• Title/Summary/Keyword: ex vitro

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Studies on In Vitro Fertilizability of Mouse Oocytes Pre-exposed to Dibutyryl Cyclic AMP (Dibutyryl Cyclic AMP로 처리된 생쥐난자의 수정능에 관한 연구)

  • 강해묵;이영기;조완규
    • The Korean Journal of Zoology
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    • v.31 no.1
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    • pp.21-28
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    • 1988
  • The present study was carried out to examine the fertilizability of the mouse oocytes pre-ex-posed to dbcAMP which is a well-known inhibitor of the oocyte maturation. The oocytes once cultured in the dbcMP-containing medium for a certain length of, time were cultivated in the dbcMp-free medium to induced the maturation, then mixed with sperms, and observed following culture for 24 hours. The fertilization rate of cocytes was judged by the index of the number of 2-cell embryo developed 24hr following insemination. The fertilization rate of the oocyte previously incubated with dbcAMP (100 g/ml) for 2, 4, 8 16 hours was 32.3, 14.5, 4.7 and 8.8%, respectively, while that of the control was 53.3% indicating that the fertilizability was decreased as a function of time exposed to dbcAMP. The pretreatment of dbcMP, however, didn't affect the process of sperm penetration to egg. In addition, there is no prominent changes in the morphological architecture of fertielized eggs which has been exposed to dbcAMP as revealed by electron microscopic observation. Consequendy, it can be concluded that the mouse cocytes once inhibited their maturation by dbcMP may retain, in some extent, the fertilizability, although most of the fertilized egg may not proceed to further development because of the failure of pronucleus formation.

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Inhibitory Effect of Petroleum Ether Extract of Panaz Ginseng Root against Growth of Human Cancer Cells (고려인삼 중 Petroleum Ether 추출물의 인체 암세포 증식억제효과)

  • 이선희;황우익
    • Journal of Ginseng Research
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    • v.10 no.2
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    • pp.141-150
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    • 1986
  • This study was attempted to screen the cytotoxic activity of petroleum ether ex- tract from panax ginseng root against human colon cancer cells. Two extracts of panax ginseng root, crude and partially purified, were used for this experiment. The crude extract was prepared by extraction with petroleum ether using Soxhlet aparatus for 12 to 15 hours from panax ginseng and the extract was partially purified by silicic acid column with mixture of petroleum ether: ethyl ether (70 : 30, v/v). Three species of human colon cancer cells, HRT-18, HCT-48 and HT-29, were maintained in DMEM (Dulbecco's modified Eagle medium), and the cells were cultured in DMEM containing serial concentration of the crude or partially purified fraction to observe the cytotoxic activity of the both extracts. The effects of incubation time and concentration of the both extracts in culture medium against the growth of the each cancer cell were determined. The results obtained are summarized as follows: 1. The doubling times of the HRT-18, HCT-48 and HT-29 cells were about 20, 24 and 22 hours, respectively. 2, The inhibitory effects of the crude extract on the growth of cancer cells were increased according to the rise of concentration of the extract and incubation time. 3. The inhibitory effect of partially purified fraction on the growth of HRT-18 cell was about 4 times stronger than that of the crude extract under same experimental condition. 4 The inhibitory effects of the crude and purified fraction on the growth of each cancer cell were shown difference by the kind of the cancer cell. In view of the above results, it could be said that the petroleum ether extract of panax ginseng root inhibited the division of the human colon cancer cell, in vitro.

