• 제목/요약/키워드: ex vitro

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Extracellular Signal-regulated Kinase Activation Is Required for Serine 727 Phosphorylation of STAT3 in Schwann Cells in vitro and in vivo

  • Lee, Hyun-Kyoung;Jung, Jun-Yang;Lee, Sang-Hwa;Seo, Su-Yeong;Suh, Duk-Joon;Park, Hwan-Tae
    • The Korean Journal of Physiology and Pharmacology
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    • 제13권3호
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    • pp.161-168
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    • 2009
  • In the peripheral nerves, injury-induced cytokines and growth factors perform critical functions in the activation of both the MEK/ERK and JAK/STAT3 pathways. In this study, we determined that nerve injury-induced ERK activation was temporally correlated with STAT3 phosphorylation at the serine 727 residue. In cultured Schwann cells, we noted that ERK activation is required for the serine phosphorylation of STAT3 by neuropoietic cytokine interleukin-6 (IL-6). Serine phosphorylated STAT3 by IL-6 was transported into Schwann cell nuclei, thereby indicating that ERK may regulate the transcriptional activity of STAT3 via the induction of serine phosphorylation of STAT3. Neuregulin-1 (NRG) also induced the serine phosphorylation of STAT3 in an ERK-dependent fashion. In contrast with the IL-6 response, serine phosphorylated STAT3 induced by NRG was not detected in the nucleus, thus indicating the non-nuclear function of serine phosphorylated STAT3 in response to NRG. Finally, we determined that the inhibition of ERK prevented injury-induced serine phosphorylation of STAT3 in an ex-vivo explants culture of the sciatic nerves. Collectively, the results of this study show that ERK may be an upstream kinase for the serine phosphorylation of STAT3 induced by multiple stimuli in Schwann cells after peripheral nerve injury.

Plant Extracts and Plant-Derived Compounds: Promising Players in Countermeasure Strategy Against Radiological Exposure: A Review

  • Kma, Lakhan
    • Asian Pacific Journal of Cancer Prevention
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    • 제15권6호
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    • pp.2405-2425
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    • 2014
  • Radiation exposure leads to several pathophysiological conditions, including oxidative damage, inflammation and fibrosis, thereby affecting the survival of organisms. This review explores the radiation countermeasure properties of fourteen (14) plant extracts or plant-derived compounds against these cellular manifestations. It was aimed at evaluating the possible role of plants or its constituents in radiation countermeasure strategy. All the 14 plant extracts or compounds derived from it and considered in this review have shown some radioprotection in different in vivo, ex-vivo and or in vitro models of radiological injury. However, few have demonstrated advantages over the others. C. majus possessing antioxidant, anti-inflammatory and immunomodulatory effects appears to be promising in radioprotection. Its crude extracts as well as various alkaloids and flavonoids derived from it, have shown to enhance survival rate in irradiated mice. Similarly, curcumin with its antioxidant and the ability to ameliorate late effect of radiation exposure, combined with improvement in survival in experimental animal following irradiation, makes it another probable candidate against radiological injury. Furthermore, the extracts of P. hexandrum and P. kurroa in combine treatment regime, M. piperita, E. officinalis, A. sinensis, nutmeg, genistein and ginsan warrants further studies on their radioprotective potentials. However, one that has received a lot of attention is the dietary flaxseed. The scavenging ability against radiation-induced free radicals, prevention of radiation-induced lipid peroxidation, reduction in radiation cachexia, level of inflammatory cytokines and fibrosis, are some of the remarkable characteristics of flaxseed in animal models of radiation injury. While countering the harmful effects of radiation exposure, it has shown its ability to enhance survival rate in experimental animals. Further, flaxseed has been tested and found to be equally effective when administered before or after irradiation, and against low doses (${\leq}5Gy$) to the whole body or high doses (12-13.5 Gy) to the whole thorax. This is particularly relevant since apart from the possibility of using it in pre-conditioning regime in radiotherapy, it could also be used during nuclear plant leakage/accidents and radiological terrorism, which are not pre-determined scenarios. However, considering the infancy of the field of plant-based radioprotectors, all the above-mentioned plant extracts/plant-derived compounds deserves further stringent study in different models of radiation injury.

