• Title/Summary/Keyword: In vivo

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The Genotoxicity Study of Molinate, an Herbicide, in Bacterial Reversion, in vitro and in vivo Mammalian System

  • Kim, Youn-Jung;Ryu, Jae-Chun
    • Molecular & Cellular Toxicology
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    • v.2 no.3
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    • pp.176-184
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    • 2006
  • The controversy on genotoxicity of molinate, an herbicide, has been reported in bacterial system, and in vitro and in vivo mammalian systems. To clarify the genotoxicity of molinate, we performed bacterial gene mutation test, in vitro chromosome aberration and mouse lymphoma $tk^{+/-}$ gene assay, and in vivo micronucleus assay using bone marrow cells and peripheral reticulocytes of mice. In bacterial gene mutation assay, no mutagenicity of molinate ($12-185{\mu}g/plate$) was observed in Salmonella typhimurium TA 98, 100, 1535 and 1537 both in the absence and in the presence of S-9 metabolic activation system. The clastogenicity of molinate was observed in the presence ($102.1-408.2\;{\mu}g/mL$) of metabolic activation system in mammalian cell system using Chinese hamster lung fibroblast. However, no clastogenicity was observed in the absence ($13.6-54.3\;{\mu}g/mL$) of metabolic activation system. It is suggested that the genotoxicity of molinate was derived some metabolites by metabolic activation. Molinate was also subjected to mouse lymphoma L5178Y $tk^{+/-}$ cells using microtiter cloning technique. In the absence of S-9 mixture, mutation frequencies (MFs) were revealed $1.4-1.9{\times}10^{-4}$ with no statistical significance. However, MFs in the presence of metabolic activation system revealed $3.2-3.4{\times}10^{-4}$ with statistical significance (p<0.05). In vivo micronucleus (MN) assay using mouse bone marrow cells, molinate revealed genotoxic potential in the dose ranges of 100-398 mg/kg of molinate when administered orally. Molinate also subjected to acridine orange MN assay with mouse peripheral reticulocytes. The frequency of micronucleated reticulocytes (MNRETs) induced 48 hr after i.p. injection at a single dose of 91, 182 and 363 mg/kg of molinate was dose-dependently increased as $10.2{\pm}4.7,\;14.6{\pm}3.9\;and\;28.6{\pm}6.3\;(mean{\pm}SD\;of\;MNRETs/2,000\;reticulocytes)$ with statistical significance (p<0.05), respectively. Consequently, genotoxic potential of molinate was observed in in vitro mammalian mutagenicity systems only in the presence of metabolic activation system and in vivo MN assay using both bone marrow cells and peripheral reticulocytes in the dose ranges used in this experiment. These results suggest that metabolic activation plays a critical role to express the genotoxicity of molinate in in vitro and in vivo mammalian system.

In Vitro/In Vivo Development after Thawing of Vitrified Mouse Blastocysts by Culture Condition and Embryo Transfer Method (초자화 동결된 생쥐 배반포기배의 융해 후 배양조건과 수정란 이식방법에 따른 체외/체내발달)

