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The global desire to improve the performance of road pavements and move towards a sustainable transportation system has immensely encouraged the usage of fibers in asphalt paving materials. In this study, glass fibers trademarked as ESGFIBER produced by the ESG Industry company Limited from Daejeon, Korea were added in dense-graded asphalt mix. The purpose of this study was to evaluate effects that fibers have on volumetric properties, mechanical properties, and long-term performance of asphalt concrete mixes. ESGFIBER were mixed together with aggregates and asphalt binder in asphalt mix and five different asphalt mixes with different dosage of fibers were evaluated in this study. The Marshall mix design method was used for designing all asphalt mixes, and laboratory tests indirect tensile strength test, deformation strength test and Hamburg wheel tracking test were conducted to evaluate moisture susceptibility, fatigue cracking behavior and rutting resistance of asphalt concrete mixes. The results showed that when ESGFIBER were added in asphalt mix moisture susceptibility, fatigue cracking and rutting resistance were both improved. The usage of ESGFIBER in asphalt concrete mixes can be very beneficial since the mechanical and long-term performance were improved upon the addition of fibers.

The Korean rockfish, Sebastes schlegeli, is most commonly farmed in sea cages along the coast of Korea; however, detailed information on intestinal microbiota regarding this fish is not readily available. In this study, comparison of the seasonal changes of microbial communities in the intestine between farmed and wild through the amplicon sequencing approach was conducted. The composition of major species in the intestine of this fish was very simple compared to that of other marine fish species, with members affiliated with the family Vibrionaceae hyper-dominating and comprising on average 97.6% of microbiota. However, the composition at the genus or species level and the pattern of seasonal changes of diversity indices showed significant differences between farmed and wild fish. In the farmed fish, Photobacterium phophoreum was most dominant throughout the year, accounting for 58.8% of the total. Aliivibrio fisherii and/or Aliivibrio finisterrensis also were dominant in the fall to winter but substituted by Photobacterium damselae during spring to summer. In the wild fish, on the other hand, opportunistic pathogens in the genera Aliivibrio or Vibrio were dominant in most of the samples. The analysis of shared species between gut microbiome, feed microbiota, and seawater microbiota indicated that the intestinal microbial diversity of farmed fish was affected more by microbiota of seawater than that of feed in spring and winter seasons. Additionally, the proportion of potential pathogenic Vibrio spp. in the gut showed a negative correlation with plasma glucose levels of the host. This study and following studies will be helpful in understanding the interaction between microbiome hosts and the development of techniques to enhance production of healthy Korean rockfish.

An anaerobic, rod-shaped, mesophilic, chemolithoautotrophic, sulfate-reducing bacterial strain IOR2T was isolated from a newly found deep-sea hydrothermal vent (OVF, Onnuri Vent Field) area in the central Indian Ocean ridge (11°24'88″ S 66°25'42″ E, 2021 m water depth). The 16S rRNA gene sequence analysis revealed that the strain IOR2T was most closely related to Desulfovibrio senegalensis BLaC1T (96.7%). However, it showed low similarity with the members of the family Desulfovibrionaceae, such as Desulfovibrio tunisiensis RB22T (94.0%), D. brasiliensis LVform1T (93.9%), D. halophilus DSM 5663T (93.7%), and Pseudodesulfovibrio aespoeensis Aspo-2T (93.2%). The strain IOR2T could grow at 23-42°C (optimum 37°C), pH 5.0-8.0 (optimum pH 7.0) and with 0.5-6.5% (optimum 3.0%) NaCl. The strain could use lactate, pyruvate, H2, and glycerol as electron donors and sulfate, thiosulfate, and sulfite as electron acceptors. The major fatty acids of the strain IOR2T were iso-C15:0, iso-C17:0, ante-iso-C15:0, and summed feature 9 (C16:0 methyl/iso-C17:1ω9c). Both the strains IOR2T and BLaC1T could grow with CO2 and H2 as the sole sources of carbon and energy, respectively. Genomic evidence for the Wood-Ljungdahl pathway in both the strains reflects chemolithoautotrophic growth. The DNA G + C content of the strain IOR2T and BLaC1T was 58.1-60.5 mol%. Based on the results of the phylogenetic and physiologic studies, Paradesulfovibrio onnuriensis gen. nov., sp. nov. with the type strain IOR2T (= KCTC 15845T = MCCC 1K04559T) was proposed to be a member of the family Desulfovibrionaceae. We have also proposed the reclassification of D. senegalensis as Paradesulfovibrio senegalensis comb. nov.

Ferredoxin-dependent metabolic engineering of electron transfer circuits has been developed to enhance redox efficiency in the field of synthetic biology, e.g., for hydrogen production and for reduction of flavoproteins or NAD(P)+. Here, we present the bioconversion of carbon monoxide (CO) gas to formate via a synthetic CO:formate oxidoreductase (CFOR), designed as an enzyme complex for direct electron transfer between noninteracting CO dehydrogenase and formate dehydrogenase using an electron-transferring Fe-S fusion protein. The CFOR-introduced Thermococcus onnurineus mutant strains showed CO-dependent formate production in vivo and in vitro. The formate production rate from purified CFOR complex and specific formate productivity from the bioreactor were 348 ± 34 μmol/mg/min and 90.2 ± 20.4 mmol/g-cells/h, respectively. The CO-dependent CO2 reduction/formate production activity of synthetic CFOR was confirmed, indicating that direct electron transfer between two unrelated dehydrogenases was feasible via mediation of the FeS-FeS fusion protein.

