• Title/Summary/Keyword: Invitro

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Neuroprotection of Dopaminergic Neurons by Hominis Placenta Herbal Acupuncture in in vitro and in vivo Models of Parkinson's Disease Induced by MPP+/MPTP Toxicity

  • Jun, Hyung Joon;Nam, Sang Soo;Kim, Young Suk
    • Journal of Acupuncture Research
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    • v.32 no.1
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    • pp.23-36
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    • 2015
  • Objectives : This study was designed to investigate the neuroprotective effects of Hominis-Placenta (HP)on dopaminergic neurons. Methods : We examined the effect of invitro administration of HP against 1-methyl-4-phenylpyridinium( MPP+)-induced dopaminergic cell loss in primary mesencephalic culture and also used behavioral tests and performed analysis in the striatum and the substantia nigra of mouse brain, to confirm the effect of HP on dopaminergic neurons in an invivo 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(MPTP)-induced PD mouse model. Animals were assigned to four groups: (1) Group 1(vehicle-treatedgroup), (2) Group 2(MPTPonlytreated group), (3) Group 3(MPTP+ saline-treated/$ST_{36}$ group), and (4) Group 4(MPTP+HP-treated/$ST_{36}$ group). HP at $20{\mu}L$ of 48 mg/kg dose was injected at $ST_{36}$ for 4 weeks at 2-day intervals. MPTP in saline was injected intraperitoneally each day for 5 days from the $8_{th}$ treatment of HP. We performed the pole test and rota-rod test on the first and seventh day after the last MPTP injection. To investigate the effect of HP on dopaminergic neurons, we performed analysis in the striatum and the substantia nigra of mouse brain after treatment with HP and/or MPTP. Results : Treatment with HP had no influence on cell proliferation and caused no cell toxicity in $PC_{12}$ and $HT_{22}$ cells. Our study showed that HP significantly prevented cell loss and protected neurites against MPP+ toxicity. Although the invivo treatment of HP herbal acupuncture at $ST_{36}$ showed a tendency to improve movement ability and protected dopaminergic cells and fibers in the substantia nigra and the striatum, it did not show significant changes compared with the MPTP treated group. Conclusions : These data suggest that HP could be a potential treatment strategy in neurodegenerative diseases such as Parkinson's disease.

Effect of the Synchronization of Meiotic Resumption in Bovine Oocytes Matured In Vitro (감수분열 재개 동기화가 소 미성숙 난자의 체외 성숙에 미치는 영향)

  • 박종임;장유니
    • Journal of Embryo Transfer
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    • v.18 no.3
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    • pp.263-267
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    • 2003
  • This study was carried out to test the efficacy of pharmacological inhibitors of the cell cycle transition in keeping bovine oocytes at the germinal vesicle(GV) stage and the reversibility of this inhibition. Bovine oocytes were incubated for 22∼24 hrs in the presence of various inhibitors : cycloheximide (2$\mu\textrm{g}$/$m\ell$), 6-DMAP (2 mM), and roscovitine (50$\mu$M). Bovine oocytes cultured with any of the inhibitors were significantly blocked at the GV stage. Reversibility of pharmacological inhibitors was assessed by culturing oocytes an additional 22∼24 hours in inhibitor-free medium. Examination of oocytes revealed that the inhibitory effect was fully reversible and effect of resuming meiotic progression on nuclear maturation varied according to the various inhibitors. This study suggests that cycloheximide, 6-DMAP and roscovitine can be applied to control meiotic arrest and resumption in maturation culture of bovine oocytes in vitro. More investigations are needed to better understand how the cell cycle of oocyte is blocked without problems to future developmental competence.

