• Title/Summary/Keyword: phase-contrast

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Radiation-induced Apoptosis is Differentially Modulated by PTK Inhibitors in K562 Cells (K562 백혈병 세포주에서 방사선에 의해 유도되는 Apoptosis에 미치는 PTK Inhibitors의 영향)

  • Lee Hyung Sik;Moon Chang Woo;Hur Won Joo;Jeong Su Jin;Jeong Min Ho;Lee Jeong Hyeon;Lim Young kin;Park Heon Joo
    • Radiation Oncology Journal
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    • v.18 no.1
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    • pp.51-58
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    • 2000
  • Purpose :The effect of PTK inhibitors (herbimycin A and genistein) on the induction of radiation-induced apoptosis in Ph-positive KS62 leukemia cell line was investigated. Materials and Methods :K562 cells in exponential growth phase were irradiated with a linear accelerator at room temperature. For 6 MV X-ray irradiation and drug treatment, cultures were initiated at 2×106 cells/mL. The cells were irradiated with 10 Gy. Stock solutions of herbimycin A and genistein were prepared in dimethyl sulphoxide (DMSO). After incubation at 37$^{\circ}C$ for 0$\~$48 h, the extent of apoptosis was determined using agarose gel electrophoresis and TUNEL assay. The progression of cells through the cell cycle after irradiation and drug treatment was also determined with flow cytometry. Western blot analysis was used to monitor bel-2, bel-X$_{L}$ and bax protein levels. Results :Treatment with 10 Gy X-irradiation did not result in the induction of apoptosis. The HMA alone (500 nM) also failed to induce apoptosis. By contrast, incubation of K562 cells with HMA after irradiation resulted in a substantial induction of nuclear condensation and fragmentation by agarose gel electro-phoresis and TUNEL assay. Genistein failed to enhance the ability of X-irradiation to induce DNA fragmentation. Enhancement of apoptosis by HMA was not attributable to downregulation of the bel-2 or bel-X$_{L}$ anti-apoptotic proteins. When the cells were irradiated and maintained with HMA, the percentage of cells in G2/M phase decreased to 30$\~$40$\%$ at 48 h. On the other hand, cells exposed to 10 Gy X-irradiation alone or maintained with genistein did not show marked cell cycle redistribution. Conclusion : We have shown that nanomolar concentrations of the PTK inhibitor HMA synergize with X-irradiation in inducing the apoptosis in Ph (+) K562 leukemia cell line. While, genistein, a PTK inhibitor which is not selective for p210$^{bcr/abl}$ failed to enhance the radiation induced apoptosis in KS62 cells. It is unlikely that the ability of HMA to enhance apoptosis in K562 cells is attributable to bel-2 family. It is plausible that the relationship between cell cycle delays and cell death is essential for drug development based on molecular targeting designed to modify radiation-induced apoptosis.

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Effect of Water Activity and Temperature on Growth, Germination, Sporulation, and Utilization of Carbon Source of Penicillium oxalicum (PENOX) as a Biocontrol Agent(BCA) for control of Clover(Trifolium repens L.) (토끼풀(Trifolium repens L.) 방제용 생물제제 Penicillium oxalicum (PENOX)의 발아, 생장, 포자생성 및 탄소원이용에 미치는 수분활성 및 온도의 영향)