Ferredoxin-dependent metabolic engineering of electron transfer circuits has been developed to enhance redox efficiency in the field of synthetic biology, e.g., for hydrogen production and for reduction of flavoproteins or NAD(P)+. Here, we present the bioconversion of carbon monoxide (CO) gas to formate via a synthetic CO:formate oxidoreductase (CFOR), designed as an enzyme complex for direct electron transfer between non-interacting CO dehydrogenase and formate dehydrogenase using an electron-transferring Fe-S fusion protein. The CFOR-introduced Thermococcus onnurineus mutant strains showed CO-dependent formate production in vivo and in vitro. The maximum formate production rate from purified CFOR complex and specific formate productivity from the bioreactor were 2.2 ± 0.2 μmol/mg/min and 73.1 ± 29.0 mmol/g-cells/h, respectively. The CO-dependent CO2 reduction/formate production activity of synthetic CFOR was confirmed, indicating that direct electron transfer between two unrelated dehydrogenases was feasible via mediation of the FeS-FeS fusion protein.

To date, NAD(P)H, ferredoxin, and coenzyme F(420) have been identified as electron donors for thioredoxin reductase (TrxR). In this study, we present a novel electron source for TrxR. In the hyperthermophilic archaeon Thermococcus onnurineus NA1, the frhAGB-encoded hydrogenase, a homolog of the F(420)-reducing hydrogenase of methanogens, was demonstrated to interact with TrxR in coimmunoprecipitation experiments and in vitro pulldown assays. Electrons derived from H(2) oxidation by the frhAGB-encoded hydrogenase were transferred to TrxR and reduced Pdo, a redox partner of TrxR. Interaction and electron transfer were observed between TrxR and the heterodimeric hydrogenase complex (FrhAG) as well as the heterotrimeric complex (FrhAGB). Hydrogen-dependent reduction of TrxR was 7-fold less efficient than when NADPH was the electron donor. This study not only presents a different type of electron donor for TrxR but also reveals new functionality of the frhAGB-encoded hydrogenase utilizing a protein as an electron acceptor. IMPORTANCE This study has importance in that TrxR can use H(2) as an electron donor with the aid of the frhAGB-encoded hydrogenase as well as NAD(P)H in T. onnurineus NA1. Further studies are needed to explore the physiological significance of this protein. This study also has importance as a significant step toward understanding the functionality of the frhAGB-encoded hydrogenase in a nonmethanogen; the hydrogenase can transfer electrons derived from oxidation of H(2) to a protein target by direct contact without the involvement of an electron carrier, which is distinct from the mechanism of its homologs, F(420)-reducing hydrogenases of methanogens.

Volume 86, no. 6, AEM02630-19, 2020, [https://doi.org/10.1128/AEM.02630-19][1]. Page 9: The following reference should read as shown. 11. Susanti D, Loganathan U, Mukhopadhyay B. 2016. A novel F420-dependent thioredoxin reductase gated by low potential FAD: a tool for redox regulation in an

To date, NAD(P)H, ferredoxin, and coenzyme F420 have been identified as electron donors for thioredoxin reductase (TrxR). In this study, we present a novel electron source for TrxR. In the hyperthermophilic archaeon Thermococcus onnurineus NA1, the frhAGB-encoded hydrogenase, a homolog of the F420-reducing hydrogenase of methanogens, was demonstrated to interact with TrxR in coimmunoprecipitation experiments and in vitro pulldown assays. Electrons derived from H2 oxidation by the frhAGB-encoded hydrogenase were transferred to TrxR and reduced Pdo, a redox partner of TrxR. Interaction and electron transfer were observed between TrxR and the heterodimeric hydrogenase complex (FrhAG) as well as the heterotrimeric complex (FrhAGB). Hydrogen-dependent reduction of TrxR was 7-fold less efficient than when NADPH was the electron donor. This study not only presents a different type of electron donor for TrxR but also reveals new functionality of the frhAGB-encoded hydrogenase utilizing a protein as an electron acceptor.IMPORTANCE This study has importance in that TrxR can use H2 as an electron donor with the aid of the frhAGB-encoded hydrogenase as well as NAD(P)H in T. onnurineus NA1. Further studies are needed to explore the physiological significance of this protein. This study also has importance as a significant step toward understanding the functionality of the frhAGB-encoded hydrogenase in a nonmethanogen; the hydrogenase can transfer electrons derived from oxidation of H2 to a protein target by direct contact without the involvement of an electron carrier, which is distinct from the mechanism of its homologs, F420-reducing hydrogenases of methanogens.

Protein disulfide oxidoreductases are redox enzymes that catalyze thiol-disulfide exchange reactions. These enzymes include thioredoxins, glutaredoxins, protein disulfide isomerases, disulfide bond formation A (DsbA) proteins, and Pyrococcus furiosus protein disulfide oxidoreductase (PfPDO) homologues. In the genome of a hyperthermophilic archaeon, Thermococcus onnurineus NA1, the genes encoding one PfPDO homologue (TON_0319, Pdo) and three more thioredoxin- or glutaredoxin-like proteins (TON_0470, TON_0472, TON_0834) were identified. All except TON_0470 were recombinantly expressed and purified. Three purified proteins were reduced by a thioredoxin reductase (TrxR), indicating that each protein can form redox complex with TrxR. SurR, a transcription factor involved in the sulfur response, was tested for a protein target of a TrxR-redoxin system and only Pdo was identified to be capable of catalyzing the reduction of SurR. Electromobility shift assay demonstrated that SurR reduced by the TrxR-Pdo system could bind to the DNA probe with the SurR-binding motif, GTTttgAAC. In this study, we present the TrxR-Pdo couple as a redox-regulator for SurR in T. onnurineus NA1.

