To clarify the geographical distribution of scrub typhus vectors in Korea, the first survey of chigger mites was conducted from 2005 to 2007 by collecting wild small mammals twice a year (spring and autumn) at 24 sites nationwide. The two predominant mite species were Leptotrombidium pallidum (52.6%) and L. scutellare (27.1%). However, the proportions of L. scutellare in southern areas, including endemic provinces such as Chungcheongnam-Do, Jeollabuk-Do, Jeollanam-Do, and Gyeongsangnam-Do, were relatively higher than in central Korean regions where L. pallidum was predominant. In autumn, the ratio of L. scutellare increased to 42% while the ratio of L. pallidum decreased. The geographical distribution map of the L. scutellare chigger index was identical to the incidence pattern of scrub typhus, whereas those of overall mites and L. pallidum showed no relationship with case incidence patterns. Distribution mapping analysis shows an identical geographical distribution of L. scutellare and epidemic incidence of scrub typhus in South Korea. The second periodical survey was performed from 2011 to 2013. The result suggests that the distribution of L. scutellare has not been changed remarkably in comparison to the first survey.

Leptotrombidium pallidum is the major vector mites for Orientia tsutsugamushi, the causative agent of scrub typhus. To understand the molecular mechanism of L. pallidum, we sequenced the whole genome using Illumina sequencing technology. Totally four genomic libraries with different insert sizes ranging from 280 bp to 8 kb were used to generate 45.1 Gb of genome in the combination of paired-end and mate-pairs sequencing reads. Quality filtering and correction of paired-end reads for very small and/or bad-quality sequences yielded 26.9 Gb of high-quality sequences, which are used to estimate the genome size as 175 Mbusing kmer methods and assembled into a 193.7 Mb genomic sequence scaffolds with N50 length of 92,945 bp. Furthermore, 94% of CEGMA completeness score were obtained from genome scaffold assembly. To facilitate gene annotation, we used a combination of de novo and homology based tools to predict gene models in the chigger mite genome. A combination of evidence-based and de novo approaches predicted 15,842 high-confidence protein-coding genes with an average transcript length of 1,511 bp and 2.4 exons per gene which corresponds to about 12.4% total gene length. Bacterial endosymbiosis are very common in mite species and can range from mutualistic to pathogenic associations. Henceforth, the endosymbionts in L. pallidum were predicted using the NCBI microbial draft genomes and mitochondrial genome. Besides, this L. pallidum draft genome can be used as a significant reference for comparative genomic studies across mite species.

Climate change by global warming is predicted to affect on public health including increasing incidence of vector borne diseases. Vector borne diseases are transmitted by arthropod vectors, such as mosquitoes, chigger mites and ticks, and are highly sensitive to climate changes. The surveillance and R&D of infectious disease vectors are becoming important for climate change preparedness in Korea. So far, 10 regional vector surveillance centers (Incheon, Gyeonggi, Gangwon, Chungbuk, Chungnam, Jeonbuk, Jeonnam, Gyeongbuk, Gyeongnam, and Jeju) have been established to monitor vectors and their pathogens against endemic diseases (japanese encephalitis, malaria, scrub typhus and SFTS) and imported diseases (dengue fever, west nile fever and yellow fever). The information on geographical distribution as well as real-time monitoring of vectors and their pathogens will be presented by this surveillance system, ‘VectorNet’. The R&D plan on vectors and their pathogens is now discussing in government-wide R&D committee on infectious diseases. The R&D areas will be included: 1) establishment of nationwide monitoring system, 2) study on biological and molecular characteristics, 3) development of vector identification and pathogen diagnosis methods, 4) development of vector control techniques, 5) management and conservation of vector resources.

