• Proceedings of the Korean Environmental Sciences Society Conference
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• 1998.10a
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• pp.2-4
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• 1998

Proliferation of Nocardia amarae cells in activated sludge has often been associated with the generation of nuisance foams. Despite intense research activities in recent years to examine the causes and control of Nocardia foaming in activated sludge, the foaming continued to persist throughout the activated sludge treatment plants in United States. In addition to causing various operational problems to treatment processes, the presence of Nocardia may have secondary effects on the fate of heavy metals that are not well known. For example, for treatment plants facing more stringent metal removal requirements, potential metal removal by Nocardia cells in foaming activated sludge would be a welcome secondary effect. In contrast, with new viosolid disposal regulations in place (Code o( Federal Regulation No. 503), higher concentration of metals in biosolids from foaming activated sludge could create management problems. The goal of this research was to investigate the metal sorption property of Nocardia amarae cells grown in batch reactors and in chemostat reactors. Specific surface area and metal sorption characteristics of N. amarae cells harvested at various growth stages were compared. Three metals examined in this study were copper, cadmium and nickel. Nocardia amarae strain (SRWTP isolate) used in this study was obtained from the University of California at Berkeley. The pure culture was grown in 4L batch reactor containing mineral salt medium with sodium acetate as the sole carbon source. In order to quantify the sorption of heavy metal ions to N amarae cell surfaces, cells from the batch reactor were harvested, washed, and suspended in 30mL centrifuge tubes. Metal sorption studies were conducted at pH 7.0 and ionlc strength of 10-2M. The sorption Isotherm showed that the cells harvested from the stationary and endogenous growth phase exhibited significantly higher metal sorption capacity than the cells from the exponential phase. The sequence of preferential uptake of metals by N. amarae cells was Cu>Cd>Ni. The specific surFace area of Nocardia cells was determined by a dye adsorption method. N.amarae cells growing at ewponential phase had significantly less specific surface area than that of stationary phase, indicating that the lower metal sorption capacity of Nocardia cells growing at exponential phase may be due to the lower specific surface area. The growth conditions of Nocardia cells in continuous culture affect their cell surface properties, thereby governing the adsorption capacity of heavy metal. The comparison of dye sorption isotherms for Nocardia cells growing at various growth rates revealed that the cell surface area increased with increasing sludge age, indicating that the cell surface area is highly dependent on the steady-state growth rate. The highest specific surface area of 199m21g was obtained from N.amarae cell harvested at 0.33 day-1 of growth rate. This result suggests that growth condition not only alters the structure of Nocardia cell wall but also affects the surface area, thus yielding more binding sites of metal removal. After reaching the steady-state condition at dilution rate, metal adsorption isotherms were used to determine the equilibrium distributions of metals between aqueous and Nocardia cell surfaces. The metal sorption capacity of Nocardia biomass harvested from 0.33 day-1 of growth rate was significantly higher than that of cells harvested from 0.5- and 1-day-1 operation, indicatng that N.amarae cells with a lower growth rate have higher sorpion capacity. This result was in close agreement with the trend observed from the batch study. To evaluate the effect of Nocardia cells on the metal binding capacity of activated sludge, specific surface area and metal sorption capacity of the mixture of Nocardia pure cultures and activated sludge biomass were determined by a series of batch experiments. The higher levels of Nocardia cells in the Nocardia-activated sludge samples resulted in the higher specific surface area, explaining the higher metal sorption sites by the mixed luquor samples containing greater amounts on Nocardia cells. The effect of Nocardia cells on the metal sorption capacity of activated sludge was evaluated by spiking an activated sludge sample with various amounts of pre culture Nocardia cells. The results of the Langmuir isotherm model fitted to the metal sorption by various mixtures of Nocardia and activated sludge indicated that the mixture containing higher Nocardia levels had higher metal adsorption capacity than the mixture containing lower Nocardia levels. At Nocardia levels above 100mg/g VSS, the metal sorption capacity of activate sludge increased proportionally with the amount of Noeardia cells present in the mixed liquor, indicating that the presence of Nocardia may increase the viosorption capacity of activated sludge.