  • Lee, Hyang-Burm;Kim, Chang-Jin
    • The Korean Journal of Pesticide Science
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    • v.4 no.3
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    • pp.68-74
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    • 2000
  • Penicillium oxalicum (PENOX) has shown the potential as a biocontrol agent(5CA) for control of a weed, clover(Trifolium repens L.) in grass plots. The bioherbicidal activity may be due to germinative and growth capacities and substrate availability of the agent over a range of environmental factors. The influences of different water activities($0.94{\sim}0.995\;a_w$) and temperatures($18{\sim}30^{\circ}C$) on mycelial growth, conidial germination, sporulation oil 2% MEA(malt extract agar) adjusted to different water activities with glycerol, and carbon source utilization using BIOLOG GN MicroPlate were determined in vitro. Decreases in $a_w$ on MEA caused a reduction in mycelial growth and conidial germination depending on temperature. The mycelial growth of PENOX was greatest at $30^{\circ}C/0.995\;a_w$. At some lowered water activity($0.97\;a_w$), the growth was similar between 25 and $30^{\circ}C$, and considerably decreased at lowered temperature($20^{\circ}C$). The germination rate was also greatest at $30^{\circ}C/0.995\;a_w$. Lag phase times for PENOX at $18^{\circ}C$ on MEA were >6hrs at tile whole $a_w$ level tested, and at 18 and $25^{\circ}C$ they were >18hrs and >12hrs at $0.94\;a_w$, respectively. However, its sporulation was some better at $0.97\;a_w$ than $0.995\;a_w$ or $0.94\;a_w$, and better at $20^{\circ}C$ than $30^{\circ}C$. In contrast, the number of carbon sources(niche size) utilized by PENOX varied with $a_w$ and temperature. Under some water stress condition($0.95\;a_w$), the agent utilized smaller number of carbon sources than $0.995\;a_w$ depending on temperature. The niche size at 0.995 and $0.95\;a_w$ were highest at $25^{\circ}C$, and showed 86 and 65, respectively. At $30^{\circ}C$, the niche size at 0.995 and $0.95\;a_w$ showed 84 and 50, respectively. There was no carbon source utilized by PENOX at $0.90\;a_w$ regardless of temperature. These information of tile fungal ecophysiology will be useful for the effective development of BCA.

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Properties of Dissolved Organic Carbon (DOC) released by Three Species of Blue- green Algae (남조류에 의해 배출된 용존유기탄소의 특성)

  • Choi, Kwang-Soon;Imai, Akio;Kim, Bom-Chul;Matsushige, Kazuo
    • Korean Journal of Ecology and Environment
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    • v.34 no.1 s.93
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    • pp.20-29
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    • 2001
  • The amount, chemical composition and optical property of extracellular dissolved organic carbon (EOC) by phytoplankton were examined using axenic cultures of Microcystis aeruginosa, Anabaena flos-aquae, and Oscillatoria agardhii. The extracellular organic matter was categorized into five fractions (hydrophobic acids; AHSs, hydrophobic neutrals; HoNs, hydrophilic acids; HiAs, hydrophilic bases; HiBs, and hydrophilic neutrals; HiNs) using three adsorbent resins(XAD-8, cation, and anion). The release pattern and chemical composition of EOC varied with algal species and their growth phases. Percentage of extracellular release increased with age in all cultures. HiAs were the dominant component of EOC in all cultures, whereas the proportion of HiAs decreased with age in all cultures. In contrast, the proportions of HiBs and HiNs increased as cultures aged. In particular, the HiN fraction increased from 0% to 44% of EOC in M. aeruginosa and from 3.0% to 28% in A. flos-aquae, respectively. The proportion of AHSs was higher in the cultures of A. flos-aquae(7.5${\sim}$16%) and O. agardhii (8.7${\sim}$16%) than M. aeruginosa(0.2${\sim}$2.5%). The proportions of AHSs increased with culture age in M. aeruginosa and O. agardhii, but decreased in A. flos-aquae. The specific UV absorbance also varied among species; 1.9 Abs${\cdot}$cm$^{-1}$/mgC${\cdot}$L$^{-1}$ for M. aeruginosa, 3.7 Abs${\cdot}$cm$^{-1}$/mgC${\cdot}$L$^{-1}$ for A. flos-aquae, and 13.0 Abs${\cdot}$cm$^{-1}$/mgC${\cdot}$L^{-1}$ for O. agardhii. The results of this study indicates that DOC excreted by three blue-green algae differed with species and the growth phase.

