Ionic liquid (IL), asymmetric chemical consist of bulky cations and tiny-mobile anions, has been known as promising DNA extraction, separation and preservation agent due to its strong interaction with DNA. However, the interaction underlying DNA-IL complex forming mechanism remains to be elucidated. Herein, we employed three types of ILs (EMIM-Cl, BMIM-Cl, and OMIM-Cl) to investigate the changes of DNA morphology upon the alkyl chain length of ILs by using solid-state nanopore technology combining with atomic force microscopy (AFM). The results of AFM show the different forms of DNA, including aggregate, stretching, and bundling shapes in terms of EMIM-Cl, BMIM-Cl, and OMIM-Cl, respectively, assuming that the shape of DNA-IL complexes is responding to the alkyl chain length of ILs. In DNA translocation experiment. From the alteration of blockade current signals during the DNA pass through the nanopore, we estimate that the shapes of DNA are changed due to the treatment with BMIM-Cl, and OMIM-Cl, which not only increased the blockade current signals about 2-4 times in the case of OMIM, but also decrease the event showing translocation of DNA folding, implying that the alkyl chain affect to DNA stretching and bundling. The results indicate the length of hydrophobic alkyl group of IL plays an important role in determination of DNA morphology, providing their further application in nanopore technique for slowing DNA translocation speed toward discovering protein-DNA interaction or DNA sequencing.

Due to the globalization of food supply have been growing, there have been a great demands for food safety and quality assuarance for on-site detection. On-site detetction isuue is the process should be fast, simple, user-friendly and require minimal equipments. Herein, we developed a Radial chromatography (RC) biosensor integrated with the immuno-gold nanoparticles-coated magnetic nanoparticle (AuNPs@Fe3O4) for specific separation and detection of the target bacteria, E. coli O157:H7, in sample. The immuno-AuNPs@Fe3O4 specifically binds to E.coli O157:H7 creating AuNP@Fe3O4-E.coli complexes and captured bacteria were concentrated by magnet. The complex can be identified with inner ring derived from the difference of mobility of free AuNPs@Fe3O4 on the RC sensor. Our results show that AuNPs@Fe3O4 based RC sensor has high sensitivity to the target bacteria over non-target bacteria with a detection limit of 103 CFU/ml. Our system offers a rapid and sensitive means of detecting E.coli O157:H7 with naked eyes, which can be applied to the field diagnosis.

Rapid, simple, and sensitive detection of pathogen bacteria is a highly topical research area due to increasingly concerning of food safety and public health. Surface-enhanced Raman spectroscopy (SERS) is a promising and attractive technique offering fast, sensitive, comparatively low-cost, and in-suit detection of pathogenic bacteria. However, this technique requires the preparation step for reducing the noise derived from heterogeneous matrixes of food sample. Immunomagnetic separation (IMS) is widely used technique enabling separation and concentration of the target analyte. It can be used not only laboratory scale but also field diagnosis easily. Here, we synthesized gold-shelled starch magnetic microparticles (GS@SMMPs) for effective separation and concentration of Escherichia coli O157:H7, which were subsequently subjected to SERS integrated with gold-coated 3D-well substrate for bacterial detection in aqueous solution. GS@SMMPs were labelled by Anti-E. coli O157 monoclonal antibody through gold binding protein and staphylococcal protein G (GBP-SPG) fusion protein. In IMS experiment, the immuno-GS@SMMPs showed high capture efficiency over 90% to E. coli O157:H7, which resulted in 10 times decrease in detection limit in PCR assay. Through SERS assay, E. coli O157:H7 concentrated by immuno-GS@SMMPs were successfully detected even at an extremely low concentration of 101 CFU/ml the subjected to SERS. Moreover, by using sandwich method using SERS reporter consisting of GBP-SPG, we found that E. coli O157:H7 were able to be detected by SERS quantitatively through measuring the SERS intensity of GBP-SPG. This novel strategy combining SERS and IMS could be meaningful for extending the application in SERS for in-suit sensitive detection of pathogenic bacteria.

