Vitamin K1 (VK1) has been widely used as a coumarin antagonist and for the treatment of hemorrhagic disease in veterinary practice. However, the potential mechanism of adverse reaction after VK1 injection has been not fully elucidated. In this study, two cases of anaphylactic reactions after subcutaneous VK1 injection were presented, and then an experimental study was performed to further characterize the anaphylactic reactions. Two dogs developed anaphylactic reactions after subcutaneous VK1 injections and were promptly treated with antihistamines and glucocorticoids, after which abnormal signs related to anaphylaxis disappeared. Subsequently, a study was undertaken to ascertain the nature of the adverse reactions to subcutaneous VK1 injection. Six healthy laboratory beagle dogs received subcutaneous VK1 administrations once daily for eight days. They were monitored for clinical signs after each injection, and blood samples were collected for the measurement of plasma histamine and immunoglobulin E concentrations using enzyme-linked immunosorbent assay. All six dogs showed mild angioedema after the VK1 injections. The dogs also displayed clinical signs including sneezing, coughing, skin reddening, excess salivation, pawing the ground, and somnolence on days 4, 6, and 8. Plasma histamine and immunoglobulin E concentrations were significantly increased by the repeated injections. In summary, this study describes anaphylactic reactions resulting from subcutaneous VK1 administration in dogs. Clinicians should be aware that the repeated subcutaneous injection of VK1 can trigger an anaphylactic reaction in dogs.

After the permanent shutdown of K1 in 2017, decommissioning processes have attracted great attention. According to the current decommissioning roadmap, the dismantling of the activated components of K1 may start in 2026, following the removal of its spent fuel. Since the reactor vessel (RV) and reactor vessel internal (RVI) of K1 contain massive components and are relatively highly activated, their decommissioning process should be conducted carefully in terms of radiological and industrial safety. For achieving maximum efficiency of nuclear waste management processes for K1, we present activation analysis of the segmentation process and waste classification of the RV and RVI components of K1. For RVI, the active fuel regions and some parts of the upper and lower active regions are classified as intermediate-level waste (ILW), while other components are classified as low-level waste (LLW). Due to the RVI’s complex structure and high activation, we suggest various underwater segmentation techniques which are expected to reduce radiation exposure and generate approximately nine ILW and nineteen very low level waste (VLLW)/LLW packages. For RV, the active fuel region and other components are classified as LLW, VLLW, and clearance waste (CW). In this case, we suggest in-situ remote segmentation in air, which is expected to generate approximately forty-two VLLW/LLW packages.

K1A1 tank commander’s primary thermal sight is a device that enables tank commanders to detect, identify, aim and track the target by observing targets in all directions during day, night and in situations of smokescreen and fog through 360° rotation independent from the gunner’s primary thermal sight and stabilizing the line of sight even under the vibrations occurring when the tank is standstill and moving. The main function of this device is to detect and process visible and thermal images and deliver the final images to the tank commander. One of the core parts to that end is the observation window (daytime/ thermal image window). This core part is mounted at the entrance of the optical path for observing the target and plays the role of making visible light during the daytime and infrared light during the night pass through the target and transmitting the resultant images to the internal optical system of the tank commander’s primary thermal sight. Such core parts have been selected as depot maintenance items so that they are replaced by new parts instead of being recycled when they are subjected to maintenance in most cases. That is, the military budget is wasted because such parts are replaced by new parts despite that they can be recycled for maintenance. Therefore, this study proposed a mounting tool for polishing and coating observation windows (daytime and thermal image window) using planar polishing equipment and DLC (Diamond-Like Carbon) coating equipment. In addition, this study presented an amendment (proposal) of the Depot Maintenance Work Request (DMWR) already published to verify the performance of recycled products including the establishment of inspection standards for recycling processes.