• Journal of Korean Society of Environmental Engineers
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• v.31 no.11
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• pp.965-974
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• 2009

This study evaluated the degradation efficiency of malodorous sulfurized-organic compounds by utilizing N- and Sdoped titanium dioxide under visible-light irradiation, and examined the catalyst deactivation and regeneration. Catalyst surface was characterized by employing Fourier-Transform-Infrared-Red (FTIR) spectra. The visible-light-driven photocatalysis techniques were able to efficiently degrade low-level dimethyl sulfide (DMS) and dimethyl disulfide (DMDS) with degradation efficiencies exceeding 97%, whereas they were not effective regarding the removal of high-level DMS and DMDS, with degradation efficiencies of 84 and 23% within 5 hrs of photocatalytic processes. As compared with DMS, DMDS which containes one more sulfur element revealed quick catalyst deactivation. Catalyst deactivation was confirmed by the equality between input and output concentrations of DMD or DMDS, the obsevation of no $CO_2$ generation during a photocatalytic process, and the FTIR spectrum peaks related with sulfur ion compounds, which are major byproducts formed on catalyst surfaces. The mineralization efficiency of DMS at 8 ppm, which was a peak value during a photocatalytic process, was calculated as 144%, exceeding 100%. The catalyst regenerated by high-temperature calcination exhibited higher catalyst recovery efficiency (53 and 58% for DMDS and DMS, respectively) as compared with dry-air and humid-air regeneration processes. However, even the calcined method was unable to totally regenerate deactivated catalysts.

• 한국균학회소식:학술대회논문집
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• 2016.05a
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• pp.19-20
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• 2016

