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Functional Analysis of the Stress-Inducible Soybean Calmodulin Isoform-4 (GmCaM-4) Promoter in Transgenic Tobacco Plants

  • Park, Hyeong Cheol;Kim, Man Lyang;Kang, Yun Hwan;Jeong, Jae Cheol;Cheong, Mi Sun;Choi, Wonkyun;Lee, Sang Yeol;Cho, Moo Je;Kim, Min Chul;Chung, Woo Sik;Yun, Dae-Jin
    • Molecules and Cells
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    • v.27 no.4
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    • pp.475-480
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    • 2009
  • The transcription of soybean (Glycine max) calmodulin isoform-4 (GmCaM-4) is dramatically induced within 0.5 h of exposure to pathogen or NaCl. Core cis-acting elements that regulate the expression of the GmCaM-4 gene in response to pathogen and salt stress were previously identified, between -1,207 and -1,128 bp, and between -858 and -728 bp, in the GmCaM-4 promoter. Here, we characterized the properties of the DNA-binding complexes that form at the two core cis-acting elements of the GmCaM-4 promoter in pathogen-treated nuclear extracts. We generated GUS reporter constructs harboring various deletions of approximately 1.3-kb GmCaM-4 promoter, and analyzed GUS expression in tobacco plants transformed with these constructs. The GUS expression analysis suggested that the two previously identified core regions are involved in inducing GmCaM-4 expression in the heterologous system. Finally, a transient expression assay of Arabidopsis protoplasts showed that the GmCaM-4 promoter produced greater levels of GUS activity than did the CaMV35S promoter after pathogen or NaCl treatments, suggesting that the GmCaM-4 promoter may be useful in the production of conditional gene expression systems.

Effects of Porphyromonas gingivalis extracts on the function of mouse calvarial primary osteoblastic cells (Porphyromonas gingivalis 추출물이 마우스 두개골 일차 조골세포의 기능에 미치는 효과)

  • Yun, Jeong-Ho;Choi, Seong-Ho;Cho, Kyoo-Sung;Chai, Jung-Kiu;Kim, Chong-Kwan;Kim, Chang-Sung
    • Journal of Periodontal and Implant Science
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    • v.33 no.4
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    • pp.585-597
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    • 2003
  • Porphyromonas gingivalis has been implicated as an important periodontophathic bacterium in the etiology and progression of periodontal diseases. It has been reported that P.gingivalis may mediate periodontal destruction not only directly through its virulence factors, but also indirectly by including complex host mediated inflammatory reponses. The purpose of this study was t o evaluate the effects of P.gingivalis on the bone formation and resorption by osteoblasts. For this purpose, after determining the concentration below which sonicated P.gingivalis extracts (SPEs) have no cytotoxicity on mouse calvarial primary osteoblastic (POB) cells, we investigated the effects of SPEs on the alkaline phosphatase (ALP) activity, matrix metalloproteinase (MMP) expression (MMP-2, -9, 13), and prostaglandin $E_2$ ($PGE_2$) release in POB cells by treatment with SPEs below that concentration. The results were as follows; 1. SPEs showed no cytotoxic effect on POB cells up to a concentration of 1 ${\mu}m$/ml. 2. The treatment with SPEs reduced ALP activity in a dose-dependent manner in POB cells, In addition, when we investigated the effect of SPEs (1 ${\mu}m$/ml) on ALP activity for different exposure periods, statistically significant inhibition of ALP activity was shown at 2 days of exposure, and further significant inhibition occurred by extending the periods of exposure. 3. The treatment with SPEs stimulated the gene expression of MMP-9 in POB cells. 4. The pre-treatment with SPEs increased the amount of $PGE_2$ released in POB cells. In summary, the present study shows that P.gingivalis could inhibit osteogenesis and stimulate bone resorption not only by reducing ALP activity but also by increasing MMP-9 mRNA expression in osteoblasts, possibly through an endogenous $PGE_2$ pathway. In addition, our results suggest that if P.gingivalis affects osteoblasts in early differentiation stage, such effects by P. gingivalis could be irreversible.