Lymphoblastosis Inhibition and Plaque-forming Cell Response of Several Anti-inflammatory Steroids in Mice

  • Choi, Hong-Pil;Kim, Kilhyoun;Kim, Hyun-Pyo
    • Archives of Pharmacal Research
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    • v.15 no.2
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    • pp.169-175
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    • 1992
  • Anti-inflammatory glucocorticoid (GC) derivatives have been clinically used in immune-malfunctional diseases for their immunosuppressive activity. However, there is still a lack of knowledge on the relationship between anti-inflammatory and immunosuppressive activities. In order to compare immunosuppressive activities with the known anti-inflammatory activities of the GC derivatives, eight clinically used GC derivatives including hydrocortisone, prednislone, 6$\alpha$-methyl prednisolone, triamcinolone, dexamethasone, betamethasone, triamcinolone acetonide and fluocinolone acetonide were selected, and lymphoblastosis inhibition and plaque-forming cell (PFC) response in mice were studied as immunological parameters. In Con A-induced lymphoblatosis inhibition invitro, all derivatives showed potent inhibition $IC_{50}$ values of the derivaties except methyl prednisolone and triamcinolone were less than $10^{-7}$M and good dose dependency was obtained. This result was well correlated with that of their anti-inflammatory potencies obtained. This result was well correlated with that of their anti-inflammatory potencies and their receptor binding affinities. However, in PFC response, consistent result were not obtained. Total numbers of PFCs per spleen were decreased by some derivatives, but numbers of PFCs per $10^6$ cells were not decreased by systemic administration of but numbers of PFCs per $10^6$ cells were not decreased by systemic administration of GC at the dose of 0.05 mg/mouse. Furthermore, at the dose of 0.1 mg/mouse, numbers of PFCs per $10^6$ cells were found to be increased, although total PFCs per spleen were decreased.

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The Studies on Embryo Production from Hanwoo Cows with High Offspring Meat Quality Grade and Embryo Transfer (후대의 육질등급이 우수한 한우 암소의 수정란 생산 및 이식에 관한 연구)

  • Lee, Hae-Lee;Park, Jae-Hee;Kim, Yong-Su;Kim, Jong-Gug
    • Journal of Animal Reproduction and Biotechnology
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    • v.34 no.1
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    • pp.50-56
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    • 2019
  • These studies were conducted to evaluate developmental competence of follicular oocyte collected from the ovaries of Hanwoo cows with the high offspring meat quality (1++ and 1+ grade). Cumulus oocyte complexes from individual cows were matured, fertilized and cultured using protocols of in-vitro maturation (IVM), invitro fertilization (IVF) and in-vitro culture (IVC). The rates of blastocyst development from Hanwoo cows with the offspring meat quality grades of 1++ and 1+ were 18.6 and 21.2%, respectively. The rates of blastocyst development were 26.3, 20.7, 20.7, 17.2 and 31.2% from Hanwoo cows with the meat quality grades of 1++, 1+, 1, 2 and 3, respectively. Fiftyseven transferable embryos were recovered from 11 Hanwoo donor cows (5.2/head) with the high offspring meat quality grades of 1++ and 1+ in vivo, and the pregnancy rate after embryo transfer was 61.1%. In conclusion, these results suggest that in vitro embryo production from the ovaries of cows with the high meat quality grades using individual culture system can be used an efficient method for livestock improvement. In addition, for the successful industrialization of embryo transfer, conception rate should be improved.

Suppress Effects of Euiiin-tang(yìyĭrén-tāng) Aqueous Extracts on Collagen Induced Arthritic(CIA) DBA/1 Mice (Collagen으로 유발된 마우스의 관절염에 대한 의이인탕(薏苡仁湯) 추출물의 억제 효과)