Leptotrombidium pallidum is the major vector mite for Orientia tsutsugamushi, the causative agent of scrub typhus, in Asian countries, including Korea. The genome size of L. pallidum was previously estimated to be 191 ± 7 Mb (Kim et al., 2014). Genomic DNA (gDNA) was extracted from a single female from a 9-generation inbred L. pallidum colony and used for whole genome amplification (WGA). The resulting amplified gDNA was used for the construction of paired-end and mate-pair libraries and sequenced using Illumina platforms (HiSeq2000 and MiSeq). An unamplified gDNA sample extracted from 20 female mites was also used for sequencing in parallel. More than 45Gb sequence reads from both paired-end and mate-pair libraries of the WGA gDNA were trimmed and then de novo assembled using the CLC Asembly Cell v.4.0 for contig assembly and SSPACE for scaffolding. The assembly generated approximately 6,545 scaffolds with N50 value of 92,945 and total size of ~193Mb, which was in a good agreement with our previous estimation. Repeat analysis showed that about 30% of genome (~58Mb) was masked as repeats, most of which were unclassified novel elements. For gene predictions, generated were the PASA models based on genomic alignments of RNA-seq reads from 4 different chigger mite samples (i.e. male, female, larva, and protonymph) and the GeneWise models based on genomic alignments of protein sequences from 4 closely related species with chigger mite. Independently, ab initio gene predictions were performed with AUGUSTUS and FgeneSH with custom trained matrices optimized for L. pallidum and GENEID with pre-trained matrix for Acyrthopsiphon pisum. By combining all together, 15,842 genes were predicted finally. Manual curation is in progress for various groups of genes, including chemosensory receptor genes, immune-related genes, acaricide target genes, etc.

Nanocomposites comprised of graphene oxide (GO) nanosheets and magnesium oxide (MgO) nanoparticles were synthesized by a sol-gel process. The synthesized samples were studied by X-ray powder diffraction, atomic force microscopy, transmission electron microscopy, and energy-dispersive X-ray analysis. The results show that the MgO nanoparticles, with an average diameter of 70 nm, are decorated uniformly on the surface of the GOs. By controlling the concentration of the MgO precursors and reaction cycles, it was possible to control the loading density and the size of the resulting MgO particles. Because the MgO particles are robustly anchored on the GO structure, the MgO/GOs nanocomposites will have future applications in the fields of adsorption and chemical sensing.

Interferon induced transmembrane protein-1 (Ifitm-1) has been reported to have an important role in primordial germ cell formation, and it has expressed in female reproductive organ. In the present study, Ifitm-1 gene expression was identified in testes and all part of epididymis using western immunoblot and immunohistochemistry. Interestingly, Ifitm-1 expression was observed on the head of spermatozoa. To investigate the role of Ifitm-1 gene expression in behavior of spermatozoa after acrosome reaction, fresh sperm was incubated with calcium ionophore to induce acrosome reaction, whereas the expression of Ifitm-1 was not altered after the acrosome reaction. Then to identify the effect of Ifitm-1 in sperm motility and other seminal parameters, different concentration of Ifitm-1 antibody was incubated with spermatozoa, and seminal parameters were assessed using computer-assisted semen analysis (CASA). Interestingly, motility, progressive, and VAP were increased in the sperm with Ifitm-1 antibody treated compared to rabbit serum, however other parameters such as straightness were not changed. In order to identify the functional significance of Ifitm-1 in fertilization, capacitated spermatozoa were pre-incubated with anti- Ifitm-1 antibody and subsequently examined the ability to adhere to mouse oocytes. However, any defection or alteration in sperm-egg fusion was not found, Ifitm-1 antibody treated or non-treated spermatozoa showed a normal penetration. Although the precise role of Ifitm-1 in sperm motility and following fertilization need to be elucidated, this study suggests that the activation of Ifitm-1 on the sperm may enhance the motility of spermatozoa in mice.

