.68 to 9.15 BCE/cm2 in these surfaces by cultivation and real-time PCR, respectively. Six biofilm hotspots were identified, including four in CC and two in XY. Among the three processing environments, the amplicon sequence variants (ASVs) of Proteobacteria, Bacteroidetes, and Actinobacteria decreased with improved processing hygiene, while Firmicutes showed a decrease in the four most abundant phyla. The most prevalent bacteria belonged to genera Psychrobacter, Acinetobacter, and Pseudomonas, demonstrating the significant differences and alteration in bacterial community composition during different environments. From the biofilm hotspots, 15 isolates with strong biofilm forming ability were identified, including 7 Pseudomonas, 7 Acinetobacter, and 1 Psychrobacter. The Pseudomonas isolates exhibited the highest production of EPS components and three strong motilities, whose characteristics were positively correlated. Thus, this study verified the presence of multi-species biofilms in seafood processing environments, offering preliminary insights into the diversity of microbial communities during processing. It highlights potential contamination sources and emphasizes the importance of understanding biofilms composition to control biofilms formation in seafood processing environments.
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Bacterias , Biopelículas , Manipulación de Alimentos , Microbiología de Alimentos , Microbiota , Alimentos Marinos , Biopelículas/crecimiento & desarrollo , Alimentos Marinos/microbiología , Bacterias/clasificación , Bacterias/genética , Bacterias/aislamiento & purificación , Bacterias/crecimiento & desarrollo , Higiene , Contaminación de Alimentos/análisisRESUMEN
Psychrotrophic Pseudomonas as the dominant spoilage bacteria, have biofilm forming ability, increasing persistence and contamination in the chilled food. Biofilm formation of spoilage Pseudomonas at cold temperature was documented, however, role of extracellular matrix in mature biofilm and stress resistance of psychrotrophic Pseudomonas are much less abundant. The aim of this study was to investigate the biofilm forming characteristics of three spoilers P. fluorescens PF07, P. lundensis PL28, and P. psychrophile PP26 at 25 °C, 15 °C and 4 °C, and to explore their stress resistance to chemical and thermal treatments of mature biofilms. The results showed that biofilm biomass of three Pseudomonas at 4 °C was significantly higher than that at 15 °C and 25 °C. The secretion of extracellular polymeric substances (EPS) greatly increased in those Pseudomonas under low temperature, of which extracellular protein constituted about 71.03%-77.44%. Compared to 25 °C, the mature biofilms were observed to more aggregation and thicker spatial structure at 4 °C ranging from 42.7 to 54.6 µm, in contrast to 25.0-29.8 µm at 25 °C, especially strain PF07. These Pseudomonas biofilms switched into moderate hydrophobicity, and their swarming and swimming were significantly inhibited at low temperature. Furthermore, the resistance to NaClO and heating at 65 °C apparently enhanced for mature biofilm formed at 4 °C, indicating the difference in EPS matrix production influenced the stress resistance of biofilm. In addition, three strains contained alg and psl operons for exopolysaccharide biosynthesis, and biofilm related genes of algK, pslA, rpoS, and luxR were significantly up-regulated, while flgA gene was down-regulated at 4 °C compared to 25 °C, consistent with the above phenotype changes. Thus, the dramatic increase of mature biofilm and their stress resistance in psychrotrophic Pseudomonas were associated with large secretion and protection of extracellular matrix under low temperature, which provide a theoretical basis for subsequent biofilm control during cold chain.
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Frío , Pseudomonas , Biopelículas , Matriz Extracelular de Sustancias Poliméricas , Matriz ExtracelularRESUMEN
In this paper, an integrated visible light communication and positioning (VLCP) code division multiple access (CDMA) system is proposed for recovering original user data and obtaining positioning information simultaneously. A generalized modification method for balanced bipolar code set is presented, which can be applied to VLCP-CDMA system with intensity modulation. Both modified bipolar code sets, zero correlation zone (ZCZ) code set and Walsh code set, are employed and evaluated in VLCP-CDMA systems. When considering synchronous and quasi-synchronous (QS) systems, it is demonstrated that the modified ZCZ code set with ideal zero correlation zone properties performs better communication and positioning performance than the modified Walsh code set by system simulations and experiments. As a result, a bit error rate (BER) of 1.8×10-3 and an average positioning error of 1.50 cm are successfully obtained by adopting modified ZCZ code set in a 4-user real-time VLCP-CDMA system. It will offer a promising solution to meet both communication and positioning requirements of future intelligent systems.