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Exploring the Temporal Relationship Between Traffic Information Web/Mobile Application Access and Actual Traffic Volume on Expressways (웹/모바일-어플리케이션 접속 지표와 TCS 교통량의 상관관계 연구)

  • RYU, Ingon;LEE, Jaeyoung;CHOI, Keechoo;KIM, Junghwa;AHN, Soonwook
    • Journal of Korean Society of Transportation
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    • v.34 no.1
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    • pp.1-14
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    • 2016
  • In the recent years, the internet has become accessible without limitation of time and location to anyone with smartphones. It resulted in more convenient travel information access both on the pre-trip and en-route phase. The main objective of this study is to conduct a stationary test for traffic information web/mobile application access indexes from TCS (Toll Collection System); and analyzing the relationship between the web/mobile application access indexes and actual traffic volume on expressways, in order to analyze searching behavior of expressway related travel information. The key findings of this study are as follows: first, the results of ADF-test and PP-test confirm that the web/mobile application access indexes by time periods satisfy stationary conditions even without log or differential transformation. Second, the Pearson correlation test showed that there is a strong and positive correlation between the web/mobile application access indexes and expressway entry and exit traffic volume. In contrast, truck entry traffic volume from TCS has no significant correlation with the web/mobile application access indexes. Third, the time gap relationship between time-series variables (i.e., concurrent, leading and lagging) was analyzed by cross-correlation tests. The results indicated that the mobile application access leads web access, and the number of mobile application execution is concurrent with all web access indexes. Lastly, there was no web/mobile application access indexes leading expressway entry traffic volumes on expressways, and the highest correlation was observed between webpage view/visitor/new visitor/repeat visitor/application execution counts and expressway entry volume with a lag of one hour. It is expected that specific individual travel behavior can be predicted such as route conversion time and ratio if the data are subdivided by time periods and areas and utilizing traffic information users' location.

Renal Sympathetic Denervation Induces Acute Myocardial Inflammation through Activation of Caspase-1 and Interleukin-1β (교감신경절제술(RDN) 후 caspase-1과 interleukin-1β 활성화로 인해 유발된 염증성 급성심근손상)

  • Lee, Dong Won;Kim, Il Young;Kwak, Ihm Soo
    • Journal of Life Science
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    • v.28 no.2
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    • pp.240-246
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    • 2018
  • Efferent and afferent sympathetic nerves are closely related to the development of hypertension and heart failure. Catheter-based renal sympathetic denervation (RDN) is implemented as a strategy to treat resistant hypertension. We investigated whether RDN procedure causes inflammatory damage on myocardium in the early phase of sympathetic denervation. Twenty-five female swine were divided into 3 groups: normal control (Normal, n=5), sham-operated control (Sham, n=5), and RDN groups (RDN, n=15). The RDN group was further subdivided into 3 subgroups according to the time of sacrifice: immediately (RDN-0, n=5), 1 week (RDN-1, n=5), and 2 weeks (RDN-2, n=5) after RDN. There were no significant changes in the clinical parameters between the normal control and sham-operated group using contrast-media. In the myocardium, inflammatory cytokines, $IL-1{\beta}$ and $TNF-{\alpha}$ increased at the first week, and then decreased at the second week after RDN. Anti-inflammatory cytokine, IL-10 increased immediately, and then decreased at the second week after RDN. Caspase-1 activity and apoptosis-associated speck-like protein containing a caspase recruitment domain (ASC) expression increased immediately after RDN until the second week. However, nucleotide-binding oligomerization domain, leucine rich repeat and pyrin domain containing protein 3 (NLRP3) expression did not show any significant differences among the groups. The RDN can cause acute myocardial inflammation through activation of caspase-1 and $IL-1{\beta}$. We should pay attention to protecting against early inflammatory myocardial damage after RDN.