Vitamin K1 is a fat-soluble vitamin essential for human health. We used the Association of Official Analytical Chemists(AOAC) method 999.15, validated by the International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use(ICH) Guideline, to monitor vitamin K1 content in infant formula in the Republic of Korea. The Certified Reference Materials (CRM), NIST 1849a, were used for validation of the test method. The parameters of validation were linearity, range, specificity, accuracy, precision, LOD, LOQ and recovery. The linearity, R2 was above 0.999 and the analytical range was 0.001 to 0.22 μg/mL. The specificity was confirmed by the retention time (RT) and positive/negative sample test. The accuracy, precision and recovery values were 101.13 ± 0.13 %, 3.15 % and 81.60 % - 99.78 %. The LOD and LOQ values were 0.582 ng/mL and 1.76 ng/mL, respectively. Five livestock products inspection agencies voluntarily participated in the collaborative study. The HORRAT value was 0.6, which was within the reference value 0.5 – 2.0. The validated test method was applied for the monitoring of vitamin K1 in infant formula in the Republic of Korea. The results also conformed to 'The Standard for Livestock Products of Processing and Ingredient Specifications' and 'The Standard for Livestock Products Labeling'.

Vitamin K (phylloquinone) is an essential cofactor in the synthesis of active blood-clotting factors II, VII, IX and X. Deficiency of vitamin K leads to inadequate activity of these factors, resulting in bleeding. In this study, we investigated vitamin K1 content of agricultural products that are widely and specifically grown in Korea including 9 leaves and vegetables, 16 fruits, and 11 cereals and specialty crops. Vitamin K1 analysis of the agro-samples was by a validated, modified, reversed phase-HPLC method with fluorescence detection after post-column derivatization. The vitamin K1 content ranged from 1.83 to 682.73 μg/100 g in leaves and vegetables, 0.17 to 28.22 μg/100 g in fruits, and ND to 279 μg/100 g in cereals and specialty crops. Among the 36 samples, high content of vitamin K1 were found in Gugija (Lycium chinense Miller) leaves (average 682.73 μg/100 g) and Hansan ramie leaves (average 423.12 μg/100 g); however, mushroom, amaranth and Chinese artichoke showed no detectable levels. The results of ourstudy provide reliable vitamin K1 content of Korean grown agricultural products that expand nutritional information and food composition database.