Solubilization and Reconstitution of ${\Delta}^{5}-3{\beta}$-Hydroxy Steroid Acyl Transferase from the Rat Brain (쥐 뇌의 ${\Delta}^{5}-3{\beta}$-Hydroxy Steroid Acyl 전이효소의 용해 및 Liposome에서의 재구성)

  • Ko, Kyu-Jung;Park, In-Ho;Han, Beom-Ku;Jo, Do-Hyun
    • Applied Biological Chemistry
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    • v.38 no.4
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    • pp.320-324
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    • 1995
  • Solubilization of microsomal ${\Delta}^{5}-3{\beta}$-hydroxy steroid acyl transfearse(${\Delta}^{5}-3{\beta}$-OH-SAT) of rat brain and its reconstitution into liposomes were investigated. Among the detergents utilized for the solubilization, deoxycholic acid was superior to Tween 80 or Triton X-100 for the reconstituted activity of ${\Delta}^{5}-3{\beta}$-OH-SAT. The enzyme activity was shown to be affected by the nature of phospholipids used for the preparation of the liposome. Phosphatidylcholines from egg yolk and soybean showed the highest activity of ${\Delta}^{5}-3{\beta}$-OH-SAT and phosphatidylethanolamine came next. However phosphatidylserine and phosphatidic acid showed a lower activity than those obtained before the reconstitution. This study suggests that the presence of quaternary ammonium salt or amine group in the phospholipids stimulates the activity of ${\Delta}^{5}-3{\beta}$-OH-SAT. However the presence of a carboxylic group or the absence of the amine group may have an inhibitory effect on the ${\Delta}^{5}-3{\beta}$-OH SAT.

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A Meta-analysis of Effectiveness of Kigong Programs for the Physical Health of the Elderly (노인 신체건강을 위한 기공(氣功)프로그램 효과성 메타분석)

  • Kim, Ilsik;Kim, Gyeryung;Lee, Su-kyung;Seo, Hochan
    • 한국노년학
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    • v.38 no.1
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    • pp.65-82
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    • 2018
  • The purpose of this study is to statistically summarize the effect sizes using meta-analysis of various Kigong programs implemented on the subject of physical health of the elderly. Including 23 academic journal studies and 11 thesis studies reported domestically until 2016, a total of 34 studies were selected and analyzed to verify the effect sizes of programs, program types, measuring variables, and activity elements. The findings are as follows. First, it was found that the programs were midium effective with overall average effect size showing .748(p<.001). A program of Tai-Chi indicated the largest effect size of .836(p<.001). In terms of measuring variables, physical strength showed the largest effect size of .806(p<.001). In terms of gender by group, it was found that groups consisting only of men indicated the largest effect size of 2.388(p<.001). As for effect size by group of disease state, normal group with showed the highest effect size of .804(p<.001). Second, in activity elements, 31~60 by the group size has the largest effect size of .873(p<.001), total number of sessions for 31~40 sessions indicated the largest effect value of .829(p<.001). As for number of sessions per week, 5 sessions showed largest effect value of .918(p<.001), in terms of activity time, 50 minutes showed large effect value of 1.391(p<.001). As summarized above, Kigong programs are midium effective for physical health of the elderly, as an activity elements to consider when configuring the program, for group size 31~60, 31~40 sessions in total, 5 sessions per week and 50 minutes per sessions were recommended.

Thiobarbituric Acid Reactive Substances Levels in Brain Tissue of Aldh2 Knockout Mice Following Ethanol Exposure for 8 Weeks (Aldh2 knockout 마우스에서 8주간 에탄올 노출에 따른 뇌조직의 thiobarbituric acid reactive substances 농도)