There have been great efforts to develop a rapid and sensitive detection method to monitor the presence of pathogenic bacteria in food. While a number of methods have been reported for bacterial detection with a detection limit to a single digit, most of them are suitable only for the bacteria in pure culture or buffered solution. On the other hand, foods are composed of highly complicated matrices containing carbohydrate, fat, protein, fibers, and many other components whose composition varies from one food to the other. Furthermore, many components in food interfere with the downstream detection process, which significantly affect the sensitivity and selectivity of the detection. Therefore, isolating and concentrating the target pathogenic bacteria from food matrices are of importance to enhance the detection power of the system. The present review provides an introduction to the representative sample preparation strategies to isolate target pathogenic bacteria from food sample. We further describe the nucleic acidbased detection methods, such as PCR, real-time PCR, NASBA, RCA, LCR, and LAMP. Nucleic acid-based methods are by far the most sensitive and effective for the detection of a low number of target pathogens whose performance is greatly improved by combining with the sample preparation methods.

Ionic liquids (ILs) are organic salts with low melting point by asymmetric ionic strength between cation and anion. They have been known as promising DNA extraction, separation and preservation agent due to their hydrophilic, hydrophobic interaction with DNA. However, few studies have been performed about how DNA-ILs complexes form and their mechanism. Herein, we present three types of ionic liquids (EMIM-Cl, BMIM-Cl, and OMIM-Cl) change the DNA structure depend on alkyl chain length of ionic liquids. Structural changes of DNA by ionic liquids are observed by Atomic force microscopy, gel electrophoresis, zeta potential and solid-state nanopore technology. The results of AFM show the different structures of DNA, including aggregate, stretching, and bundling shapes in terms of EMIM-Cl, BMIM-Cl, and OMIM-Cl respectively. In DNA translocation experiment, DNA/EMIM-Cl show rare translocation signal due to aggregated structure by neutralized surface charge. DNA/BMIM-Cl and DNA/OMIM-Cl show slowing down the translocation speed due to changes of DNA net charge and structure. Especially, OMIM-Cl make slowing down the DNA translocation speed about 102~104 times compared to translocation speed of bare DNA by unzipping the bundling shape of complex. In conclusion, the morphology of DNA could be modified by the incorporation with different alkyl chain length of ILs, providing their further application in nanopore technique for slowing DNA sequencing or understanding protein-DNA interaction.

Starch is an abundant, renewable, and low cost material that has been extensively studied for its role in crystallization. The aim of this study is to develop a convenient and green approach to synthesize starch nanoparticles (StNPs). Short glucan chains were successfully prepared by using pullulanase that could debranch the amylopectin obtained from waxy maize starch. StNPs were prepared via the self-association of short glucan chains, of which the crystallinity structure changed from A-type (native starch) to B-type (starch nanoparticles) through the enzymatic hydrolysis and reassembly process at 4°C. Scanning electron microscopy (SEM), X-ray diffraction (XRD), dynamic light scattering (DLS) and differential scanning calorimetry (DSC) were used to characterize the morphology and crystalline structure of StNPs. The results showed that the diameter of StNPs ranged from 300 nm to 1.5 μm, depending on the initial concentration of short glucan chains and self-assembly time. The developed approach could produce well-defined and uniform starch nanoparticles that could readily be employed to encapsulate various functional guest molecules in biocompatible starch based nanoparticles in food industry.

Amylose is carbohydrate polymer defined as a linear natural polysaccharide composed of α(1→4) bound glucose units. Due to its abundance, renewable nature, low cost, and biodegradability, this polymer is regarded as a promising green material for producing crystals and particles of different sizes ranging from the nanometer scale to the micrometer scale. Herein, short amylose chains and dextran-coated iron oxide magnetic nanoparticles (Dex@MNPs) were introduced to fabricate individual superparamagnetic amylose microparticles (SAMPs), which have a well-defined spherical shape and a uniform size of about 1 μm. We found that the aggregation of SAMPs can be mediated by the introduced Dex@MNPs in a concentration-dependent manner, indicating that Dex@MNPs, as the seed crystals, play an important role in self-assembly of SAMPs. By using streptococcal protein G tagged with maltose binding protein (MBP-SPG), specific antibody against Escherichia coli O157:H7 was successfully immobilized on the surface of SAMPs. The Ab-functionalized SAMPs showed a high capture efficiency (>90%) comparable to the commercial immunomagnetic microparticles regardless of suspending agents (1X PBS and milk). Moreover, SAMPs exhibited excellent recyclability, in which the Ab immobilized on the surface of SAMPs can be refreshed by using the maltose elution buffer along with the unchanged capture efficiency. In addition, SAMPs were assembled into the linear rod-shape microstructure by the introduced magnetic field during the amylose-mediated precipitation process. The convenient self-assembly of SAMPs with the well-defined size and shape, biocompatibility, tolerance to environmental variances, high magnetic response behavior, and excellent recyclability in the functionalization make these magnetic microparticles promising for many potential applications such as bio-sensing, labeling, and smart delivery of active compounds.

