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Establishment of an Analytical Method for Novobiocin in Livestock Products Using HPLC-UVD (HPLC-UVD를 이용한 축산식품 중 Novobiocin의 시험법 확립)

  • Park, Hee-Ra;Kwon, Chan-Hyeok;Lee, Jong-Goo;Kim, Hyung-Soo;Chae, Young-Sik;Oh, Jae-Ho;Kwon, Ki-Sung
    • Korean Journal of Food Science and Technology
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    • v.44 no.3
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    • pp.263-268
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    • 2012
  • Novobiocin is a coumarin-containing antibiotic, and has a longer half-life in various animals than other veterinary medicines. A simple and rapid high-performance liquid chromatography assay for the determination of residual novobiocin levels in chicken, beef and milk has been developed and validated. The separation condition for HPLC/UVD was optimized by a MG II $C_{18}$ (4.6 mm $ID{\times}250$ mm, 5 ${\mu}m$) column with 0.1% formic acid in $H_2O$/0.1% formic acid in Acetonitrile (40/60, v/v) as the mobile phase at a flow rate of 1.0 mL/min and the detection wavelength was set at 340 nm. Residues were extracted from tissue by blending with methanol. After liquid-liquid partitioning, lipid materials were removed with n-hexane and purification as Silica (1 g, 6 mL) cartridge with 10 mL acetone/dichloromethane (10/90, v/v). Limit of quantification and linearity performed by the analytical method were 0.02 mg/kg and 0.999 ($r^2$), and the recovery range was $88.8{\pm}5.6-100.3{\pm}4.4$, $88.8{\pm}7.2-97.0{\pm}3.2$ and $88.1{\pm}4.3-92.8{\pm}3.6%$. It is expected that this analytical method with regards to novobiocin in chicken, beef and milk could be applied as an official method to administer food safety on veterinary medicines.

Adrenal and Testicular Androgens in Serum of Men after Physical Endurance Training (격심한 운동후 남성 혈청 내 부신 및 정소 Androgen 변화)

  • Yoon, Yong-Dal;Lee, Chang-Joo;Lee, Joon-Yeong
    • Development and Reproduction
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    • v.5 no.1
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    • pp.73-79
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    • 2001
  • Recently many studies have reported that total and bioavailable androgens reduced in male and female athletes and that physical exercise reduces the body weight and increases the reproductive abnormalities such as oligomenorrhea, anovulation, inadequate luteal phase, and delayed puberty in women by the inhibition of the hypothalamic-pituitary-gonadal (HPG) axis . In addition, high mileage endurance 겨nning, psychological stress, and military endurance training in men also reduce the secretion of reproductive hormones. To investigate the efffcts of physical endurance exercise on the secretion of reproductive hormones in men, androgenic hormones from adrenal glands and testis were measured in serum by the conventional radioimmunoassays after long-term (more than3 months), short-term (1 week), and acute (1${sim}$2 hours) physical exercises. Androgenic hormones from adrenal glands and testis such as total testosterone (TT), free testosterone (fT), dehydroepiandrosterone (DHEA), and androstenedione (A) decreased after thesestrenuous endurance trainings, whereas ACTH, cortisol, and dehydroepiandrosterone sulfetes (DHEAS) increased. Conadotropins (LH and FSH) were not idluenced by the physical exercises. Based upon the present results, we assume that the decrease in adrenal and testicular androgens by physical endurance exercises might be associated with the reproductive abnormalities in athletes by unknown factor(s) in addition to the HPG axis disturbance.

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Effects of Creep Feed and Milk Replacer and Nursery Phase-feeding Programs on Pre- and Post-weaning Growth of Pigs (돼지에서 입붙이 사료와 대용유 급여 및 이유자돈 사양프로그램이 이유 전과 이유 후의 성장에 미치는 영향)