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In this paper, a novel adaptive feedback threshold (AFT) based demodulation for mobile visible light communication and positioning (VLCP) integrated system is proposed. The AFT can vary with the received signal without excessive fluctuation and support communication and positioning in mobile environment. For the VLCP integrated system with single light emitting diodes (LED) and multiple photodetectors (PDs), maximal ratio combining (MRC) and received optical power ratio (ROPR) based on the AFT is further achieved for communication and positioning with high performance, respectively. It is demonstrated that high credible communication and high accuracy positioning for the mobile integrated VLCP system can be realized by using the proposed AFT based demodulation. As a result, the implemented VLCP system with a moving speed of 1 m/s is evaluated experimentally. Average positioning error of 3.43 cm with 800 mA current and the bit error ratio (BER) with different currents are also obtained.
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Shewanella baltica is a typical specific spoilage organism causing the deterioration of seafood, but the exact regulation of its adaptive and competitive dominance in diverse environments remains undefined. In this study, the regulatory function of two sigma factors, RpoS and RpoN, in environmental adaptation and spoilage potential were evaluated in S. baltica SB02. Two in-frame deletion mutants, ΔrpoS and ΔrpoN, were constructed to explore the roles in their motility, biofilm formation, stress response and spoilage potential, as well as antibiotics by comparing the phenotypes and transcription with those of wild type (WT) strain. Compared with WT strain, the ΔrpoN showed the slower growth and weaker motility due to loss of flagella, while swimming of the ΔrpoS was increased. Deletion of rpoN significantly decreased biofilm biomass, and production of exopolysaccharide and pellicle, resulting in a thinner biofilm structure, while ΔrpoS formed the looser aggregation in biofilm. Resistance of S. baltica to NaCl, heat, ethanol and three oxidizing disinfectants apparently declined in the two mutants compared to WT strain. The ΔrpoN mutant decreased sensory score, accumulation of trimethylamine, putrescine and TVB-N and protease activity, while a weaker effect was observed in ΔrpoS. The two mutants had significantly higher susceptibility to antibiotics than WT strain, especially ΔrpoN. Deficiency of rpoN and rpoS significantly repressed the activities of two diketopiperazines related to quorum sensing (QS). Furthermore, transcriptome analyses revealed that RpoN was involved in the regulation of the expression of 143 genes, mostly including flagellar assembly, nitrogen and amino acid metabolism, ABC transporters. Transcript changes of seven differentially expressed coding sequences were in agreement with the phenotypes observed in the two mutants. Our findings reveal that RpoN, as a central regulator, controls the fitness and bacterial spoilage in S. baltica, while RpoS is a key regulatory factor of stress response. Characterization of these two sigma regulons in Shewanella has expanded current understanding of a possible co-regulatory mechanism with QS for adaptation and spoilage potential.
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Proteínas Bacterianas/metabolismo , Perciformes/microbiología , Shewanella/fisiología , Factor sigma/metabolismo , Adaptación Fisiológica , Animales , Proteínas Bacterianas/genética , Biopelículas , Contaminación de Alimentos/análisis , Regulación Bacteriana de la Expresión Génica , Percepción de Quorum , Regulón , Shewanella/genética , Factor sigma/genéticaRESUMEN
The Clostridium perfringens alpha toxin (CPA), encoded by the plc gene, is the causative pathogen of gas gangrene, which is a lethal infection. In this study, we used an E. coli system for the efficient production of recombinant proteins and developed a bicistronic design (BCD) expression construct consisting of two copies of the C-terminal (247-370) domain of the alpha toxin (CPA-C) in the first cistron, followed by Cholera Toxin B (CTB) linked with another two copies of CPA-C in the second cistron that is controlled by a single promoter. Rabbits were immunized twice with purified proteins (rCPA-C rCTB-CPA-C) produced in the BCD expression system, with an inactivated recombinant E. coli vaccine (RE), C. perfringens formaldehyde-inactivated alpha toxoid (FA-CPA) and C. perfringensl-lysine/formaldehyde alpha toxoid (LF-CPA) vaccines. Following the second vaccination, 0.1 mL of pooled sera of the RE-vaccinated rabbits could neutralize 12× mouse LD100 (100% lethal dose) of CPA, while that of the rCPA-C rCTB-CPA-C-vaccinated rabbits could neutralize 6× mouse LD100 of CPA. Antibody titers against CPA were also assessed by ELISA, reaching titers as high as 1:2048000 in the RE group; this was significantly higher compared to the C. perfringens alpha toxoid vaccinated groups (FA-CPA and LF-CPA). Rabbits from all vaccinated groups were completely protected from a 2× rabbit LD100 of CPA challenge. These results demonstrate that the recombinant proteins are able to induce a strong immune responses, indicating that they may be potentially utilized as targets for novel vaccines specifically against the C. perfringens alpha toxin.