  • Kim, M.K.;Kim, E.Y.;Yi, B.K.;Yoon, S.H.;Park, S.P.;Chung, K.S.;Lim, J.H.
    • Clinical and Experimental Reproductive Medicine
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    • v.24 no.3
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    • pp.347-353
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    • 1997
  • This study was to test whether in vitro/in vivo survival of vitrified mouse blastocysts was influenced by culture conditions and ET method. Mouse blastocysts were obtained from in vitro fertilization and cultured for 4 days in M16 medium, and they were vitrified in EFS40 which contained 40% ethlyene glycol, 18% Ficoll and 0.5 mol sucrose in PBS. In experiment I, in vitro and in vivo survival rate of these embryos were evaluated in different culture condition after thawing. When thawed embryos were cultured in M16 medium as a control, m-CR1 medium contained 20 amino acids (2% BME amino acis and 1% MEM non-essential amino acids solution) and 4 mg/ml BSA and cumulus monolayer cell co-cultured condition in mCR1 medium (10% FBS), their in vitro survival at 24 hr after thawing was not affected by culture condition (75.6, 83.1, 82.4%). However, in vivo survival rates of implantation in m-CR1 medium (80.4%) were significantly higher than those of M16 medium (51.2%), co-culture (57.1%) condition, although there was no difference in live fetuses rates on day 15 gestation (39.0, 49.0, 38.1%). In experiment II, the in vivo development potential of embryos by ET methods was examined. When blastocysts were transferred to the day 2, 3 pseudopregnant recipient without culture soon after thawing, no pregnant recipient was obtained on the day 2 pseudopregnancy, and 50% of pregnancy rates and 15.4% of live fetus rates were obtained on the day 3 pseudopregnant recipients. These results were significantly lower than those of transferred group (day 3 pseudopregnant recipients) after culture for 16 hr post thawing (73.5, 57.1%) (p<0.05). In experiment III, to elevate usability of delayed embryos in vitro/in vivo survival of vitrified embryos (day 4 early, day 5 early and expanding blastocyst) were examined. in vivo survival rates (live fetus, total implantation) were higher in day 4 early blastocysts (33.3, 66.7%) than in day 5 expanding blastocysts (29.0, 38.7%), although the highest in vitro survival rates were obtained in the day 5 expanding brastocysts (78.3%). Therefore, these results suggest that the in vitro/in vivo survival rates of vitrified embryos could be improve by the culture condition and ET method and that the in vivo development rates of delayed embryos were decreased with longer culture duration in vitro. It means that more effective cryopreservation was obtained in day 4 early blastocysts than in day 5 expanding blastocysts.

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In Vitro Maturation, Fertilization and Development of Mouse Oocytes Derived from In Vitro Grown Preantyal Follicles (체외성장된 Preantral Follicle에서 유래된 생쥐난자의 체외성숙, 수정 및 발달)

  • Kim, D.H.;Lee, H.J.;K.S. Chung;Lee, H.T.
    • Korean Journal of Animal Reproduction
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    • v.26 no.2
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    • pp.133-142
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    • 2002
  • The objective of this study was to examine maturation, fertilization and developmental rate of the in vitro-grown mouse oocytes, and to compare these results with those of oocytes grown and matured in vivo. The preantral follicles isolated from 12-day-old mice were cultured on Transwell-COL membrane inserts. After in vitro growth and maturation, 72.5 % of oocytes grown in vitro produced polar body which can be comparable to in vivo growth (70.5 %). However, the mean oocyte diameter of the in vitro group (69.6$\pm$2.1$\mu$m) was smaller than that of the in vivo group (73.3$\pm$3.0$\mu$m). The fertilization rate was significantly lower (p<0.05) in the in vitro group (76.5%) than in the in vivo group (90.2%), however, there was no difference in the percentage of monospermic and polyspermic oocytes between two groups. The capacities of in vitro grown ova to cleave and develop to blastocyst were (57.8 and 14.4%, respectively) significantly lower (p<0.001) than those of the in vivo counterpart (84.4 and 56.6%, respectively). Moreover, the mean number of cells per blastocyst was significantly lower (p<0.05) in the in vitro group (39.0$\pm$10.8) than in the in vivo group (60.5$\pm$12.5). Live young were produced from transferred 2-cell embryos derived from in vitro-grown and matured oocytes. In conclusion, the results show that in vitro-grown oocytes did not achieve the developmental capacity of in vitro-grown oocytes.

The Ultrasonographic Images of Normal Uterus in Korean Native Cow (한우 정상 자궁의 초음파적 관찰)