  • Ha, Duck-Min;Jang, Kyoung-Soon;Won, Hye-Sook;Ha, Seung-Ho;Park, Man-Jong;Kim, Sung-Woo;Lee, C.-Young
    • Journal of Animal Science and Technology
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    • v.53 no.4
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    • pp.333-339
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    • 2011
  • The present study was performed to investigate the effects of pre- and post-weaning feeding programs on growth of pigs. A total of 24 litters (avg. 10.6 piglets/litter) born from multiparous (Yorkshire ${\times}$ Landrace) dams ${\times}$ Duroc sires were provided with neither creep feed (CF) nor milk replacer ("CON"), 200 gm CF/litter/d from the 15th day of lactation ("MIN"), or CF and milk replacer ad libitum for 7 h during the daytime from the 7th day ("MAX") through weaning at d 21 of age. Sixty-eight weanling pigs selected randomly from each of CON and MIN were provided with phases 1, 2 and 3 nursery diets for 7, 14, and 13 days, respectively, in two pens; an equal number of piglets from MAX received the same diets for 6, 10, and 18 days, respectively. Subsequently, all pigs were fed grower 1 and 2 diets sequentially up to d 95 and 135, respectively. The entire pre- and post-weaning feeding trial was repeated three times under a split-plot design of experiment. Initial and final weights and ADG of the suckling pigs did not differ between MAX/MIN and CON. However, final wt of MAX adjusted for initial wt, which was 0.17-kg less in MAX than in CON, was greater than that of CON by 0.31kg, whereas the difference between MIN and CON in final wt barely changed after the adjustment. Growth of the animals during the nursery and growing phases was not affected by the feeding program, whereas d 55 and 135 BW, as well as d 6 BW, were highly correlated with weaning weight (r=0.81, 0.57, and 0.76; P<0.001, <0.05, and <0.001, respectively). In conclusion, results suggest that provision of creep feed and milk replacer from early lactation may be effective for increasing weight gain of light piglets, but that limited provision of creep feed during late lactation or extension of the duration of phases 1 and 2 vs. 3 nursery diets for several days is unlikely to influence the growth of pigs during the corresponding and subsequent periods.

Annually Reproductive Cycles of Gonadotropic Cells, Endocrine Materials and Plasma Components in Special Relation to Oogenesis in Rainbow Trout, Oncorhynchus mykiss (번식주기에 있어서 자성무지개송어 (Oncorhynchus mykiss) 뇌하수체의 생식소자극호르몬 분비세포와 난형성에 특이하게 작용하는 내분비물질 및 혈장성분의 연중변화)

  • Yoon, Jong-Man;Kim, Gye-Woong;Park, Kwan-Ha
    • Applied Microscopy
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    • v.31 no.1
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    • pp.19-35
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    • 2001
  • Outlines for plasma $estradiol-17\beta$, components, electrophoretic patterns, and ultrastructural changes were obtained in female rainbow trout (Oncorhynchus mykiss) during the seasonal reproductive cycles. Plasma $estradiol-17\beta$ under the natural conditions, exhibited distinct seasonal variation, peaking very late in vitellogenic season during September, decreasing gradually the halt of spawning in December, and ultimately falling during the early stages of seasonal ovarian recrudescence in February and March. This change in $estradiol-17\beta$ appeared to stimulate vitellogenin production as evidenced by increases in plasma calcium, phosphorus, glucose, albumin and total protein levels. The electrophoretic patterns of late maturing or spawning oocytes were stained more intensively than those of late perinucleolus oocytes (molecular weights of approximately 70,000 and 200,000). Two protein bands were found in the SDS-PAGE separation, coincident with the $estradiol-17\beta$ hormone peak. Gonadosomatic indices (GSI) significantly increased from October to January, and showed the highest peak in January, coinciding with the numerically abrupt increase of ripe ova in female. A positive correlation (r=0.701, p<0.01) was established between plasma $estradiol-17\beta$ levels and the gonadosomatic index during the prespawning. The highest level of hepatosomatic index (HSI) observed in December. During the breeding season (December), the gonadotropes were large and filled with GTH-containing inclusions such as granules and globules. The vitellogenic phase began as late perinurleolus oocytes became transformed into early maturing oocytes through the accumulation of yolk, and oocytes reached the late maturing stages as the ooplasm was completely packed with yolk. Marked ultrastructural changed in the granulosa cells during nuclear migration involve the dilation of the rough endoplasmic reticulum and the appearance of the rod-shaped mitochondria with tubular cristae. Microvilli (finger-like projections), from the zona radiata and from the oocyte grew, and made contact with each other in the pore canals of the zona radials during vitellogenesis, but were withdrawn as the zona radiata became more compact and devoid of pore canals during oocyte maturation. The zona radiata grew to a tripartite structure such as an outer thin homogeneous layer, and two inner thick helicoidal layers (zona radials interna and zona radiata externa). Under the normal conditions, the ovarian follicle influenced the histological development and periodical secretion of the hormones , sufficient for a oogenesis and gonadal steroid production.