  • Cho, Jung-Hyun;Kwon, O-Gon;Woo, Chang-Hoon;An, Hee-Duk
    • Journal of Korean Medicine Rehabilitation
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    • v.20 no.1
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    • pp.37-59
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    • 2010
  • Objectives : The object of this study was to observe the favorable anti arthritic effects of Euiiin-tang($y{\grave{i}}y{\breve{i}}r{\acute{e}}n-t{\bar{a}}ng$) aqueous extract(EIITe), has been traditionally used in Korean medicine for treating rheumatoid arthritis on collagen induced arthritic(CIA) DBA/1 mice. Methods : In the present study, effects of EIITe on the releases of human tumor necrosis factor(TNF)-${\alpha}$, interleukin(IL)-$1{\beta}$, matrix metalloproteinase(MMP)-13 and production of Nitric oxide(NO) were observed by in vitro. In addition, to observe the effects on the CIA mice, three different dosages of EIITe, 300, 150 and 150 mg/kg were orally administered once a day for 18 days from 24hrs after antigen challenges(type II collagen) on 21 days after immunization using Type II collagen Freund's complete adjuvant. Six groups, each of 8 DBA/1 mice per group were used in the present study as follows. Changes on the body weights, macroscopic arthritis scores, splenic weights, splenic TNF-${\alpha}$ and IL-6 contents, articular cartilage(femur and tibia) collagen and glycosaminoglycans-chondroitin sulphate, sulphate and hyaluronic acid contents, histopathological observations(microscopic arthritis scores, thicknesses of femur and tibia cartilage thicknesses were monitored, compared to that of dexamethasone, a potent anti inflammatory agents, 1 mg/kg treated mice. Results : As results of collagen challenges, marked decreases of body weights and gains, articular cartilage collagen and glycosaminoglycan - chondroitin sulphate, sulphate and hyaluronic acid contents were observed with increases of macroscopic arthritis scores, splenic weights, splenic TNF-${\alpha}$ and IL-6 contents, articular cartilage(in the both femur and tibia) loss and damages. However, these CIA signs were significantly and dosages dependently inhibited by treatment of EIITe 300 and 150 mg/kg as compared with CIA control, respectively. In addition, the releases of TNF-${\alpha}$, IL-$1{\beta}$, NO and MMP-13 were markedly and dose dependently inhibited by treatment of EIITe, invitro. Although CIA were more favorably inhibited by treatment of dexamethasone 1 mg/kg as compared with EIITe 300 mg/kg, marked decreases of body weights were detected in dexamethasone 1 mg/kg treated mice. Conclusions : The results obtained in this study suggest that over 150 mg/kg of EIITe showed favorable anti arthritic effects on the CIA mediated by immunomodulatory and/or anti oxidative effects. However, detail mechanism studies should be conduced in future with the screening of the biological active compounds in this herb. lthough CIA were more favorably inhibited by treatment of dexamethasone 1 mg/kg as compared with EIITe 300 mg/kg, marked decreases of body weights were detected in dexamethasone 1 mg/kg treated mice, in the present study.

Positive Effect of Musa paradisiaca Peel Ethanolic Extract on Antioxidant Activity and Melanin Synthesis (바나나 껍질 에탄올 추출물이 멜라닌 합성에 미치는 영향)

  • Kim, JaeRyeon;Kim, Moon-Moo
    • Journal of Life Science
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    • v.28 no.7
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    • pp.802-810
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    • 2018
  • Aging is accompanied by changes in the body, such as graying hair, wrinkles, and black spots composed of lipid peroxides and proteins. Melanin is a polymer substance produced by an oxidation polymerization reaction from tyrosine, and it determines the color of hair and skin. It has been reported that melanin is synthesized by melanocyte, and its excessive production by reactive oxygen species is associated with aging. The purpose of this study was to determine the direct effects of Musa paradisiaca peel ethanolic extract (MPEE) on antioxidative activity and melanin synthesis. It was observed that the antioxidant activity of MPEE was similar to that of vitamin C, a positive control, in both DPPH radical scavenging assay and reducing power assay. In order to examine cytotoxicity prior to cell experimentation, 3-(4,5-dimethythiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was performed for B16F1 cells. MPEE was not cytotoxic at $32{\mu}g/ml$ or less. In addition, MPEE increased melanin synthesis in live cells in addition to tyrosinase activity and melanin synthesis in dihydroxyphenylalanine (DOPA)-oxidation assay in vitro. Moreover, MPEE increased melanin synthesis in cells aged by pretreatment with $H_2O_2$. The expression levels of tyrosinase-related protein (TRP)-1, TRP-2, and superoxide dismutase (SOD)-2 by western blot analysis were increased in the presence of MPEE. These results suggest that MPEE could promote the melanin synthesis as an antioxidative substance.

