에너지 음료는 카페인을 주성분으로 타우린, 비타민 같은 다른 energy-enhancing 성분을 함유하고 있다. 미국과 유럽에서는 글루쿠로노락톤이 에너지 음료에 첨가될수 있으나, 국내에서 의약품으로는 허가되어 있다. 따라서 식품 첨가물로는 그 사용이 금지 되어 있어, 지속적으로 수입 및 유통 음료에서 시험검사를 하여 규제하고 있다. 현재 분석법으로 사용하는 LC-PDA 법은 복잡한 유도체화 과 정을 거치고, 음료 중에 당류들이 위양성 결과를 나타내 기도 한다. 이런 기존 방법의 단점을 개선하기 위해 HILICESI- MS/MS (hydrophilic interaction liquid chromatography coupled to electrospray ionization tandem mass spectrometry) 를 이용한 분석법을 개발하고, 선택성, 직선성, 검출한계, 정량한계, 정밀도, 정확성, 재현성에 대하여 분석법 유효성 검증을 수행했고, AOAC, EURACHEM 가이드라인에 부합되는 결과를 얻었다.

The mammary gland is a complex organ, made up of various cell types that work together for mil synthesis. A previous study had established a clonal cell line from primary bovine mammary alveolar cells (MAC-T) for the study of bovine milk production and synthesis. In this study, we transplanted MAC-T bovine alveolar cell line to BALB/C nude mice for regeneration of bovine mammary gland alveolar ducts. The MAC-T cells were suspended with matrigel and injected into the dorsal tissue of 8 weeks old male BALB/C nude mice. After 6 weeks, the injected cells were showed typical morphology of the tubuloalveolar female glands by histological analysis. It was made the branching ducts that were surrounded by smooth muscle with small alveoli budding off the ducts. In addition, the epithelial markers CK14 and CK18 were expressed within the duct-like structure. Prolactin was detected in the duct interior in these CK14 and CK18 positive cells but not in the non-transplanted MAC-T cells. These results showed that duct-like tissue had been successfully formed after 6 weeks of transplantation of the CK14 and CK18 positive MAC-T cells into mice dorsal tissue. This mouse model will be a useful tool for further research on the bovine mammary gland.

Oral exposure of humans by excess amounts of arsenic may cause disturbances of the reproductive system. In the present study, such exposure was modelled in rats, with the support of sperm principal parameters and histopa-thological observations. Male Sprague–Dawley rats were randomly divided into three groups where the group I was served as a normal control, group II was received sodium meta-arsenite as arsenic (10 mg/kg b.w/day) and a combi-nation of sodium meta- arsenite and sodium selenite (3 mg/kg b.w/day) in group III. After 6 weeks, there was no significant change in testis weight and in total motility of all the three experimental groups, whereas, rapid moving spermatozoa, moderately moving spermatozoa and slow moving spermatozoa were significantly decreased in arsenic treated rats as compared to control rats. The other sperm principal parameters like progressiveness, average path velocity, straightness linear velocity (VSL), curvilinear velocity (VCL), straightness, linearity sperm head elongation ratio, area, linearity amplitude of lateral head department (ALH) and beat cross frequency (BCF) were found to be reduced in arsenic intoxicated rats. These results are not correlated with the histological studies. On oral admini-stration of selenium ameliorated the adverse effects of arsenic as compared to arsenic alone treated rats. Our findings clearly demonstrate that administration of selenium could prevent some of the deleterious effects of arsenic in the testis.