돼지 수정란의 체외 생산 효율성 향상을 위해서는 배발생율과 더불어 고품질의 배를 조기에 선별해야 한다. 체외 배 발생율에 대한 보고는 많지만, 고품질의 배를 선별할 수 있는 기술에 대한 연구는 거의 없었다. 본 연구에서는 돼지 난포란 유래 수정란의 체외배양에 있어서 배반포로의 배 발달과 생존에 미치는 Vitamin K1(vit K1) 첨가 농도, 시기 및 시간의 효과를 검토하였다. 1.0 μM, 3.0 μM 및 6.0 μM vit K1을 배양 1일째 24시간 첨가한 결과, 배반포 발달율이 시험군이 14.5 ± 4.3, 0.0 및 0.0%로써 대조군의 35.5 ± 3.2%에 비하여 유의하게 낮았고(p<0.05), 배반포의 생존율도 대조군이 31.8 ± 2.6%로써 시험군의 22.2 ± 2.9, 0.0 및 0.0%에 비하여 유의하게 높았다(p<0.05). 상기 첨가 농도에서 첨가 시간을 달리한 결과, 1.0 μM 농도에서 6시간 처리군의 배반포 발달율과 생존율이 각각 26.5 ± 2.9% 및 47.2 ± 2.8%로써 가장 높았고 특히, 12시간 처리군보다 유의하게 높았다(p<0.05). 3.0 μM 농도에서는 대조군의 배발달율이 36.4 ± 3.1%로 가장 높았으나, 생존율은 3.0시간 첨가군이 41.7 ± 3.2%로 대조군에 비하여 유의하게 높았다(p<0.05). 6.0 μM 농도에서도 배발달율은 대조군(32.0 ± 2.8%), 생존율은 0.5시간 첨가군(42.9 ± 1.8%)이 가장 높았다. 각각의 vit K1 첨가 농도와 시간을 기준으로 서로 다른 배양 시기에 첨가한 결과, 1.0 μM 6시간 첨가군에서는 배반포 발달율은 배양 4일째 첨가군, 생존율은 배양 2일째 첨가군이 가장 높았다. 한편, 3.0 μM 3.0시간 및 6.0 μM 0.5시간 첨가군에서는 배양 4일째 첨가군의 배반포 발달율(59.5 ± 4.1% 및 50.0 ± 3.6%)과 생존율(72.7 ± 5.4% 및 79.2 ± 4.0%)이 대조군과 다른 시험군에 비하여 유의하게 높았다(p<0.05). 한편, vit K1 첨가에 따른 배반포의 세포 수를 조사한 결과, 첨가군(1.0 μM 6시간 배양 2일째, 3.0 μM 3.0시간 배양 4일째 및 6.0 μM 0.5시간 배양 6일째)이 53.4 ± 5.8, 49.4 ± 3.8 및 51.5 ± 4.5개로써 대조군의 40.2 ± 2.3개에 비하여 유의하게 많았다(p<0.05). 그러나 사멸세포 수는 시험군이 3.2 ± 0.9∼3.7 ± 2.1개로써 대조군의 4.2 ± 1.2개보다 적었으나, 유의차는 없었다. 세포 사멸 유도 유전자인 Bax mRNA 발현은 처리군과 대조군은 비슷하였으나, 세포 사멸 억제 유전자인 Bcl-xL mRNA 발현은 처리군이 대조군보다 높았고 특히, 6.0 μM 0.5시간 배양 4일째 첨가군이 가장 높았다. 이상의 결과로부터 돼지 미성숙 난포란 유래 수정란의 체외 배양에 vit K1의 첨가는 배반포의 생존율과 세포수 증가에 효과적이었다. 그 이유에 대해서는 아직 많은 부분이 밝혀져야 되겠지만, 고품질의 배반포 조기 선발에는 활용이 가능할 것으로 생각된다.

Mamestra brassicae (cabbage moth) is a common European moth of the order Lepidoptera and the family Noctuidae. The larval stage is highly polyphagous and is known to feed on more than 70 species of host plants from 22 families, including Brassica species, lettuce, onion, potato, pea, tomato and apple. M. brassicae has become a significant pest also in Asia due to the damage caused to agriculturally and economically important Brassica crops. It is difficult to control M. brassicae using chemical insecticide because of its rapid development of resistance. The objective of our study, therefore, was the mass production and formulation of a local strain of M. brassicae nucleopolyhedrovirus-K1 (MabrNPV-K1) for the development of viral insecticide to control it. In production efficiency of MabrNPV-K1 using M. brassicae larvae, the mortality of the 3rd instar larvae was 100% when inoculated with 1.0 × 105 PIBs/larva and the yield of MabrNPV-K1 was maximal. Regarding the mortality, yield of polyhedra, inoculation doses and required time, the 1.0 × 104 PIBs/larva at 30°C was determined as optimal conditions producing polyhedra efficiently. To formulate MabrNPV-K1, feeding toxicities of various supplements including spreader and ultraviolet (UV) -protectant were determined. Tinopal UNPA-GX which is UV-protectants was effective for protection of polyhedra from UV and showed the increased mortality when added with 1% concentration. Other supplements did not influence significantly the mortality of MabrNPV-K1. Formulated MabrNPV-K1 with several supplements showed higher pathogencity than un-formulated MabrNPV-K1.

