RESUMO
BACKGROUND: Early identification of patients at risk of potential death and timely transfer to appropriate healthcare facilities are critical for reducing the number of preventable trauma deaths. This study aimed to establish a cutoff value to predict in-hospital mortality using the reverse shock index multiplied by the Glasgow Coma Scale (rSIG). METHODS: This multicenter retrospective cohort study used data from 23 emergency departments in South Korea between January 2011 and December 2020. The outcome variable was the in-hospital mortality. The relationship between rSIG and in-hospital mortality was plotted using the shape-restricted regression spline method. To set a cutoff for rSIG, we found the point on the curve where mortality started to increase and the point where the slope of the mortality curve changed the most. We also calculated the cutoff value for rSIG using Youden's index. RESULTS: A total of 318,506 adult patients with trauma were included. The shape-restricted regression spline curve showed that in-hospital mortality began to increase when the rSIG value was less than 18.86, and the slope of the graph increased the most at 12.57. The cutoff of 16.5, calculated using Youden's index, was closest to the target under-triage and over-triage rates, as suggested by the American College of Surgeons, when applied to patients with an rSIG of 20 or less. In addition, in patients with traumatic brain injury, when the rSIG value was over 25, in-hospital mortality tended to increase as the rSIG value increased. CONCLUSIONS: We propose an rSIG cutoff value of 16.5 as a predictor of in-hospital mortality in adult patients with trauma. However, in patients with traumatic brain injury, a high rSIG is also associated with in-hospital mortality. Appropriate cutoffs should be established for this group in the future.
Assuntos
Lesões Encefálicas Traumáticas , Ferimentos e Lesões , Adulto , Humanos , Escala de Coma de Glasgow , Estudos Retrospectivos , Mortalidade Hospitalar , Serviço Hospitalar de EmergênciaRESUMO
Microbial production of various TCA intermediates and related chemicals through the reductive TCA cycle has been of great interest. However, rumen bacteria that naturally possess strong reductive TCA cycle have been rarely studied to produce these chemicals, except for succinic acid, due to their dependence on fumarate reduction to transport electrons for ATP synthesis. In this study, malic acid (MA), a dicarboxylic acid of industrial importance, was selected as a target chemical for mass production using Mannheimia succiniciproducens, a rumen bacterium possessing a strong reductive branch of the TCA cycle. The metabolic pathway was reconstructed by eliminating fumarase to prevent MA conversion to fumarate. The respiration system of M. succiniciproducens was reconstructed by introducing the Actinobacillus succinogenes dimethylsulfoxide (DMSO) reductase to improve cell growth using DMSO as an electron acceptor. Also, the cell membrane was engineered by employing Pseudomonas aeruginosa cis-trans isomerase to enhance MA tolerance. High inoculum fed-batch fermentation of the final engineered strain produced 61 g/L of MA with an overall productivity of 2.27 g/L/h, which is the highest MA productivity reported to date. The systems metabolic engineering strategies reported in this study will be useful for developing anaerobic bioprocesses for the production of various industrially important chemicals.
Assuntos
Mannheimia , Engenharia Metabólica , Animais , Mannheimia/genética , Mannheimia/metabolismo , Dimetil Sulfóxido/metabolismo , Elétrons , Fumaratos/metabolismoRESUMO
In this study, the levels of polycyclic aromatic hydrocarbons (PAHs) and polybrominated diphenyl ethers (PBDEs) were characterized in firefighters' personal protective equipment (PPE) (i.e., jackets, pants, hoods, and gloves) and vehicle dust wipe samples to assess the exposure and potential risk of firefighters to these combustion-related toxic pollutants. The mean levels of ∑PBDEs in the fire vehicle dust samples (778 and 449 pg/cm2 for pump trucks and command cars, respectively) were significantly higher than those in the private vehicles (31.2 pg/cm2) (Kruskal-Wallis test, p < 0.05), which was similar to the ∑PAH levels (521, 185, and 46.8 pg/cm2 for pump trucks, command cars, and private vehicles, respectively). In the case of firefighters' PPE, the levels of ∑PBDEs and ∑PAHs in used jackets and pants were found to be, respectively, 70- to 2242-folds and 11- to 265-folds higher than those in their unused counterparts. Biomass/petroleum combustion was found to be the main source of PAH contamination in fire vehicle dust and used PPE in the present study. Both carcinogenic and noncarcinogenic risks via vehicle dust ingestion and dermal absorption from wearing of PPE were within permissible limits, although the relative risk evaluation showed that PAH/PBDE absorption via wearing of PPE could pose a higher likelihood of carcinogenic and noncarcinogenic risks than the ingestion of pollutants via fire vehicle dust, warranting the need for appropriate management of firefighters' personal protective ensembles.