The Effect of Excess Dietary Vitamin A on Vitamin K-dependent Carboxylation in Rat Liver Microsomes (비타민 A 과량 섭취가 흰쥐의 간 Microsome의 비타민 K-dependent Carboxylation에 미치는 영향)

  • Lilha Lee
    • Journal of Nutrition and Health
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    • v.25 no.6
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    • pp.492-500
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    • 1992
  • The rate of vitamin K-dependent carboxylation of endogenous liver microsomal proteins and an exogenous peptide substrate for carboxylase were measured to test the effects of excess vitamin A on vitamin K function in rats. In vitro vitamin A incubation in normal rat microsomes of vitamin K-sufficient ras did not influence the carboxylation rates of either endogenous prothrombin precursors or a peptide substrate added, Similarly vitamin A incubation in micro-somes from control and excess vitamin A-fed rats that were on vitamin K-free diet did not change the rate significantly within the respectively groups ; however the rates of endogenous protein carboxylation from excess vitamin A-fed rats tended to be increased by the in vitro vitamin A addition compared to that of control rats. Excess vitamin A-fed rats had 2- to 3- fold higher carboxylase activites of endogenous protein carboxylation either with or without the invitro vitamin A incubation than did control rats. In an in vivo study carboxyalase activites with an added exogenous peptide substrate were not influenced by excess intake of vitamin excess vitamin A-fed rats than for control rats. Carboxylase activites tended to be increased amounts of vitamin A on endogenous protein carboxylation appeareed as early as one week post-initiation of the diet. The results of this study indicate that excess vitamin A produces toxic effect rapidly and that excess dietary vitamin A increase the rate of carboxylation of endogenous protein mainly prothrombin precursors which is an indication of vitamin K defi-ciency.

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Polyamine, Cytochrome c and Enzymes Related to the Utilization of Methanol in Methylobacterium extorquens AMI Growing at Different pHs (상이한 수소이온농도에서 성장하는 Methylobacterium extorquens AM1의 메탄올 이용 관련효소와 Cytochrome c 및 폴리아민)

  • 박기정;이순희;김영민
    • Korean Journal of Microbiology
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    • v.30 no.6
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    • pp.533-538
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    • 1992
  • The generation time of Methylobacterium extorquens AMI growing on methanol at pH 5.5 and 7.0 was found to be 23 hand 8.3 h. respectively. The bacterium grown at pH 7.0 were found to contain more amounts of spermidine and putrescine than the cell grown at pH 5.5. Cells grown at both conditions exhibited strong methanol dehydrogenase (MDH) activity at the mid-exponential growth phase. The amounts of MDH. however. were found to be almost equal through all gro~1h phases. Cells growing at the stationary phase contained large amounts of cytochrome c. The cytochrome c content was higher in cells growing at pH 7.0 than the cells growing at pH 5.5. Cells growing at pH 5.5 in the presence of putrescine or spermidine contained increased amounts of putrescine. The level of spermine, however. was decreased and that of spermidine was not changed. Spermine added into the medium was found to have no effect on the level of cellular polyamines. Putrescine or spermidine added into the medium stimulated MDH and hydroxypyruvate reductase activities. but did not affect the contents of MDH and cytochrome c. It was found that preincubation of cell-free extracts with polyamines does not stimulate MDH and hydroxypyruvate reductase activities.