To date, NAD(P)H, ferredoxin, and coenzyme F

A strictly anaerobic, dissimilatory Fe(III)-reducing hyperthermophilic archaeon, designated as strain IOH1T, was isolated from a new deep-sea hydrothermal vent (Onnuri Vent Field) area in the Central Indian Ocean ridge. Strain IOH1T showed > 99% 16S rRNA gene sequence similarity with Thermococcus celericrescens TS2T (99.4%) and T. siculi DSM 12349T (99.2%). Additional three species T. barossii SHCK-94T (99.0%), T. celer Vu13T (98.8%), and T. piezophilus (98.6%) showed > 98.6% of 16S rRNA gene sequence similarity, however, the maximum OrthoANI value is 89.8% for the genome of T. celericrescens TS2T. Strain IOH1T cells are coccoid, 1.2-1.8 μm in diameter, and motile by flagella. Growth was at 70-82°C (optimum 80°C), pH 5.4-8.0 (optimum pH 6.0) with 2-4% (optimum 3%) NaCl. Growth of strain IOH1T was enhanced by starch, pyruvate, D(+)-maltose and maltodextrin as a carbon sources, and elemental sulfur as an electron acceptor; clearly different from those of related species T. celecrescens DSM 17994T and T. siculi DSM 12349T. Strain IOH1T, T. celercrescence DSM 17994T, and T. siculi DSM 12349T reduced soluble Fe(III)-citrate present in the medium, whereas the amount of total cellular proteins increased with the concomitant accumulation of Fe(II). We determined a circular chromosome of 2,234 kb with an extra-chromosomal archaeal plasmid, pTI1, of 7.7 kb and predicted 2,425 genes. The DNA G + C content was 54.9 mol%. Based on physiological properties, phylogenetic, and genome analysis, we proposed that strain IOH1T (= KCTC 15844T = JCM 39077T) is assigned to a new species in the genus Thermococcus and named Thermococcus indicus sp. nov.

In this study, we developed a rapid and sensitive enzymatic assay for 2,3-butanediol (2,3-BDO) detection. The concentration of 2,3-BDO was determined by measuring the reduction of NADP(+) using Clostridium ljungdahlii 2,3-butanediol dehydrogenase (CL-Bdh). The enzymatic assay could detect as low as 0.01 mM of 2,3-BDO, while the high-performance liquid chromatography (HPLC) method required a much higher concentration than 0.15 mM. Gratifyingly, the developed method was 15 times more sensitive than the HPLC method. When the enzymatic assay was applied to high-throughput screening, the enzymatic assay detected 14 positive samples out of 23 tested, as compared to 8 by the HPLC method. These results suggest that the enzymatic assay is an effective screening method for the detection of 2,3-BDO-producing microbes in a microtiter plate-based format. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s13205-019-1705-9) contains supplementary material, which is available to authorized users.

The biosynthetic pathway of 2,3-butanediol (2,3-BDO) production from pyruvate under anaerobic conditions includes three enzymes: acetolactate synthase (ALS), acetolactate decarboxylase (ALDC), and acetoin reductase (AR). Recently, in anaerobic hyperthermophilic Pyrococcus furiosus, it has been reported that acetoin, a precursor of 2,3-BDO, is produced from pyruvate by ALS through a temperature-dependent metabolic switch. In this study, we first attempted to produce 2,3-BDO from Thermococcus onnurineus NA1 using a simple biosynthetic pathway by two enzymes (ALS and AR) at a high temperature. Two heterologous genes, acetolactate synthase (alsS) from Pyrococcus sp. NA2 and alcohol dehydrogenase (adh) from T. guaymacensis, were introduced and expressed in T. onnurineus NA1. The mutant strain produced approximately 3.3 mM 2,3-BDO at 80 °C. An acetyl-CoA synthetase IIIα (TON_1001) was further deleted to enhance 2,3-BDO production, and the mutant strain showed a 25% increase in the specific production of 2,3-BDO. Furthermore, when carbon monoxide (CO) gas was added as a reductant, specific production of 2,3-BDO increased by 45%. These results suggest a new biosynthetic pathway for 2,3-BDO and demonstrate the possibility of T. onnurineus NA1 as a platform strain for 2,3-BDO production at high temperatures.

PURPOSES : Performance evaluation of four types of asphalt concrete overlays for deteriorated national highways. METHODS : Pavement distress surveys for crack rate and rut depth have been conducted annually using an automated pavement survey vehicle since 2007. Linear and non-linear performance prediction models of the asphalt concrete overlays were developed for 43 sections. The service life of the asphalt overlays was defined as the number of years after which a crack rate of 30% or rut depth of 15mm is observed. RESULTS : The service life of the asphalt overlays was estimated as 17.4 years on an average. In 90.7% of the sections, the service life of the overlays was 15 years or more which is 1.5 times the life of conventional asphalt concrete overlays used in national highways. The performance of the overlays was dependent on the type of asphalt mixture, traffic volume levels, and environmental conditions. CONCLUSIONS : The usage of stone mastic asphalt (SMA) and polymer-modified asphalt (PMA) for the overlays provided good resistance to cracking and rutting development. It is recommended that appropriate asphalt concrete overlays must be applied depending on the type of existing pavement distress.