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Anticuerpos Antibacterianos/sangre , Toxinas Bacterianas , Proteínas de Unión al Calcio , Proteínas Recombinantes , Fosfolipasas de Tipo C , Animales , Toxinas Bacterianas/biosíntesis , Toxinas Bacterianas/genética , Toxinas Bacterianas/inmunología , Toxinas Bacterianas/aislamiento & purificación , Vacunas Bacterianas , Proteínas de Unión al Calcio/biosíntesis , Proteínas de Unión al Calcio/genética , Proteínas de Unión al Calcio/inmunología , Proteínas de Unión al Calcio/aislamiento & purificación , Toxina del Cólera/genética , Clonación Molecular , Clostridium perfringens/genética , Clostridium perfringens/metabolismo , Escherichia coli/genética , Ratones , Conejos , Proteínas Recombinantes/biosíntesis , Proteínas Recombinantes/genética , Proteínas Recombinantes/inmunología , Proteínas Recombinantes/aislamiento & purificación , Fosfolipasas de Tipo C/biosíntesis , Fosfolipasas de Tipo C/genética , Fosfolipasas de Tipo C/inmunología , Fosfolipasas de Tipo C/aislamiento & purificación , Vacunación/métodosRESUMEN
The use of baker's yeast to reduce ethyl 2-oxo-4-phenylbutyrate (EOPB) in conventional biphasic systems is hindered by low productivities due to mass transfer resistance between the biocatalyst and the substrate partitioned into two different phases. To overcome the limitation, a new reaction-separation coupling process (RSCP) was configured in this study, based on the novel thermosensitive ionic liquids (ILs) with polyoxyethylene-tail. The solubility of ILs in common solvents was investigated to configure the unique thermosensitive ionic liquids-solvent biphasic system (TIBS) in which the reduction was performed. [(CH3)2N(C2H5)(CH2CH2O)2H][PF6] (c2) in 1,2-dimethoxyethane possesses the thermosensitive function of homogeneous at lower temperatures and phase separating at higher temperatures. The phase transformation temperature (PTT) of the mixed system of c2/1,2-dimethoxyethane (v/v, 5:18) was about 33 °C. The bioreaction takes place in a "homogeneous" liquid phase at 30 °C. At the end of each reduction run, the system temperature is increased upon to the PTT, while c2 is separated from 1,2-dimethoxyethane with turning the system into two phases. The enantiomeric excesses (e.e.) of ethyl (R)-2-hydroxy-4-phenylbutyrate ((R)-EHPB) increased about 25~30% and the yield of ethyl-2-hydroxy-4-phenylbutyrate (EHPB) increased 35% in TIBS, compared with the reduction in 1,2-dimethoxyethane. It is expected that the TIBS established in this study could provide many future opportunities in the biocatalysis.
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Líquidos Iónicos , Fenilbutiratos/metabolismo , Polietilenglicoles/química , Saccharomyces cerevisiae/metabolismo , Biocatálisis , Catálisis , Cloruros/química , Tecnología Química Verde , Solubilidad , Solventes , Estereoisomerismo , Temperatura , AguaRESUMEN
In this paper, a new construction of optical zero correlation zone (OZCZ) code set pair is proposed for quasi-synchronous code division multiple access (QS-CDMA) visible light communication (VLC) system. Owing to its excellent correlation properties within zero correlation zone and greater code weight, the multi-user QS-CDMA-VLC system using the proposed code construction can obtain both good performance in illumination and communication. An experimental QS-CDMA-VLC system with two optical channels and two experiment setups is designed to evaluate the performance of the new OZCZ code set pair. When the free-space transmission distance difference is 1 m, we successfully achieve the 2-user and 14-user CDMA-VLC for 32.5 Mb/s and 24.1 Mb/s overall bit rate respectively, under the 7% forward-error-correction (FEC) limit of 3.8×10 -3. The experimental results indicate that, comparing to the other existing optical code sets, the proposed OZCZ construction can effectively reduce the impact of non-perfect synchronous problem, so that the bit error rate (BER) performance is improved. And dimming values and overall bit rates of the system can be significantly improved.