Assuntos
Poluentes Ambientais , Bombeiros , Exposição Ocupacional , Hidrocarbonetos Policíclicos Aromáticos , Humanos , Exposição Ocupacional/análise , Éteres Difenil Halogenados/análise , Poeira/análise , Equipamento de Proteção Individual , Medição de Risco , República da CoreiaRESUMO
Measuring the daily use of an affected limb after hospital discharge is crucial for hemiparetic stroke rehabilitation. Classifying movements using non-intrusive wearable sensors provides context for arm use and is essential for the development of a home rehabilitation system. However, the movement classification of stroke patients poses unique challenges, including variability and sparsity. To address these challenges, we collected movement data from 15 hemiparetic stroke patients (Stroke group) and 29 non-disabled individuals (ND group). The participants performed two different tasks, the range of motion (14 movements) task and the activities of daily living (56 movements) task, wearing five inertial measurement units in a home setting. We trained a 1D convolutional neural network and evaluated its performance for different training groups: ND-only, Stroke-only, and ND and Stroke jointly. We further compared the model performance with data augmentation from axis rotation and investigated how the performance varied based on the asymmetry of movements. The joint training of ND + Stroke yielded an increased F1-score by a margin of 31.6% and 10.6% compared to ND-only training and Stroke-only training, respectively. Data augmentation further enhanced F1-scores across all conditions by an average of 11.3%. Finally, asymmetric movements decreased the F1-score by 25.9% compared to symmetric movements in the Stroke group, indicating the importance of asymmetry in movement classification.
Assuntos
Aprendizado Profundo , Reabilitação do Acidente Vascular Cerebral , Acidente Vascular Cerebral , Dispositivos Eletrônicos Vestíveis , Humanos , Atividades Cotidianas , Acidente Vascular Cerebral/diagnósticoRESUMO
Reduction of extracellular acceptors requires electron transfer across the periplasm. In Geobacter sulfurreducens, three separate cytoplasmic membrane cytochromes are utilized depending on redox potential, and at least five cytochrome conduits span the outer membrane. Because G. sulfurreducens produces 5 structurally similar triheme periplasmic cytochromes (PpcABCDE) that differ in expression level, midpoint potential, and heme biochemistry, many hypotheses propose distinct periplasmic carriers could be used for specific redox potentials, terminal acceptors, or growth conditions. Using a panel of marker-free single, quadruple, and quintuple mutants, little support for these models could be found. Three quadruple mutants containing only one paralog (PpcA, PpcB, and PpcD) reduced Fe(III) citrate and Fe(III) oxide at the same rate and extent, even though PpcB and PpcD were at much lower periplasmic levels than PpcA. Mutants containing only PpcC and PpcE showed defects, but these cytochromes were nearly undetectable in the periplasm. When expressed sufficiently, PpcC and PpcE supported wild-type Fe(III) reduction. PpcA and PpcE from G. metallireducens similarly restored metal respiration in G. sulfurreducens. PgcA, an unrelated extracellular triheme c-type cytochrome, also participated in periplasmic electron transfer. While triheme cytochromes were important for metal reduction, sextuple ΔppcABCDE ΔpgcA mutants grew near wild-type rates with normal cyclic voltammetry profiles when using anodes as electron acceptors. These results reveal broad promiscuity in the periplasmic electron transfer network of metal-reducing Geobacter and suggest that an as-yet-undiscovered periplasmic mechanism supports electron transfer to electrodes. IMPORTANCE Many inner and outer membrane cytochromes used by Geobacter for electron transfer to extracellular acceptors have specific functions. How these are connected by periplasmic carriers remains poorly understood. G. sulfurreducens contains multiple triheme periplasmic cytochromes with unique biochemical properties and expression profiles. It is hypothesized that each could be involved in a different respiratory pathway, depending on redox potential or energy needs. Here, we show that Geobacter periplasmic cytochromes instead show evidence of being highly promiscuous. Any of 6 triheme cytochromes supported similar growth with soluble or insoluble metals, but none were required when cells utilized electrodes. These findings fail to support many models of Geobacter electron transfer, and question why these organisms produce such an array of periplasmic cytochromes.