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A Study on the Formation of Lamellar Liquid Crystalline Using Skin Mimicking Surfactant (피부모사체 계면활성제를 사용한 라멜라 액정의 생성에 관한 연구)

  • Kim, In-Young;Nam, Eun-Hee;Shin, Moon-Sam
    • Journal of the Korean Applied Science and Technology
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    • v.37 no.3
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    • pp.484-495
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    • 2020
  • This study is a mixed surfactant (MimicLipid-MSM1000) that forms the same structure as that of the stratum corneum, sucrose distearate, polyglyceryl-2 dioleate, fermented squalane, ergosterol, 10-hydroxystearic acid, mixture consisting of was synthesized. When using 2~5 wt% of this mixed surfactant, it was possible to make an artificial skin mimetic that forms a multi-layer lamellar structure of 5~30 layers. An emulsion was prepared using this mixed surfactant, and a multi-layered lamellar phase was formed and analyzed mechanically. The appearance of this surfactant was a light brown hard wax, the hydrophilic lipophilic balance (HLB) was 12.53, the critical parameter value was 0.987, and the acid value was 0.13. Stability according to pH change was also stable in acidic (3.8), neutral (7.2) and alkaline (10.8). The particle size of the liquid crystal was found to be the most stable maltese cross lamellar crystalline droplet at 5~25mm. The size of the emulsified particles according to the change in the speed of the homo agitator is 2500 rpm (17.9mm±2.6mm), 3500rpm (12.5mm±2.1mm), 4500rpm (6.2mm±1.8mm) particles were formed. Liquid crystal forming particles were observed through a polarization microscope, and the formation structure of the liquid crystal was precisely analyzed with a scanning electron microscope (cryo-SEM). As an application field, it is expected that it will be widely applicable to the development of various prescriptions, such as various skin care cosmetics, makeup care cosmetics, and scalp protection cosmetics, by using a skin-mimicking surfactant.

Deposition of Poly-$Si_{1-x}Ge_x$ Thin Film by RTCVD (RTCVD에 의한 다결정 $Si_{1-x}Ge_x$ 박막 증착)

  • Kim, Jae-Jung;Lee, Seung-Ho;So, Myeong-Gi
    • Korean Journal of Materials Research
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    • v.5 no.6
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    • pp.690-698
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    • 1995
  • The Poly-S $i_{1-x}$G $e_{x}$ thin films were deposited on oxidized Si wafer by RTCVD(rapid thermal chemical vapor deposition) using Si $H_4$and Ge $H_4$, at 450 ~5$50^{\circ}C$. The variation of Ge mole fraction and the deposition rate of S $i_{1-x}$G $e_{x}$ thin film were studied as a function of the deposition temperature and the Ge $H_4$/Si $H_4$input ratio, and the crystal phase and the surface roughness were studied by XRD and AFM(atomic force microscopy), respectively. The experimental results showed that the activation energy for the deposition of poly-S $i_{1-x}$G $e_{x}$ was about 32~37Kca /mol and the deposition rate of S $i_{1-x}$G $e_{x}$ thin films was increased with increasing the deposition temperature and the input ratio. From the analysis of composition, it was known that the Ge mole fraction within the poly-S $i_{1-x}$G $e_{x}$ thin film was decreased with decreasing the input ratio and increasing the deposition temperature. As-deposited S $i_{1-x}$G $e_{x}$ thin films were polycrystalline over the entire experimental range. But those were amorphous at the deposition temperature of 450, 475$^{\circ}C$ and the input ratio of 0.05. By adding the Ge $H_4$, poly-S $i_{1-x}$G $e_{x}$ thin film were deposited at relatively lower deposition temperatures($\leq$ 5$50^{\circ}C$) than those of conventional poly-Si(>$600^{\circ}C$). From surface roughness measurement of poly-S $i_{1-x}$G $e_{x}$ it was found that the surface roughness( $R_{i}$ ) increased with increasing the deposition temperature and input ratio.and input ratio.