Cracking is an inevitable fact of asphalt concrete pavements and plays a major role in pavement deterioration. Pavement cracking is one of the main factors determining the frequency and method of repair. Cracks can be treated with a number of preventative maintenance actions, including overlay surface treatments such as slurry sealing, crack sealing, or crack filling. Pavement cracks can show up as one or all of the following types: transverse, longitudinal, fatigue, block, reflective, edge, and slippage. Crack sealing is a frequently used pavement maintenance treatment because it significantly extends the pavement service life. However, crack sealant often fails prematurely due to a loss of adhesion. Because current test methods are mostly empirical and only provide a qualitative measure of the bond strength, they cannot accurately predict the adhesive failure of the sealant. This study introduces a laboratory test aimed at assessing the bonding of hot-poured crack sealant to the walls of pavement cracks. A pneumatic adhesion tensile testing instrument (PATTI) was adopted to measure the bonding strength of the hot-poured crack sealant as a function of the curing time and temperature. Based on a limited number of test results, the hot-poured crack sealants have very different bonding performances. Therefore, this test method can be proposed as part of a newly developed performance-based standard specification for hot-poured crack sealants for use in the future. PURPOSES : The purpose of this study was to evaluate both the adhesion and failure performance of a crack sealant as a function of its curing time and curing temperature. METHODS: A pneumatic adhesion tensile testing instrument (PATTI) was adopted to measure the adhesion performance of a crack sealant as a function of the curing time and curing temperature. RESULTS: With changes in the curing time, curing temperature, and sealant type, the bond strengths were found to be significantly different. Also, higher bond strengths were measured at lower temperatures. Different sealant types produced completely different bond strengths and failure behaviors. CONCLUSIONS: The bonding strength of an evaluated crack sealant was shown to differ depending on various factors. Two sealant types, which were composed of different raw materials, were shown to perform differently. The newly proposed test offers the possibility of evaluating and differentiating between different crack sealants. Based on alimited number of test results, this test method can be proposed as part of a newly developed performance-based standard specification for crack sealants or as part of a guideline for the selection of hot-poured crack sealant in the future.

PURPOSES : Recently, crack, rutting, and stripping problems from the surface of asphalt pavements in National highway are observed and they affect the drivers to feel uncomfortable on the road. Surface treatments are recommended to use in distressed pavements due to cost-effective, and improvement of surface performance. The purpose of this study is to evaluate the performance of micro-surfacing and polymer slurry seal treatments for distressed asphalt pavements. METHODS : Surface conditions and friction resistance are evaluated for asphalt pavements treated with micro-surfacing and polymer slurry seal mixes in National highway 30 line and 34 line. Visual observation is conducted and surface performance is measured by PES (Performance Evaluation Surveyor) in terms of crack ratio, rutting and IRI(International Roughness Index). BPN(British Pendulum Number) is measured by BPT(British Pendulum Tester) to evaluate the friction resistance in the field. RESULTS : The surface evaluation results are presented for asphalt pavement treated with micro-surfacing and polymer slurry seal treatments in National highway 30 line and 34 line. Based on the visual observation, micro-surfacing and polymer slurry seal treatments show better improvements in terms of cracks and stripping. Based on the surface conditions measured by PES vehicle, the surface performance of micro-surfacing treatments improves from 53.3% to 54.2% and the surface performance of polymer slurry seal treatments improves from 21.6% to 59.7%. However, the friction resistance of both micro-surfacing and polymer slurry seal treatments decreases from 2.5% to 6.7%. Further, it should be verified to produce the surface exposed with aggregates during the construction process of both treatment methods in the field. CONCLUSIONS : Based on the performance evaluation results in the filed, the surface performance of asphalt pavement treated with micro-surfacing and polymer slurry seal treatments improves from 21.6% to 59.7%. While, the friction resistance of asphalt pavement treated with micro-surfacing and polymer slurry seal treatments does not improve. It can be concluded that current micro-surfacing and polymer slurry seal treatments would improve surface performance but would not improve the friction resistance.

Among four basic mechanisms for biological hydrogen (H2) production, dark fermentation has been considered to show the highest hydrogen evolution rate (HER). H2 production from one-carbon (C1) compounds such as formate and carbon monoxide (CO) is promising because formate is an efficient H2 carrier, and the utilization of CO-containing syngas or industrial waste gas may render the industrial biohydrogen production process cost-effective. A variety of microbes with the formate hydrogen lyase (FHL) system have been identified from phylogenetically diverse groups of archaea and bacteria, and numerous efforts have been undertaken to improve the HER for formate through strain optimization and bioprocess development. CO-dependent H2 production has been investigated to enhance the H2 productivity of various carboxydotrophs via an increase in CO gas-liquid mass transfer rates and the construction of genetically modified strains. Hydrogenogenic CO-conversion has been applied to syngas and by-product gas of the steel-mill process, and this low-cost feedstock has shown to be promising in the production of biomass and H2. Here, we focus on recent advances in the isolation of novel phylogenetic groups utilizing formate or CO, the remarkable genetic engineering that enhances H2 productivity, and the practical implementation of H2 production from C1 substrates.

PURPOSES : An conventional method for electric power generation is converting thermal energy into mechanical energy then to electrical energy. Due to environmental issues such as global warming related with emission etc., were the limiting factor for the energy resources which resulting in extensive research and novel technologies are required to generate electric power. Thermal energy harvesting using thermoelectric generator is one of energy harvesting technologies due to diverse advantages for new green technology. This paper presents a possibility of application of the thermoelectric generator`s application in the direct exchange of waste solar energy into electrical power in road space. METHODS : To measure generated electric power of the thermoelectric generator, data logger was adopted as function of experimental factors such as using cooling sink, connection methods etc. Also, the thermoelectric generator、s behavior at low ambient temperature was investigated as measurement of output voltage vs. elapsed times. RESULTS : A few temperature difference between top an bottom of the thermoelectric generator is generated electric voltage. Components of an electrical circuit can be connected in various ways. The two simplest of these are called series and parallel and occur so open. Series shows slightly better performance in this study. An installation of cooling sink in the thermoelectric generator system was enhanced the output of power voltage. CONCLUSIONS : In this paper, a basic concepts of thermoelectric power generation is presented and applications of the thermoelectric generator to waste solar energy in road is estimated for green energy harvesting technology. The possibility of usage of thermoelectric technology for road facilities was found under the ambient thermal gradient between two surfaces of the thermoelectric module. An experiment results provide a testimony of the feasibility of the proposed environmental energy harvesting technology on the road facilities.