  • Moon, Sun-In;Eom, Sang-Yong;Kim, Jung-Hyun;Yim, Dong-Hyuk;Kim, Hyong-Kyu;Kim, Yong-Dae;Kim, Heon
    • Journal of Life Science
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    • v.21 no.8
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    • pp.1163-1167
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    • 2011
  • Excessive alcohol consumption causes various degenerative brain diseases including Alzheimer's disease and Parkinson's disease. Absorbed ethanol is metabolized to acetaldehyde and acetic acid by alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH). Acetaldehyde is well known as a toxicant through generation of reactive oxygen species (ROS). Therefore, ALDH2 activity may play important roles in the pathogenesis of alcohol-induced brain diseases. In this study, we demonstrated the effects of ALDH2 enzyme activity on lipid peroxidation in brain tissues and urine of mice exposed to ethanol for 8 weeks. Five male, 8-week old Aldh2 (+/+) and Aldh2 (-/-) mice (C57BL/6J strain) in each group were exposed to ethanol for 8 weeks (2 g/kg wt./day) using gavage, and those in the control group received 0.9% saline alone. Thiobarbituric acid reactive substances (TBARS) level, a marker for lipid peroxidation, was measured in whole brain tissue and urine by high performance liquid chromatography. As a result, chronic ethanol treatment did not show any statistical change on the TBARS level of brain tissue in both Aldh2 (+/+) mice and in Aldh2 (-/-) mice. However, following ethanol exposure for 8 weeks in Aldh2 (-/-) mice, the urinary TBARS levels were significantly increased to more than double compared to the pretreatment group. This result was not observed in Aldh2 (+/+) mice. These results suggest that although ALDH2 enzyme activity plays a role in the generation of ROS in the whole body, it does not seem to be important in the pathogenesis of alcohol induced degenerative brain diseases.

The impact of functional brain change by transcranial direct current stimulation effects concerning circadian rhythm and chronotype (일주기 리듬과 일주기 유형이 경두개 직류전기자극에 의한 뇌기능 변화에 미치는 영향 탐색)

  • Jung, Dawoon;Yoo, Soomin;Lee, Hyunsoo;Han, Sanghoon
    • Korean Journal of Cognitive Science
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    • v.33 no.1
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    • pp.51-75
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    • 2022
  • Transcranial direct current stimulation (tDCS) is a non-invasive brain stimulation that is able to alter neuronal activity in particular brain regions. Many studies have researched how tDCS modulates neuronal activity and reorganizes neural networks. However it is difficult to conclude the effect of brain stimulation because the studies are heterogeneous with respect to the stimulation parameter as well as individual difference. It is not fully in agreement with the effects of brain stimulation. In particular few studies have researched the reason of variability of brain stimulation in response to time so far. The study investigated individual variability of brain stimulation based on circadian rhythm and chronotype. Participants were divided into two groups which are morning type and evening type. The experiment was conducted by Zoom meeting which is video meeting programs. Participants were sent experiment tool which are Muse(EEG device), tdcs device, cell phone and cell phone holder after manuals for experimental equipment were explained. Participants were required to make a phone in frount of a camera so that experimenter can monitor online EEG data. Two participants who was difficult to use experimental devices experimented in a laboratory setting where experimenter set up devices. For all participants the accuracy of 98% was achieved by SVM using leave one out cross validation in classification in the the effects of morning stimulation and the evening stimulation. For morning type, the accuracy of 92% and 96% was achieved in classification in the morning stimulation and the evening stimulation. For evening type, it was 94% accuracy in classification for the effect of brain stimulation in the morning and the evening. Feature importance was different both in classification in the morning stimulation and the evening stimulation for morning type and evening type. Results indicated that the effect of brain stimulation can be explained with brain state and trait. Our study results noted that the tDCS protocol for target state is manipulated by individual differences as well as target state.

Effect of the Oriental Medicine Gi-Gong Exercise on the Brain Power, HRV, Live Blood Condition in the Youth (한방기공체조가 두뇌력, 심박변이율, 생혈액형태에 미치는 영향)

