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Our study mainly focused on diffusible hydrogen in aluminum-silicon-coated hot-stamped boron steel during a hot press forming process and in pre-treatment sequential lines of the automotive manufacturing process using a thermal desorption spectroscopy (TDS) technique. First, in the hot stamping procedure, as the soaking time increased in the heating furnace at a specific dew point when austenitizing, a high concentration of diffusible hydrogen was absorbed into the hot-stamped boron steel. Based on the TDS analysis of hydrogen absorbed from hot stamping, the activation energy value of hydrogen trapping in 1.8 GPa grade steel is lower than that of 1.5 GPa grade steel. This means that diffusible hydrogen can be more easily diffused into defective sites of the microstructure at a higher level of the tensile strength grade. Second, in sequential pre-treatment lines of the automotive manufacturing process, additional hydrogen did not flow into the surface, and an electro-deposition process, including a baking procedure, was effective in removing diffusible hydrogen, which was similar to the residual hydrogen of the as-received state (i.e., initial cold rolled blank). Based on these results, the hydrogen absorption was facilitated during hot press forming, but the hydrogen was sequentially desorbed during automotive sequential lines on aluminum-coated hot-stamped steel parts.
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BACKGROUND: Bioethanol is one of the most representative eco-friendly fuels developed to replace the non-renewable fossil fuels and is the most successful commercially available bio-conversion technology till date. With the availability of inexpensive carbon sources, such as cellulosic biomass, bioethanol production has become cheaper and easier to perform, which can facilitate the development of methods for converting ethanol into higher value-added biochemicals. In this study, a bioconversion process using Pseudomonas putida as a biocatalyst was established, wherein ethanol was converted to mevalonate. Since ethanol can be converted directly to acetyl-CoA, bypassing its conversion to pyruvate, there is a possibility that ethanol can be converted to mevalonate without producing pyruvate-derived by-products. Furthermore, P. putida seems to be highly resistant to the toxicity caused by terpenoids, and thus can be useful in conducting terpenoid production research. RESULTS: In this study, we first expressed the core genes responsible for mevalonate production (atoB, mvaS, and mvaE) in P. putida and mevalonate production was confirmed. Thereafter, through an improvement in genetic stability and ethanol metabolism manipulation, mevalonate production was enhanced up to 2.39-fold (1.70 g/L vs. 4.07 g/L) from 200 mM ethanol with an enhancement in reproducibility of mevalonate production. Following this, the metabolic characteristics related to ethanol catabolism and mevalonate production were revealed by manipulations to reduce fatty acid biosynthesis and optimize pH by batch fermentation. Finally, we reached a product yield of 0.41 g mevalonate/g ethanol in flask scale culture and 0.32 g mevalonate/g ethanol in batch fermentation. This is the highest experimental yield obtained from using carbon sources other than carbohydrates till date and it is expected that further improvements will be made through the development of fermentation methods. CONCLUSION: Pseudomonas putida was investigated as a biocatalyst that can efficiently convert ethanol to mevalonate, the major precursor for terpenoid production, and this research is expected to open new avenues for the production of terpenoids using microorganisms that have not yet reached the stage of mass production.
Asunto(s)
Acetilcoenzima A/metabolismo , Etanol/metabolismo , Ingeniería Metabólica/métodos , Ácido Mevalónico/metabolismo , Microorganismos Modificados Genéticamente , Pseudomonas putida , Terpenos/metabolismo , Biocombustibles , Microorganismos Modificados Genéticamente/genética , Microorganismos Modificados Genéticamente/metabolismo , Pseudomonas putida/genética , Pseudomonas putida/metabolismo , Ácido Pirúvico/metabolismoRESUMEN
Currently, only a few states in the USA (e.g., Colorado and Ohio) require mandatory baseline groundwater sampling from nearby groundwater wells prior to drilling a new oil or gas well. Colorado is the first state to regulate groundwater testing before and after drilling, which requires one pre-drilling sample and two additional post-drilling samples within 6-12 months and 5-6 years of drilling. However, the monitoring method is limited to the state's regulatory agency and to ex situ sampling, which offers only a snapshot in time. To overcome the limitations and increase monitoring performance, a new groundwater monitoring system, Colorado Water Watch (CWW), was introduced as a decision-making tool to support the state's regulatory agency and also to provide real-time groundwater quality data to both the industry and the public. The CWW uses simple in situ water quality sensors based on the surrogate sensing technology that employs an event detection system to screen the incoming data in near real-time.