Assuntos
Geobacter , Geobacter/genética , Geobacter/metabolismo , Periplasma/metabolismo , Compostos Férricos/metabolismo , Elétrons , Transporte de Elétrons , Citocromos/genética , Citocromos/química , Citocromos/metabolismo , OxirreduçãoRESUMO
BACKGROUND: Person-centred care has the potential to improve the patient experience in the intensive care unit (ICU). However, the relationship between person-centred care perceived by critically ill patients and their ICU experience has yet to be determined. OBJECTIVES: The aim of this study was to investigate the relationship between person-centred care and the ICU experience of critically ill patients. METHODS: This study was a multicentre, cross-sectional survey involving 19 ICUs of four university hospitals in Busan, Korea. The survey was conducted from June 2019 to July 2020, and 787 patients who had been admitted to the ICU for more than 24 hours participated. We measured person-centred care using the Person-Centered Critical Care Nursing perceived by Patient Questionnaire. Participants' ICU experience was measured by the Korean version of the Intensive Care Experience Questionnaire that consists of four subscales. We analysed the relationship between person-centred care and each area of the ICU experience using multivariate linear regression. RESULTS: Person-centred care was associated with 'awareness of surroundings' (ß = 0.29, p < .001), 'frightening experiences' (ß = -0.31, p < .001), and 'satisfaction with care' (ß = 0.54, p < .001). However, there was no significant association between person-centred care and 'recall of experience'. CONCLUSIONS: We observed that person-centred care was positively related to most of the ICU experiences of critically ill patients except for recall of experience. Further studies on developing person-centred nursing interventions are needed.
Assuntos
Estado Terminal , Unidades de Terapia Intensiva , Humanos , Estudos Transversais , Cuidados Críticos , Assistência Centrada no PacienteRESUMO
This study investigated the occupational exposure of Korean firefighters to a suite of combustion-related pollutants. Exposure to polybrominated diphenyl ethers (PBDEs) and polycyclic aromatic hydrocarbons (PAHs) was assessed by measurement of their levels in serum and metabolites in urine (i.e., monohydroxylated PAHs, OH-PAHs). The mean level of ∑PBDEs in the serum of firefighters (17.1 ng/g lipid weight (lw)) was significantly higher than that of the general population (1.39 ng/g lw) (Mann-Whitney U Test, p < 0.05), which is similar to the ∑PAH levels (1286 ng/g lw for firefighters and 1016 ng/g lw for the general population). Individual OH-PAH levels showed 2.1- to 4.2-fold increases in postfire urine samples compared to the control urine samples, with the mean ∑OH-PAHs being significantly higher in postfire urine samples (22,658 ng/g creatinine) than in the control urine samples (10,253 ng/g creatinine) (Mann-Whitney U test, p < 0.05). It was found that ∑PBDEs correlated with firefighters' length of service and years dedicated to on-site dispatch, while ∑OH-PAHs was strongly associated with firefighters' exposure duration, age, length of service, and years dedicated to on-site dispatch. Indeed, the results of the present study indicate that Korean firefighters are prone to elevated risk of exposure to toxic combustion-related pollutants compared with the general population.