식용식물의 항산화효과 검색 (Screening for Antioxidant Activity of Edible Plants)

  • 조세연;한용봉;신국현
    • 한국식품영양과학회지
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    • 제30권1호
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    • pp.133-137
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    • 2001
  • 21종 식용식물의 부위별 MeOH Ex. DPPH와 TBA법으로 항산화 효과를 조사한 결과 DPPH를 이용한 유리라디칼 소거 효과는 참취잎, 쑥갓잎 및 머위잎에서 특이적으로 높은 효과가 관찰되었으며 시료추출물의 buffer에 의해 유발된 과산화지질 억제효과는 참외껍질의 억제 효과 강도는 $IC_{50}$ 값이 $10\mu\textrm{g}/mL$로 매우 강하였다. $S_2O_z$로 유발시킨 지질과산화에 대한 억제효과는 0.25 mg 농도에서 L-ascorbic acid의 과산화 억제율과 비교하면 가지 잎, 머위 잎, 감자껍질의 항산화 억제 효과가 가장 높았으며 $FeSO_4$로 유발시킨 지질과산화에 대한 억제효과는 쑥갓잎의 $IC_{50}$가 0.08 mg/mL로 특이적으로 높았다.

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Novel Anti-Angiogenic Activity in Rubus coreanus Miquel Water Extract Suppresses VEGF-Induced Angiogenesis

  • Kim, Eok-Cheon;Kim, Hye Jin;Kim, Tack-Joong
    • 대한의생명과학회지
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    • 제20권4호
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    • pp.209-220
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    • 2014
  • Vascular endothelial growth factor (VEGF) is a key factor involved in the induction of angiogenesis and has become an attractive target for anti-angiogenesis therapies. The purpose of this study was to elucidate the anti-angiogenic activity of Rubus coreanus Miquel water extract (RCME). Rubus coreanus Miquel has long been employed as a traditional medicine, and recent studies have demonstrated that it has measureable biological activities. Thus, we investigated for the first time the effect of RCME on angiogenesis and its underlying signaling pathways. The effects of RCME were tested on in vitro models of angiogenesis, namely, proliferation, migration, invasion and tube formation of human umbilical vein endothelial cells as well as an ex vivo model of vessel sprouting from the rat aorta in response to VEGF. We observed that VEGF-induced angiogenesis was strongly suppressed by RCME treatment compared to that of the control group. Moreover, we found that RCME inhibited VEGF-induced activation of matrix metalloproteinases and phosphorylation of extracellular signal-regulated kinase and p38, and also effectively inhibited phosphorylation of VEGF receptor 2. These results indicated that RCME inhibits angiogenesis by suppressing phosphorylation of the VEGF receptor and may be useful for the treatment of angiogenesis-dependent diseases such as cancer and diabetic retinopathy.

계지복령환의 효능에 대한 국내외 실험연구 동향 분석 (The Analysis of the Experimental Research Trend of Efficacy of Gyejibokryeong-hwan)

  • 장새별;백선은;최경희;유정은
    • 대한한방부인과학회지
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    • 제29권2호
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    • pp.99-112
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    • 2016
  • Objectives: Gyejibokryeong-hwan (GBH), a traditional Korean herbal medicine, has been widely used for the treatment of the blood stasis syndrome. This study is purposed to analyze the experimental research trend of GBH in Korea for developing further research plan. Methods: A search of Korean research database-Oasis, RISS and KISS- and Pubmed was carried out for publications until 2015, for the words, 'Guizhifulingwan', ‘Gyejibokryeonghwan’, or ‘Keishibukuryogan’. Then study selection is conducted according to PRISMA guidelines, studies not related or using modified formula or administered for human are excluded, 48 studies are included in this review, finally. We analyzed studies by research method, subject, outcome measure, and result of the study. Results: There were 31 in vivo studies about the effect of GBH on platelet aggregation, anti-oxidant, blood viscosity, and hypercholesterolemia, etc. 12 in vitro studies were about the effect of GBH on the cervical carcinoma, chronic kidney disease, uterine myoma, hepatocarcinoma, atherosclerosis, cancer chemo-prevent. 9 ex vivo studies were about the effect of GBH on the platelet aggregation, chronic kidney disease, ovaulatory disorder, and rheumarthritis.Conclusions: We proposed the translational research of GBH involving scientific discoveries and developing practical applications by investigating the concept of blood stasis syndrome in terms of current physiopathological mechanism.