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호장근 부탄올 분획의 비만 예방 및 치료 효과

  • Kim, Jin-Suk
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2010.10a
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    • pp.7-7
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    • 2010
  • 전 세계적으로 폭발적으로 증가하고 있는 비만은 만병의 근원이다. 동시에 대사성질환의 발병을 증가시키는 결정적인 역할을 하고 있다. 이러한 상황을 극복할 수 있는 안전하고 효능이 우수한 약의 개발이 매우 시급하다. 시판되는 약들은 예견할 수 없는 지방변으로 실제생활을 매우 불편하게 하는 부작용과 우울증 및 자살충동 등의 심각한 부작용을 유발시키고 있다. 특히 생명을 위협하는 약은 시판이 금지되었다. 200여종의 한약재들을 In vitro screening (pancreatic lipase inhibition, PDE inhibition, c-AMP activity), ex vivo screening (lipolytic action on fat pad), short term animal screening(혈중 TG 함량 분석)을 토대로 long term animal model에서비만 예방 및 치료 효능을 검증하기 위하여 호장근 부탄올 분획을 선정하였다. 고지방 사료로 비만을 유도한 rat (Diet induced obesty (DIO) rat)에서 비만 치료 효능이, 고지방 사료로 비만을 유도한 ICR-mice에서 비만 예방 효능이 우수함을 입증하였다 (체중감소, 지방세포의 크기억제, 지방간 예방/치료(간무게, TG함량, 간 색상, 고지혈증 증상억제), 혈중 TNF-a, IL-6, leptin, adponectin 등, 간 조직에서의 pAMPK, SOCS, NF-${\kappa}B$ DNA biding activity, ACC level, FAS expression, CPT-1 activity의 정상화). 호장근 부탄올 분획의 이러한 효능은 AMPK 작용과 CPT-1 작용을 활성화하고 동시에 지방산 합성 억제와 지방산 산화를 촉진함으로 인함임을 규명하였다. 동시에 비만으로 인한 pancreatic beta cell의 파괴를 예방함으로 인슐린 내성을 예방(치료)함을 입증하였다. 이는 AMPK 활성화와 SOCS-3 단백질 억제와 NF-${\kappa}B$-DNA 결합 억제로 인함임을 증명하였다. 3T3-L1 지방 세포주에서 lipogenesis 예방(치료) 및 lipolytic effect에 관여하는 인자들의 변화를 확인하였다. 이는 Multi-compounds-multi-targets에 의한 시너지 효과임을 알 수 있었다.

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Anti-oxidation and Fat Accumulation Inhibitory Effects of Silbi-san (실비산의 항산화 및 지방축적 억제 효과)

  • Kim, Geon-Woo;Woo, Chang-Hoon;Kim, Young-Jun;An, Hee-Duk
    • Journal of Korean Medicine Rehabilitation
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    • v.30 no.4
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    • pp.41-53
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    • 2020
  • Objectives The purpose of this study was to investigate the effects of Silbi-san on the antioxidant and fat accumulation inhibition and to analyze the anti-obesity effect by analyzing the changes in serum lipid composition in obese mice. Methods We compared contents of phytochemicals like total polyphenols and total flavonoid and antioxidant activities such as 2,2-dipheny-1-picrylhydrazyl and 2.2'-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) radical scavenging activity. After Silbi-san in 3T3-L1 cells in vitro and mouse adipose tissue ex vivo, we quantified intracellular triglyceride accumulation and lipolysis. Moreover, the anti-obesity activity though inhibiting pancreatic lipase were analyzed. In 3T3-L1 cells, morphological changes showed that control cells had many lipid while cells treated with Silbi-san had less lipid accumulation. 30% EtOH Silbi-san treatment also suppressed the fat absorption by inhibiting the activity of pancreatic lipase and led to high lipolysis through promoting glycerol release. The experimental group was divided into four groups: Normal group fed normal feed, Control group fed 60% high fat diet (HFD) and distilled water, drug group fed 60% high fat diet and 200 mg/kg of Silbi-san water extract, drug group fed 60% HFD and 200 mg/kg of Silbi-san 30% ethanol extract. Results Serum total cholesterol content and serum low density lipoprotein-cholesterol content were significantly decreased in the Silbi-san extract group compared to the control group, serum high density lipoprotein-cholesterol content was significantly increased in Silbi-san extract group. Conclusions In this study, the antioxidant and fat accumulation inhibitory effects of Silbi-san were confirmed.

Factors influencing Adventitious Shoot induction from Leaves of Populus deltoides Bartr (미류나무 잎절편체 조직배양을 통한 식물체 재분화에 미치는 요인)

  • 신동일;설일환
    • Journal of Life Science
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    • v.7 no.2
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    • pp.134-141
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    • 1997
  • We have investigated several factors influencing adventitious shoot induction from the Populus deltodies bartr. leaf segments. To obtain in vitro materials, at first, stem segments from ex vivro were tested for axillary bud breaking on the five macronutrients levels of two differenr media. WPM was better both in bud breaking and leaf expansion than MS medium. The amount of NH$_{4}$NO$_{3}$ added in the medium did not seem to affect axillary bud breaking, significantly and subsequent shoot elongation from the stem segments of P. deltodies. However, other components in WPM might have played important roles in axillary bud breaking and shoot elongation. Regenerability from three sections (the distal, the middle and the base) of leaves cultured on WPM supplemented with TDZ and NAA combinations appeared to be different; middle and basal sections of leaves produced more organogenic sites than those of top section on WPM and those sesctions produced the highest ogranogenic sites on the same medium supplemented with 0.01 mg/l TDZ and 0.02mg.l NAA. Among 3 carbon sources tested for adventitious shoot elongation, fructose seemed to be stimulating the elongation of adventitious shoots. Sucrose and glucose added in the medium resulted in the necrosis which caused dying of adventitious shoots, eventually.