Assuntos
Bombeiros , Exposição Ocupacional , Hidrocarbonetos Policíclicos Aromáticos , Monitoramento Ambiental , Éteres Difenil Halogenados/análise , Humanos , Exposição Ocupacional/análise , Hidrocarbonetos Policíclicos Aromáticos/análise , República da CoreiaRESUMO
To address climate change and environmental problems, it is becoming increasingly important to establish biorefineries for the production of chemicals from renewable non-food biomass. Here we report the development of Escherichia coli strains capable of overproducing a four-carbon platform chemical 4-hybroxybutyric acid (4-HB). Because 4-HB production is significantly affected by aeration level, genome-scale metabolic model-based engineering strategies were designed under aerobic and microaerobic conditions with emphasis on oxidative/reductive TCA branches and glyoxylate shunt. Several different metabolic engineering strategies were employed to develop strains suitable for fermentation both under aerobic and microaerobic conditions. It was found that microaerobic condition was more efficient than aerobic condition in achieving higher titer and productivity of 4-HB. The final engineered strain produced 103.4g/L of 4-HB by microaerobic fed-batch fermentation using glycerol. The aeration-dependent optimization strategy of TCA cycle will be useful for developing microbial strains producing other reduced derivative chemicals of TCA cycle intermediates.
Assuntos
Vias Biossintéticas/genética , Ciclo do Ácido Cítrico/genética , Escherichia coli/fisiologia , Melhoramento Genético/métodos , Hidroxibutiratos/metabolismo , Engenharia Metabólica/métodos , Proteínas de Escherichia coli/genética , Proteínas de Escherichia coli/metabolismo , Regulação Bacteriana da Expressão Gênica/fisiologia , Glicerol/metabolismo , Hidroxibutiratos/isolamento & purificação , Redes e Vias Metabólicas/genéticaRESUMO
Succinic acid (SA) is a four carbon dicarboxylic acid of great industrial interest that can be produced by microbial fermentation. Here we report development of a high-yield homo-SA producing Mannheimia succiniciproducens strain by metabolic engineering. The PALFK strain (ldhA-, pta-, ackA-, fruA-) was developed based on optimization of carbon flux towards SA production while minimizing byproducts formation through the integrated application of in silico genome-scale metabolic flux analysis, omics analyses, and reconstruction of central carbon metabolism. Based on in silico simulation, utilization of sucrose would enhance the SA production and cell growth rates, while consumption of glycerol would reduce the byproduct formation rates. Thus, sucrose and glycerol were selected as dual carbon sources to improve the SA yield and productivity, while deregulation of catabolite-repression was also performed in engineered M. succiniciproducens. Fed-batch fermentations of PALFK with low- and medium-density (OD600 of 0.4 and 9.0, respectively) inocula produced 69.2 and 78.4g/L of homo-SA with yields of 1.56 and 1.64mol/mol glucose equivalent and overall volumetric SA productivities of 2.50 and 6.02g/L/h, respectively, using sucrose and glycerol as dual carbon sources. The SA productivity could be further increased to 38.6g/L/h by employing a membrane cell recycle bioreactor system. The systems metabolic engineering strategies employed here for achieving homo-SA production with the highest overall performance indices reported to date will be generally applicable for developing superior industrial microorganisms and competitive processes for the bio-based production of other chemicals as well.