In vitro evaluation of the bond strength between various ceramics and cobalt-chromium alloy fabricated by selective laser sintering

  • Bae, Eun-Jeong;Kim, Hae-Young;Kim, Woong-Chul;Kim, Ji-Hwan
    • The Journal of Advanced Prosthodontics
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    • 제7권4호
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    • pp.312-316
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    • 2015
  • PURPOSE. This study aimed to present the clinical applicability of restorations fabricated by a new method, by comparing the bond strength of between ceramic powder with different coefficient of thermal expansion and alloys fabricated by Selective laser sintering (SLS). MATERIALS AND METHODS. Fifty Co-Cr alloy specimens ($25.0{\times}3.0{\times}0.5mm$) were prepared by SLS and fired with the ceramic ($8.0{\times}3.0{\times}0.5mm$) (ISO 9693:1999). For comparison, ceramics with different coefficient of thermal expansion were used. The bond strength was measured by three-point bending testing and surfaces were observed with FE-SEM. Results were analyzed with a one-way ANOVA (${\alpha}$=.05). RESULTS. The mean values of Duceram Kiss ($61.18{\pm}6.86MPa$), Vita VM13 ($60.30{\pm}7.14MPa$), Ceramco 3 ($58.87{\pm}5.33MPa$), Noritake EX-3 ($55.86{\pm}7.53MPa$), and Vintage MP ($55.15{\pm}7.53MPa$) were found. No significant difference was observed between the bond strengths of the various metal-ceramics. The surfaces of the specimens possessed minute gaps between the additive manufactured layers. CONCLUSION. All the five powders have bond strengths higher than the required 25 MPa minimum (ISO 9693); therefore, various powders can be applied to metal structures fabricated by SLS.

In vitro Expansion of Umbilical Cord Blood Derived Mesenchymal Stem Cells (UCB-MSCs) Under Hypoxic Conditions

  • Yang, Jungyun;Kwon, Jihye;Kim, Miyeon;Bae, Yunkyung;Jin, Hyejin;Park, Hohyun;Eom, Young Woo;Rhee, Ki-Jong
    • 대한의생명과학회지
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    • 제21권1호
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    • pp.40-49
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    • 2015
  • Mesenchymal stem cells (MSCs) have the ability to self-renew and differentiate into multi-lineage cells, thus highlighting the feasibility of using umbilical cord blood-derived MSCs (UCB-MSCs) for cell-therapy and tissueengineering. However, the low numbers of UCB-MSC derived from clinical samples requires that an ex vivo expansion step be implemented. As most stem cells reside in low oxygen tension environments (i.e., hypoxia), we cultured the UCBMSCs under 3% $O_2$ or 21% $O_2$ and the following parameters were examined: proliferation, senescence, differentiation and stem cell specific gene expression. UCB-MSCs cultured under hypoxic conditions expanded to significantly higher levels and showed less senescence compared to UCB-MSCs cultured under normoxic conditions. In regards to differentiation potential, UCB-MSCs cultured under hypoxic and normoxic conditions both underwent similar levels of osteogenesis as determined by ALP and von Kossa assay. Furthermore, UCB-MSCs cultured under hypoxic conditions exhibited higher expression of OCT4, NANOG and SOX2 genes. Moreover, cells expanded under hypoxia maintained a stem cell immnunophenotype as determined by flow cytometry. These results demonstrate that the expansion of human UCB-MSCs under a low oxygen tension microenvironment significantly improved cell proliferation and differentiation. These results demonstrate that hypoxic culture can be rapidly and easily implemented into the clinical-scale expansion process in order to maximize UCB-MSCs yield for application in clinical settings and at the same time reduce culture time while maintaining cell product quality.