Ionic liquids (ILs) have been used in DNA extraction/separation, DNA preservation and PCR based on their characteristic affinity to DNA. However, few studies have been performed about how DNA-IL complex forms and its mechanism which would be essential to understand the role of ILs over the range of applications. Herein, we present that the differences in the structure of the DNA- IL complex are associated with the alkyl chain length of IL. The assumption was evidenced by Atomic force microscopy, DNA specific dye staining, gel-electrophoresis and real-time electrical measurement. We observed unique electrical signals with altered duration time and amplitude when DNA- ILs complexes pass through solid-state nanopore. We examined three types of ILs (EMIM-Cl, BMIM-Cl, and OMIM-Cl) for their characteristics to form DNA-ILs complexes. The results indicated that the length of hydrophobic alkyl group in respective ILs determines the form of DNA-IL complex. In conclusion, the morphology of DNA could be modified by the incorporation with different alkyl chain length of ILs, providing their further application in biosensor such as nanopore technique for DNA sequencing or understanding protein-DNA interaction.

Various methods for the detection of E. coli O157:H7 in food have been developed in the past decades. However, current detection methods require specialized instruments and lengthy preparation time. In an effort to achieve a rapid and sensitive detection, we developed a radial chromatography (RC) biosensor integrated with gold nanoparticle (AuNP) conjugated with antibody for the detection of E. coli O157:H7. The immuno-AuNP binds to E. coli O157:H7 creating AuNP-E. coli O157:H7 complexes by specific antigen-antibody interaction. The AuNP-E. coli O157:H7 complexes can be identified clearly from free AuNP by RC based on their mobility on porous matrix. Thus, the AuNP complexed with target bacteria, E. coli O157:H7 could be discriminated from free AuNP by radial chromatography. The results showed that the developed RC biosensor is highly selective to E. coli O157:H7 over non-target bacteria, including Bacillus cereus, Staphylococcus aureus and Vibrio cholerae with a detection limit of 105 CFU/ml. When combined with a pre-concentration step using immunomagnetic beads, we could further enhance the detection limit down to 103 CFU/ml. In this study, we developed a novel method that is rapid, sensitive and applicable for qualitative and quantitative detection of E. coli O157:H7. The detection procedure is simple and the results can be easily determined by naked eyes, suggesting that this system is practical and can be applied to the field diagnosis in food industry for the detection of pathogenic bacteria.

Heavy metals are typical contaminant in water and need to be removed because they are non-biodegradable and can accumulate in human body. To remove metal and other contaminants from water, chemical absorbents are widely used due to their low cost. Herein, hybrid materials comprised of amylose and single wall carbon-nanotube (swCNT) was developed as the absorbent for water purification. A high adsorption properties of carbon-nanotubes have been utilized in designing effective absorbent but its poor dispersity in water is a limiting factor for practical use. Single wall carbon-nanotube was hybridized with amylose chain produced by enzyme reaction of amylosucrase and their tendency to self-assemble in aqueous environment. The amylose-swCNT microparticles were characterized by FE-SEM and turned out to be spherical structure with CNTs embedded throughout the amylose matrix. The ability of amylose-CNT microparticles to remove copper was examined. Concentration of copper was decreased after reaction with amylose-swCNT microparticles. Although amount of decreasing copper was less than swCNT on same total mass, Adsorption efficiency of amylose-swCNT microparticles was good because their swCNT content was only 10% of total mass.