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Inhibitory Effects of Ethanolic Extracts from Aster glehni on Xanthine Oxidase and Content Determination of Bioactive Components Using HPLC-UV (섬쑥부쟁이 에탄올 추출물의 잔틴산화효소 저해 효능 및 HPLC-UV를 이용한 유효성분의 함량 분석)

  • Kang, Dong Hyeon;Han, Eun Hye;Jin, Changbae;Kim, Hyoung Ja
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.45 no.11
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    • pp.1610-1616
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    • 2016
  • This study aimed to establish an optimal extraction process and high performance liquid chromatography-ultraviolet (HPLC-UV) analytical method for determination of 3,5-dicaffeoylquinic acid (3,5-DCQA) as a part of materials standardization for the development of a xanthine oxidase inhibitor as a health functional food. The quantitative determination method of 3,5-DCQA as a marker compound was optimized by HPLC analysis using a Luna RP-18 column, and the correlation coefficient for the calibration curve showed good linearity of more than 0.9999 using a gradient eluent of water (1% acetic acid) and methanol as the mobile phase at a flow rate of 1.0 mL/min and a detection wavelength of 320 nm. The HPLC-UV method was applied successfully to quantification of the marker compound (3,5-DCQA) in Aster glehni extracts after validation of the method with linearity, accuracy, and precision. Ethanolic extracts of A. glehni (AGEs) were evaluated by reflux extraction at 70 and $80^{\circ}C$ with 30, 50, 70, and 80% ethanol for 3, 4, 5, and 6 h, respectively. Among AGEs, 70% AGE at $70^{\circ}C$ showed the highest content of 3,5-DCQA of $52.59{\pm}3.45mg/100g$ A. glehni. Furthermore, AGEs were analyzed for their inhibitory activities on uric acid production by the xanthine/xanthine oxidase system. The 70% AGE at $70^{\circ}C$ showed the most potent inhibitory activity with $IC_{50}$ values of $77.01{\pm}3.13{\sim}89.96{\pm}3.08{\mu}g/mL$. The results suggest that standardization of 3,5-DCQA in AGEs using HPLC-UV analysis would be an acceptable method for the development of health functional foods.

Organization and function of shoot apical meristem affecting growth and development in plants (식물의 생장과 발달에 영향을 미치는 슈트 정단분열조직의 체제와 기능)

  • Lee, Kyu Bae
    • Journal of Plant Biotechnology
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    • v.41 no.4
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    • pp.180-193
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    • 2014
  • In plants, a shoot apex has a small region known as the shoot apical meristem (SAM) having a group of dividing (initiating) cells. The SAM gives rise to all the groundabove structures of plants throughout their lifetime, and thus it plays important role in growth and development of plants. This review describes theories to explain the SAM organization and function developed over the last 250 years. Since in 1759 German botanist C. F. Wolff has described firstly the SAM, in 1858 Swiss botanist C. N${\ddot{a}}$geli proposed the apical cell theory from the observation of a large single apical cell in the SAM of seedless vascular plants: however, this view was recognized to be unsuitable to seed plants. In 1868, German botanist J. Hanstein suggested the histogen theory: this concept subdividing the SAM into dermatogen, periblem, and plerome was unable to generally apply to seed plants. In 1924, German botanist A. Schmidt proposed the tunica-corpus theory from the examination of angiosperm SAM in which two parts show different planes of cell division: this theory was proved to be not suitable to gymnosperm SAM, not have stable surface tunica layer. In 1938, American botanist A. Foster described zones in gymnosperm SAM based on the cytohistologic differentiation and thus called it a cytohistological zonation theory. With works by E. Gifford, in 1954, this zonation pattern was demonstrated to be also applicable to angiosperm SAM. As another theory, in 1952 French botanist R. Buvat proposed the m${\acute{e}}$rist${\grave{e}}$me d'attente (waiting meristem) theory: however, this concept was confuted because of its negation of function during vegetative growth phase to central initial cells. Rescent studies with Arabidopsis thaliana have found that formation and maintenance of the SAM are under the control of selected genes: SHOOTMERISTEMLESS (STM) gene forms the SAM, and WUSCHEL (WUS) and CLAVATA (CLV) genes function in maintaining the SAM; signaling between WUS and CLV genes act through a negative feedback loop.