Two entomopathogenic bacteria, Xenorhabdus nematophila and Photorhabdus temperata subsp. temperata, are known to be potent against the diamondback moth, Plutella xylostella, when the bacteria are injected into the hemocoel. This study investigated any pathogenic effect of their culture broth on P. xylostella by oral administration. Only culture broth of both bacterial species did not give enough pathogenic effects by the oral administration. However, when the culture broth was orally treated together with Bacillus thuringiensis (Bt), both cell-free culture broth significantly enhanced Bt pathogenicity against the 3rd instar larvae of P. xylostella. The culture broth was then fractionated into hexane, ethyl acetate, and aqueous extracts. Most synergistic effect on Bt pathogenicity was found in ethyl acetate extracts of both bacterial species. Thin layer chromatography of these extracts clearly showed that ethyl acetate extracts of both bacterial culture broths possessed metabolites that were different to those of hexane and aqueous extracts. These results suggest that the both entomopathogenic bacteria produce and secrete different factors to give significant synergistic effect on Bt pathogenicity.

Bacillus thuringiensis, an entomopathogenic bacterium belonging to the B. cereus group, harbors numerous extra-chromosomal DNA molecules whose sizes range from 2 to 250 kb. In this study, we used a plasmid capture system (PCS) to clone three small plasmids from B. thuringiensis subsp. kurstaki K1 using PCS which were not found in B. thuringiensis subsp. kurstaki HD-1, and determined the complete nucleotide sequence of plasmid pK1S-1 (5.5 kb). Of the six putative open reading frames (ORF2-ORF7) in pK1S-1, ORF2 (MobK1) showed approximately 90% aa identity with the Mob-proteins of pGI2 and pTX14-2, which are rolling circle replicating group VII (RCR group VII) plasmids from B. thuringiensis. In addition, a putative origin of transfer (oriT) showed 95.8% identity with those of pGI2 and pTX14-2. ORF3 (RepK1) showed relatively low aa identity (17.8-25.2%) with the Rep protein coded by RCR plasmids, however. The putative double-strand origin of replication (dso) and single-strand origin of replication (sso) of pK1S-1 exhibited approximately 70% and 64% identities with those of pGI2 and pTX14-2. ORF6 and 7 showed greater than 50% similarities with alkaline serine protease, which belongs to the subtilase family. The other 2 ORFs were identified as hypothetical proteins. To determine the replicon of pK1S-1, seven subclones were contructed in the B. t huringiensis ori-negative pHT1K vector and were electroporated into a plasmid cured B. thuringiensis strain. The 1.6 kb region that included the putative ORF3 (Rep1K), dso and ORF4, exhibited replication ability. These findings identified pK1S-1 as a new RCR group VII plasmid, and determined its replication region.

도둑나방(Mamestra brassicae)은 배추, 무, 양배추 등 45종 107과의 식물을 가해하는 광식성 해충이며 연 2회 발생하나, 현재까지 국내에서는 등록된 적절한 약제나 천적이 없는 난방제해충이다. 이러한 도둑나방의 방제법을 개발하고자 도둑나방에 대하여 높은 병원성을 보이는 도둑나방 핵다각체병 바이러스(M. brassicae nucleopolyhedrovirus: MbNPV)를 국내에서 분리하고 바이러스 살충제의 개발을 위한 생물학적 및 분자생물학적 기초 특성을 조사하였다. MbNPV-K1의 다각체는 직경 약 1.8㎛의 부정형이고, 하나의 envelope에 다수의 nucleocapsid가 존재하는 MNPV(Multiple nucleocapsid nucleopolyhedrovirus)형태였다. 다각체 단백질은 Autographa californica 다각체 단백질의 분자량과 동일한 약 31 kDa의 단일 단백질로 나타났으며, 그 유전자의 염기서열을 확인한 결과 기존에 보고된 MbNPV와 동일한 염기서열을 보였다. 살충력을 조사하기 위한 생물검정은 외국에서 개발되어 상용화되고 있는 MbNPV 살충제인 Mamestrin을 대조구로 하여 도둑나방 3령충에 대하여 비교 조사하였다. 그 결과 MbNPV-K1의 반수치사농도(LC50)는 3.9 X 103 PIBs/larva로, 6.0 X 104 PIBs/larva 인 Mamestrin 보다 15배 높은 병원성을 나타냈다. 또한 반수치사일수(LT50)도 MbNPV-K1은 4.4~6.1일, Mamestrin 은 4.1~8.6일로 나타났다.