Artificial insemination (AI) has been performed widely in swine industry using fresh liquid sperm instead of frozen type of sperm. However fresh sperm are not able to preserve more than three days with optimal motility and other sperm parameters for the successful fertilization, since in vitro stored sperm has an oxidative stress that resulted increase of abnormality and acrosome reation. To overcome these major problems, novel preservative formulation is needed to neutralize the oxidative stress and to provide suitable physiological environment for sperm in in vitro. In this study, naturally derived substances such as Poncirus trifoliate (Trifoliate orange), Garcinia mangostana (Mangosteen), pig placenta and testis extracts were tested as sperm preservative agents. Placenta extracts (PE), trifoliate orange extracts (TOE), testes extracts (TE) and mangosteen extracts (ME) were applied to analyze specific parameters for sperm motion characteristics individually and combinatorial. Each individual extract treatment can accelerate the sperm motility but noticeably TOE, TE and ME treatments exhibited the considerable and significant preservation of sperm motility. PE, TE and ME showed a significant (p<0.05) increase in ALH after one week. Further we evaluated the five different combinations of these extracts on sperm motility and its motion characteristics. Surprisingly even after one week ME, TOE and TE combination significantly preserved the sperm motility about 75%. It is noteworthy that unlike individual extract treatment, combination of ME, TOE and TE simultaneously protect the sperm motility and its motion characteristics. Taken together these data conclude that addition of ME, TOE and TE can be effective for preservation of pig sperm.

INTRODUCTION In rodent, molecular markers of spermatogonia, spermatocyte, spermatid and sperm have been identified. It has been reported that DBA, PGP 9.5 and NanpG can be the markers for spermatogonia in pig. For further understanding the spermatogenesis of the pig on morphological and molecular level, we report identification of testicular cells in neonatal and pubertal pig testis, and a putative marker for spermatogonia and spermatid in pig testis. MATERIALS AND METHODS Neonatal (3 day) and pubertal testis (150 day) was cut and fixed in Bouin’s solution for immunohistochemistry (IHC). Gonocytes were isolated from neonatal testis for immunocytochemistry (ICC). Based on references (Phillips et al., 2010), thirteen antibodies (VASA, Oct4, NanoG, PGP 9.5, DAZL, SCF, GFR-alpha 1, PLZF, c-kit, integrin-beta1, Thy1, Sohlh1 and CD9) were used for IHC and ICC. Paraffin section was performed for IHC. Gonocytes were attached to the APS-coated slides for ICC. HRP-conjugated and florescent-labeled secondary antibody was used for IHC and ICC, respectively. RESULTS In histological analysis, spermatogonia and Sertoli cells, which are enclosed by seminiferous tubule and connective tissue, were observed in neonatal testis. However, complete spermiogenesis, including spermatocyte, spermatid and spermatozoa, was not observed in neonatal testis. Spermatocyte, spermatid and elongated spermatid were observed in pubertal testis but matured spermatozoa were not observed. As a result, two antibodies (PGP 9.5 and GFR-alpha1) of thirteen antibodies were available for IHC and ICC. As reported in other studies, PGP 9.5 protein was detected in spermatogonia of ne-onatal in IHC. In addition, it was observed in spermatogonia of pubertal testis. GFR- alpha1 protein was detected in spermatogonia of neonatal testis and spermatid of pubertal testis. In ICC, PGP 9.5 protein was detected in gonocytes as reported in other studies. GFR-alpha1 was also observed in gonocyte isolated from pig testis. In this study, we found that 1) only spermatogonia exist in neonatal pig testis (3 day), 2) GFR-alpha1 is a new marker for spermatogenesis in pig testis.

Interferon induced transmembrane protein-1 (IFITM1) is one of transmembrane protein which is differentially expressed in uterus during estrus cycle and pregnancy, that IFITM1 gene is highly expressed in estrus stage by the effect of estrogen, and in parturition by the effect of PGF2 alpha. This genes are also up-regulated in cells with hyperactivation of the WNT/β-catenin signaling pathway. In this study, to identify a function of IFITM1, the binding partner of IFITM1 were determined using immunoprecipitation and LC- MASSMASS methods. 1, 3 and 5 ug of polyclonal anti-IFITM1 antisera were used for immunoprecipitation, and the 75 kDa of specific band was detected in silver stained polyacylamide gel. This band were chracterized using LC-MASS-MASS, and revealed this band is glucose regulated protein 75 (GRP75) which binds to p53 and inhibits the p53 action in nucleus. To identify the localization of GRP75 in cells, immunocytochemical approach has been applied, and GRP75 is expressed in mitochondria of L929 murine connective tissue cells. Co-localization study between IFITM1 and GRP75 in L929 cell identified that these two proteins were closely expressed in mitochondria. Although the role of the interaction of these two protein need to be clarified in various biological phenomena, this data suggest that close interaction of IFITM1 and GRP75 may regulate cellular functions in uterus on sets of estrus cycle and pregnancy.