Aquaporin Z and bacteriorhodopsin known as membrane protein transport water molecules and protons, respectively, in and out of the cell in highly selective manner. The selective character of these biomolecules can be applied to water filtration or other industry. However, transmembrane protein like as aquaporin Z and bacteriorhodopsin has many problems for using associated with expression, purification, and activation of protein character. To solve these problems, we developed purification system and characterization method for transmembrane protein. Green Fluorescence Protein (GFP)-fused aquaporin Z were expressed in inducible expression vector system, and purified by Ni-NTA affinity chromatography. GFP-fused aquaporin Z proteins were incorporated into the liposome for optical observation of water flux, and tested in hypertonic external condition by osmotic pressure to check water molecule selective transport. Bacteriorhodopsin was acquired by same method for aquaporin Z purification, but need to activation process by retinal cofactor because Escherichia coli has not retinal synthesis system. According to the absorbance spectra analysis, we confirmed that the purified bacteriorhodopsin was activated by retinal. This study will help to understanding of aquaporin Z and bacteriorhodopsin character and industrial application of transmembrane protein.

We developed a Amylose magnetic beads (AMBs) based detection system for high efficient separation, concentration and detection of E. coli O157:H7 in real sample. AMBs were synthesized by amylosucrase from Deinococcus geothermalis (DgAS) with iron-oxide nanoparticle (NP). The design of amylose magnetic beads (AMBs) have studied by an enzymatic synthesis with optimized reaction condition such as substrate, sucrose, and iron-oxide NP. AMBs have specific feature. AMBs decorated with functional fusion protein, which consists in a maltose binding protein (MBP) and a streptococcal protein G (SPG). Amylose chains has maltose, thus MBP-SPG binds to the AMB. In addition, SPG specifically binds to the Fc part of antibody. That was used as a linker to immobilize antibody to the surface of AMBs. The resulting AMBs were efficiently separated and concentrated target bacteria, E. coli O157:H7. Concentrated sample is qualitatively analyzed by PCR. Our studies demonstrated that AMB-based PCR significantly reduced the limitation of detection as low as 10 1 CFU/mL, compared to that of conventional PCR. The principle of this system can be served as a high efficiency for detection method of any pathogenic bacteria. In addition, AMBs and MBP-SPG cross-linker protein developed in this study is expected to be applicable to the portable food based biological processing monitoring system.

In order to promote the prevention of microbial and enzymatic spoilage and to retain the freshness, sweet persimmons harvested in Gyeongsangnam-do were treated with grapefruit seed extract(GFSE)-CaCO₃ mixture and stored in the proper packaging conditions. A low concentration of GFSE showed effective growth inhibition of plant pathological bacteria and fungi, Enterobacter pyrinus and Fusarium sp., which were involved in the decay of fruits and vegetables. GFSE was stable to heat treatment; its antimicrobial activity was not changed by heat treatment upto 100℃. However, when the temperature was raised to 120℃, about 90% of total activity was retained within 30 min. GFSE was also highly stable to broad pH changes; its activity was not changed in the range of pH 2.0 to pH 12.0. The physiological function of cell membrane in the spores of Bacillus cereus and the hyphae of Fusarium sp. was destroyed by treating with GFSE. It was observed that treating sweet persimmons with GFSE mixture and storing them in strech-wrapped packages could prolong the freshness of sweet persimmons and reduce quality deterioration.

This technical paper deals the practical transformation algorithms between several lunar reference frames which will be used for Korea pathfinder lunar orbiter (KPLO) flight operation. Despite of various lunar reference frame definitions already exist, use of a common transformation algorithm while establishing lunar reference frame is very important for all members related to KPLO mission. This is because use of slight different parameters during frame transformation may result significant misleading while reprocessing data based on KPLO flight dynamics. Therefore, details of practical transformation algorithms for the KPLO mission specific lunar reference frames is presented with step by step implementation procedures. Examples of transformation results are also presented to support KPLO flight dynamics data user community which is expected to give practical guidelines while post processing the data as their needs. With this technical paper, common understandings of reference frames that will be used throughout not only the KPLO flight operation but also science data reprocessing can be established. It is expected to eliminate, or at least minimize, unnecessary confusion among all of the KPLO mission members including: Korea Aerospace Research Institute (KARI), National Aeronautics and Space Administration (NASA) as well as other organizations participating in KPLO payload development and operation, or further lunar science community world-wide who are interested in KPLO science data post processing.