Assuntos
Proteínas de Bactérias/genética , Glicerol/metabolismo , Mannheimia/fisiologia , Engenharia Metabólica/métodos , Ácido Succínico/metabolismo , Sacarose/metabolismo , Reatores Biológicos/microbiologia , Vias Biossintéticas/genética , Melhoramento Genético/métodos , Redes e Vias Metabólicas/genética , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Ácido Succínico/isolamento & purificaçãoRESUMO
Succinic acid (SA) is one of the fermentative products of anaerobic metabolism, and an important industrial chemical that has been much studied for its bio-based production. The key to the economically viable bio-based SA production is to develop an SA producer capable of producing SA with high yield and productivity without byproducts. Mannheimia succiniciproducens is a capnophilic rumen bacterium capable of efficiently producing SA. In this study, in silico genome-scale metabolic simulations were performed to identify gene targets to be engineered, and the PALK strain (ΔldhA and Δpta-ackA) was constructed. Fed-batch culture of PALK on glucose and glycerol as carbon sources resulted in the production of 66.14 g/L of SA with the yield and overall productivity of 1.34 mol/mol glucose equivalent and 3.39 g/L/h, respectively. SA production could be further increased to 90.68 g/L with the yield and overall productivity of 1.15 mol/mol glucose equivalent and 3.49 g/L/h, respectively, by utilizing a mixture of magnesium hydroxide and ammonia solution as a pH controlling solution. Furthermore, formation of byproducts was drastically reduced, resulting in almost homo-fermentative SA production. This allowed the recovery and purification of SA to a high purity (99.997%) with a high recovery yield (74.65%) through simple downstream processes composed of decolorization, vacuum distillation, and crystallization. The SA producer and processes developed in this study will allow economical production of SA in an industrial-scale. Biotechnol. Bioeng. 2016;113: 2168-2177. © 2016 Wiley Periodicals, Inc.
Assuntos
Melhoramento Genético/métodos , Mannheimia/genética , Mannheimia/metabolismo , Engenharia Metabólica/métodos , Ácido Succínico/isolamento & purificação , Ácido Succínico/metabolismo , Simulação por Computador , Glucose/metabolismo , Glicerol/metabolismo , Mannheimia/classificação , Análise do Fluxo Metabólico , Modelos Biológicos , Complexos Multienzimáticos/genética , Complexos Multienzimáticos/metabolismo , Especificidade da EspécieRESUMO
Justification in the field of radiology refers to the appropriate use of radiologic imaging modalities, and may be achieved by establishing clinical imaging guidelines (CIGs). Recently, CIGs have been shown to be useful in selecting the proper medical imaging modality, resulting in the reduction of inappropriate radiologic examinations, thereby enhancing justifications. However, the development of CIGs is both time-consuming and difficult as the methodology of evidence-based medicine should be adhered to. Thus, although the radiologic societies in developed countries such as the United Kingdom and USA are already developing and implementing CIGs in their clinical practices, CIGs are not yet readily available in many other countries owing to differences in medical circumstances and resources. In this review, we assess the role and limitations of CIGs by examining the current status of CIGs in developed countries, and also describe the specific efforts made to establish CIGs in Korea.
Assuntos
Diagnóstico por Imagem , Guias como Assunto , Medicina Baseada em Evidências , Humanos , Proteção Radiológica , Sociedades Científicas , Tomografia Computadorizada por Raios XRESUMO
Due to the growing concerns on the climate change and sustainability on petrochemical resources, DOE selected and announced the bio-based top 12 building blocks and discussed the needs for developing biorefinery technologies to replace the current petroleum based industry in 2004. Over the last 10 years after its announcement, many studies have been performed for the development of efficient technologies for the bio-based production of these chemicals and derivatives. Now, ten chemicals among these top 12 chemicals, excluding the l-aspartic acid and 3-hydroxybutyrolactone, have already been commercialized or are close to commercialization. In this paper, we review the current status of biorefinery development for the production of these platform chemicals and their derivatives. In addition, current technological advances on industrial strain development for the production of platform chemicals using micro-organisms will be covered in detail with case studies on succinic acid and 3-hydroxypropionic acid as examples.