Sesquiterpenoids are defined as $C_{15}$ compounds derived from farnesyl pyrophosphate (FPP), and their complex structures are found in the tissue of many diverse plants (Degenhardt et al. 2009). FPP's long chain length and additional double bond enables its conversion to a huge range of mono-, di-, and tri-cyclic structures. A number of cyclic sesquiterpenes with alcohol, aldehyde, and ketone derivatives have key biological and medicinal properties (Fraga 1999). Fungi, such as the wood-rotting Polyporus brumalis, are excellent sources of pharmaceutically interesting natural products such as sesquiterpenoids. In this study, we investigated the biosynthesis of P. brumalis sesquiterpenoids on modified medium. Fungal suspensions of 11 white rot species were inoculated in modified medium containing $C_6H_{12}O_6$, $C_4H_{12}N_2O_6$, $KH_2PO_4$, $MgSO_4$, and $CaCl_2$ for 20 days. Cultivation was stopped by solvent extraction via separation of the mycelium. The metabolites were identified as follows: propionic acid (1), mevalonic acid lactone (2), ${\beta}$-eudesmane (3), and ${\beta}$-eudesmol (4), respectively (Figure 1). The main peaks of ${\beta}$-eudesmane and ${\beta}$-eudesmol, which were indicative of sesquiterpene structures, were consistently detected for 5, 7, 12, and 15 days These results demonstrated the existence of terpene metabolism in the mycelium of P. brumalis. Polyporus spp. are known to generate flavor components such as methyl 2,4-dihydroxy-3,6-dimethyl benzoate; 2-hydroxy-4-methoxy-6-methyl benzoic acid; 3-hydroxy-5-methyl phenol; and 3-methoxy-2,5-dimethyl phenol in submerged cultures (Hoffmann and Esser 1978). Drimanes of sesquiterpenes were reported as metabolites from P. arcularius and shown to exhibit antimicrobial activity against Gram-positive bacteria such as Staphylococcus aureus (Fleck et al. 1996). The main metabolites of P. brumalis, ${\beta}$-Eudesmol and ${\beta}$-eudesmane, were categorized as eudesmane-type sesquiterpene structures. The eudesmane skeleton could be biosynthesized from FPP-derived IPP, and approximately 1,000 structures have been identified in plants as essential oils. The biosynthesis of eudesmol from P. brumalis may thus be an important tool for the production of useful natural compounds as presumed from its identified potent bioactivity in plants. Essential oils comprising eudesmane-type sesquiterpenoids have been previously and extensively researched (Wu et al. 2006). ${\beta}$-Eudesmol is a well-known and important eudesmane alcohol with an anticholinergic effect in the vascular endothelium (Tsuneki et al. 2005). Additionally, recent studies demonstrated that ${\beta}$-eudesmol acts as a channel blocker for nicotinic acetylcholine receptors at the neuromuscular junction, and it can inhibit angiogenesis in vitro and in vivo by blocking the mitogen-activated protein kinase (MAPK) signaling pathway (Seo et al. 2011). Variation of nutrients was conducted to determine an optimum condition for the biosynthesis of sesquiterpenes by P. brumalis. Genes encoding terpene synthases, which are crucial to the terpene synthesis pathway, generally respond to environmental factors such as pH, temperature, and available nutrients (Hoffmeister and Keller 2007, Yu and Keller 2005). Calvo et al. described the effect of major nutrients, carbon and nitrogen, on the synthesis of secondary metabolites (Calvo et al. 2002). P. brumalis did not prefer to synthesize sesquiterpenes under all growth conditions. Results of differences in metabolites observed in P. brumalis grown in PDB and modified medium highlighted the potential effect inorganic sources such as $C_4H_{12}N_2O_6$, $KH_2PO_4$, $MgSO_4$, and $CaCl_2$ on sesquiterpene synthesis. ${\beta}$-eudesmol was apparent during cultivation except for when P. brumalis was grown on $MgSO_4$-free medium. These results demonstrated that $MgSO_4$ can specifically control the biosynthesis of ${\beta}$-eudesmol. Magnesium has been reported as a cofactor that binds to sesquiterpene synthase (Agger et al. 2008). Specifically, the $Mg^{2+}$ ions bind to two conserved metal-binding motifs. These metal ions complex to the substrate pyrophosphate, thereby promoting the ionization of the leaving groups of FPP and resulting in the generation of a highly reactive allylic cation. Effect of magnesium source on the sesquiterpene biosynthesis was also identified via analysis of the concentration of total carbohydrates. Our current study offered further insight that fungal sesquiterpene biosynthesis can be controlled by nutrients. To profile the metabolites of P. brumalis, the cultures were extracted based on the growth curve. Despite metabolites produced during mycelia growth, there was difficulty in detecting significant changes in metabolite production, especially those at low concentrations. These compounds may be of interest in understanding their synthetic mechanisms in P. brumalis. The synthesis of terpene compounds began during the growth phase at day 9. Sesquiterpene synthesis occurred after growth was complete. At day 9, drimenol, farnesol, and mevalonic lactone (or mevalonic acid lactone) were identified. Mevalonic acid lactone is the precursor of the mevalonic pathway, and particularly, it is a precursor for a number of biologically important lipids, including cholesterol hormones (Buckley et al. 2002). Farnesol is the precursor of sesquiterpenoids. Drimenol compounds, bi-cyclic-sesquiterpene alcohols, can be synthesized from trans-trans farnesol via cyclization and rearrangement (Polovinka et al. 1994). They have also been identified in the basidiomycota Lentinus lepideus as secondary metabolites. After 12 days in the growth phase, ${\beta}$-elemene caryophyllene, ${\delta}$-cadiene, and eudesmane were detected with ${\beta}$-eudesmol. The data showed the synthesis of sesquiterpene hydrocarbons with bi-cyclic structures. These compounds can be synthesized from FPP by cyclization. Cyclic terpenoids are synthesized through the formation of a carbon skeleton from linear precursors by terpene cyclase, which is followed by chemical modification by oxidation, reduction, methylation, etc. Sesquiterpene cyclase is a key branch-point enzyme that catalyzes the complex intermolecular cyclization of the linear prenyl diphosphate into cyclic hydrocarbons (Toyomasu et al. 2007). After 20 days in stationary phase, the oxygenated structures eudesmol, elemol, and caryophyllene oxide were detected. Thus, after growth, sesquiterpenes were identified. Per these results, we showed that terpene metabolism in wood-rotting fungi occurs in the stationary phase. We also showed that such metabolism can be controlled by magnesium supplementation in the growth medium. In conclusion, we identified P. brumalis as a wood-rotting fungus that can produce sesquiterpenes. To mechanistically understand eudesmane-type sesquiterpene biosynthesis in P. brumalis, further research into the genes regulating the dynamics of such biosynthesis is warranted.