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Monitoreo del Ambiente , Industria Procesadora y de Extracción , Agua Subterránea/química , Yacimiento de Petróleo y Gas , Contaminantes Químicos del Agua/análisis , Colorado , Gas Natural , Ohio , Agua , Calidad del Agua , Abastecimiento de Agua , Pozos de AguaRESUMEN
The potential impact of rapid development of unconventional oil and natural gas resources using hydraulic fracturing and horizontal drilling on regional groundwater quality has received significant attention. Major concerns are methane or oil/gas related hydrocarbon (such as TPHs, BTEX including benzene, toluene, ethybenzene and xylene) leaks into the aquifer due to the failure of casing and/or stray gas migration. Previously, we investigated the relationship between oil and gas activity and dissolved methane concentration in a drinking water aquifer with the major finding being the presence of thermogenic methane contamination, but did not find detectable concentrations of TPHs or BTEX. To understand if aqueous and gas phases from the producing formation were transported concurrently to drinking water aquifers without the presence of oil/gas related hydrocarbons, the ionic composition of three water groups was studied: (1) uncontaminated deep confined aquifer, (2) suspected contaminated groundwater - deep confined aquifer containing thermogenic methane, and (3) produced water from nearby oil and gas wells that would represent aqueous phase contaminants. On the basis of quantitative and spatial analysis, we identified that the "thermogenic methane contaminated" groundwater did not have similarities to produced water in terms of ionic character (e.g. Cl/TDS ratio), but rather to the "uncontaminated" groundwater. The analysis indicates that aquifer wells with demonstrated gas phase contamination have not been contacted by an aqueous phase from oil and gas operations according to the methodology we use in this study and the current groundwater quality data from COGCC. However, the research does not prove conclusively that this the case. The results may provide insight on contamination mechanisms since improperly sealed well casing may result in stray gas but not aqueous phase transport.
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Monitoreo del Ambiente , Agua Subterránea/análisis , Metano/análisis , Yacimiento de Petróleo y Gas/química , Contaminantes Químicos del Agua/análisis , Colorado , Iones/análisis , Pozos de AguaRESUMEN
The objectives of this study were to monitor total phosphorus concentrations and loads along the Cache la Poudre River in Northern Colorado as it flows from a pristine area through urban regions and, finally, through mixed land uses. The study attempted to evaluate the sources and influences of total phosphorus under different hydrologic conditions. Nine sampling events were completed from April 2010 to May 2011 to assess the influence of various hydrologic conditions on aqueous and riverbed sediment total phosphorus concentrations. Total phosphorus concentrations and loads exceeded the in-stream limits proposed by the Colorado Department of Public Health and Environment in all observed hydrologic conditions, and nonpoint sources were significant in high-flow conditions. Reducing nutrients only at water resource recovery facilities (WRRFs) could not achieve the in-stream limits without substantial reduction of non-point-source loads. The study exposed a need for flexibility in WRRF discharge limits based on the overall total phosphorus load in the river from other sources.
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Monitoreo del Ambiente/métodos , Sedimentos Geológicos/química , Fósforo/análisis , Ríos/química , Contaminantes Químicos del Agua/análisis , Colorado , Fenómenos Geológicos , Hidrología , Movimientos del AguaRESUMEN
Excess nutrients are among the leading sources of water quality impairment in the Unites States, and the USEPA has been working with state regulatory agencies to develop nutrient criteria for wastewater treatment plants (WWTPs). The Colorado Department of Public Health and Environment is scheduled to establish nutrient regulations in 2013, and stream total P (TP) concentration standards of 0.16 mg L in warm water and 0.11 mg L in cold water have been proposed for the rivers in the state. The objectives of this study were to monitor TP concentrations and loads along the Cache La Poudre River as it flows from the pristine upstream area through urban regions and finally through a mixture of agricultural and urban land uses. The study attempts to evaluate the sources and influences of TP under different hydrologic conditions. Twelve sampling events were completed from April 2010 to August 2011 to assess the influence of various flow and precipitation conditions on aqueous TP concentrations. During midrange flows and dry conditions, WWTPs were the major sources of TP, but other sources were more significant under high-flow and wet conditions according to a load analysis. The analysis indicates that reducing the TP load from WWTPs will only marginally affect the TP load in the river, and therefore it appears that other sources (e.g., stormwater and agricultural runoff) need to be addressed before the aquatic life-based stream standard can be achieved.
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Fósforo , Aguas Residuales , Colorado , Monitoreo del Ambiente , Ríos , Contaminantes Químicos del AguaRESUMEN
The Colorado Department of Public Health and Environment (CDPHE) proposed the in-stream numeric nutrient criteria as 2 mg TN per L and 0.16 mg TP per L for warm surface waters and 0.40 mg TN per L and 0.11 mg TP per L for cold surface waters. Consequently the department presented the nutrient limits for the municipal wastewater treatment plants (WWTPs) as annual averages of 0.7 mg TP per L and 5.7 mg TIN per L and quarterly averages of 1.0 mg TP per L and 9.0 mg TIN per L. Implementing stringent nutrient reduction at point sources is unlikely to result in improvements to the environment without non-point source controls. In this study, total nitrogen (TN) load inputs from known point source, WWTPs, and other non-point sources at six sub-basins of the Cache La Poudre (CLP) River Basin were estimated and compared under various hydrologic conditions. Significant loading exceedance from the proposed limits was observed during lower flow conditions and other sources dominated during events when the exceedance was observed except for one point. The point receives direct TN inputs from a WWTP which has the highest TN concentration in its effluent among all WWTPs in the study area; however, TN loads entered the point from other sources were significant during higher flow conditions. TN loads in the CLP River were simulated to determine whether the loads meet the proposed in-stream limits in a case in which all WWTPs comply with the proposed regulations for WWTPs. From this study, it was observed that reducing TN concentrations only at WWTPs merely impacts total TN loads in the river.