Assuntos
Ácido Láctico/análogos & derivados , Engenharia Metabólica/métodos , Ácido Succínico/metabolismo , Ácido Láctico/biossínteseRESUMO
Differentiated somatic cells can be reprogrammed into pluripotent stem cells by transduction of exogenous reprogramming factors. After induced pluripotent stem (iPS) cells are established, exogenous genes are silenced. In the pluripotent state, retroviral genes integrated in the host genome are kept inactive through epigenetic transcriptional regulation. In this study, we tried to determine whether exogenous genes remain silenced or are reactivated upon loss of pluripotency or on differentiation using an in vitro system. We induced differentiation of iPS cells into neural stem cells (NSCs) in vitro; the NSCs appeared morphologically indistinguishable from brain-derived NSCs and stained positive for the NSC markers Nestin and Sox2. These iPS cell-derived NSCs (iPS-NSCs) were also capable of differentiating into all three neural subtypes. Interestingly, iPS-NSCs spontaneously formed aggregates on long-term culture and showed reactivation of the Oct4-GFP marker, which was followed by the formation of embryonic stem cell-like colonies. The spontaneously reverted green fluorescent protein (GFP)-positive (iPS-NSC-GFP(+) ) cells expressed high levels of pluripotency markers (Oct4 and Nanog) and formed germline chimeras, indicating that iPS-NSC-GFP(+) cells had the same pluripotency as the original iPS cells. The reactivation of silenced exogenous genes was tightly correlated with the downregulation of DNA methyltransferases (Dnmts) during differentiation of iPS cells. This phenomenon was not observed in doxycycline-inducible iPS cells, where the reactivation of exogenous genes could be induced only by doxycycline treatment. These results indicate that pluripotency can be regained through reactivation of exogenous genes, which is associated with dynamic change of Dnmt levels during differentiation of iPS cells.
Assuntos
Diferenciação Celular , Células-Tronco Pluripotentes Induzidas/citologia , Células-Tronco Neurais/citologia , Animais , DNA (Citosina-5-)-Metiltransferases/metabolismo , Células-Tronco Embrionárias/citologia , Células-Tronco Embrionárias/metabolismo , Inativação Gênica , Genes Virais , Células-Tronco Pluripotentes Induzidas/metabolismo , Camundongos , Células-Tronco Neurais/metabolismo , Retroviridae/genética , TransgenesRESUMO
Growing concerns over limited fossil resources and associated environmental problems are motivating the development of sustainable processes for the production of chemicals, fuels and materials from renewable resources. Metabolic engineering is a key enabling technology for transforming microorganisms into efficient cell factories for these compounds. Systems metabolic engineering, which incorporates the concepts and techniques of systems biology, synthetic biology and evolutionary engineering at the systems level, offers a conceptual and technological framework to speed the creation of new metabolic enzymes and pathways or the modification of existing pathways for the optimal production of desired products. Here we discuss the general strategies of systems metabolic engineering and examples of its application and offer insights as to when and how each of the different strategies should be used. Finally, we highlight the limitations and challenges to be overcome for the systems metabolic engineering of microorganisms at more advanced levels.
Assuntos
Bactérias , Engenharia Metabólica/métodos , Compostos Orgânicos , Organismos Geneticamente Modificados , Biologia Sintética/métodos , Biologia de Sistemas/métodos , Bactérias/enzimologia , Bactérias/genética , Evolução Molecular Direcionada , Microbiologia Industrial , Compostos Orgânicos/química , Compostos Orgânicos/metabolismoRESUMO
Comprehensive studies simultaneously investigating the occurrence of chemicals of concern are limited. In this study, sediments and bivalves were collected from 24 locations along the Korean coast to evaluate the relative distribution, contamination characteristics, and ecological risks of legacy/emerging persistent organic pollutants (POPs) and polycyclic aromatic hydrocarbons (PAHs). Our findings reveal that the concentrations of these contaminants were comparable to or lower than historical levels in the same Korean coast and other Asian countries. Notably, PAHs exhibited the highest distribution in sediments (84 %), whereas short-chain chlorinated paraffins (SCCPs) were dominant in bivalves (91 %). This study highlighted significant correlations in the sediment levels of each legacy pollutants, suggesting similar sources and geochemical behaviors. However, SCCPs displayed unique contamination patterns. Ecologically, PAHs and SCCPs presented low risks in sediments compared to Canadian Sediment Quality Guidelines, however 100 % and 33 % of bivalves, respectively, exceeded US EPA/Canadian Fish Tissue Guidelines.