It has been reported that light-emitting diodes(LED) can be used in the treatment of oral diseases. Although bio-stimulatory effects of LED irradiation such as promotes stimulation of wound healing have been well known, there are few reports about molecular mechanism associated with cell cycle by LED irradiation. The purpose of present study was to examine the molecular event in cell cycle of LED irradiation on primary human gingival fibroblast(hGF) in vitro. The source of light for irradiation was a continuous-wave LED emitting at a wavelength of 635nm, and manufactured that energy density was 5mW/cm2 on sample surface. The hGF were irradiated for 1 hour at 37℃ in 5% CO2 humidified chamber. Experimental samples were acquired at 0 (right after irradiation), 8 and 24 hour after irradiation. To investigate the molecular mechanisms associated with cell cycle, growth phase was determined by flow cytometry and mRNA expression of cyclin A, cyclin B, cyclin D1, cyclin E, cdc2, PCNA, p18, p27, p21, and p53 were determined by real time RT-PCR. Flow cytometric analysis demonstrated the percentage of cells in the G1 and S phase were decreased, but the G2 phase increased, which showed cells irradiated by LED were transitioned from S to G2 phase. For mRNA expression, cyclin B, cdc2, PCNA and p53 were increased at 0 hour after irradiation, and most of cell cycle molecules were increased at 8 hour after irradiation. At 24 hour after irradiation, cyclin A, cyclin E, PCNA and p18 were increased. Taken together, LED irradiation induced proliferation of hGF cells through transition from S to G2 phase.

The purpose 01' pl'esent study was to examine the molecular events in apoptosis by CoCl2, mimicking hypoxic cond ition and recovering effects by LED ir l'adiation on Human SH-SY5Y neuroblastoma cells The SOUl'ce 0 1' light for ir l'adiation was a continuous-wave LED emitting at a wavelenl양h of 590 nm, and manufactured that ene rgy density was 5 mW!cm2 on sample surface, After ir l'adiation, cell viabi lity was measured with BrdU , cell morphol ogy was examined with Diff- Quik staining, cell signaling was monitored with various apoptosis-related molecules using RNase Pl'otection Assay(RPA) , W11en treated with CoC12, apoptotic induction was found in the SH-SY5Y cells in a concentration-dependent and time-dependent manner , Diff-Quik s taining was revealed that DNA fragmentation re presented apoptosis was examined in CoC12-tl'eated group, Moreover, RPA assay of SH-SY5Y cclls lIs ing val'iolls apoptosis-related molecllles showed that the apoptotic cell population was mcreased J-loweve. there was sorne signifïcant change in LED irradiatied cells aftel' treatement of CoC12 The main mechanism for Lhese a poptosis appearecl to be mito c hondriεt - m ecliated pathway, such as cytochrome- c‘ caspase-9, caspase-3, pro-apototic protein ßax, anti-apototic protein Bcl-2, and death receptor• mediated pathway, such as Fas, cas pase- 8, a ncl TNFRl These results demonstrate that CoCI2 induce apoptosis in SH-SY5Y via different dual apop tosis pathway through death receptor pathway as well as mitochondria- dependent pathway and LED irradiation can recl llces the CoCl2-induced apoptosis by blocking their internal signaling pathway