SERPINB3 (also known Squamous cell carcinoma antigen 1, SCCA1) is involved in apoptosis, immune response, cell migration and invasiveness of cells. It has been investigated in various types of squamous cell carcinoma. Therefore we investigated the functional role of SERPINB3 gene in human epithelial ovarian cancer (EOC) using laying hens, the most relevant animal model. In 136 laying hens, EOC was found in 10 (7.4%). We compared the expression and localization of SERPINB3 using RT-PCR, quantitative RT-PCR, in situ hybridization and immunohistochemistry, and SERPINB3 activation was detected in chicken and human ovarian cancer cell lines using immunofluorescence microscopy. Thereafter, we examined the prognostic value of SERPINB3 expression in patients with EOC by multivariate linear logistic regression and Cox’ proportional hazard analyses. In present study, SERPINB3 mRNA was induced in cancerous ovaries (p< 0.01), and it was only expressed in the glandular epithelium(GE) of cancerous ovaries of laying hens. SERPINB3 protein was localized predominantly to the nucleus of glandular epithelium in cancerous ovaries of laying hens, and it was abundant in the nucleus of both chicken and human ovarian cancer cell lines. In 109 human patients with EOC, 15 (13.8%), 66 (60.6%) and 28 (25.7%) of those patients showed weak, moderate and strong expression of SERPINB3 protein, respectively. Strong expression of SERPINB3 protein was a prognostic factor for platinum resistance (adjusted OR, 5.94; 95% Confidence Limits, 1.21-29.15). Therefore SERPINB3 may play an important role in ovarian carcinogenesis and be a novel biomarker for predicting platinum resistance and a poor prognosis for survival in patients with EOC. This research was funded by the World Class University (WCU) program (R31-10056), Basic Science Research Program (2010- 0013078) through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science, and Technology and by the Next-Generation BioGreen 21 Program (No.PJ008142), Rural Development Administration, Republic of Korea.

하천수사용 관리를 적정하게 관리하기 위해서는 객관적인 하천수사용량 자료가 수집되어야 한다. 그러나, 하천수 사용자의 취수여건을 고려하여 유량계 외에 수문조작 등의 간접적인 계측방법을 인정하고 있고, 사용자의 자발적인 보고에 의존함에 따라 자료의 객관화가 어려운 실정이다. 이에 한강홍수통제소에서는 농업용수 사용 비중이 커 하천유량 파악이 어려운 만경강의 고산~봉동 수위관측소 구간에 위치한 어우보 취수로에 V-ADCP를 이용한 계측시설을 설치 및 운영하고 있다. 본 연구에서는 V-ADCP로 측정된 유속을 이용하여 실시간 하천수사용량 산정을 위한 유속분포법의 적 용성을 평가하였다. 이를 위해 Chiu의 2차원 유속분포식의 매개변수 민감도를 분석하고, 실측유량 자료에 기초한 최적 매개변수를 산정하였다. 또한 수위-유량관계법, 지표유속법과 비교 평가하여 유속분포법의 특성을 분석하였다.