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Nitrógeno/análisis , Eliminación de Residuos Líquidos/legislación & jurisprudencia , Aguas Residuales/química , Contaminantes Químicos del Agua/análisis , Calidad del Agua/normas , Monitoreo del Ambiente , Política Ambiental , Ríos/química , Contaminación Química del Agua/legislación & jurisprudencia , Contaminación Química del Agua/estadística & datos numéricosRESUMEN
Deep blue emitting materials with a new core structure containing indenopyrazine have been synthesized; a non-doped device using one of these materials as a blue emitter was found to exhibit high external quantumn efficiency of 4.6% and excellent color purity of (0.154, 0.078) as well as narrow emission band of 47 nm FWHM.
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Iridovirus is a causative agent of epizootics among cultured rock bream (Oplegnathus fasciatus) in Korea. Here, we report the complete genomic sequence of rock bream iridovirus (RBIV). The genome of RBIV was 112080 bp long and contained at least 118 putative open reading frames (ORFs), and its genome organization was similar to that of infectious spleen and kidney necrosis virus (ISKNV). Of the RBIV's 118 ORFs, 85 ORFs showed 60-99% amino acid identity to those of ISKNV. Phylogenetic analysis of major capsid protein (MCP), DNA repair protein RAD2, and DNA polymerase type-B family indicated that RBIV is closely related to red sea bream iridovirus (RSIV), Grouper sleepy disease iridovirus (GSDIV), Dwarf gourami iridovirus (DGIV), and ISKNV. The genome sequence provides useful information concerning the evolution and divergence of iridoviruses in cultured fish.
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ADN Viral/genética , Genoma Viral , Iridovirus/genética , Perciformes/virología , Animales , Replicación del ADN , Bases de Datos de Proteínas , Iridovirus/clasificación , Iridovirus/patogenicidad , Iridovirus/fisiología , Datos de Secuencia Molecular , Filogenia , Especificidad de la Especie , Proteínas Virales/genéticaRESUMEN
Anti-4-1BB (CD137) monoclonal antibody (mAb) has been reported to suppress immune responses and to have the potential for use as a therapeutic agent to block autoimmune diseases. Previously, the authors prepared an antagonistic anti-human 4-1BB (CD137) mAb, BBK2. Here the authors report the humanization of BBK2 using a phage display library. Four humanized single-chain Fv (scFv) fragments were selected from a combinatorial library expressing a phage-displayed humanized scFv. They were found to retain the epitope specificity of the original mAb and to have affinities higher than those of the original. Both the soluble and bound forms of the humanized scFv suppressed the proliferation of human peripheral blood mononuclear cells, similar to the original mAb. These results suggest that humanized anti-human 4-1BB scFvs can be used as a valuable reagent for clinical application.
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Anticuerpos Monoclonales/química , Técnicas Inmunológicas , Biblioteca de Péptidos , Receptores de Factor de Crecimiento Nervioso/química , Receptores del Factor de Necrosis Tumoral/química , Secuencia de Aminoácidos , Antígenos CD , Unión Competitiva , División Celular , Separación Celular , ADN/química , Relación Dosis-Respuesta Inmunológica , Ensayo de Inmunoadsorción Enzimática , Citometría de Flujo , Biblioteca de Genes , Humanos , Fragmentos de Inmunoglobulinas , Inmunoterapia , Cinética , Leucocitos Mononucleares/metabolismo , Datos de Secuencia Molecular , Unión Proteica , Homología de Secuencia de Aminoácido , Miembro 9 de la Superfamilia de Receptores de Factores de Necrosis TumoralRESUMEN
Given the key role 4-1BB plays in the stimulation of T cells, humanization of agonistic anti-human 4-1BB monoclonal antibody (mAb) may have important clinical applications. In this paper, we present the humanization of agonistic anti-human 4-1BB mAb, BBK-4, using a phage display library. We first prepared the combinatorial library by incorporating murine and human alternative at positions representing buried residues that might affect the structural integrity of the antigen binding site. Six humanized single chain Fv (scFv) fragments were selected from the combinatorial library expressing phage-displayed humanized scFv. They were found to retain the epitope specificity of the original mAb but had affinities of lower than 1/10 of the original. In spite of the lower affinity, the humanized scFv coated on the surface expanded human peripheral blood mononuclear cells (PBMCs) in MLR similarly to the original mAb in the presence of anti-CD3 mAb. These results suggest that humanized anti-human 4-1BB scFvs can be used as a valuable reagent for clinical application.