Background : Unlike Dioscorea japonica or Dioscore opposita, Dioscore alata is a high-temperature crop mainly cultivated in Africa and subtropical regions. Due to recent climate change, its cultivation area is expanding in Korea. In 2016, the Institute for Bioresource Research clarified optimal harvesting stage to increase its tuber yield and ensure safe storability. Methods and Results : As the seed-tubers for this study, Dioscorea alata tubers were cut into a size of 50 g, disinfected with lime powder, dentated on the seed bed with electrothermal wire installed on March 20th, and then planted on the main field on April 15th. The planting distance was 120 ㎝ in width, where the furrows of 90 ㎝ were cladded with black vinyl, in two rows with a column spacing of 25 ㎝. The amounts of applied fertilizers were 2,000 ㎏ compost, 34 ㎏ N2, 28 ㎏ P2O6, and 28 ㎏ K2O. The compost and P2O5 were used in full, where as N2 and K2O were used as original fertilizer 14 (10 ㎏) and additional fertilizer 20 (18 ㎏), respectively. The crop was harvested in six times from September 20th to November 9th, in an interval of 10 days. While harvesting, the ground fresh weight yield and underground tuber yield were investigated. The underground tuber yield was 836, 1,744 and 2,975 ㎏ low at the early harvests on September 20th, 30th, and October 10th, respectively, while it was 3,622, 3,828, 3,818 ㎏ high on October 20th, 30th, and November 9th, respectively. The ground fresh weight yield remained the same. To clarify tuber storability by harvest time, they were stored at 15℃after harvest, and then healthy and corrupt tubers were studied on March 23th. The corruption ratio by harvest time was highest at 39.1% and 29.3% on October 30th and November 9th, respectively, when frost and low temperature damaged harvest. It was 59.0% and 42.8% at the early harvests on September 20th and 30th, respectively. The corruption ratio was lowest on October 20th and October 10th at 29.3% and 39.1%, respectively. Conclusion : Ford. Dioscore alata cultivation, Korea presents a premature and disadvantageous environment with lower temperature and pre-harvest frost at the time of planting. After raising seeding on an electrically heated hot bed, the optimal harvesting stage should be from planting in the middle of April where temperature rises above 1 5℃ to harvesting in the period October 10th - 20th so as to achieve the maximum tuber yield and high storability.

Background : The study about ginseng cultured roots have been reported mainly ginsenosides in saponins family. Other phytochemical such as non-saponins of fatty acid has been revealed its bioactive activity including anti-oxidation, whitening, anti-cancer. Supercritical extraction (SE) process mainly refer to the extraction with CO2, is usually from a solid matrix, is a sample preparation step for analytical purposes. SE produce no residual solvent and possess high stability of the extract component, which is advantageous for fatty acid analysis. Methods and Results : Fermented ginseng cultured roots used in the experiment were used for fermentation using Pediococcus pentosaceus. SE performed at different temperature, pressure and extraction time using non-fermented and fermented ginseng roots. Further we fractionated from fermented ginseng using Methanol, Hexane, Ethanol, Ethyl acetate and Butanol. We compared fatty acids contents ginseng extractions by GC analysis. Methyl linoleate contents was 44% of fatty acids supercritical extraction contained. The contents of Methyl linoleate was the most dominant component among 37 types of fatty acids by SE and other extractions solvent. Total fatty acids contents obtained by SE process from fermented ginseng (1325.61ppm) was twice than from non-fermented ginseng (618.47ppm). Conclusion : Fatty acids contents by SE was increased at high pressure. The best condition for fatty acids contents extraction was 60℃, 350bar and 3h.

Background : Perilla frutescens L. is valuable as a medicinal plant as well as a natural medicine and functional food. Limonene perilla collected from various places showed 60% limonene compounds. However biological activity of these accession has not been reported before. Therefore, this study was conducted to investigate the biological activity of limonene perilla. Methods and Results : Fractional solvent extracts were obtained by using organic solvents such as n-hexane, chloroform, ethyl acetate, n-BuOH, and aqueous solvent from different parts of limonene perilla extracted initially in 70% EtOH. We investigated the effects of limonene perilla on total phenol and flavonoid contents, FRAP (Ferric Reducing Antioxidant Power), total saponin contents and tyrosinase inhibition activity. Leaves of limonene perilla produced the highest total phenolic contents (29.88 mg·CAE/g), flavonoid (8.39 mg·QE/g) and saponin contents (47.77 mg·GIE/g) than stems and roots of limonene perilla. FRAP of leaves was 823.00±3.58 μM·FeSO4·E/mg. Tyrosinase inhibition activity rate was 40.31% in 70% ethanol extracts from leaves of limonene perilla. Conclusion : This results suggest that leaf of limonene perilla fractions has significant antioxidant activity. Also, limonene perilla could be used as a functional biomaterial in developing cosmetics and functional foods.