• Proceedings of the Korean Vacuum Society Conference
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• 2013.08a
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• pp.205-205
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• 2013

테라헤르쯔(terahertz: THz)파는 0.1~10 THz 의 범위로 적외선과 방송파 사이에 광대역 주파수 스펙트럼을 차지하고 있으며 직진성, 투과성, 그리고 낮은 에너지 (meV)를 가지고 있어 비 파괴적이고 무해한 장점을 지니고 있다. Ti:sapphire laser와 같은 femto-pulse source 등이 많은 발전이 되어 현재 많은 연구와 발전이 이루어지고 있다. femto-pulse source를 이용한 THz 응용에서는 높은 저항, 큰 전자 이동도, 그리고 아주 짧은 전하수명의 기판을 요구하는데 저온에서 성장한 (low-temperature grown : LT) GaAs는 격자 내에 Gallium 자리에 Arsenic이 치환 하면서 AsGa antisite가 발생하여 전하수명을 짧아지는 것을 응용하여 가장 많이 이용되고 있다. 현재 THz 응용분야에서 보다 작고 가격경쟁력이 있는 광통신을 이용한 THz photomixer등이 활발히 연구 하고 있다. 광섬유 내에서 손실과 분산이 최소값을 가지는 부분이 1.55 ${\mu}m$ 부근이고 In0.53Ga0.47As 기판을 이용하였을 때 여기에 완벽하게 만족하게 된다. 하지만 LT-InGaAs 의 경우 AsGa antisite로 인하여 carrier lifetime은 짧아지지만 높은 n-type 전하밀도를 가지게 된다. 이때 Be을 doping하여 전하밀도를 보상하여 높은 저항을 유지해야 하는데 Be의 활성화를 위해서는 열처리를 필요로 한다. 하지만 열처리를 하면 carrier lifetime이 길어지기 때문에 carrier lifetime과 저항을 적절히 조율해야 한다. 이는 물질자체의 특성이기 때문에 InGaAs는 GaAs보다 낮은 amplitude와 짧은 cut-off frequency를 가진다. 본 연구에서는 보다 높은 저항을 얻기 위하여 molecular beam epitaxy를 이용하여 semi-insulating InP:Fe 기판위에 격자 정합된 InGaAs:Be/InAlAs multi quantum well (MQW)를 온도별 ($250{\sim}400^{\circ}C$), 주기별 (50~150)로 성장을 하였고 이때 InGaAs layer의 Be doping level은 $2{\times}1018\;cm^{-3}$, Ex-situ annealing은 $550^{\circ}C$에서 10분으로 고정 하였다. THz 발생 실험에서는 InGaAs/InAlAs MQW은 4000 pA로 1,000 pA를 가지는 InGaAs epilayer보다 4배 높은 전류 신호를 얻을 수 있었고 모든 샘플이 2 THz에서 cut-off frequency를 가지고 있었다. THz 검출 실험에서는 LT-InGaAs:Be epilayer LT-InGaAs:Be/InAlAs, HT-InGaAs/InAlAs 샘플이 각각 180, 9000, 12000 pA의 전류신호를 가지고 있었고 모든 샘플이 2 THz에서 cut-off frequency를 가지고 있었다. HT-InGaAs/InAlAs MQW를 이용한 검출실험에서는 InGaAs layer가 defect free이지만 LT-InGaAs:Be/ InAlAs MQW 보다 높은 전류 신호를 얻을 수 있었다. 이는 InAlAs layer가 저항만 높이는 것뿐만 아니라 carrier trapping layer로써의 역할도 하는 것으로 사료된다.