PURPOSES : The objective of this study is to evaluate the tack-coating material`s properties using the bitumen bond strength(BBS) test and damping test as function of changed curing times. In this study, bonding strength tests were performed according to the curing time of tack coating materials. METHODS : In order to investigate bonding characteristic of tack coating materials, the Pneumatic Adhesion tensile Testing Instrument(PATTI) device is used to measure the bond strength between the tack coating materials and aggregate substrate based on the AASHTO TP-91. Also, damping test as in situ test was used to determine an appropriate traffic openting time for construction vehicle. Four different tack-coating materials were used in this study. The BBS tests were performed a one hour curing and testing temperatures of , , and . Damping test was conducted at 30min, 60min, 90min, and 120 min of curing times with temperatures of and . RESULTS and CONCLUSIONS : The BBS test results show various bond strength as function of tack coat materials. At the same testing condition, A tack coat material shows almost two times higher than D tack coat materials although both materials are satisfied the criteria of material`s physical properties. Also, Dampting test results shows similar trend with BBS test result. The damping test result was significantly changed as function of tack coat materials. Based on this study, the tack coating material`s curing time is very important. Therefore, both curing time and the bond strength`s characteristic has to be considered in standard specification.

Transverse rumble strips were commonly installed on approaches to intersections, toll plazas, and horizontal curves. The main function is to alert drivers by creating noise and vibration. This TRS has been demonstrated to be effective in reducing traffic accident due to sleeping. However, there are some disadvantages in its utilization, such as traffic noise produced by TRS. The objective of this study was to quantify the level of traffic noise; to verify the effects of speed and TRS shapes. Two types of vehicles [sedan and minivan] were driven over four different patterns of TRS and over cement concrete pavement at four different speeds. Researchers measured the produce noise at 7.5m distance with 1.2 m height from center of the vehicle lane at each test section. Results indicate that vehicle type, speed, TRS shape affects created traffic noise. The sound level of Type A was statistically highest among others.

In Korea, recently, surface treatments are recommended to use in distressed pavements due to cost-effective, extension of pavement life, and improvement of surface performance such as friction and noise. In this study, microsurfacing and polymer slurry seal are selected to evaluate assessment of field application and their performance. The test section using both surface treatments were built on hot-mix asphalt (HMA) surfaced national highway at the same time and then their performance are monitored right after the construction. To evaluate the field performance of surface treatments, friction, noise, crack and IRI values are measured using noise-measuring vehicle, British pendulum tester (BPT), and pavement evaluation surveyor (PES) before and after applying surface treatments. On the basis of the field performance of surface treatments, pavement surface treated with microsurfacing and polymer slurry seal shows better fiction resistance and noise reduction compared to pavement surface without preservation treatment. It is postulated that the surface characteristics using surface treatments can be improved but long-term performance should be monitored over the service life.

Hydrogenogenic CO oxidation (CO + H 2 O → CO 2 + H 2 ) has the potential for H 2 production as a clean renewable fuel. Thermococcus onnurineus NA1, which grows on CO and produces H 2 , has a unique gene cluster encoding the carbon monoxide dehydrogenase (CODH) and the hydrogenase. The gene cluster was identified as essential for carboxydotrophic hydrogenogenic metabolism by gene disruption and transcriptional analysis. To develop a strain producing high levels of H 2 , the gene cluster was placed under the control of a strong promoter. The resulting mutant, MC01, showed 30-fold-higher transcription of the mRNA encoding CODH, hydrogenase, and Na + /H + antiporter and a 1.8-fold-higher specific activity for CO-dependent H 2 production than did the wild-type strain. The H 2 production potential of the MC01 mutant in a bioreactor culture was 3.8-fold higher than that of the wild-type strain. The H 2 production rate of the engineered strain was severalfold higher than those of any other CO-dependent H 2 -producing prokaryotes studied to date. The engineered strain also possessed high activity for the bioconversion of industrial waste gases created as a by-product during steel production. This work represents the first demonstration of H 2 production from steel mill waste gas using a carboxydotrophic hydrogenogenic microbe.

PURPOSES: This study is to investigate the Hot In-Place recycling asphalt mixture in Korea using field produced materials. METHODS: Hot In-Place reclaimed asphalt mixture was investigated to evaluate the mixture properties based on various test results such as Marshall Test, Indirect Tensile Test, TSR, and Wheel Tracking Test. These test values were compared with domestic standard specification. RESULTS: The result of the laboratory experiment indicates that the Hot In-Place Reclaimed(HIR) asphalt mixture produced at the field constrution site was satisfied all of the test criteria such as Indirect tensile test, Marshall and TSR test, and wheel tracking test. During the test, the research team found that current HIR system is required an extention of mixing time to improve quality and to reduce variation of sample to sample. Although the current HIR mixture was passed the test criteria, there is a potential capability to enhance the mixture properties as extend mixting time. CONCLUSIONS: Based on these laboratory test results, It would be concluded that domestic HIR mixture`s properties were satisfied all standard specification related with evaluation of recycling asphalt mixtures. Based on this case study result, there is a chance to save construction cost and increase the usage of reclaimed asphalt concrete in the future.