Background : study of long yam and short yam (Dioscorea. opposita) are cultivated in temperate regions but tropical yam (Dioscorea alata) are mainly grown in Africa and sub-tropical regions cultivated crops. Recent tropical yam cultivation area of Korea increased but lack the proper cultivation techniques to climate warming. This study was conducted to elucidate the effect of the timely harvest on Tropical yam at 2015 in Institute of Bioresources Research, GBARES. Methods and Results : Tuber of tropical yam were cut as 40 g, and dust-coating sterilized with lime. Seedlings were grown in heating wire installed seed bed from 11. March to 16. May. Experimental field were fertilized 2,000 kg compost, 34 kg N2, 28 kg P2O5, and 28 kg K2O. All amount of compost and P2O5 treated as basal fertilizer. N2 and K2O treated 14 and 10 kg of basal fertilizer, and 20 and 18 kg of additional fertilizer, respectively. Tillage, covering black PE film with 60×25 cm spacing holes on 120 cm row were conducted. Emergence days from 6. April to 18 May until 28 days to 36 days but 15. June and 9. June were each 19 and 7 days. Dry weigh in early stage due to seedling date, while seedling date according from 6. April to 18. May had no difference. seedling date was not effective on total number of tuber but number of marketable tuber (over 200 g). Marketable tuber number of seedling date according to 20. April was increased 35~132% as 4,028 number per 10 a. And tuber with 20. April was 13~73 g heavier. Tuber yield and marketable tuber yield of seedling date as affected by 20. April were respectively 2,518 and 1,273 kg per 10 a compared to 6. April and from 4. May to 15. June as 1,743~2,457 kg and 484~1,027 kg. Conclusion : Tropical yam is low temperature in the cultivation of Korea but also adverse environmental conditions. Frost is not mature enough off before harvesting. Yam had significant increased marketable tuber (over 200 g) yield due to seedling in mid-April Requires cultivation technology.

Freesia is one of the most popular flowers over the world including Korea, due to the fragrance and beauty of the plant flower. The first domestic freesia cultivar ‘Shiny Gold’ was developed by NIHHS, RDA, in 2003, which has yellow double and large petals and strong fragrance. Ten years have passed since ‘Shiny Gold’ was cultivated at floral farms, and the deterioration of cut flower quality and yield are reported from the farms. Virus infection causes a reduction in the quantity and quality of the cut freesia flowers and is one of the most serious problems in Korea. Virus detection was carried by reverse transcription polymer chain reaction (RT-PCR) for FreMV, FreSV, BYMV, CMV, and TRV, as known to infect freesia. FreMV, FreSV, BYMV, and TRV were detected single or multiply, and CMV was not found in the freesia leaves collected from the farms. To produce virus-free freesia, meristem culture of ‘Shiny Gold’ was conducted in MS medium added ribavirin at different concentration. As the increased of ribavirin concentration, the growth of ‘Shiny Gold’ plantlets was inhibited in freesia’Shiny Gold’. The plantlets produced by meristem culture in ‘Shiny Gold’ were virus free at the enzyme-linked immunosorbent assay (ELISA) level.

Fusarium crown root rot (FCRR) is a severe fungal disease caused by Fusarium oxysporum f. sp. radicis-lycopersici (FORL) in tomato. Resistance to FORL is conferred by single dominant locus Frl on chromosome 9, but its precise genomic location is not clearly determined. In this study, detailed location of Frl was assessed by using a set of molecular markers physically anchored on Chr.9 and F2 and RIL population derived from FORL-resistant inbred AV107-4 (S.lycopersicum) x susceptible L3708 (S. pimpinellifolium). Bioassay of the two populations with a FORL strain isolated from Korea resulted in single dominant heritance of the resistance. Two SCAR and 11 CAPS markers encompassing 3.6Mb~72Mb of Chr.9 were developed from the Tomato-EXPEN 2000 map and SolCAP SNP-array analysis. These markers were genotyped on 345 F2 plants. A high level of cosegregation with the resistance were observed for 5 markers which were mapped at a large physical interval of 5.1Mb (T1212) to 46.4Mb (SSR237), indicating that genetic recombination was highly suppressed in this region. Cosegregation of these markers with Frl was confirmed by using 126 RILs. The results implied that, in contrast with the previously reported long arm, Frl is present on a pericentromeric region of short arm of Chr. 9, in which crossing-over is severely suppressed. The marker set was further tested on 12 FORL-resistance or susceptibility commercial cultivars. Unlike the biparental populations, frequent linkage break was observed for T1212 and D4 in commercial cultivars. T1212 and D4 showed 50% and 100% match with the phenotype, respectively. D4, a CAPS, was converted to a high resolution melting (HRM) marker and tested on 55 breeding lines from private seed companies (Fig.3). All breeding lines showed the HRM genotype for resistance allele, indicating that D4 can be useful for selecting FORL-resistance tomato plants.