The ground tracking support is a critical factor for the navigation performance of spacecraft orbiting around the Moon. Because of the tracking limit of antennas, only a small number of facilities can support lunar missions. Therefore, case studies for various ground tracking support conditions are needed for lunar missions on the stage of preliminary mission analysis. This study analyzes the ground supporting condition effect on orbit determination (OD) of Korea Pathfinder Lunar Orbiter (KPLO) in the lunar orbit. For the assumption of ground support conditions, daily tracking frequency, cut-off angle for low elevation, tracking measurement accuracy, and tracking failure situations were considered. Two antennas of deep space network (DSN) and Korea Deep Space Antenna (KDSA) are utilized for various tracking conditions configuration. For the investigation of the daily tracking frequency effect, three cases (full support, DSN 4 pass/day and KDSA 4 pass/day, and DSN 2 pass/day and KDSA 2 pass/day) are prepared. For the elevation cut-off angle effect, two situations, which are 5 deg and 10 deg, are assumed. Three cases (0%, 30%, and 50% of degradation) were considered for the tracking measurement accuracy effect. Three cases such as no missing, 1-day KDSA missing, and 2-day KDSA missing are assumed for tracking failure effect. For OD, a sequential estimation algorithm was used, and for the OD performance evaluation, position uncertainty, position differences between true and estimated orbits, and orbit overlap precision according to various ground supporting conditions were investigated. Orbit prediction accuracy variations due to ground tracking conditions were also demonstrated. This study provides a guideline for selecting ground tracking support levels and preparing a backup plan for the KPLO lunar mission phase.

In this study, the observational arc-length effect on orbit determination (OD) for the Korea Pathfinder Lunar Orbiter (KPLO) in the Earth-Moon Transfer phase was investigated. For the OD, we employed a sequential estimation using the extended Kalman filter and a fixed-point smoother. The mission periods, comprised between the perigee maneuvers (PM) and the lunar orbit insertion (LOI) maneuver in a 3.5 phasing loop of the KPLO, was the primary target. The total period was divided into three phases: launch–PM1, PM1–PM3, and PM3–LOI. The Doppler and range data obtained from three tracking stations [included in the deep space network (DSN) and Korea Deep Space Antenna (KDSA)] were utilized for the OD. Six arc-length cases (24 hrs, 48 hrs, 60 hrs, 3 days, 4 days, and 5 days) were considered for the arc-length effect investigation. In order to evaluate the OD accuracy, we analyzed the position uncertainties, the precision of orbit overlaps, and the position differences between true and estimated trajectories. The maximum performance of 3-day OD approach was observed in the case of stable flight dynamics operations and robust navigation capability. This study provides a guideline for the flight dynamics operations of the KPLO in the trans-lunar phase.

This paper analyzes delta-Vs to maintain an extremely low altitude on the Moon and investigates the possibilities of performing a CubeSat mission. To formulate the station-keeping (SK) problem at an extremely low altitude, current work has utilized real-flight performance proven software, the Systems Tool Kit Astrogator by Analytical Graphics Inc. With a high-fidelity force model, properties of SK maneuver delta-Vs to maintain an extremely low altitude are successfully derived with respect to different sets of reference orbits; of different altitudes as well as deadband limits. The effect of the degree and order selection of lunar gravitational harmonics on the overall SK maneuver strategy is also analyzed. Based on the derived SK maneuver delta-V costs, the possibilities of performing a CubeSat mission are analyzed with the expected mission lifetime by applying the current flight-proven miniaturized propulsion system performances. Moreover, the lunar surface coverage as well as the orbital characteristics of a candidate reference orbit are discussed. As a result, it is concluded that an approximately 15-kg class CubeSat could maintain an orbit (30–50 km reference altitude having ±10 km deadband limits) around the Moon for 1–6 months and provide almost full coverage of the lunar surface.