Assuntos
Bivalves , Monitoramento Ambiental , Sedimentos Geológicos , Poluentes Orgânicos Persistentes , Hidrocarbonetos Policíclicos Aromáticos , Poluentes Químicos da Água , Hidrocarbonetos Policíclicos Aromáticos/análise , Poluentes Químicos da Água/análise , República da Coreia , Animais , Sedimentos Geológicos/químicaRESUMO
Airway management is a fundamental and intricate process that involves a sequence of integrated tasks. Situations requiring emergency airway management may occur in emergency department, intensive care units, and various other spaces. Emergency airway management can face a variety of challenges during preparation, intubation, and post-intubation, and may result in significant complications for the patients. Therefore, many countries are establishing step-by-step systemization and detailed guidelines, and updating the content based on the latest research. This clinical review introduces the current trends in emergency airway management, such as emergency airway management algorithms, comparison of video and direct laryngoscopy, rapid sequence intubation, pediatric airway management, pre-hospital airway management, surgical airway management, and airway management education.
RESUMO
γ-Butyrolactone (GBL) is an important four carbon (C4) chemical, which has a wide range of industrial applications. GBL can be produced by acid treatment of 4-hydroxybutyric acid (4-HB), which is a derivative of succinic acid. Heterologous metabolic pathways were designed and established in succinic acid overproducing Mannheimia succiniciproducens LPK7 (ldhA pflD pta ackA mutant) by the introduction of heterologous genes that encode succinyl-CoA synthetase, CoA-dependent succinate semialdehyde dehydrogenase, and either 4-hydroxybutyrate dehydrogenase in LPK7 (p3S4CD) or succinate semialdehyde reductase in LPK7 (p3SYCD). Fed-batch cultures of LPK7 (p3S4CD) and LPK7 (p3SYCD) resulted in the production of 6.37 and 6.34 g/L of 4-HB (molar yields of 0.143 and 0.139), respectively. Finally, GBL was produced by acid treatment of the 4-HB obtained from the fermentation broth with molar yield of 0.673. This study demonstrates that 4-HB, and potentially other four carbon platform chemicals, can be produced by the engineered rumen bacterium M. succiniciproducens.
Assuntos
4-Butirolactona , Proteínas de Bactérias , Hidroxibutiratos , Mannheimia , Engenharia Metabólica , Mutação , 4-Butirolactona/biossíntese , 4-Butirolactona/genética , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Hidroxibutiratos/metabolismo , Hidroxibutiratos/farmacologia , Mannheimia/enzimologia , Mannheimia/genéticaRESUMO
Fumaric acid is a naturally occurring organic acid that is an intermediate of the tricarboxylic acid cycle. Fungal species belonging to Rhizopus have traditionally been employed for the production of fumaric acid. In this study, Escherichia coli was metabolically engineered for the production of fumaric acid under aerobic condition. For the aerobic production of fumaric acid, the iclR gene was deleted to redirect the carbon flux through the glyoxylate shunt. In addition, the fumA, fumB, and fumC genes were also deleted to enhance fumaric acid formation. The resulting strain was able to produce 1.45 g/L of fumaric acid from 15 g/L of glucose in flask culture. Based on in silico flux response analysis, this base strain was further engineered by plasmid-based overexpression of the native ppc gene, encoding phosphoenolpyruvate carboxylase (PPC), from the strong tac promoter, which resulted in the production of 4.09 g/L of fumaric acid. Additionally, the arcA and ptsG genes were deleted to reinforce the oxidative TCA cycle flux, and the aspA gene was deleted to block the conversion of fumaric acid into L-aspartic acid. Since it is desirable to avoid the use of inducer, the lacI gene was also deleted. To increase glucose uptake rate and fumaric acid productivity, the native promoter of the galP gene was replaced with the strong trc promoter. Fed-batch culture of the final strain CWF812 allowed production of 28.2 g/L fumaric acid in 63 h with the overall yield and productivity of 0.389 g fumaric acid/g glucose and 0.448 g/L/h, respectively. This study demonstrates the possibility for the efficient production of fumaric acid by metabolically engineered E. coli.