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Plant Regeneration via in Vitro Culture of Ovule Obtain by Intergeneric Crossing Between Citrus junos Sieb. et Tanaka and Poncirus trifoliata Raf. (유자와 탱자의 속간교잡후 배주배양에 의한 식물체 유기)

  • 이만상;남궁승박
    • Korean Journal of Plant Tissue Culture
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    • v.22 no.6
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    • pp.317-322
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    • 1995
  • As a basic research for breeding new varieties, reciprocal -intergeneric crosses between Citrus junos and P.trifoliata were made. F$_1$ hybrid production using in vitro ovule culture, gametogenesis, and fertilization phenomena were investigated. Frequency of fruit set resulting from crossing of Citrus junos and Poncirus Trifoliata was 16.6% while that of Poncirus Trifoliata and Citrus junos was 11.7%. Callus formation occurred well when ovules at the 6th week after pollination were cultured on MT (Murashige and Tucker) medium supplemented with zeatin 0.5 mg/L and NAA 1.0 or 3.0 mg/L. Immature ovules developed into mature embryos of the MT medium supplemented with 2,4-D 0.1 or 3.0 mg/L. Immature ovules developed into mature embryos of the MT medium supplemented with 2,4 D 0.1 or 0.5 mg/L. The invitro germination rates of 20-week-old ovules set C. junos $\times$ P. Trifoliata and P. Trifoliata $\times$ C. junos were 54.5% and 48.6%, respectively. The emergence ratios of trifoliate hybrids obtained by C. junos $\times$ P. Trifoliata and P. Trifoliata $\times$ C. junos were 56.7% and 100%, respectively. The chromosome number of C. junos and P. Trifoliata was n = 9 or 2n = 18, and the sizes of their pollen grain were 33.75 $\mu$ and 25.0 $\mu$. The length and width of embryo sac in C. junos and P. Trifoliata were 69.38~79.23 $\mu$ and 27.50~38.56 $\mu$, and those of egg cells were 17.50~41.50 $\mu$ and 6.25~8.12$\mu$. Fertilization of C. junos and P. trifoliata terminated 72 h after pollination.

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The Effects of Diagnostic Radiology Image on Radiopharmaceutical Testing (방사성의약품 검사 시 진단(CT)영상에 미치는 영향)

  • Lee, Eun-Hye;Lee, Ye-Seul;Kim, Gha-Jung;Choi, Jun-Gu
    • Korean Journal of Digital Imaging in Medicine
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    • v.12 no.2
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    • pp.113-117
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    • 2010
  • This research attempts to qualitatively evaluate the intensity change by radiopharmaceuticals and obtain computed tomography using phantom injected with various nuclide. Cylindrical phantom is used for comparing and analysing the effect on diagnosis image during radiopharmaceuticals inspection. Inside of the phantom, water is injected and computed tomography image is scanned. During nuclear medicine invitro, frequently used radiopharmaceuticals, $^{99m}TcO_4$ 20 mCi and $^{18}F$ 14 mCi, is diluted in the water phantom and scanned in the same method. Traverse image obtained by CT scan is divided into six traverse image in the same slice of each scanned image. CT-number(HU) value of 10 measuring point is measured in 2 cm interval based on the center of the phantom. Measured HU value, based on the water phantom, is compared with the image after injecting $^{99m}TcO_4$ and $^{18}F$. Average scale of water is 2.8~1.6 HU, $^{99m}TcO_4$ is 3.0~1.6 HU and $^{18}F$ is 1.2~0 HU. Average of water is $2.3{\pm}0.17$ HU, $^{99m}TcO_4$ is $2.2{\pm}0.85$ HU and F-18 is $0.7{\pm}0.95$ HU. Based on water, reduced value of about 0.1 HU and about 0.5 HU is acquired from $^{99m}TcO_4$ and F-18. Radionuclide used in nuclear medicine inspection utilizes 100~200 KeV energy and obtains image through scintillation camera and PET-CT utilizes 511 KeV positron annihilation energy to obtain image. What we learned from this research is that gamma rays from these energies used in CT scan for diagnosis purpose or radioactive therapy plan can change the intensity of the image. The nuclear medicine inspection for reducing the effect of emitted gamma ray diagnosis image should be obtained after a period of time considering half-life which would be reduced distortion or changed in image.

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