• Clean Technology
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• v.25 no.3
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• pp.223-230
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• 2019

This research was performed to evaluate the characteristics of food waste from 5 areas in Gangwon Province, Korea and to predict the $CH_4$ gas production rate. Food wastes were sampled in July and September, 2017. The amount of methane gas generation was evaluated through the biochemical methane potential (BMP) test and a calculation method using chemical composition. Average bulk density and pH of the food wastes were in the range of $0.758{\sim}0.850g\;cm^{-3}$ and 4.29 ~ 4.75, respectively. By physical composition, vegetables were the highest with 56.43 ~ 72.81% with fruits recording 5.31 ~ 8.95%, cereals 1.60 ~ 18.73%, fish and meat 4.47 ~ 12.11%, and filtrate 1.76 ~ 3.64%. The average water content was 69.30 ~ 75.87%, and VS and ash content were 22.50 ~ 27.98% and 1.63 ~ 2.48%, respectively. In addition, $BOD_5$, $COD_{Cr}$, and $COD_{Mn}$ were in the ranges of $17,690.3{\sim}33,154.9mg\;L^{-1}$, $106,212.3{\sim}128,695.5mg\;L^{-1}$, and $51,266.1{\sim}63,426.3mg\;L^{-1}$, respectively. The NaCl content ranged from 0.81 to 1.17%. The results of elemental analysis showed that the contents of C, H, O, N, and S were 44.87 ~ 48.1%, 7.12 ~ 7.57%, 40.13 ~ 43.78%, 3.22 ~ 4.14%, and 0.00 ~ 0.02%, respectively. In a comparison of the methane production yield per VS mass of food waste, there was no significant difference between the cumulative amount (${0.303{\sim}0.354m_{CH4}}^3\;{kg_{VS}}^{-1}$) by the BMP test and the theoretical amount (${0.294{\sim}0.352m_{CH4}}^3\;{kg_{VS}}^{-1}$) calculated by chemical composition.

• Journal of the Korean Chemical Society
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• v.19 no.3
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• pp.186-192
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• 1975

In the course of anthracite briquet combustion, air draft is usually controlled to continue burning of definite amount of briquet in the conventional hollow clay cylinder with air inlet hole open for given time, so that a large amount of CO tends to be produced. Therefore, it is necessary to establish an improved combustion process to depress the yielding rate of CO and for this purpose, we performed a basic experiment in which combustion rate of CO was measured in the mixture of $N_2, O_2 $and CO gas with or without the presence charcoal at the various temperature. The observed results showed that the burning temperature of CO is about 680${\sim}700^{\circ}C$, further burning rate of it was increased with increasing the amount of draft. From these facts, longer combustion time and low CO generation are thus contradictory to each other and it has been long desired to make those two compatible somehow. The purpose of the present investigation lies in designing an effective new briquet stove to meet the above requirements. The essential feature of the new briquet stove consisted in the use of two hollow iron cylinders with different inside diameter. A cylindrical air jacket thus formed served as a path through which small amount of secondary air run from the bottom of the stove to the upper vent holes. Heat exchange occurred between the upgoing secondary air and the burning briquet, which lowered the combustion temperature of the briquet. The results observed were selfevident as anticipated. It was confirmed that the combustion time was increased tolerably due to the heat loss from the combustion zone and that CO in the flue gas was reoxidized at the upper portion of the stove by the upgoing hot secondary air. By this reoxidation reaction the concentration of CO in the flue gas was found to be about 1/20 of that in case the conventional clay cylinder was used as briquet jacket.