Formate-dependent proton reduction to H 2 (HCOO − + H 2 O → HCO 3 − + H 2 ) has been reported for hyperthermophilic Thermococcus strains. In this study, a hyperthermophilic archaeon, Thermococcus onnurineus strain NA1, yielded H 2 accumulation to a partial pressure of 1 × 10 5 to 7 × 10 5 Pa until the values of Gibbs free energy change (Δ G ) reached near thermodynamic equilibrium (−1 to −3 kJ mol −1 ). The bioenergetic requirement for the metabolism to conserve energy was demonstrated by Δ G values as small as −5 kJ mol −1 , which are less than the biological minimum energy quantum, −20 kJ mol −1 , as calculated by Schink (B. Schink, Microbiol. Mol. Biol. Rev. 61:262-280, 1997). Considering formate as a possible H 2 storage material, the H 2 production potential of the strain was assessed. The volumetric H 2 production rate increased linearly with increasing cell density, leading to 2,820 mmol liter −1 h −1 at an optical density at 600 nm (OD 600 ) of 18.6, and resulted in the high specific H 2 production rates of 404 ± 6 mmol g −1 h −1 . The H 2 productivity indicates the great potential of T. onnurineus strain NA1 for practical application in comparison with H 2 -producing microbes. Our result demonstrates that T. onnurineus strain NA1 has a highly efficient metabolic system to thrive on formate in hydrothermal systems.

The incidence of upper gastrointestinal bleeding has decreased in the general population, but increased for the geriatric patients worldwide. The aim of this study was to evaluate the clinical characteristics and outcomes of geriatric patients (aged 65 or older) with upper gastrointestinal bleeding and compare with non-geriatric patients (less than 65 years old). Methods: We conducted a retrospective study of 288 patients with upper gastrointestinal bleeding from January 2007 to August 2010. We compared the clinical characteristics and outcomes of geriatric patients (n=148, 51.4%) with those of non-geriatric patients (n=140, 48.6%). Results: Cardiovascular disease, hypertension, and cerebrovascular accident were more prevalent in the geriatric group, compared with the non-geriatric group (p 0.05). The length of hospital stay and length of ICU stay in the geriatric patients were significantly longer than the non-geriatric group (p

The oxidation of formate to carbon dioxide and hydrogen is a common reaction in microorganisms in anaerobic environments, but it releases little energy and had not been shown to sustain growth in an isolated species. Now Kim et al. have discovered that that several hyperthermophilic archaea of the Thermococcus genus are indeed capable of using formate oxidation for growth. These organisms thrive at above 80 °C, a habitat that may give a competitive advantage to organisms using what is one of the simplest forms of anaerobic respiration so far described. The oxidation of formate and water to bicarbonate and H2 is relatively common in microorganisms under anaerobic conditions. But can this reaction sustain growth in an isolated species? Here it is shown that several individual Thermococcus species can use formate oxidation for growth. Moreover, the biochemical basis of this ability is delineated. Although a common reaction in anaerobic environments, the conversion of formate and water to bicarbonate and H2 (with a change in Gibbs free energy of ΔG° = +1.3 kJ mol−1) has not been considered energetic enough to support growth of microorganisms. Recently, experimental evidence for growth on formate was reported for syntrophic communities of Moorella sp. strain AMP and a hydrogen-consuming Methanothermobacter species and of Desulfovibrio sp. strain G11 and Methanobrevibacter arboriphilus strain AZ1. The basis of the sustainable growth of the formate-users is explained by H2 consumption by the methanogens, which lowers the H2 partial pressure, thus making the pathway exergonic2. However, it has not been shown that a single strain can grow on formate by catalysing its conversion to bicarbonate and H2. Here we report that several hyperthermophilic archaea belonging to the Thermococcus genus are capable of formate-oxidizing, H2-producing growth. The actual ΔG values for the formate metabolism are calculated to range between −8 and −20 kJ mol−1 under the physiological conditions where Thermococcus onnurineus strain NA1 are grown. Furthermore, we detected ATP synthesis in the presence of formate as a sole energy source. Gene expression profiling and disruption identified the gene cluster encoding formate hydrogen lyase, cation/proton antiporter and formate transporter, which were responsible for the growth of T. onnurineus NA1 on formate. This work shows formate-driven growth by a single microorganism with protons as the electron acceptor, and reports the biochemical basis of this ability.

The ATP synthase of many archaea has the conserved sodium ion binding motif in its rotor subunit, implying that these A1AO-ATP synthases use Na(+) as coupling ion. However, this has never been experimentally verified with a purified system. To experimentally address the nature of the coupling ion, we have purified the A1AO-ATP synthase from T. onnurineus. It contains nine subunits that are functionally coupled. The enzyme hydrolyzed ATP, CTP, GTP, UTP, and ITP with nearly identical activities of around 40 units/mg of protein and was active over a wide pH range with maximal activity at pH 7. Noteworthy was the temperature profile. ATP hydrolysis was maximal at 80 °C and still retained an activity of 2.5 units/mg of protein at 45 °C. The high activity of the enzyme at 45 °C opened, for the first time, a way to directly measure ion transport in an A1AO-ATP synthase. Therefore, the enzyme was reconstituted into liposomes generated from Escherichia coli lipids. These proteoliposomes were still active at 45 °C and coupled ATP hydrolysis to primary and electrogenic Na(+) transport. This is the first proof of Na(+) transport by an A1AO-ATP synthase and these findings are discussed in light of the distribution of the sodium ion binding motif in archaea and the role of Na(+) in the bioenergetics of archaea.

Genomic analysis of a hyperthermophilic archaeon Thermococcus sp. NA1 revealed the presence of an 885-bp open reading frame encoding a protein of 295 amino acids with a calculated molecular mass of 32,981 Da. Analysis of the deduced amino acid sequence showed that amino acid residues important for catalytic activity and the metal binding ligands conserved in all of methionyl aminopeptidases (MetAP) were also conserved and belonged to type IIa MetAP. The protein, designated TNA1_MetAP (Thermococcus sp. NA1 MetAP), was cloned and expressed in Escherichia coli. The recombinant enzyme was a Mn(2+)-, Ni(2+)-, Fe(2+)-, or Co(2+)-dependent metallopeptidase. Optimal MetAP activity against L: -methionine p-nitroanilide (Met-pNA) (K (m) = 0.68 mM) occurred at pH 7.0 and 80 to 90 degrees C. The MetAP was very unstable compared to Pyrococcus furiosus MetAP, which was completely inactivated by heating at 80 degrees C for 5 min. It seemed likely that the cysteine residue (Cys53) played a critical role in regulating the thermostability of TNA1_MetAP.