RESUMEN
In this paper, we propose a new construction of optical zero correlation zone (OZCZ) code set for quasi-synchronous code division multiple access (QS-CDMA) visible light communication (VLC) system. The proposed code set includes a pair of unipolar and bipolar code sets, which is more suitable for intensity modulation and direct detection (IM/DD) and maintains dimming values at 50% regardless of user numbers for CDMA-VLC system. A QS-CDMA-VLC system adopting the proposed code set is investigated by numerical simulation and experimental verification. The system bit error rate (BER) performance is evaluated with different user numbers, sample rates, delay user numbers and transmission distances. In a 32-user CDMA-VLC system, the 250 MS/s transmission at a distance of 1.5 m can be achieved successfully with the BER less than the forward-error-correction (FEC) threshold. And the BER varies slightly taking account of small delay time. The results indicate that the new OZCZ code construction can effectively overcome non-perfect synchronous problem when the time delay among users does not exceed the zero correlation zone length.
RESUMEN
Developmentally programmed genome rearrangement accompanies differentiation of the silent germline micronucleus into the transcriptionally active somatic macronucleus in the ciliated protozoan Tetrahymena thermophila. Internal eliminated sequences (IES) are excised, followed by rejoining of MAC-destined sequences, while fragmentation occurs at conserved chromosome breakage sequences, generating macronuclear chromosomes. Some macronuclear chromosomes, referred to as non-maintained chromosomes (NMC), are lost soon after differentiation. Large NMC contain genes implicated in development-specific roles. One such gene encodes the domesticated piggyBac transposase TPB6, required for heterochromatin-dependent precise excision of IES residing within exons of functionally important genes. These conserved exonic IES determine alternative transcription products in the developing macronucleus; some even contain free-standing genes. Examples of precise loss of some exonic IES in the micronucleus and retention of others in the macronucleus of related species suggest an evolutionary analogy to introns. Our results reveal that germline-limited sequences can encode genes with specific expression patterns and development-related functions, which may be a recurring theme in eukaryotic organisms experiencing programmed genome rearrangement during germline to soma differentiation.
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Proteínas Protozoarias/metabolismo , Tetrahymena thermophila/genética , Transposasas/metabolismo , Cromosomas/genética , Exones , Reordenamiento Génico , Heterocromatina/genética , Secuencias Invertidas Repetidas , Macronúcleo/genética , Micronúcleo Germinal , Proteínas Protozoarias/genética , Interferencia de ARN , Transposasas/genéticaRESUMEN
OBJECTIVES: Low-intensity pulsed ultrasound (LIPUS) combined with porous scaffolds can be used as a new therapy to treat bone defect repair. The aim of this study was to evaluate the effects of 1 and 3.2 MHz LIPUS on osteogenesis on porous Ti64 alloy scaffolds for both in vitro and in vivo studies. METHODS: Scaffolds were randomly divided into the high-frequency ultrasound group, low-frequency ultrasound group, and control group. Mouse pre-osteoblast cells were cultured with porous Ti-6Al-4V scaffolds in vitro to evaluate cell proliferation and differentiation. In addition, scaffolds were implanted into rabbit mandibular defects in vivo. The effects of LIPUS on bone regeneration were evaluated by observing the micro-computed tomography (micro-CT), toluidine blue staining, and von Kossa staining. RESULTS: The results revealed no significant difference in the cell counting kit-8 values between the ultrasound groups and control groups (P > .05). Compared with the control group, ultrasound promoted alkaline phosphatase activity and osteocalcin levels of the cells on the scaffolds (P < .05), but there was no significant difference between the two frequencies. In addition, histomorphologic analyses revealed that the volume and amount of new bone formation increased and that bone maturity improved in the ultrasound groups compared with the control group, but no significant difference was noted between the two frequencies. CONCLUSIONS: Under the present experimental conditions, LIPUS promoted osteoblast differentiation and promoted bone maturity on porous Ti64 scaffolds. No significant differences were noted between the two frequencies.