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Thermal changes during implant site preparation with a digital surgical guide and slot design drill: an ex vivo study using a bovine rib model

  • Choi, Yoon-Sil;Oh, Jae-Woon;Lee, Young;Lee, Dong-Woon
    • Journal of Periodontal and Implant Science
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    • v.52 no.5
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    • pp.411-421
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    • 2022
  • Purpose: In this study, we aimed to evaluate the degree of heat generation when a novel drill design with an irrigation slot was used with metal sleeve-free (MF) and metal sleeve-incorporated (MI) surgical guides in an environment similar to that of the actual oral cavity. Methods: A typodont with a missing mandibular right first molar and 21 bovine rib blocks were used. Three-dimensional-printed MF and MI surgical guides, designed for the placement of internal tapered implant fixtures, were used with slot and non-slot drills. The following groups were compared: group 1, MI surgical guide with slot drill; group 2, MI surgical guide with a non-slot drill; and group 3, MF surgical guide with a slot drill. A constant-temperature water bath at 36℃ was used. The drilling was performed in 6 stages, and the initial, highest, and lowest temperatures of the cortical bone were measured at each stage using a non-contact infrared thermometer. Results: There were no temperature increases above the initial temperature in any drilling procedure. The only significant difference between the non-slot and slot groups was observed with the use of the first drill in the MI group, with a higher temperature in the non-slot group (P=0.012). When the heat generation during the first and the second drilling was compared in the non-slot group, the heat generation during the first drilling was significantly higher (P<0.001), and there was no significant difference in heat generation between the drills in the slot group. Conclusions: Within the limitations of this study, implant-site preparation with the surgical guide showed no critical increase in the temperature of the cortical bone, regardless of whether there was a slot in the drill. In particular, the slotted drill had a cooling effect during the initial drilling.

Immunostimulatory effects of dairy probiotic strains Bifidobacterium animalis ssp. lactis HY8002 and Lactobacillus plantarum HY7717

  • Ju-Yeon, Kim;Joo Yun, Kim;Hyeonji, Kim;Eun Chae, Moon;Keon, Heo;Jae-Jung, Shim;Jung-Lyoul, Lee
    • Journal of Animal Science and Technology
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    • v.64 no.6
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    • pp.1117-1131
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    • 2022
  • Previous studies reported that Bifidobacterium animalis ssp. lactis HY8002 (HY8002) improved intestinal integrity and had immunomodulatory effects. Lactobacillus plantarum HY7717 (HY7717) was screened in vitro from among 21 other lactic acid bacteria (LAB) and demonstrated nitric oxide (NO) production. The aims of this study were to investigate the individual and combined ex vivo and in vivo effects of LAB strains HY8002 and HY7717 at immunostimulating mice that have been challenged with an immunosuppressant drug. The combination of HY8002 and HY7717 increased the secretion of cytokines such as interferon (IFN)-γ, interleukin (IL)-12, and tumor necrosis factor (TNF)-α in splenocytes. In a cyclophosphamide (CTX)-induced immunosuppression model, administration of the foregoing LAB combination improved the splenic and hematological indices, activated natural killer (NK) cells, and up-regulated plasma immunoglobulins and cytokines. Moreover, this combination treatment increased Toll-like receptor 2 (TLR2) expression. The ability of the combination treatment to upregulate IFN-γ and TNF-α in the splenocytes was inhibited by anti-TLR2 antibody. Hence, the immune responses stimulated by the combination of HY8002 and HY7717 are associated with TLR2 activation. The preceding findings suggest that the combination of the HY8002 and HY7717 LAB strains could prove to be a beneficial and efficacious immunostimulant probiotic supplement. The combination of the two probiotic strains will be applied on the dairy foods including yogurt and cheese.