Genomic analysis of a hyperthermophilic archaeon, Thermococcus sp. NA1, revealed the presence of an 1,497 bp open reading frame, encoding a protein of 499 amino acids. The deduced amino acid sequence was similar to thermostable carboxypeptidase 1 from Pyrococcus furiosus, a member of peptidase family M32. Five motifs, including the HEXXH motif with two histidines coordinated with the active site metal, were conserved. The carboxypeptidase gene was cloned and overexpressed in Escherichia coli. Molecular masses assessed by SDS-PAGE and gel filtration were 61 kDa and 125 kDa respectively, which points to a dimeric structure for the recombinant enzyme, designated TNA1_CP. The enzyme showed optimum activity toward Z-Ala-Arg at pH 6.5 and 70-80 degrees C (k(cat)/K(m)=8.3 mM(-1) s(-1)). In comparison with that of P. furiosus CP (k(cat)/K(m)=667 mM(-1) s(-1)), TNA1_CP exhibited 80-fold lower catalytic efficiency. The enzyme showed broad substrate specificity with a preference for basic, aliphatic, and aromatic C-terminal amino acids. This broad specificity was confirmed by C-terminal ladder sequencing of porcine N-acetyl-renin substrate by TNA1_CP.

Genomic analysis of a hyperthermophilic archaeon, Thermococcus sp. strain NA1, revealed the presence of a 1,068-bp open reading frame encoding a protein consisting of 356 amino acids with a calculated molecular mass of 39,714 Da (GenBank accession no. DQ144132). Sequence analysis showed that it was similar to the putative aminopeptidase P (APP) of Thermococcus kodakaraensis KOD1. Amino acid residues important for catalytic activity and the metal binding ligands conserved in bacterial, nematode, insect, and mammalian APPs were also conserved in the Thermococcus sp. strain NA1 APP. The archaeal APP, designated TNA1_APP ( Thermococcus sp. strain NA1 APP), was cloned and expressed in Escherichia coli . The recombinant enzyme hydrolyzed the amino-terminal Xaa-Pro bond of Lys( N ε -Abz)-Pro-Pro-pNA and the dipeptide Met-Pro ( K m , 0.96 mM), revealing its functional identity. Further enzyme characterization showed the enzyme to be a Co 2+ -, Mn 2+ -, or Zn 2+ -dependent metallopeptidase. Optimal APP activity with Met-Pro as the substrate occurred at pH 5 and a temperature of 100°C. The APP was thermostable, with a half-life of >100 min at 80°C. This study represents the first characterization of a hyperthermophilic archaeon APP.

A gene homologous to rpoS was cloned from a fatal human pathogen, Vibrio vulnificus . The functional role of rpoS in V. vulnificus was accessed by using an rpoS knockout mutant strain. This mutant was impaired in terms of the ability to survive under oxidative stress, nutrient starvation, UV irradiation, or acidic conditions. The increased susceptibility of the V. vulnificus mutant in the exponential phase to H 2 O 2 was attributed to the reduced activity of hydroperoxidase I (HPI). Although σ S synthesis was induced and HPI activity reached the maximal level in the stationary phase, the mutant in the stationary phase showed the same susceptibility to H 2 O 2 as the wild-type strain in the stationary phase. In addition, HPII activity, which is known to be controlled by σ S in Escherichia coli , was not detectable in V. vulnificus strains under the conditions tested. The mutant in the exponential phase complemented with multiple copies of either the rpoS or katG gene of V. vulnificus recovered both resistance to H 2 O 2 and HPI activity compared with the control strain. Expression of the katG gene encoding HPI in V. vulnificus was monitored by using a katG :: luxAB transcriptional fusion. The expression of this gene was significantly reduced by deletion of σ S in both the early exponential and late stationary phases. Thus, σ S is necessary for increased synthesis and activity of HPI, and σ S is required for exponentially growing V. vulnificus to develop the ability to survive in the presence of H 2 O 2 .

Implicit finite element analysis of the flat surface-straight edge hemming process is performed by using a commercial code ABAQUS/Standard. Methods of finite element modeling for springback simulation and contact pair definition are discussed. An optimal mesh system is chosen through the error analysis that is based on the smoothing of discontinuity in the state variables. This study has focused on the investigation of the influence of process parameters in flanging, pre-hemming and main hemming on final hem quality, which can be defined by turn-down, warp and roll-in. The parameters adopted in this parametric study are flange length, flange angle, flanging die corner radius, face angle and insertion angle of pre-hemming punch, and over-stroke of pre-hemming and main hemming punches.

Thermococcus onnurineus NA1 is known to grow by the anaerobic oxidation of formate to CO2 and H2, a reaction that operates near thermodynamic equilibrium. Here we demonstrate that this reaction is coupled to ATP synthesis by a transmembrane ion current. Formate oxidation leads to H(+) translocation across the cytoplasmic membrane that then drives Na(+) translocation. The ion-translocating electron transfer system is rather simple, consisting of only a formate dehydrogenase module, a membrane-bound hydrogenase module, and a multisubunit Na(+)/H(+) antiporter module. The electrochemical Na(+) gradient established then drives ATP synthesis. These data give a mechanistic explanation for chemiosmotic energy conservation coupled to formate oxidation to CO2 and H2. Because it is discussed that the membrane-bound hydrogenase with the Na(+)/H(+) antiporter module are ancestors of complex I of mitochondrial and bacterial electron transport these data also shed light on the evolution of ion transport in complex I-like electron transport chains.