Isolation and Characterization of a Marine Bacterium, Pseudomonas sp. YJ-1 with Anti-Methicillin Resistant Staphylococcus aureus Activity (항 Methicillin Resistant Staphylococcus aureus 활성을 가지는 해양미생물 Pseudomonas sp. YJ-1의 분리와 특성)

  • Woo, Ye-Ju;Jeong, Seong-Yun
    • Korean Journal of Environmental Biology
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    • v.35 no.4
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    • pp.694-705
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    • 2017
  • The aim of this study was to isolate and identify marine bacterium with anti-methicillin-resistant Staphylococcus aureus (MRSA) activity, and to purify the anti-MRSA compound, as well as to determine its activity and synergistic effects. Among the marine bacteria isolated in this study, the YJ-1 isolate had the strongest anti-MRSA activity. The YJ-1 isolate was identified on the basis of its biochemical characteristics and an analysis of 16S rRNA gene sequences. The YJ-1 isolate showed over 99.2% homology with Pseudomonas stutzeri, and was designated as a Pseudomonas sp. YJ-1. The optimal culture conditions were $25^{\circ}C$ and initial pH 7.0. For the purification of the anti-MRSA compounds, the YJ-1 was cultured in Pa PES-II medium, and the culture filtrates were extracted by ethyl acetate, hexane, and 80% MeOH. The 80% MeOH fraction was separated by a $C_{18}$ ODS column, silica gel chromatography and a reverse phase HPLC, to yield three anti-MRSA agents, the MR1, MR2, and MR3 compounds. When the MR1 compound of $250{\mu}g\;mL^{-1}$ concentration was applied to the MRSA cells, over 95% of bacterial cells was killed within 48 hr. Compared with vancomycin and ampicillin, the MR1 compound showed significant anti-MRSA activity. In addition, the anti-MRSA activity was increased by dose and time dependent manners. Furthermore, the combination of an MR1 compound with vancomycin produced a more rapid decrease in the MRSA cells than did the MR1 compound alone. Taken together, our results suggest that the Pseudomonas sp. YJ-1 and its anti-MRSA compounds could be employed as a natural antibacterial agent in MRSA infections.

Analytical Validation of Rosmarinic Acid in Water Extract of Perilla frutescens Britton var. acuta Kudo as Functional Health Ingredient (건강기능식품 기능성 원료로써 장흥 차조기 열수 추출물의 지표성분인 로즈마린산 분석법 검증)

  • Park, Sung-Yong;Kim, Jung-Eun;Choi, Chul-Yung;Lee, Dong-Wook;Kim, Ki-Man;Yoon, Goo;Yoon, In-Su;Moon, Hong-Seop;Cho, Seung-Sik
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.44 no.1
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    • pp.85-88
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    • 2015
  • This study attempted to establish an HPLC analysis method for determination of marker compounds as a part of material standardization for the development of health functional food materials from Perilla frutescens Britton var. acuta Kudo. The quantitative determination method of rosmarinic acid as a marker compound of P. frutescens Britton var. acuta Kudo extract (PFE) was optimized by HPLC analysis using a C18 column ($4.6{\times}150mm$, $5{\mu}m$) with 0.1% acetic acid as the elution gradient and methanol as the mobile phase at a flow rate of 1 mL/min and detection wavelength of 280 nm. The HPLC/UV method was applied successfully to quantification of the marker compound in PFE after validation of the method with linearity, accuracy, and precision. The method showed high linearity in the calibration curve at a coefficient of correlation ($R^2$) of 0.9995, and the limit of detection and limit of quantitation were $0.36{\mu}g/mL$ and $1.2{\mu}g/mL$, respectively. Relative standard deviation (RSD) values of data from intra- and inter-day precision were less than 3.21% and 1.43%, respectively. Recovery rate test at rosmarinic acid concentrations of 12.5, 25 and $50{\mu}g/mL$ scored between 97.04~98.98% with RSD values from 0.25~1.97%. These results indicate that the established HPLC method is very useful for the determination of marker compound in PFE to develop a health functional material.