The Chinese Hamster Ovarian cells CHOK1 are one of the most extensively used cells for the evaluation of gene expression and toxicology. However, these cells are frequently used for biomedical research without consideration of their cytogenetic characteristics. Therefore, we carried out to investigate the karyologic profiles, the frequency and type of chromosome aberration, and the distribution of telomeric DNA on chromosomes of the CHOK1 cells. The GTGbanding and fluorescence in situ hybridization on CHOK1 cells were performed to characterize the karyotype and the distribution of telomeric DNA. The present study revealed that the chromosome modal number of CHOK1 cells was 2n=20; eight chromosomes appeared to be identical with those of the normal Chinese hamster, whereas the remaining 12 chromosomes were shown to be translocated, deleted, inversed, or rearranged from Chinese hamster chromosomes. The telomeric DNA on CHOK1 chromosomes was intensively distributed at the centromeres rather than the ends of chromosomes. In addition, three chromosomes had interstitial telomeres and one marker chromosome entirely consisted of telomeric DNAs. The frequency and type of chromosome aberrations in CHOK1 cells were examined. Of the 822 metaphase spreads, 68 (8.3%) cells resulted in chromosome aberrations of which the chromosome breakage was the most frequently occurred.

세로토닌 수용체는 세로토닌과 반응하여 세포막의 G단백질을 통해 중개단백질 (adenylyl cyclase, phospholipase C, cGMP phosphodiesterase, ion channel)을 활성화시켜, 이뇨, 기억, 발생 등의 다양한 생리적 반응에 관여한다. 곤충세포인 Schneider2 (S2)와 척추동물 세포인 Chinese hamster ovary (CHO)-Kl에서 Aedes 5-HT 수용체 유전자 발현을 비교하기 위해, Aedes 5-HT 수용체 유전자를 형질이입시켰다. 선발된 세포주들(Tr-S2, Tr-CHO)에서 세로토닌 수용체 유전자의 발현은 reverse transcription-PCR, Western blot, immunocytochemistry를 이용하여 확인하였다. 세로토닌 농도증가에 대한 Aedes 5-HT수용체의 기능을 세포 내 cAMP수준을 통해 조사한 결과,Tr-CHO 세포주는 Tr-S2 세포주보다 9배 이상 cAMP수준이 높게 나타났으며, 농도에 의존적이었다. 이 결과는 수용체 유전자가 세포에서 발현되었으나, 세포의 종류와 세포막에 존재하는 G단백질 차이에 따라 중개단백질 활성 차이가 있다는 것을 보여주었다. CHO-Kl 세포에서 Aedes 5-HT 수용체의 기능이 S2 세포보다 더 효율적이며, Aedes 5-HT 수용체를 발현하는 Tr-CHO 세포주는 동력제 또는 대립제 검정에 활용될 수 있을 것으로 기대된다. 것으로 기대된다.

Equine chorionic gonadotropin (eCG) is a unique molecule that elicits the response characteristics of both follicle- stimulating hormone (FSH) and luteinizing hormone (LH) in other species. Previous studies from this laboratory had demonstrated that recombinant eCG (rec-eCG) from Chinese hamster ovary (CHO-K1) cells exhibited both FSH- and LH-like activity in rat granulosa and Leydig cells. In this study, we analyzed receptor internalization through rec-eCGs, wild type eCG (eCGβ/α) and mutant eCG (eCGβ/αΔ56) with an N-linked oligosaccharide at Asn56 of the α-subunit. Both the rec-eCGs were obtained from CHO-K1 cells. The agonist activation of receptors was analyzed by measuring stimulation time and concentrations of rec-eCGs. Internalization values in the stably selected rat follicle-stimulating hormone receptor (rFSHR) and rat luteinizing/ chorionic gonadotropin receptor (rLH/CGR) were highest at 50 min after stimulation with 10 ng of rec-eCGβ/α. The dose-dependent response was highest when 10 ng of rec-eCGβ/α was used. The deglycosylated eCGβ/αΔ56 mutant did not enhance the agonist-stimulated internalization. We concluded that the state of activation of rFSHR and rLH/CGR could be modulated through agonist-stimulated internalization. Our results suggested that the eLH/CGRs are mostly internalized within 60 min by agonist-stimulation by rec-eCG. We also suggested that the lack of responsiveness of the deglycosylated eCGβ/ αΔ56 was likely because the site of glycosylation played a pivotal role in agonist-stimulated internalization in cells expressing rFSHR and rLH/CGR.