• Proceedings of the Korean Vacuum Society Conference
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• 2012.08a
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• pp.154-155
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• 2012

The promise of nano-crystalites (nc) as a technological material, for applications including display backplane, and solar cells, may ultimately depend on tailoring their behavior through doping and crystallinity. Impurities can strongly modify electronic and optical properties of bulk and nc semiconductors. Highly doped dopant also effect structural properties (both grain size, crystal fraction) of nc-Si thin film. As discussed in several literatures, P atoms or radicals have the tendency to reside on the surface of nc. The P-radical segregation on the nano-grain surfaces that called self-purification may reduce the possibility of new nucleation because of the five-coordination of P. In addition, the P doping levels of ${\sim}2{\times}10^{21}\;at/cm^3$ is the solubility limitation of P in Si; the solubility of nc thin film should be smaller. Therefore, the non-activated P tends to segregate on the grain boundaries and the surface of nc. These mechanisms could prevent new nucleation on the existing grain surface. Therefore, most researches shown that highly doped nc-thin film by using conventional PECVD deposition system tended to have low crystallinity, where the formation energy of nucleation should be higher than the nc surface in the intrinsic materials. If the deposition technology that can make highly doped and simultaneously highly crystallized nc at low temperature, it can lead processes of next generation flexible devices. Recently, we are developing a novel CVD technology with a neutral particle beam (NPB) source, named as neutral beam assisted CVD (NBaCVD), which controls the energy of incident neutral particles in the range of 1~300eV in order to enhance the atomic activation and crystalline of thin films at low temperatures. During the formation of the nc-/pm-Si thin films by the NBaCVD with various process conditions, NPB energy directly controlled by the reflector bias and effectively increased crystal fraction (~80%) by uniformly distributed nc grains with 3~10 nm size. In the case of phosphorous doped Si thin films, the doping efficiency also increased as increasing the reflector bias (i.e. increasing NPB energy). At 330V of reflector bias, activation energy of the doped nc-Si thin film reduced as low as 0.001 eV. This means dopants are fully occupied as substitutional site, even though the Si thin film has nano-sized grain structure. And activated dopant concentration is recorded as high as up to 1020 #/$cm^3$ at very low process temperature (＜ $80^{\circ}C$) process without any post annealing. Theoretical solubility for the higher dopant concentration in Si thin film for order of 1020 #/$cm^3$ can be done only high temperature process or post annealing over $650^{\circ}C$. In general, as decreasing the grain size, the dopant binding energy increases as ratio of 1 of diameter of grain and the dopant hardly be activated. The highly doped nc-Si thin film by low-temperature NBaCVD process had smaller average grain size under 10 nm (measured by GIWAXS, GISAXS and TEM analysis), but achieved very higher activation of phosphorous dopant; NB energy sufficiently transports its energy to doping and crystallization even though without supplying additional thermal energy. TEM image shows that incubation layer does not formed between nc-Si film and SiO2 under later and highly crystallized nc-Si film is constructed with uniformly distributed nano-grains in polymorphous tissues. The nucleation should be start at the first layer on the SiO2 later, but it hardly growth to be cone-shaped micro-size grains. The nc-grain evenly embedded pm-Si thin film can be formatted by competition of the nucleation and the crystal growing, which depend on the NPB energies. In the evaluation of the light soaking degradation of photoconductivity, while conventional intrinsic and n-type doped a-Si thin films appeared typical degradation of photoconductivity, all of the nc-Si thin films processed by the NBaCVD show only a few % of degradation of it. From FTIR and RAMAN spectra, the energetic hydrogen NB atoms passivate nano-grain boundaries during the NBaCVD process because of the high diffusivity and chemical potential of hydrogen atoms.

• Journal of the Microelectronics and Packaging Society
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• v.27 no.4
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• pp.83-89
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• 2020