Background : This study aimed to determine the use of proper organic fertilizer through side dressing application in the environmental cultivation of Adenophora triphylla var. japonica Hara. to improve farming income. Methods and Results : In the cultivation of eco-friendly and environmental medicinal plant like Adenophora triphylla var. japonica Hara., it is necessary to use organic instead of chemical fertilizer. Organic fertilizers such as HL, CK, OK were applied three (3) times during the last ten days of July, August, and September. There was no significant difference between CF and NT, in terms of the number of rooting per 1㎡ which was 12.1 -12.9. There were also no significant differences in plant length, leaf length, leaf width and dry weight. The root length was 19.9 ㎝, which was the longest in CK, and 16.8 ㎝, the shortest in ST, but described as no significant difference. The root width was 14.8 – 5.5 ㎜, and the number of rootlet was 5.7 - 7.1, but was considered not a significant difference. The dry weight was 16.6 g in HL, 16.3 g in CK and ST, and 15.6 g in OK, but still there were no significant differences observed. However, it was 14.6 g in NT, which was considered the lowest, thus there was a significant difference. The number of dry root per 10 a was 149 ㎏ which is considered the most in HL, 141 ㎏ in CK, 140 ㎏ in ST, and 138kg in OK yet there were no significant differences observed. While NT only had 123 ㎏ hence, there was a significant difference on the number of dry root per 10a as compared with the group applied with the three organic fertilizers. Conclusion : In conclusion, it is proper to use Organic Fertilizer HL and CK through side dressing application in growing Adenophora triphylla var. japonica Hara.

Background : This project was conducted to determine proper seed harvest time and seed storage methods of Adenophora triphylla var. japonica Hara. Methods and Results : When Adenophora triphylla var. japonica Hara. transplanted this year, seeding can only be done the following year. Therefore, the seeds harvested in the interval of ten (10) days were investigated for seed harvest amount, germination rate, seed storage temperature and storing on container, between the period late August and mid-October. It is when the seed fructification started and ended. The seeds harvested from the first ten (10) days of October to mid October were 87.9% in whole seeds. The proper seed harvesting time was mid October which has a germination rate of 75% in whole seed harvest time. The lowest germination rate of 27.3% was at the germination temperature of 15℃. As the temperature rises, the germination rate increases. In 29℃, the germination rate was at 79% which is the highest; and from 30℃, the germination rate decreased to 71%. When the storage temperature was at 0℃ after storage of 5 months, the germination rate was at 83%. However, after storage of 12 months, it decreased by 7% that was 76%. When in 4℃, after storage of 5 months the germination rate was at 85%. However, after storage of 12 months it decreased by 8% which was 77%. In room temperature after storage of 5 months, germination rate was at 77%, but after storage of 12 months it decreased by 22% which was 55%. When seeds were stored in vinyl container, it was observed that the germination rate was at 86% after 5 months of storage, which was 9% higher than seed stored in paper which was 77%. After 12 months storage, seed in vinyl container had a germination rate of 71%, which was 3% higher than seed in paper at 68%. Conclusion : It is concluded that the proper time of seed gathering of Adenophora triphylla var. japonica Hara. is during the first ten (10) days of October to mid-October. When the long time storage is needed, below zero temperature with vinyl container is more appropriate.

Background : Adenophora triphylla var. japonica Hara. is one of the important oriental medicinal herbs being used to cure lung/respiratory diseases. It belongs to the five Sam (ginseng) including Panax ginseng, Salvia miltiorrhiza, Sophora flavescens, and Scrophularia buergeriana. In 2015, Korea produced 72 tons of medicinal herbs in a 14-hectare area and imported 24 tons valued at US$95,000. This study was carried out to investigate the effect of several biopesticides to control leaf spot of A. triphylla var. japonica Hara. Methods and Results : One year plants at the Department of Herbal Crop Research, NIHHS, RDA were used. The treatments used biopesticides such as plant extracts (BP), microbial agent (DM), microbial agent (PJ), sulfur powder (HS), and controlled chemicals (ST), while non treatment (NT) was used as control. After the early part of disease outbreak, the plants were treated four times with 7 days interval. The incidence of leaf spot, efficacy, phytotoxicity and control value were investigated. Based on the efficacy test, incidence of leaf spot is lowest at 15.6% in plant extracts (BP). Other biopesticides showed higher incidence - 16.7% for microbial agent (DM), 17.8% for microbial agent (PJ), 18.9% for sulfur powder (HS) and 71.1% for non treatment (NT), compared with 11.1% in controlled chemicals (ST). The microbial agent (PJ), sulfur powder (HS), plant extracts (BP) and microbial agent (DM) were not damaged in the phytotoxicity test. In the test of application dose, time and methods, the control value was highest in plant extracts (BP) at 78.1%, followed by microbial agent (DM) at 76.6%, microbial agent (PJ) at 75.0%, and sulfur powder (HS) at 73.4% , compared with controlled chemicals (ST) at 84.4%. Conclusion : It is possible to use biopesticides at the proper time to prevent leaf spot in Adenophora triphylla var. japonica Hara.