확대머리 SD600 고강도 인장철근으로 단부 정착된 SFRC 깊은보의 전단성능을 평가하기 위해 전단 실험을 수행하였다. 실험 변수는 주인장 철근의 단부 정착방법(확대머리 철근, 일자형 철근), 단부 정착길이, 전단보강근 유무 등이다. 전단경간비는 1을 가지는 실험체에 대한 전단실험결과, 모든 실험체는 초기 휨 균열이 발생한 후 경사균열이 진행되면서 최종적으로 압축전단파괴되었다. 확대머리 철근으로 기계적 정착된 실험체들이 일자형 철근 정착에 비하여 5.6∼22.4% 더 큰 전단강도를 나타내었다. 확대머리 철근으로 기계적 정착된 실험체들에 대하여 최대하중의 75%까지는 지압응력이 전체 정착응력의 0.9~17.2%에 도달하였으나, 최대하중 시점에서 지압응력이 전체 정착응력의 22.4%~46%에 도달하여 큰 응력 부담률을 나타내었다. 이를 통하여 확대머리 지압응력에 의한 정착응력 증가가 전단강도에 큰 영향을 미침을 알 수 있다. 실험 전단강도가 실용식에 의한 전단강도의 2.68~4.65 배로 평가되어, 실용식이 전단내력을 안전측으로 평가하였다.

In this study, we investigated the in vitro anti-biofilm activities of plant extracts of chives (Allium tuberosum), garlic (Allium sativum), and radish (Raphanus sativus L.) against environment harmful bacteria (gram-positive Staphylococcus aureus and, gram-negative Salmonella typhimurium and Escherichia coli O157:H7). In the paper disc assay, garlic extracts exhibited the highest anti-biofilm activity. The Minimal Inhibitory Concentration (MIC) of all plant extracts was generally higher for gram-negative bacteria than it was for gram-positive bacteria. Gram-negative bacteria were more resistant to plant extracts. The tetrazolium dye (XTT) assay revealed that, each plant extract exhibited a different anti-biofilm activity at the MIC value depending on the pathogen involved. Among the plant extracts tested, garlic extracts (fresh juice and powder) effectively reduced the metabolic activity of the cells of food-poisoning bacteria in biofilms. These anti-biofilm activities were consistent with the results obtained through light microscopic observation. Though the garlic extract reduced biofilm formation for all pathogens tested, to elucidate whether this reduction was due to antimicrobial effects or anti-biofilm effects, we counted the colony forming units of pathogens in the presence of the garlic extract and a control antimicrobial drug. The garlic extract inhibited the E. coli O157:H7 biofilm effectively compared to the control antimicrobial drug ciprofloxacin; however, it did not inhibit S. aureus biofilm significantly compared to ciprofloxacin. In conclusion, garlic extracts could be used as natural food preservatives to prevent the growth of foodborne pathogens and elongater the shelf life of processed foods.

In this study, orbit determination (OD) simulation for the Korea Pathfinder Lunar Orbiter (KPLO) was accomplished for investigation of the observational arc-length effect using a sequential estimation algorithm. A lunar polar orbit located at 100 km altitude and 90° inclination was mainly considered for the KPLO mission operation phase. For measurement simulation and OD for KPLO, the Analytical Graphics Inc. Systems Tool Kit 11 and Orbit Determination Tool Kit 6 software were utilized. Three deep-space ground stations, including two deep space network (DSN) antennas and the Korea Deep Space Antenna, were configured for the OD simulation. To investigate the arc-length effect on OD, 60-hr, 48-hr, 24-hr, and 12-hr tracking data were prepared. Position uncertainty by error covariance and orbit overlap precision were used for OD performance evaluation. Additionally, orbit prediction (OP) accuracy was also assessed by the position difference between the estimated and true orbits. Finally, we concluded that the 48-hr-based OD strategy is suitable for effective flight dynamics operation of KPLO. This work suggests a useful guideline for the OD strategy of KPLO mission planning and operation during the nominal lunar orbits phase.