For the past few decades, as part of efforts to protect the environment where fossil fuels, which have been a key energy resource for mankind, are becoming increasingly depleted and pollution due to industrial development, ecofriendly secondary batteries, hydrogen generating energy devices, energy storage systems, and many other new energy technologies are being developed. Among them, the lithium-ion battery (LIB) is considered to be a next-generation energy device suitable for application as a large-capacity battery and capable of industrial application due to its high energy density and long lifespan. However, considering the growing battery market such as eco-friendly electric vehicles and drones, it is expected that a large amount of battery waste will spill out from some point due to the end of life. In order to prepare for this situation, development of a process for recovering lithium and various valuable metals from waste batteries is required, and at the same time, a plan to recycle them is socially required. In this study, we introduce a nanoscale pattern transfer printing (NTP) process of Li2CO3, a representative anode material for lithium ion batteries, one of the strategic materials for recycling waste batteries. First, Li2CO3 powder was formed by pressing in a vacuum, and a 3-inch sputter target for very pure Li2CO3 thin film deposition was successfully produced through high-temperature sintering. The target was mounted on a sputtering device, and a well-ordered Li2CO3 line pattern with a width of 250 nm was successfully obtained on the Si substrate using the NTP process. In addition, based on the nTP method, the periodic Li2CO3 line patterns were formed on the surfaces of metal, glass, flexible polymer substrates, and even curved goggles. These results are expected to be applied to the thin films of various functional materials used in battery devices in the future, and is also expected to be particularly helpful in improving the performance of lithium-ion battery devices on various substrates.

• Journal of the korean academy of Pediatric Dentistry
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• v.26 no.2
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• pp.377-398
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• 1999

Calcium fluoride, created by topical fluoride application, is the reservoir for fluoride ion regulated by pH in the oral environment. Therefore, the amount and the maintenance of calcium fluoride have an important role in preventing dental caries. The aim of this study is to evaluate the effect of Nd:YAG laser irradiation on the generation of calcium fluoride and the acid resistance of tooth enamel. The bovine anterior permanent teeth were prepared (n=276), and divided into following groups : no treatment (control) fluoride application alone, laser irradiation alone, laser irradiation after fluoride application, and fluoride application after laser irradiation. And each group was subdivided based on the application time of 1.23% acidulated phosphate fluoride (APF) (5 min and 30 min) and the irradiation energy of Nd:YAG laser ($20J/cm^2\;and\;40J/cm^2$). In case of fluoride application, each group was divided according to KOH treatment. Twenty three treatment conditions were made for this experiment and twelve specimens were assigned to each treatment condition. In each treatment condition, ten specimens were used for chemical analysis and two specimens were observed under SEM. In groups without treating KOH, fluoride content and the depth of enamel dissolved were measured using enamel biopsy technique. In groups with treating KOH, the amount of calcium fluoride was measured by the treatment with 1 M KOH for 24 hours and enamel biopsy was performed after KOH treatment. The results were analyzed by the fluoride content and the depth of enamel dissolved by enamel biopsy, amount and thickness of calcium fluoride, and the surface structures of enamel. The results are as follows: 1. In groups without treating KOH, the fluoride content of removed enamel showed a positive relationship with the energy density of laser when the laser irradiated before fluoride application 2. In groups without treating KOH, the depth of enamel dissolved decreased more with the combined laser and fluoride treatment than with laser or fluoride treatment, except for the case of $20J/cm^2$ laser irradiation after 5 minute fluoride application (p<0.05). 3. The amount of calcium fluoride did not increased by laser treatment with no statistical significance(p>0.05). 4. The particle size of calcium fluoride increased in case of fluoride treatment after laser irradiation, compared with fluoride application alone. In case of laser treatment after fluoride application, the particle size of calcium fluoride increased and some of the particles fused as well. 5. There were no significant differences in the fluoride content of dissolved enamel between groups without treating KOH and control group, except for the case of laser irradiation after treatment of APF for 30 minutes (p>0.05). 6. In groups with treating KOH, depth of removed enamel in the groups of combined treatment with laser and fluoride was shallower than that in fluoride application groups (p<0.05). 7. In groups without treating KOH, the relationship between fluoride content and the depth of enamel dissolved showed more negative (Spearman correlation coefficient: -0.6281) than in groups with treating KOH (Spearman correlation coefficient: -0.3792). The greater amount of calcium fluoride could be found in case where there was a significant differences of the depth of enamel dissolved between groups with and without treating KOH. From these results, it can be concluded that laser seems to be a little effects on the amount of calcium fluoride formation, but has some effect on the lowering the solubility of calcium fluoride. As the combined treatment of laser and fluoride application showed more effective acid-resistant property, more extended recall period for fluoride application can be achieved with this combined treatment in the clinic.