Background : Adenophora triphylla var. japonica Hara. is used to cure respiratory/ lung diseases, which makes it an important oriental medicinal herb. It is one of the five Sam (ginseng), which includes Panax ginseng, Salvia miltiorrhiza, Sophora flavescens, and Scrophularia buergeriana. In 2015, Korea imported 24 tons of medicinal herbs worth US$95,000. This study was carried out to investigate the effect of several mulching materials on the growth and root yield of A. triphylla var. japonica using organic fertilizer. Methods and Result : Seeds of A. triphylla var. japonica were collected at the nursery of the Department of Herbal Crop Research, NIHHS, RDA in 2015. Mulching materials used are black plastic, black woven fabric, and biodegradable plastic. Hand weeding was done to serve as control. The soil was fertilized two months before planting. Seeds of A. triphylla var. japonica were sowed in tray in early March and were allowed to grow for two months. It was planted in early May with planting density of 30 ㎝ x 15 ㎝ and was covered with various mulching materials. Parameters investigated are survival rate, aerial part and root growth characteristics, and yield at harvest time. Results showed that biodegradable plastic is most favorable for plant growth with height of 17.2cm, leaf length of 13.4cm and leaf width of 5.6 ㎝. Dry plant weight was heaviest in biodegradable plastic treatment at 2.8 g, and lightest at 2.5 g when planted without mulch. Biodegradable plastic has the highest dry root weight of 19.3 g, followed by black vinyl (18.4 g), woven fabric (18.3 g) and without mulch (17.9 g). The yield obtained per 10 a was highest in biodegradable plastic treatment at 174 ㎏, while black vinyl and woven fabric treatments produced 169 ㎏ and 160 ㎏, respectively. Plants without mulch had the lowest yield of 157 ㎏. Conclusion : In the above results, biodegradable plastic and woven fabric are considered suitable mulching materials for the cultivation of A. triphylla var. japonica.

“Jecy Gold” is a new kiwifruit variety developed at the National Institute of Subtropical Agriculture (NISA), RDA in 2003, which was for the utilization as fruit with yellow flesh and high soluble solids. This variety was selected from seeds obtained from the cross breeding between Actinidia chinensis cv. “Golden Yellow” and A. chinensis cv. “Songongu” with yellow flesh in 1997. Seedling and line selections were conducted from 1998 to 1999, and characteristic trials were carried out from 2000 to 2002. After developed, it was also conducted to the test of adaptability in kiwifruit orchards of Jeju island from 2003 to 2006. The branch of “Jecy Gold” sprouts strongly and the color of leaf is slightly dark green. The shape of fruit is obovoid, and the color of peel is yellowish-brown without hair. The average weight of fruit is 128.4 g. The core is middle and soft, the percicarp is golden yellow, and the texture is fragile and juicy. Soluble solids concentration is 14.8 oBrix. The total acid content is 0.69% and the fruit yield is higher about 30% than that of A. deliciosa cv. “Hayward”. The content of vitamin C is higher than that of A. deliciosa cv. “Hayward”. The harvesting time is from late October to early November and the fruit can be preserved about 90 days at 2°C. This variety can be planted bellow 100 m sea level and should be grown under shelter to prevent from disease and cold. This variety will be cultivated about 100 ha by 2011 in the southern part of Korea and has been supplied about 10 ha in 2007.