  • Park, Chul-Ho;Kim, Jae-Hong;Mun, Byeong-Gwon;Kim, Hee-Su;Kim, Bang-Sil;Lee, Ju-Hwan;Park, In-Chul;Kim, Jong-Taek;Suh, Guk-Hyun;Oh, Ki-Seok;Son, Chang-Ho
    • Journal of Embryo Transfer
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    • v.24 no.3
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    • pp.159-167
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    • 2009
  • Ultrasonographic examination was performed to observe the ultrasonographic image of Korean native cows' normal uterus in condition of in vitro and in vivo. The experiment was done 28 slaughtered cows' uterus using immersed in water in vitro, and 41 healthy breeding cows taken rectal ultrasonography in vivo. Ultrasonographic examination of uterine was taken on the reference of cross section of intercornual ligaments' cranial. Each uterus on the experiments was compared by estrous cycle and ultrasonographic frequency. The uterine structure using ultrasonography was 5 layers of uterine horn in vivo as well as in vitro. Uterine horn was observed to be distinguished from inside to outside as endometrium to inner echogenic layer, circular muscle layer to slightly echogenic elliptical layer, stratum vasculare to central echogenic layer, longitudinal muscle layer to slightly echogenic arched layer, and perimetrium to outer echogenic layer, respectively. According to the observation of uterus related to estrous cycle and ultrasonographic examination, uterine endometrium in vitro was constantly founded irrespective of estrous cycle and ultrasonographic frequency. On the low frequency, endometrium and circular muscle layer in estrus were prone to distinguished than in diestrus. On the high frequency, endometrium and circular muscle layer were always distinguished regardless of estrous cycle. In vivo, uterine endometrium and circular muscle layer were observed regardless of estrus and ultrasonographic frequency. On the low frequency, stratum vasculare and longitudinal muscle layer were not likely to be distinguished in diestrus, but estrus. On the high frequency, stratum vasculare and longitudinal muscle layer were observed regardless of estrous cycle. Also, every uterine structure was easily distinguished on high frequency than low frequency owing to precision of distinction in layers. The difference of results followed by the experiments conditions between in vitro and in vivo was that uterine endometrium and circular muscle layer in diestrus in vitro were difficult to be distinguished and uterine lumen was observed during whole estrous cycle. In vivo, It was founded that the distinction of stratum vasculare and logitudinal muscle layer in diestrus was complicated and uterine lumen was observed during only estrus. In view of the result so far achieved, normal uterine structure divided in 5 layers on ultrasonography was accorded with microscopic organization, uterine structure was likely to be observed during estrus than diestrus, high frequency checkup than low frequency, and uterine endometrium, circular muscle, stratum vasculare was easily observed regardless of estrous cycle and ultrasonographic frequency.

In Vitro and In Vivo Agreement of Microwave Radio-Thermometer and Needle Probe Thermometer During Therapeutic Ultrasound (초음파 가열 시 In Vitro 및 In Vivo에서 Microwave Radio-Thermometer와 탐침온도계의 일치도)

  • Lee, Su-Young;Cho, Sang-Hyun;Yi, Chung-Hwi;Kim, Jong-Man
    • Physical Therapy Korea
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    • v.10 no.1
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    • pp.15-27
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    • 2003
  • Therapeutic ultrasound is commonly applied for deep heating in physical therapy setting. However, it is difficult to determine the exact application dosage and to confirm the immediate heating effect. Microwave Radio-Thermometer (MRT) can measure the temperature by the electromagnetic energy in the microwave region of the object that emits above absolute zero temperature. MRT was used for early diagnosis of breast cancer since it was not harmful, non-invasive, and non-ionizing to the human body. The purposes of this study were to investigate how accurately 1.1 GHz RTM (RES Ltd. Russia) measures the change of average temperature in the tissue, and to determine the depth of temperature change measurement. Therapeutic ultrasound was applied (continuous wave for 5 minutes, 1 MHz, intensity of 1.5 $W/cm^2$ [in vitro] and 1.0 $W/cm^2$ [in vivo]) in four different conditions: (1) 30 cases of in vitro specimen of pork, (2) 30 cases of in vitro specimen of pork ankle joint, (3) 10 cases of in vivo canine thigh, and (4) 30 cases of in vivo human body. Intraclass Correlation Coeffients (ICC[3,1]) between average needle probe thermometer below surface and MRT temperature was revealed as followed: (1) Before ultrasound application ICCs ranges above .8 in specimen of pork (15 mm underneath the skin) and above .82 in specimen of pork ankle joint (10~30 mm underneath the skin). (2) After ultrasound application ICCs ranges above .7 in both specimens of pork and pork ankle joint. (3) Before ultrasound application ICCs ranges above .8 in canine thigh (20 mm underneath the skin). (4) After ultrasound application ICCs ranges above .82 in canine thigh. The temperature of the human body increased significantly with the mean of $15^{\circ}C$ in muscle tissue and with the mean of $3.5^{\circ}C$ in joint (p<.00). It was revealed that the average depth of temperature measurement of the tissue by MRT was in between 10 and 35 mm, and determined that the proper temperature measurement band was $36.5{\sim}37.0^{\circ}C$.