THE MORPHOLOGICAL OBSERVATION OF HUMAN PERIODONTAL LIGAMENT CELLS ATTACHMENT AND SPREADING ON THE SURFACE OF SLIDE GLASS (치주인대세포의 부착과 전개에 관한 형태학적 관찰)

  • Lee, Jin-Mi;Suh, Jo-Young;Park, Joon-Bong
    • Journal of Periodontal and Implant Science
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    • v.23 no.1
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    • pp.97-108
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    • 1993
  • One of the important initial events required for periodontal regeneration is the attachment and subsequent spreading of periodontal ligament cells on the root surface. The purposes of this study is to investigate the attachment and spreading pattern of human periodontal ligament cell on the surface of glass slides. After establishment of a cell line of the primary cell culture from the periodontal ligament of 1st premolar teeth which were extracted for the purpose of orthodontic treatment, author dispersed the cells at $5{\times}10^3\;cells/ml$ into the each 35mm culture petri-dish containing 2 glass slides. To observe the morphological changes of the cells which attached to the surfaces of glasses at every designed time schedule, author used the inverted phase contrast microscope and scanning electron microscope. During the whole experiment culture condition was at $37^{\circ}C$, 100% Humidity, 5% $CO_2$ gas incubator. The following results were obtained. Periodontal ligament cells showed spherical outline and started to attach to glass surface by basal sytoplasmic extension after 10min in culture. After 30min in culture, periodontal ligament cells were attached to glass surface by well - developed filopodia which protruded from the lamellipodia. The cell surface is covered with bubble-like structures and occasional microvillus can be seen with diffculty among these structures. After 1.5hr in culture, peridontal ligament cells shhowed radially well-spread cytoplasm and the nucleus was centered on its cytoplasm. Unspread central region of the cell was covered with numerous microvilli. The change of cell attachment and spreading pattern was manifest at 6hr in culture. At this time, periodontal ligament cell showed elongated outline and an oval-shaped nucleus. After 12hr in culture, periodontal ligament cells showed more stretched fibroblast-like appearance with polarity. Two long lamellipodia can be seen around the both terminal ends of cells. After 24hr in culture, periodontal ligament cells showed spindle shapes and an oval-shaped nucleus was slanted toward one side of the cell.

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A Polarization-based Frequency Scanning Interferometer and the Measurement Processing Acceleration based on Parallel Programing (편광 기반 주파수 스캐닝 간섭 시스템 및 병렬 프로그래밍 기반 측정 고속화)

  • Lee, Seung Hyun;Kim, Min Young
    • Journal of the Institute of Electronics and Information Engineers
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    • v.50 no.8
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    • pp.253-263
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    • 2013
  • Frequency Scanning Interferometry(FSI) system, one of the most promising optical surface measurement techniques, generally results in superior optical performance comparing with other 3-dimensional measuring methods as its hardware structure is fixed in operation and only the light frequency is scanned in a specific spectral band without vertical scanning of the target surface or the objective lens. FSI system collects a set of images of interference fringe by changing the frequency of light source. After that, it transforms intensity data of acquired image into frequency information, and calculates the height profile of target objects with the help of frequency analysis based on Fast Fourier Transform(FFT). However, it still suffers from optical noise on target surfaces and relatively long processing time due to the number of images acquired in frequency scanning phase. 1) a Polarization-based Frequency Scanning Interferometry(PFSI) is proposed for optical noise robustness. It consists of tunable laser for light source, ${\lambda}/4$ plate in front of reference mirror, ${\lambda}/4$ plate in front of target object, polarizing beam splitter, polarizer in front of image sensor, polarizer in front of the fiber coupled light source, ${\lambda}/2$ plate between PBS and polarizer of the light source. Using the proposed system, we can solve the problem of fringe image with low contrast by using polarization technique. Also, we can control light distribution of object beam and reference beam. 2) the signal processing acceleration method is proposed for PFSI, based on parallel processing architecture, which consists of parallel processing hardware and software such as Graphic Processing Unit(GPU) and Compute Unified Device Architecture(CUDA). As a result, the processing time reaches into tact time level of real-time processing. Finally, the proposed system is evaluated in terms of accuracy and processing speed through a series of experiment and the obtained results show the effectiveness of the proposed system and method.