TCF4-Targeting miR-124 is Differentially Expressed amongst Dendritic Cell Subsets

  • Sun Murray Han;Hye Young Na;Onju Ham;Wanho Choi;Moah Sohn;Seul Hye Ryu;Hyunju In;Ki-Chul Hwang;Chae Gyu Park
    • IMMUNE NETWORK
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    • v.16 no.1
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    • pp.61-74
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    • 2016
  • Dendritic cells (DCs) are professional antigen-presenting cells that sample their environment and present antigens to naïve T lymphocytes for the subsequent antigen-specific immune responses. DCs exist in a range of distinct subpopulations including plasmacytoid DCs (pDCs) and classical DCs (cDCs), with the latter consisting of the cDC1 and cDC2 lineages. Although the roles of DC-specific transcription factors across the DC subsets have become understood, the posttranscriptional mechanisms that regulate DC development are yet to be elucidated. MicroRNAs (miRNAs) are pivotal posttranscriptional regulators of gene expression in a myriad of biological processes, but their contribution to the immune system is just beginning to surface. In this study, our in-house probe collection was screened to identify miRNAs possibly involved in DC development and function by targeting the transcripts of relevant mouse transcription factors. Examination of DC subsets from the culture of mouse bone marrow with Flt3 ligand identified high expression of miR-124 which was able to target the transcript of TCF4, a transcription factor critical for the development and homeostasis of pDCs. Further expression profiling of mouse DC subsets isolated from in vitro culture as well as via ex vivo purification demonstrated that miR-124 was outstandingly expressed in CD24+ cDC1 cells compared to in pDCs and CD172α+ cDC2 cells. These results imply that miR-124 is likely involved in the processes of DC subset development by posttranscriptional regulation of a transcription factor(s).

In vitro propagation of a rare and endangered species, Echinosophora koreensis Nakai, by axillary bud culture (희귀 및 멸종위기 수종 개느삼의 액아배양을 통한 기내번식)

  • Moon, Heung-Kyu;Kim, Yong-Wook
    • Journal of Plant Biotechnology
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    • v.35 no.3
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    • pp.229-234
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    • 2008
  • An efficient micropropagation was established by using axillary bud explants from two-year-old tree(Echinosphorea koreensis Nakai), which has been known as a rare and endangered species. Among various basal media tested, DKW medium was shown to be the best for axillary shoot elongation. The addition of both BA and TDZ to the medium induced 6 to 10 shoots per explant during eight weeks of culture, without showing any abnormal morphology at the shoot proliferation stage. However, high concentration of TDZ(>0.05 mg/L) appeared to cause hyperhydration on either leaf or shoot at the later developmental stage. Approximately 20% of shoots produced roots by the addition of 1.0 mg/L NAA but not by IBA($0.2{\sim}1.0$ mg/L). Ex vitro micro-cuttings were better source for root induction; up to 58.6% of the micro-cuttings rooted when 100 mg/L IBA was applied to the soil(vermiculite). More than 90% of plantlets with roots were successfully acclimatized and grew normally in the field. Therefore, we suggest that this endangered tree species can be effectively micropropagated by axillary bud culture system developed in this study.

IN VITRO STUDY ON CARIOGENIC POTENTIAL OF SYRUP-FORM MEDICINES FOR CHILDREN (어린이용 시럽제의 우식유발능에 대한 생체 외 연구)

  • Lim, Hwa-Shin;Lee, Kwang-Hee;La, Ji-Young;An, So-Youn;Kim, Yun-Hee
    • Journal of the korean academy of Pediatric Dentistry
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    • v.38 no.2
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    • pp.146-154
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    • 2011
  • An in vitro study on cariogenic potential of 8 over-the-counter syrups for children was performed. The experimental groups were 8 over-the-counter syrups. The positive control group was 10% sucrose solution, and the negative control group was artificial saliva. The pH of each group was determined. The buffering capacity was measured by the volume of 2 N NaOH adding to equalize the pH of 20 ml of experimental solution to pH 7. The consistency was measured by the time to pass Ostwald pipette for 2 ml of the experimental solution. The experimental solutions were inoculated with S. mutans and cultured in $37^{\circ}C$ anaerobic condition for 48 hours. To estimate acid production, pH of the experimental solutions were measured before and after the culture. The primary teeth specimens were soaked in the experimental solutions for 20 minutes three times a day. Except on those hours the specimens were stored in artificial saliva. After 5 days, the microhardness changes of the specimens were measured. These results show that most of syrup-form medicines for children tend to have cariogenic potential partially in endogenous pH, buffering capacity, consistency, acid production and erosive ability of enamel. For the oral health of children, the alternative sweeteners (ex. xylitol) may be substituted for the cariogenic sweeteners of syrups. Additionally, It may be helpful that the chewable tablet replace liquid or syrup in term of dose form.