4종 창포류의 성분 패턴 및 항산화 효능 비교 (Comparative Study of Changpo(Ch$\={a}$ngp$\'{u}$) species on Antioxidant Activity and HPLC Pattern Analysis)

  • 최고야;고병섭;이미영;채성욱;김영화;육진아;백지성;이혜원
    • 대한본초학회지
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    • 제26권1호
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    • pp.13-19
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    • 2011
  • Objective : This study was to compare antioxidant activity and HPLC pattern analysis from 4 species of changpo(ch$\={a}$ngp$\'{u}$). Methods : To compare the antioxidant activity and HPLC pattern analysis from the 4 species of changpo, we performed the in vitro anti-oxidative activity assays and HPLC analysis from 70% ethanol extracts of Acorus gramineus Sol. (=AG), A. tatarinowii Schott (=AT), A. calamus L. (=AC) and Anemone altaica Fisch. ex C.A.Mey (=AA) taken in the herbal medicine market of Korea. Results : AG has the most effective anti-oxidative activity among 4 species of changpo. As the HPLC pattern analysis, AT was detected the unknown peak at retention time 14.9 min whereas AG was not showed any peak at the same retention time. These results suggest that AG could be used rather than AT when it need to be prescribed as anti-oxidative medicine. Conclusions : This result can be used as the basic data contributing to the stability of AG according to an appropriate clinical application.

Biological Control of Apple Ring Rot on Fruit by Bacillus amyloliquefaciens 9001

  • Li, Yan;Han, Li-Rong;Zhang, Yuanyuan;Fu, Xuechi;Chen, Xinyi;Zhang, Lixia;Mei, Ruhong;Wang, Qi
    • The Plant Pathology Journal
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    • 제29권2호
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    • pp.168-173
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    • 2013
  • Apple ring rot disease, caused by Botryosphaeria dothidea (Moug. ex. Fr) Ces. et de Not., is one of the most important diseases on apple fruits. In this study, strain 9001 isolated from healthy apple fruits from an infested orchard was evaluated for its biocontrol activity against apple ring rot in vitro and in vivo. Strain 9001 showed obvious antagonistic activity to B. dothidea YL-1 when plated on potato dextrose agar. Soaking healthy apples in the bacterial suspensions of strain 9001 prior to artificial inoculation of fungal pathogen resulted in a dramatic decrease in disease incidence when compared to the control. Moreover, either field application in the growth season or postharvest treatment of apples from infected orchards with bacterial suspensions of strain 9001 resulted in significantly reduced disease incidence within the storage period for 4 months at room temperature. Based on the phylogenetic analysis of 16S rRNA and the gyrA gene, strain 9001 was identified as Bacillus amyloliquefaciens. These results indicated that B. amyloliquefaciens 9001 could be a promising agent in biocontrol of apple ring rot on fruit, which might help to minimize the yield loss of apple fruit during the long postharvest period.

Water soluble tomato concentrate regulates platelet function via the mitogen-activated protein kinase pathway

  • Jeong, Dahye;Irfan, Muhammad;Saba, Evelyn;Kim, Sung-Dae;Kim, Seung-Hyung;Rhee, Man Hee
    • 대한수의학회지
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    • 제56권2호
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    • pp.67-74
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    • 2016
  • Tomato extract has been shown to exert antiplatelet activity in vitro and to change platelet function ex vivo, but with limitations. In this study, antiplatelet activity of water soluble tomato concentrate (Fruitflow I) and dry water soluble tomato concentrate (Fruitflow II) was investigated using rat platelets. Aggregation was induced by collagen and adenosine diphosphate and granule-secretion, $[Ca^{2+}]_i$, thromboxane B2, cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP) levels were examined. The activation of integrin ${\alpha}_{IIb}{\beta}_3$ and phosphorylation of signaling molecules, including mitogen-activated protein kinase (MAPK) and PI3K/Akt, were investigated by flow cytometry and immunoblotting, respectively. Prothrombin time (PT) and activated partial thromboplastin time (aPTT) were examined. Moreover, in vivo thrombus weight was tested by an arteriovenous shunt model. Fruitflow I and Fruitflow II significantly inhibited agonist induced platelet aggregation, adenosine triphosphate and serotonin release, $[Ca^{2+}]_i$, and thromboxane B2 concentration, while having no effect on cAMP and cGMP levels. Integrin ${\alpha}_{IIb}{\beta}_3$ activation was also significantly decreased. Moreover, both concentrates reduced phosphorylation of MAPK pathway factors such as ERK, JNK, P38, and PI3K/Akt. In vivo thrombus formation was also inhibited. Taken together, these concentrates have the potential for ethnomedicinal applications to prevent cardiovascular ailments and can be used as functional foods.