Biological Functions of N- and O-linked Oligosaccharides of Equine Chorionic Gonadotropin and Lutropin/Chorionicgonadotropin Receptor

  • Min, K. S.
    • Proceedings of the KSAR Conference
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    • 2000.10a
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    • pp.10-12
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    • 2000
  • Members of the glycoprotein family, which includes CG, LH, FSH and TSH, comprise two noncovalently linked $\alpha$- and $\beta$-subunits. Equine chorionic gonadotropin (eCG), known as PMSG, has a number of interesting and unique characteristics since it appears to be a single molecule that possesses both LH- and FSH-like activities in other species than the horse. This dual activity of eCG in heterologous species is of fundamental interest to the study of the structure-function relationships of gonadotropins and their receptors. CG and LH $\beta$ genes are different in primates. In horse, however, a single gene encodes both eCG and eLH $\beta$-subunits. The subunit mRNA levels seem to be independently regulated and their imbalance may account for differences in the quantities of $\alpha$ - and $\beta$ -subunits in the placenta and pituitary. The dual activities of eCG could be separated by removal of the N-linked oligosaccharide on the $\alpha$-subunit Asn 56 or CTP-associated O-linked oligosaccharides. The tethered-eCG was. efficiently secreted and showed similar LH-like activity to the dimeric eCG. Interestingly, the FSH-like activity of the tethered-eCG was increased markedly in comparison with the native and wild type eCG. These results also suggest that this molecular can implay particular models of FSH-like activity not LH-like activity in the eCG/indicate that the constructs of tethered molecule will be useful in the study of mutants that affect subunit association and/or secretion. A single-chain analog can also be constructed to include additional hormone-specific bioactive generating potentially efficacious compounds that have only FSH-like activity. The LH/CG receptor (LH/CGR), a membrane glycoprotein that is present on testicular Leydig cells and ovarian theca, granulosa, luteal, and interstitial cells, plays a pivotal role in the regulation of gonadal development and function in males as well as in nonpregnant and pregnant females. The LH/CGR is a member of the family of G protein-coupled receptors and its structure is predicted to consist of a large extracellular domain connected to a bundle of seven membrane-spanning a-helices. The LH/CGR phosphorylation can be induced with a phorbol ester, but not with a calcium ionophore. The truncated form of LHR also was down-regulated normally in response to hCG stimulation. In contrast, the cell lines expressing LHR-t63I or LHR-628, the two phosphorylation-negative receptor mutant, showed a delay in the early phase of hCG-induced desensitization, a complete loss of PMA-induced desensitization, and an increase in the rate of hCG-induced receptor down-regulation. These results clearly show that residues 632-653 in the C-terminal tail of the LHR are involved in PMA-induced desensitization, hCG-induced desensitization, and hCG-induced down-regulation. Recently, constitutively activating mutations of the receptor have been identified that are associated with familial male-precocious puberty. Cells expressing LHR-D556Y bind hCG with normal affinity, exhibit a 25-fold increase in basal cAMP and respond to hCG with a normal increase in cAMP accumulation. This mutation enhances the internalization of the free and agonist-occupied receptors ~2- and ~17-fold, respectively. We conclude that the state of activation of the LHR can modulate its basal and/or agonist-stimulated internalization. Since the internalization of hCG is involved in the termination of hCG actions, we suggest that the lack of responsiveness detected in cells expressing LHR-L435R is due to the fast rate of internalization of the bound hCG. This statement is supported by the finding that hCG responsiveness is restored when the cells are lysed and signal transduction is measured in a subcellular fraction (membranes) that cannot internalize the bound hormone.