Bacterial blight (BB) of rice, caused by Xanthomonas oryzae pv. oryzae (Xoo), is a significant disease in most rice cultivation areas. The present study was performed to identify new BB R-gene conferring resistance to Korea Xoo isolates, derived from IR65482-7-216-1-2 and to construct a physical map of the candidate gene. An F2 population derived from a cross between 11325 and Anda was used to determine the exact position of the nearest recombination event to the target region. The position of the R-gene was delimited by flanking markers, RM1233 and RM5766, on chromosome 11. Of the 56 markers designed in the flanking region, 20 were selected as anchor markers and the R-gene was mapped to a 295kb region on chromosome 11. To narrow down the interval spanning the R-gene, an additionally SSR marker, 20 STS markers, and CAPS marker between RM27320 and ID55.05-79 were developed using rice reference genome information. From the result the gene was defined by RM27320 and ID55.WA18-5 located in the BAC clone OSJNBa0036K13. The physical distance between these two markers is approximately 80kb. In a further study, gene expression analysis against listed candidate genes was investigated using semi-quantitative transcription PCR. These results will useful for future disease breeding as well as gene function studies regarding resistance genes.

조생종 벼 품종인 오대, 운광과 HR28021-AC16 계통에 대한 벼흰잎마름병 저항성 유전자분석과 K1과 K3a 균계를 접종한 후에 수량 및 품질 관련 형질들의 변이에 대하여 분석하였다. 오대는 저항성 유전자를 보유하지 않는 것으로 나타났고, 운광은 Xa3 유전자를, HR28021-AC16은 Xa21 유전자를가지는 것으로 확인되었다. 오대는 K1과 K3a 균계 모두에 이병성을 보였고, 운광은 K1에는 저항성을 K3a에는 이병성을보였다. HR28021-AC16은 K1 균계에는 이병성을 나타냈고K3a 균계에는 저항성 반응을 보였다. 균계접종에 의한 이병성 정도와 2차 감염 정도로 볼 때, K3a 균계가 K1 균계에 비해 병원성이 강한 것으로 나타났다. K1과 K3a 균계 접종에따른 생산력검정시험에서 균계 처리와 반응에 의해 변이가발생한 형질들 중에 등숙률, 정현비율, 현미수량과 완전미도정수율은 서로 간에 정의 상관관계를 나타냈으며, 이들 형질들과 현미 및 백미 단백질 함량은 부의 상관관계를 나타냈다.사미는 등숙률 및 완전미도정수율과 부의 상관관계를 나타냈다. 주성분분석에서 주성분 1을 기준으로 등숙률, 정현비율,현미수량 및 완전미도정수율과 사미, 현미 및 백미 단백질 함량이 다른 방향성을 나타냈고, 주성분 2를 기준으로는 등숙률과 사미가 다른 방향성을 나타냈다. Xa21 유전자를 가지고있는 HR28021-AC16은 K3a에는 저항성 반응을 보였으나K1 균계 접종구에서는 등숙률, 정현비율, 현미수량 및 완전미도정수율은 감소하였고 사미는 증가하는 등 이병성 반응을나타냈다. Xa21 유전자는 최근 큰 피해를 주는 K3a 균계에대해 강한 저항성을 보이나, 우리나라 우점 균계인 K1에 이병성을 보이기 때문에 K1 균계에 저항성인 다른 유전자와의집적을 통한 활용이 안정적인 저항성 증진에 효과적인 것으로 나타났다.