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Regeneración Ósea/fisiología , Enfermedades Mandibulares/terapia , Osteogénesis/fisiología , Titanio , Terapia por Ultrasonido/métodos , Ondas Ultrasónicas , Aleaciones , Animales , Técnicas de Cultivo de Célula , Diferenciación Celular , Proliferación Celular , Modelos Animales de Enfermedad , Técnicas In Vitro , Masculino , Mandíbula/diagnóstico por imagen , Enfermedades Mandibulares/diagnóstico por imagen , Ratones , Osteoblastos/fisiología , Conejos , Microtomografía por Rayos X/métodosRESUMEN
Some studies have demonstrated that acrylamide (AA) was correlated with oxidative stress, resulting in physical damage. The jackfruit flake was an immature pulp that contained a high level of antioxidant activity. This study aimed to assess the defensive efficacy of jackfruit flake in AA-induced oxidative stress before and after simulated gastrointestinal digestion. Our results indicate that the total polyphenol content of Jackfruit flake digest (Digestive products of jackfruit flake after gastrointestinal, JFG) was diminished; however, JFG had raised the relative antioxidant capacity compared to Jackfruit flake extract (JFE). Additionally, the results of High Performance Liquid Chromatography-Mass Spectrometry (HPLC-MS) implied that a proportion of compounds were degraded/converted into other unknown and/or undetected metabolites. Further, by high content analysis (HCA) techniques, JFG markedly reduced cytotoxicity and excessive production of reactive oxygen species (ROS) in cells, thereby alleviating mitochondrial disorders. In this study, it may be converted active compounds after digestion that had preferable protective effects against AA-induced oxidative damage.
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Antioxidantes/análisis , Artocarpus/química , Estrés Oxidativo/efectos de los fármacos , Acrilamida/toxicidad , Apoptosis/efectos de los fármacos , Células CACO-2 , Cromatografía Líquida de Alta Presión , Digestión , Humanos , Intestino Delgado/efectos de los fármacos , Intestino Delgado/patología , Espectrometría de Masas , Potencial de la Membrana Mitocondrial/efectos de los fármacos , Membranas Mitocondriales/efectos de los fármacos , Permeabilidad , Polifenoles/análisis , Especies Reactivas de Oxígeno/metabolismo , Estómago/efectos de los fármacosRESUMEN
The safety of food additives has been widely concerned. Using single additives in the provisions of scope is safe, but the combination of additives, may induce additive, synergy, antagonism and other joint effects. This study investigated the cytotoxicity of aspartame (AT) together with potassium sorbate (PS). Thiazolyl Blue Tetrazolium Bromide (MTT) assay indicated that AT and PS had IC50 values of 0.48 g/L and 1.25 g/L at 24 h, respectively. High content analysis (HCA) showed that both AT and PS had a negative effect on mitochondrial membrane potential (MMP), reactive oxygen species (ROS) and DNA damage while the joint group behaved more obviously. The biochemical assays revealed typical cell morphological changes and the activation of cytochrome c and caspase-3 verified apoptosis induced by AT together with PS. With dissipation of MMP and increase of cell membrane permeability (CMP), it indicated AT together with PS-induced apoptosis was mediated by mitochondrial pathway. Meanwhile, p53 were involved in DNA damage, and the ratio of Bax/Bcl-2 was increased. Moreover, excessive ROS induced by AT together with PS is a key initiating factor for apoptosis. All these results proved that p53 was involved in apoptosis via mitochondria-mediated pathway and the process was regulated by ROS.