Genetically modified (GM) papaya (Carica papaya L.) line 55-1 (55-1), which is resistant to papaya ringspot virus infection, has been marketed internationally. Many countries such as the European Union, Japan, and Korea have a mandatory safety assessment, approval and labeling regulations for GM foods. Thus, there is a need for specific methods for detecting 55-1. In this study, we established a real-time PCR detection method applicable to 55-1 for a variety of papaya products. The limit of detection was possible for fresh papaya fruit up to dilutions of 0.005% and 0.01% (weight per weight [w/w]) for homozygous SunUp and heterozygous Rainbow cultivars, respectively, in non-GM papaya. The 55-1 event-specific detection method observed parallelism (r2>0.99) between the concentration of line 55-1 cultivars and Ct values obtained in amplification plots at concentrations of 0.005-10% for SunUp DNA and 0.01-10% for Rainbow DNA. The method was applicable to the qualitative detection in various types of processed products (cocktail fruit, dried fruit, juice, etc.) containing papaya as a main ingredient. Monitoring papaya products for the presence of GM papaya were demonstrated using a P35S and T-nos real-time PCR detection method but no amplification signals were detected.

탈모방지 및 발모효과를 가지는 소재를 개발하기 위해 한의학에서 전통적으로 사용되는 23가지 한방소재를 선정하여 한방복합처방단을 개발하였다. 한방복합처방단을 구성하는 한방소재들은 예로부터 전통적으로 발모 및 탈모방지, 흰머리방지, 염증 치료 및 혈액순환개선 효과를 가진 것으로 알려져 있는 당귀, 보골지, 측백엽, 한련초, 구기자, 복분자, 상백피, 숙지황, 여정실, 적하수오, 흑지마, 고삼, 백지, 익모초, 단삼, 도인, 몰약, 감국, 유향, 인삼, 천궁,합환피, 현호색 등이다. 또한, 한방복합처방단의 발모효과를 확인하기 위해 in vitro와 in vivo 평가모델을 이용하여 모발성장 및 촉진에 미치는 영향을 실험하였다. In vitro 상에서는 모유두세포, 각질형성세포 및 섬유아세포의 증식을 확인하였다. 또한, 흰머리방지 효과와 관련하여 멜라노마 세포에서의 멜라닌 합성능력을 확인하였다. 한방복합처방단의 in vitro 상에서의 육모효과는 C57BL/6 마우스를 이용한 in vivo 상에서도 확인하였다. 연구 결과 한방복합처방단은 50 μg/mL의 농도에서 모유두 세포의 증식을 175 %까지, 섬유아세포인 NIH3T3 세포의 증식은 120 %까지 증가시켰으며, 20 μg/mL의 농도에서 각질형성세포인 HaCaT 세포의 증식을 133 %까지 증가시켰다. 멜라닌 합성의 경우, 50 μg/mL의 농도에서 154 %까지 증가시켰다. 또한, C57BL/6 마우스를 이용한 육모효과에 있어서는 한방복합처방단 처리 4주 후 98 % 이상의 육모효과를 나타내는 것을 확인하였다. 이상의 결과로 볼 때 본 연구에서 개발한 한방복합처방단은 모발의 성장 촉진에 유용하게 활용될 수 있는 처방으로 사료된다.