  • Lee, Jeong-Won;Kim, Yi-Soon;Kim, Gyeong-Cheol
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.21 no.1
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    • pp.126-135
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    • 2007
  • This study was conducted to identify the effects of the oriental medicine Qigong Exercise on the brain power, HRV, pulsation, live blood condition among young boys and girls. The study was performed with two group(control group and experimental group) in a pre-test/post-test design. The subjects were 44 young boys and girls selected by a some middle school in Busan. The oriental medicine Qigong Exercise program consists of 80-minute sessions three times a week over 5 months. All of the subjects were examined on the congnition assessment tool, stress assessment tool, oriental medicine pulsation 3-D MAC, live blood condition analyzer Prior and post surveys were measured before and after the experiment. In the cognition assessment, the amplitude of ERS were increased afer Qigong Exercise. The Success and the Concentration were significantly increased afer Qigong Exercise, the Error was significantly decreased afer Qigong Exercise. The Cognition strength was significantly increased, but the Reponse time was not significantly decreased afer Qigong Exercise. And the Workload was not significantly decreased, the Total Score was not significantly increased afer Qigong Exercise. Among the stress assessment, RRV tachogram's ‘mean RR’ was significantly increased, ‘mean HRV’ was significantly decreased afer Qigong Exercise. SDNN was not significantly increased, Complexity was not significantly increased afer Qigong Exercise. And TP(RRV power's total power) was not significantly increased, VLF and HF was significantly increased, and LF was significantly decreased afer Qigong Exercise. ANS's norm LF was not significantly decreased, but norm HF was significantly increased afer Qigong Exercise. In the RRV's Phase Plot, RMSSD and SDSD were not significantly increased, pnn50 was not significantly decreased afer Qigong Exercise. On the whole, Parasympathetic Activity and Stress Endurance were significantly increased, but Cardiac Activity and Physical Arousal were not significantly increased afer Qigong Exercise. Cardiac Aging was not significantly decreased afer Qigong Exercise. Sympathetic Activity, Autonomic Nervous System Balance and Heart-load were not significantly decreased afer Qigong Exercise. In the pulsation, press power was increased(15%), and w/t(pressurization time / pulsation time) was decreased(20%) afer Oigong Exercise. And the live blood condition was not changed afer Qigong Exercise. As mentioned above; the oriental medicine Qigong Exercise program was identified the effects of the inspiration of the brain power, heart rate and anti-stress.

2-DG Autoradiographic Imaging of Brain Activity Patterns by Electroacupuncture Stimulation in Awake Rats (전침자극(電針刺戟)에 의한 흰쥐 중추신경계(中樞神經系)내 대사활성(代謝活性) 변화(變化)의 영상화(映像化) 연구(硏究))

  • Sohn, Young-Joo;Won, Ran;Jung, Hyuk-Sang;Kim, Yong-Suk;Park, Young-Bae;Sohn, Nak-Won
    • Journal of Acupuncture Research
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    • v.18 no.3
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    • pp.56-68
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    • 2001
  • Objective : Functional brain mapping study on acupuncture stimulation using the [14C]2-deoxyglucose([14C]2-DG) autoradiography provides quantitative data and visualized pathway in central nervous system(CNS). We aimed to investigate the neural pathway and spatial distribution of metabolic activity elicited in CNS on electroacupuncture stimulation using [14C]2-DG autoradiography. Methods : The study were divided into three groups by stimulation times. 45-mins stimulation group according to Sokoloffs method, 5-mins stimulation group according to Duncun's method, and 15-mins stimulation group. ;A venous catheter was equipped into right jugular vein. The rats (Sprague-Dawley rats, 230-260g) were kept fastened loosely on a holding platform without anesthesia. Electroacupuncture stimulation (5 ms, 2 Hz, 1~3 mA) were applied on the left Zusanli (ST36) acupoint and [14C]2-DG ($25{\mu}Ci/rat$) injection was performed through the catheter. After sacrifice, the brain and the spinal cord were made to sections for film image. The film images were digitalized as the isotope concentration based upon comparison of optical densities with that of the standards and normalized by the optical density of corpus callosum. Results : 1. 15-mins stimulation group was most effective among 3 experiments. 2. On 15-mins stimulation group, medial geniculate nucleus, intetpeduncular nucleus intermedius, ventral periolivary nucleus, caudal periolivary nucleus, medial superior olive, lateral paragigantocellular nucleus, including hypothalamic arcuate nucleus were increased by more than 25% (at least, p<0.05) by electroacupuncture stimulation. 3. Especially, the metabolism in hypothalamic arcuate nucleus was increased by 90% (p<0.05). 4. The fact that arcuate nucleus of hypothalamus might play a role of interconnection area between ascending and descending pathway of acupuncture stimulation was demonstrated visually. Conclusions : Advanced study on electroacupuncture stimulation elicited significant increase of metabolic activity in various nuclei of hypothalamus will provide the important experimental basis in research of the relationship between electroacupuncture stimulation and internal visceral functions.

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