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Effect of Gal-Geun-Tang on Antigen-Specific Immune Response (갈근탕(葛根湯)이 면역조절작용에 미치는 영향)

  • Cho, Dae-Yeoun;Yun, Young-Gab;Jung, Myung;Lee, Eun-Hye;Bok, Young-Ok;Jung, Chang-Ohk;Lim, Kyu-Sang
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.29 no.3
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    • pp.134-149
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    • 2016
  • Objectives : Gal-Geun-Tang (GT) has been described from SANGHAN in Korean traditional medicine and known to act against cold, fever, hypertension, and nasal catarrh. However, little has yet been learned about the effect of GT on immune function. In the current study, in vitro and in vivo immunomodulatory activity of GT (water extract) was investigated.Methods : Water extract of GT induced in vitro proliferation of spleen cells and significantly increased their proliferative responses during anti-CD3 activation. Using purified splenic T and B cells, it was revealed that GT has a mitogenic activity to B cells and promotes their proliferation induced by lipopolysaccharide, whereas T cell proliferation was not triggered and GT was rather inhibitory to T cell activation caused by anti-CD3 antibody. In the presence of antigen presenting cells (APC), GT addition resulted in a significant increase of IFNγ and IL-4, but not IL-2, production. However, addition of high concentration (1,000㎍/㎖) of GT led to a marked reduction in T cell cytokine production and under such condition, GT facilitated apoptosis of T cells when examined by flow cytometry with propidium iodide staining.Results : In vivo immunomdulation of GT was also investigated using a mouse model. Following keyhole limpet hemocyanin (KLH) immunization, GT (1 ㎎/day) was orally administered for 9 days. Cell numbers in thymus, spleen and peripheral blood were not altered by GT administration, indicating that such dose is not immunotoxic. Cell numbers in draining lymph nodes (LN) and ex vivo Ag-specific proliferation of LN cells were significantly elevated by GT administration. However, any preferential stimulation of T or B and CD4+ or CD8+ T cell subpopulations was not observed in a flow cytometric analysis of LN cells. This result shows that GT does not promote in vivo B cell proliferation while GT enhances Ag-specific proliferation of LN cells, unlike what was observed in vitro.Conclusions : For a further understanding of in vivo immunomodulatory activity of GT, ex vivo cytokine production of LN cells obtained from KLH-immunized mice was evaluated. Ag-specific IFNγ production was significantly higher in GT-treated mice when compared to PBS-treated control mice. In contrast, IL-4 production in GT-treated group was comparable to control group unlike to in vitro data. In addition, GT administration did not result in any significant differences in serum levels of Ig (IgM, IgG1 and IgG2a) between GT-treated and control groups. Taken together, these data strongly support that GT promotes immune response, more profoundly type 1 helper T cell (Th1) activity and GT may be applicable for treatment of intracellular parasite infection such as viral diseases.

Diagnostic Assay of Toxic Zinc in an Ex Vivo Cell Using Voltammetry

  • Ly, Suw-Young;Yoo, Hai-Soo
    • Toxicological Research
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    • v.28 no.2
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    • pp.123-127
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    • 2012
  • Voltammetric detection of the toxic Zn ion was investigated using a fluorine-doped graphite pencil electrode (FPE). It is notable from the study that pencils were used as reference and working electrodes. In all the experiments, a clean seawater electrolyte solution was used to yield good results. The analytical working range was attained to 10 ${\mu}gL^{-1}$. The optimized voltammetric condition was examined to maximize the effect of the detection of trace Zn. The developed sensor was applied to an earthworm's tissue cell. It was found that the methods can be applicable to in vivo fluid or agriculture soil and plant science.