The National Highway pavement management system (PMS) in Korea is currently operated by the Ministry of Land, Infrastructure, and Transport Affairs (MOLIT) for efficient use of the budget through proper pavement maintenance. The PMS database system was developed in Excel spreadsheet for each road's history such as pavement material, maintenance method, and pavement conditions. In this study, the authors design a new PMS database system using geographic information system (GIS) technology for national highways to overcome the restrictions of the existing PMS. The newly designed PMS database system can provide pavement-related information more effectively than the previous PMS system. Also, it can secure the reliability of the pavement material for performing research and evaluating the exact pavement conditions.

In Korea 97.6% of a total length in national highway system is paved. Particularly, most of the national highway has been paved as asphalt pavement. Surface treatments are recommended to use in distressed asphalt pavements in the national highway due to cost-effective, extension of pavement life, and improvement of surface performance such as friction and noise. By applying a cost-effective surface treatment at the right time, the pavement is restored almost to its original condition. Most of the government agencies state that surface treatments are the most effective use of limited budgetary dollars to maintain roadway serviceability. Recently, polymer-modified emulsion is being used to improve the stability of surface treatment mixes. However, it is sensitive to curing time and content. In this study, influences of curing time and content on two types of polymer-modified emulsion used for microsurfacing and slurry seal mixes are evaluated using a load wheel tester (LWT) that is a traffic simulating device. It exhibits the approximate compacting effort of 1 million vehicle passes in less than 25 minutes when loaded with 125 lb (56.7 kg) of lead shot weight in the laboratory. Based on the LWT test results, the compactability and stability of microsurfacing and slurry seal mixes using polymer-modified emulsions significantly increase as curing time increases. Two polymer-modified emulsions used in microsurfacing and slurry seal mixes show superior performance compared to a conventional emulsion. It can be postulated that polymer-modified emulsions would be more flexible with less bleeding.

The F420-reducing hydrogenase has been known as a key enzyme in methanogenesis. Its homologs have been identified in non-methanogenic hyperthermophilic archaea, including Thermococcus onnurineus NA1, but neither physiological function nor biochemical properties have been reported to date. The enzyme of T. onnurineus NA1 was distinguished from those of other methanogens and the members of the family Desulfurobacteriaceae with respect to the phylogenetic distribution of the α and β subunits, organization of frhAGB genes and conservation of F420-coordinating residues. RT-qPCR and Western blot analyses revealed frhA gene is not silent but is expressed in T. onnurineus NA1 grown in the presence of sulfur, carbon monoxide, or formate. The trimeric enzyme complex was purified to homogeneity via affinity chromatography from T. onnurineus NA1 and exhibited catalytic activity toward the electron acceptors such as viologens and flavins but not the deazaflavin coenzyme F420. This is the first biochemical study on the function of the frhAGB-encoding enzyme from a non-methanogenic archaea.

As a result of the 1997 Kyoto treaty on climate changes, many countries in the world were faced with greenhouse gas reduction targets. Warm-mix asphalt (WMA) technologies are currently of interest to road construction industry around the world. WMA technology was developed in Europe and introduced into the United States in 2002. WMA technologies can decrease the energy consumption and emissions associated with conventional hot-mix asphalt (HMA) production. Based on those benefits and increasing awareness of WMA, the Korea Institute of Construction and Technology (KICT) developed WMA technology in Korea. WMA developed by KICT used a WMA additive that was developed by Korea Kumho Petrochemical Co. LTD. To evaluate the performance of WMA developed in Korea, an experimental test program was designed to evaluate rutting and crack resistance by using the wheel track test and indirect tensile strength test as a function of mixture types. In the case of rutting resistance, WMA shows better performance than HMA at high temperature. The Indirect Tensile Strength (IDT) test results of both WMA and HMA were same.

Members of the genus Thermococcus , sulfur-reducing hyperthermophilic archaea, are ubiquitously present in various deep-sea hydrothermal vent systems and are considered to play a significant role in the microbial consortia. We present the complete genome sequence and feature analysis of Thermococcus onnurineus NA1 isolated from a deep-sea hydrothermal vent area, which reveal clues to its physiology. Based on results of genomic analysis, T. onnurineus NA1 possesses the metabolic pathways for organotrophic growth on peptides, amino acids, or sugars. More interesting was the discovery that the genome encoded unique proteins that are involved in carboxydotrophy to generate energy by oxidation of CO to CO 2 , thereby providing a mechanistic basis for growth with CO as a substrate. This lithotrophic feature in combination with carbon fixation via RuBisCO (ribulose 1,5-bisphosphate carboxylase/oxygenase) introduces a new strategy with a complementing energy supply for T. onnurineus NA1 potentially allowing it to cope with nutrient stress in the surrounding of hydrothermal vents, providing the first genomic evidence for the carboxydotrophy in Thermococcus .

Genomic analysis of Thermococcus sp. NA revealed the presence of a 3,927-base-pair (bp) family B-type DNA polymerase gene, TNA1_pol. TNA1_pol, without its intein, was overexpressed in Escherichia coli, purified using metal affinity chromatography, and characterized. TNA1_pol activity was optimal at pH 7.5 and 75 degrees C. TNA1_pol was highly thermostable, with a half-life of 3.5 h at 100 degrees C and 12.5 h at 95 degrees C. Polymerase chain reaction parameters of TNA1_pol such as error-rate, processivity, and extension rate were measured in comparison with rTaq, Pfu, and KOD DNA polymerases. TNA1_pol averaged one incorrect bp every 4.45 kilobases (kb), and had a processivity of 150 nucleotides (nt) and an extension rate of 60 bases/s. Thus, TNA1_pol has a much faster elongation rate than Pfu DNA polymerase with 7-fold higher fidelity than that of rTaq.