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Apoptosis/efectos de los fármacos , Aspartame/farmacología , Mitocondrias/efectos de los fármacos , Mitocondrias/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Ácido Sórbico/farmacología , Proteína p53 Supresora de Tumor/metabolismo , Proliferación Celular/efectos de los fármacos , Citocromos c/metabolismo , Daño del ADN , Sinergismo Farmacológico , Células Hep G2 , Humanos , Potencial de la Membrana Mitocondrial/efectos de los fármacos , Proteínas Proto-Oncogénicas c-bcl-2/metabolismoRESUMEN
Microbial spoilage is associated with the regulation of quorum sensing (QS). A. salmonicida AE03 with QS mediated acylated homoserine lactones (AHLs) activity was isolated from spoiled large yellow croaker (Pseudosciaena crocea). In this study the activity and role of AHLs in spoilage phenotypes, motility and biofilm formation of AE03 were investigated. The strain AE03 could induce Chromobacterium violaceum CV026 to produce the violacein pigment both at 28⯰C and 4⯰C in a density-dependent manner. Five types of AHLs were detected in AE03 culture by LC-MS/MS analysis, and N-butanoyl-l-homoserine lactone (C4-HSL) was a major signal molecule, reaching the highest concentration when incubated for 30â¯hâ¯at 28⯰C. An asaI-mutant, constructed by a suicide plasmid, failed to produce short chain AHLs signal. Compared with wild type (WT) strain, the production of trimethylamine (TMA), biogenic amino and protease significantly increased in asaI-mutant during the exponential and stationary phase, while the growth rate did not differ. Swimming motility in asaI-mutant was comparatively stronger than that of WT strain, whereas, asaI-mutant resulted in the decrease of maturing biofilm. Furthermore, supplementation of exogenous C4-HSL restored the production of spoilage metablites, protease and biofilm formation in mutant. In accordance with the effect of asaI deletion on the spoilage phenotypes and motility, asaI-mutant was showed to significantly up-regulate the transcript levels of torA, cadA and fliR, as well as asaR, indicating that C4-HSL could be involved in the modulation of the spoilage related enzymes and flagella. Indeed, the asaI-mutant promoted the spoilage progress of fish fillets stored at 4⯰C, while exogenous C4-HSL repressed the sensory change and TVB-N accumulation. The present study highlighted that AsaI/C4-HSL was an important regulator in spoilage, motility and biofilm formation of A. salmonicida, and spoilage potential was under the negative control of AsaI/AsaR-type system.
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Aeromonas salmonicida/genética , Proteínas Bacterianas/genética , Biopelículas/crecimiento & desarrollo , Peces/microbiología , Fenotipo , Percepción de Quorum/genética , Factores de Transcripción/genética , 4-Butirolactona/análogos & derivados , 4-Butirolactona/metabolismo , Aeromonas salmonicida/aislamiento & purificación , Animales , Microbiología de Alimentos , Conservación de AlimentosRESUMEN
A boy aged 6 years and 3 months developed upper respiratory tract infection and pyrexia 2 months ago and was given oral administration of nimesulide by his parents according to directions. Half an hour later, the boy experienced convulsions and cardiopulmonary arrest, and emergency examination found hypoketotic hypoglycemia, metabolic acidosis, significant increases in serum aminotransferases and creatine kinase, and renal damage. Recovery of consciousness and vital signs was achieved after cardiopulmonary resuscitation, but severe mental and movement regression was observed. The boy had a significant reduction in free carnitine in blood and significant increases in medium- and long-chain fatty acyl carnitine, urinary glutaric acid, 3-hydroxy glutaric acid, isovalerylglycine, and ethylmalonic acid, suggesting the possibility of multiple acyl-CoA dehydrogenase deficiency. After the treatment with vitamin B2, L-carnitine, and bezafibrate, the boy gradually improved, and reexamination after 3 months showed normal biochemical parameters. The boy had compound heterozygous mutations in the ETFDH gene, i.e., a known mutation, c.341G>A (p.R114H), from his mother and a novel mutation, c.1484C>G (p.P495R), from his father. Finally, he was diagnosed with multiple acyl-CoA dehydrogenase deficiency. Reye syndrome and sudden death symptoms were caused by nimesulide-induced acute metabolic crisis. It is concluded that inherited metabolic diseases may be main causes of Reye syndrome and sudden death, and biochemical and genetic analyses are the key to identifying underlying diseases.
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Carnitina , Infecciones del Sistema Respiratorio , Síndrome de Reye , Acil-CoA Deshidrogenasa , Administración Oral , Niño , Muerte Súbita , Humanos , Masculino , SulfonamidasRESUMEN
(-)-Epicatechin (EPI) has beneficial effects on the cardiovascular disease. The human ether-a-go-go-related gene (HERG) potassium channel is crucial for repolarization of cardiac action potential. Dysfunction of the HERG channel can cause long QT syndrome type 2 (LQT2). Arsenic trioxide (As2 O3 ) has shown efficacy in the treatment of acute promyelocytic leukemia. However, As2 O3 can induce the deficiency of HERG channel and cause LQT2. In this study, we examined whether EPI could rescue the As2 O3 -induced HERG channel deficiency. We found that 3 µM EPI obviously increased protein expression and current of HERG channel. EPI was able to recover the protein expression and current of HERG channel disrupted by As2 O3 . EPI was able to increase the expression of SP1 protein and recover the expression of SP1 protein disrupted by As2 O3 . In addition, EPI significantly shortened action potential duration prolonged by As2 O3 . Our data suggest that EPI rescues As2 O3 -induced HERG channel deficiency through upregulating SP1 expression.