신물질 ASPALATPNE의 항혈전 작용

  • 한병훈
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1993.04a
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    • pp.123-123
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    • 1993
  • Acetylsalicylic acid (aspirin)와 인삼의 항산화 성분인 maltol의 축합반응으로 합성된 신물질 Aspalatene이 저용량: 장기 복용시 항산화활성과 더불어 지혈시간 연장효과가 탁월함을 보고 한 바 있다. Aspalatone은 aspirin과는 달리, 장기 복용시에도 위궤양 유발 작용이 거의 나타나지 않으므로 말초순환개선제로서의 개발이 기대된다. 본 연구에서는 Aspalatone의 항혈전작용을 in vivo, ex vivo 및 in vitro에서 연구하였다. Mouse thromboembolism test를 이용한 in vivo실험에서 Aspalatone은 aspirin과 유사하게 collagen에 의한 치사를 유의적으로 감소시켰으며 ADP에 의한 치사에는 영향을 미치지 않았다. 또한 10일간의 장기 투여 실험에서는 저용량에서 영향을 미치지 않았다. 또한 10일간의 장기 투여 실험에서는 저용량에서 항혈전 작용을 나타내며 투여 중단 4일 후에도 약효가 유지되는 것을 밝혔다. 이는 Aspalatone이 aspirin과 같이 cycloosygenase에 작용하여 항혈전 작용을 나타냄을 강력히 시사하고 있다.

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Design of Two-Dimensional Lateral Antenna for Wireless Power Transmission to In Vivo Robotic Capsule

  • Kim, Jong-Dae;Lee, Seon-Woo;Ryu, Mun-Ho;Kim, Jong-Won
    • Journal of Biomedical Engineering Research
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    • v.29 no.3
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    • pp.187-190
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    • 2008
  • This paper presents two dimensional receiving coils to provide hundreds of milli-watt power via inductive link to in vivo robotic capsules, whose orientation are practically undetermined. The wireless power transmission system consists of a transmitter powered by class E power amplifier, and a receiver with three dimensional antenna, rectifier, and voltage regulator. As the 2D lateral antenna construction is more critical for the receiving antenna, two types of 2D antennas are introduced and evaluated by theoretic and experimental analyses. Experimental results verifies that the cross-type construction show better directional performance for receiving power than the cylindrical one for the 2D antenna. The former could deliver the power homogeneously regardless of its orientation, with less than 20 % of variation from the possible maximum power.

Three newly recorded heterotrophic euglenids(Protist), Entosiphon oblongum, Euglena longa and Keelungia pulex from South Korea

  • Lee, Won Je
    • Journal of Species Research
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    • v.11 no.2
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    • pp.89-93
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    • 2022
  • Three heterotrophic euglenids from marine water column (Seodo port, Yeosu) and freshwater sediment (Seodong-chun, Incheon), Korea were identified as Entosiphon oblongum Cavalier-Smith and Vickerman, 2016; Euglena longa (Pringsheim, 1936) Marin and Melkonian, 2003; and Keelungia pulex Chan and Moestrup, 2013 based on morphological characters and 18S rDNA sequence analysis. These species are reported taxonomically for the first time from Korea and are described with micrographs. Diagnoses of these species are as follows. Entosiphon oblongum: phagotrophic, gliding, size in vivo, 23.1-29.3 ㎛ (Avg. 26.5 ㎛, n=30) long, ovate with a protrusive feeding siphon (apparatus), several deep grooves and two heterodynamic flagella. Euglena longa: osmotrophic, swimming, size in vivo, 32.3-52.2 ㎛ (Avg. 42.2 ㎛, n=26) long, elongated with many paramylum granules and two flagellar. Keelungia pulex: phagotrophic, gliding, size in vivo, 13.5-19.7 ㎛(Avg. 16.4 ㎛, n=97) long, oblong to ovoid with a hook-shaped ingestion apparatus, several dorsal ridges and two flagella.