Assuntos
Biotecnologia/métodos , Escherichia coli/genética , Escherichia coli/metabolismo , Fumaratos/metabolismo , Engenharia Metabólica/métodos , Redes e Vias Metabólicas/genética , Aerobiose , Escherichia coli/fisiologia , Deleção de Genes , Expressão Gênica , Vetores Genéticos , Glucose/metabolismo , Análise do Fluxo Metabólico , Plasmídeos , Regiões Promotoras GenéticasRESUMO
The restricted gene expression pattern of a differentiated cell can be reversed by fusion of the somatic cell with a more developmentally potent cell type, such as an embryonic stem (ES) cell. During this reprogramming process, somatic cells obtain most of the characteristics of pluripotent cells. Reactivation of an inactive X chromosome (Xi) is an important epigenetic marker confirming the pluripotent reprogramming of somatic cells. Female somatic cells contain one active X chromosome (Xa) and one Xi, and following the fusion of these cells with male ES cells, the Xi becomes activated, resulting in XaXaXaY fusion hybrid cells. To monitor Xi reactivation, transgenic female neural stem cells (fNSCs) carrying a green fluorescent protein (GFP) reporter gene expressed on the Xa (X-GFP), but not on the Xi, were used for reprogramming. XaXi(GFP) NSCs, whose GFP reporter was silenced, were fused with HM1 ES cells (XY) to induce pluripotent reprogramming. The Xi(GFP) of NSCs were found to be activated on day 4 post-fusion, indicating reactivation of the Xi. Hybrid cells showed pluripotent cell-specific characteristics cells including inactivation of the NSC marker Nestin, DNA demethylation of Oct4, DNA methylation of Nestin, and reactivation of the Xi. Following differentiation of the (GFP-positive) hybrid cells through embryoid body formation, the proportion of GFP-negative cells was found to be approximately 26 %, indicating that there was random inactivation of one of the three Xas. Here, we showed that the Xi of somatic cells is reprogrammed to the Xa state and that cellular differentiation occurs randomly, i.e., regardless of the Xa or Xi state, indicating that the memory of the Xi of somatic cells has been erased and reset to the ground state (i.e., inner cell mass-like state), indicating that random X-chromosome inactivation occurs upon differentiation.
Assuntos
Reprogramação Celular/genética , Células-Tronco Embrionárias/metabolismo , Células-Tronco Neurais/metabolismo , Cromossomo X/genética , Animais , Diferenciação Celular/genética , Fusão Celular , Células Cultivadas , Feminino , Citometria de Fluxo , Proteínas de Fluorescência Verde/genética , Proteínas de Fluorescência Verde/metabolismo , Hibridização in Situ Fluorescente , Cariótipo , Masculino , Camundongos Transgênicos , Inativação do Cromossomo XRESUMO
In this study, we investigated the occurrence and distribution of phenolic compounds, including phenol, cresols, chlorophenols, nitrophenol, and bromophenols, in freshwater environments. We also focused on phenolic compounds in crucian carp (Carassius auratus) tissues, specifically the muscle, gills, brain, blood, liver, and gonads, to assess their potential bioaccumulation in fish and human health risks. Phenolic compounds were found to be widespread in various freshwater environments throughout South Korea. Phenol was predominant in all matrices, with median concentrations of 57.0 ng/L in freshwater, 54.3 ng/g dry weight (dw) in sediment, and ranging from 71 ng/g wet weight (ww) to 621 ng/g ww in crucian carp tissues. Cresols were the second most dominant compound, with m-cresol exhibiting the highest prevalence. Most of the compounds detected in crucian carp samples were also detected in freshwater and sediment, whereas pentachlorophenol and 2,4,6-tribromophenol were exclusively detected in crucian carp tissues. A high bioaccumulation potential in the liver was observed for most phenolic compounds [median log bioconcentration factor (BCF): 3.2-3.7]. Interestingly, only m-cresol showed high bioaccumulation potential in the gills (median log BCF: 3.1). The estimated daily intake of phenolic compounds suggested that it does not pose an immediate concern for human exposure owing to crucian carp consumption. These findings enhance our understanding of the exposure status, distribution, and bioaccumulation potency of phenolic compounds in aquatic ecosystems and emphasize the importance of ongoing monitoring and risk assessment efforts.