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Arsenicales/efectos adversos , Catequina/farmacología , Canal de Potasio ERG1/metabolismo , Miocitos Cardíacos/efectos de los fármacos , Óxidos/efectos adversos , Factor de Transcripción Sp1/metabolismo , Potenciales de Acción/efectos de los fármacos , Animales , Trióxido de Arsénico , Células Cultivadas , Canal de Potasio ERG1/genética , Células HEK293 , Humanos , Cinética , Miocitos Cardíacos/metabolismo , Técnicas de Placa-Clamp , Ratas Sprague-DawleyRESUMEN
Diastereoisomers of quinidine and quinine are used to treat arrhythmia and malaria, respectively. It has been reported that both drugs block the hERG (human ether-a-go-go-related gene) potassium channel which is essential for myocardium repolarization. Abnormality of repolarization increases risk of arrhythmia. The aim of our research is to study and compare the impacts of quinidine and quinine on hERG. Results show that both drugs block the hERG channel, with quinine 14-fold less potent than quinidine. In addition, they presented distinct impacts on channel dynamics. The results imply their stereospecific block effect on the hERG channel. However, F656C-hERG reversed this stereoselectivity. The mutation decreases affinity of the two drugs with hERG, and quinine was more potent than quinidine in F656C-hERG blockage. These data suggest that F656 residue contributes to the stereoselective pocket for quinidine and quinine. Further study demonstrates that both drugs do not change hERG protein levels. In rescue experiments, we found that they exert no reverse effect on pentamidine- or desipramine-induced hERG trafficking defect, although quinidine has been reported to rescue trafficking-deficient pore mutation hERG G601S based on the interaction with F656. Our research demonstrated stereoselective effects of quinidine and quinine on the hERG channel, and this is the first study to explore their reversal potency on drug-induced hERG deficiency.
RESUMEN
Objective: Food phytochemicals as biofilm inhibitor of pathogens have been highlighted. Our study aimed to investigate the effects of resveratrol on biofilm formation of an aquatic pathogen Vibrio parahaemolyticus, and to elucidate the important regulatory genes. Methods: In the subinhibitory concentrations, the inhibition of resveratrol aganist biofilm and exopolysaccharides of V. parahaemolyticus was detected, and the differentially expressed genes were analyzed based on RNA-Seq. Four genes involved in biofilm formation was validated by qRT-PCR. Results: The minimum inhibitory concentration of resveratrol against V. parahaemolyticus was 20 µg/mL. Resveratrol at the subinhibitory concentration of 5 µg/mL and 10 µg/mL significantly decreased the biofilm development and exopolysaccharides production in V. parahaemolyticus (P < 0.05). Scanning electron microscopy micrographs showed a significant reduction of adherence and extracellular polymeric substances. RNA-Seq analysis revealed that 22.6% up-regulated and 77.4% down-regulated gene (P < 0.05) after treatment by 10 µg/mL resveratrol among 106 differential gene expressions. These differential genes in V. parahaemolyticu focused on 7 metabolic pathways, and 14 genes involved in biofilm formation were down-regulated by resveratrol, such as outer membrane protein (W, YedS, OmpK), quorum sensing (LuxS), flagellin (FlaA), fimbrial assembly protein (PilQ), hemolysin secreted protein. qRT-PCR confirmed that the expressions of luxS, trh, tlh and flaA, was significantly repressed in the presence of resveratrol, which was consistent with transcriptomics data. Conclusion: Inhibitory activity of resveratrol on biofilm formation was assicated with multiple genes and diverse cellular processes in V. parahaemolyticus. These findings suggest that resveratrol would disturb various metabolic pathways, particularly quorum sensing system, adhesion process and membrane proteins secretion, resulting in inhibition of attachment and biofilm development. The present work provided valuable information to explore molecular mechanism of resveratrol as an novel anti-biofilm compound.