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Estimation of the Lowest and Highest Astronomical Tides along the west and south coast of Korea from 1999 to 2017 (서해안과 남해안에서 1999년부터 2017년까지 최저와 최고 천문조위 계산)

  • BYUN, DO-SEONG;CHOI, BYOUNG-JU;KIM, HYOWON
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.24 no.4
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    • pp.495-508
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    • 2019
  • Tidal datums are key and basic information used in fields of navigation, coastal structures' design, maritime boundary delimitation and inundation warning. In Korea, the Approximate Lowest Low Water (ALLW) and the Approximate Highest High Water (AHHW) have been used as levels of tidal datums for depth, coastline and vertical clearances in hydrography and coastal engineering fields. However, recently the major maritime countries including USA, Australia and UK have adopted the Lowest Astronomical Tide (LAT) and the Highest Astronomical Tide (HAT) as the tidal datums. In this study, 1-hr interval 19-year sea level records (1999-2017) observed at 9 tidal observation stations along the west and south coasts of Korea were used to calculate LAT and HAT for each station using 1-minute interval 19-year tidal prediction data yielded through three tidal harmonic methods: 19 year vector average of tidal harmonic constants (Vector Average Method, VA), tidal harmonic analysis on 19 years of continuous data (19-year Method, 19Y) and tidal harmonic analysis on one year of data (1-year Method, 1Y). The calculated LAT and HAT values were quantitatively compared with the ALLW and AHHW values, respectively. The main causes of the difference between them were explored. In this study, we used the UTide, which is capable of conducting 19-year record tidal harmonic analysis and 19 year tidal prediction. Application of the three harmonic methods showed that there were relatively small differences (mostly less than ±1 cm) of the values of LAT and HAT calculated from the VA and 19Y methods, revealing that each method can be mutually and effectively used. In contrast, the standard deviations between LATs and HATs calculated from the 1Y and 19Y methods were 3~7 cm. The LAT (HAT) differences between the 1Y and 19Y methods range from -16.4 to 10.7 cm (-8.2 to 14.3 cm), which are relatively large compared to the LAT and HAT differences between the VA and 19Y methods. The LAT (HAT) values are, on average, 33.6 (46.2) cm lower (higher) than those of ALLW (AHHW) along the west and south coast of Korea. It was found that the Sa and N2 tides significantly contribute to these differences. In the shallow water constituents dominated area, the M4 and MS4 tides also remarkably contribute to them. Differences between the LAT and the ALLW are larger than those between the HAT and the AHHW. The asymmetry occurs because the LAT and HAT are calculated from the amplitudes and phase-lags of 67 harmonic constituents whereas the ALLW and AHHW are based only on the amplitudes of the 4 major harmonic constituents.

Excretroy Urography in the Minipig (미니돼지에서 배설성요로조영술의 적용)

  • Jung, Joo-Hyun;Chang, Jin-Hwa;Choi, Min-Cheol
    • Journal of Life Science
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    • v.17 no.9 s.89
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    • pp.1211-1216
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    • 2007
  • Excretory urography was performed on eight healthy minipigs to evaluate radiographic anatomical status and excretive function of the urinary tracts and to get an optimal procedure. The right and left kidneys were located in $T13{\sim}L4$and $T12{\sim}L3.5$, respectively. The left kidneys were located more cra-nially than the right. The everage lengths and widths of kidneys were $8.50{\pm}0.58$ $(mean{\pm}SD)$and $4.30{\pm}0.39$ cm on the ventrodorsal projection and $8.70{\pm}0.76$, and $4.10{\pm}0.40$ cm on the lateral projection, respectively. The lengths of kidneys were twice the width. When the lengths of kidneys were com-pared with lengths of the second lumbar vertebras, the ratios of kidney lengths to L2 lengths were $3.62{\pm}0.30$ on the ventrodorsal projection and $3.63{\pm}1.10$ on the lateral projection. The lengths and 쟝손 of renal pelvis including major calices were $4.01{\pm}0.46$, and 2.20{\pm}0.41$ cm, respectively. The lengths and 쟝손 of minor calices were $0.49{\pm}0.06$, and $0.10{\pm}0.01$ cm. The lengths and widths of the ureters were $12.25{\pm}2.05$, and 2.94{\pm}0.86$ cm. The nephrogram stage was reached in 3{\sim}5$ minutes, and the pyelogram phase in $5{\sim}15$ minutes individually after injection of contrast medium. The result of this study shows similar information and procedure of excretory urography for other laboratory ani-mals and leads to the potential use of this method for minipigs.