• Title/Summary/Keyword: Translational research

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Neurogenesis and neuronal migration of dopaminergic neurons during mesencephalon development in mice

  • Kim, Mun-ki;Lee, Si-Joon;Vasudevan, Anju;Won, Chungkil
    • Journal of Biomedical and Translational Research
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    • v.19 no.4
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    • pp.125-129
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    • 2018
  • Dopaminergic neurons are one of the major neuronal components in the brain. Mesencephalon dopamine (DA) neurogenesis takes place in the ventricular zone of the floor plate, when DA progenitors divide to generate postmitotic cells. These cells migrate through the intermediate zone while they differentiate and become DA neurons on reaching the mantle zone. However, neurogenesis and neuronal migration on dopaminergic neurons remain largely unexplored in the mesencephalon development. This study presents neurogenesis and neuronal migration patterns of dopaminergic neurons during mesencephalic development of the mouse. Neurons from embryonic day (E) 10-14 were labelled by a single injection of 5-bromodeoxyuridine and immunohistochemistry was performed. The neurogenesis occurred mainly at the E10 and E11, which was uniformly distributed in the mesencephalic region, but neurons after E13 were observed only in the dorsal mesencephalon. At the postnatal day 0 (P0), E10 generated neurons were spread out uniformly in the whole mesencephalon whereas E11-originated neurons were clearly depleted in the red nucleus region. DA neurons mainly originated in the ventromedial mesencephalon at the early embryonic stage especially E10 to E11. DA neurons after E12 were only observed in the ventral mesencephalon. At E17, E10 labelled neurons were only observed in the substantia nigra (SN) region. Our study demonstrated that major neurogenesis occurred at E10 and E11. However, neuronal migration continued until neonatal period during mesencephalic development.

The detection of Toxoplasma gondii ME49 infections in BALB/c mice using various techniques

  • Hae-Ji Kang;Jie Mao;Min-Ju Kim;Keon-Woong Yoon;Gi-Deok Eom;Ki-Back Chu;Eun-Kyung Moon;Fu-Shi Quan
    • Parasites, Hosts and Diseases
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    • v.61 no.4
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    • pp.418-427
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    • 2023
  • Toxoplasma gondii infections are primarily diagnosed by serological assays, whereas molecular and fluorescence-based techniques are garnering attention for their high sensitivity in detecting these infections. Nevertheless, each detection method has its limitations. The toxoplasmosis detection capabilities of most of the currently available methods have not been evaluated under identical experimental conditions. This study aimed to assess the diagnostic potential of enzyme-linked immunosorbent assay (ELISA), real-time polymerase chain reaction (RT-PCR), immunohistochemistry (IHC), and immunofluorescence (IF) in BALB/c mice experimentally infected with various doses of T. gondii ME49. The detection of toxoplasmosis from sera and brain tissues was markedly enhanced in mice subjected to high infection doses (200 and 300 cysts) compared to those subjected to lower doses (10 and 50 cysts) for all the detection methods. Additionally, increased B1 gene expression levels and cyst sizes were observed in the brain tissues of the mice. Importantly, IHC, IF, and ELISA, but not RT-PCR, successfully detected T. gondii infections at the lowest infection dose (10 cysts) in the brain. These findings may prove beneficial while designing experimental methodologies for detecting T. gondii infections in mice.

A Dynamic Analysis of 150 ton Winch using Ocean Environment Data (해양 환경 데이터를 이용한 150톤 윈치의 동특성 해석)

  • Lee, Chang-Ho;Min, Cheon-Hong;Kim, Hyung-Woo;Jang, Jin-Woo;Hwang, Dong-Hwan;Rhyu, Yong-Suk
    • Ocean and Polar Research
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    • v.39 no.3
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    • pp.205-211
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    • 2017
  • This paper seeks to provide a dynamic analysis of a 150 ton winch based on ocean environmental data. The winch model that was subjected to analysis was modeled from CAD to each subsystem by the commercial software DAFUL. The winch model has tree brake systems (disk brake, band brake and ratchet brake). The rotation motion of the motor and contact elements of the brake are applied to the winch model in order to analyze its dynamic characteristics. In addition, a crane-barge was modeled to apply ocean environmental data. The motion data of the crane-barge was produced by means of the RAO(Response Amplitude Operator) of the barge and wave spectrum. The reaction force of the translational joint was measured instead of the tension of the cable. The brake performance of the winch was produced and assessed based on the operating motion of the crane-barge.

Clinical and Neurobiological Relevance of Current Animal Models of Autism Spectrum Disorders

  • Kim, Ki Chan;Gonzales, Edson Luck;Lazaro, Maria T.;Choi, Chang Soon;Bahn, Geon Ho;Yoo, Hee Jeong;Shin, Chan Young
    • Biomolecules & Therapeutics
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    • v.24 no.3
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    • pp.207-243
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    • 2016
  • Autism spectrum disorder (ASD) is a neurodevelopmental disorder characterized by social and communication impairments, as well as repetitive and restrictive behaviors. The phenotypic heterogeneity of ASD has made it overwhelmingly difficult to determine the exact etiology and pathophysiology underlying the core symptoms, which are often accompanied by comorbidities such as hyperactivity, seizures, and sensorimotor abnormalities. To our benefit, the advent of animal models has allowed us to assess and test diverse risk factors of ASD, both genetic and environmental, and measure their contribution to the manifestation of autistic symptoms. At a broader scale, rodent models have helped consolidate molecular pathways and unify the neurophysiological mechanisms underlying each one of the various etiologies. This approach will potentially enable the stratification of ASD into clinical, molecular, and neurophenotypic subgroups, further proving their translational utility. It is henceforth paramount to establish a common ground of mechanistic theories from complementing results in preclinical research. In this review, we cluster the ASD animal models into lesion and genetic models and further classify them based on the corresponding environmental, epigenetic and genetic factors. Finally, we summarize the symptoms and neuropathological highlights for each model and make critical comparisons that elucidate their clinical and neurobiological relevance.

Galectin-3-independent Down-regulation of GABABR1 due to Treatment with Korean Herbal Extract HAD-B Reduces Proliferation of Human Colon Cancer Cells

  • Kim, Kyung-Hee;Kwon, Yong-Kyun;Cho, Chong-Kwan;Lee, Yeon-Weol;Lee, So-Hyun;Jang, Sang-Geun;Yoo, Byong-Chul;Yoo, Hwa-Seong
    • Journal of Pharmacopuncture
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    • v.15 no.3
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    • pp.19-30
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    • 2012
  • Objectives: Many efforts have shown multi-oncologic roles of galectin-3 for cell proliferation, angiogenesis, and apoptosis. However, the mechanisms by which galectin-3 is involved in cell proliferation are not yet fully understood, especially in human colon cancer cells. Methods: To cluster genes showing positively or negatively correlated expression with galectin-3, we employed human colon cancer cell lines, SNU-61, SNU-81, SNU-769B, SNU-C4 and SNU-C5 in high-throughput gene expression profiling. Gene and protein expression levels were determined by using real-time quantitative polymerase chain reaction (PCR) and western blot analysis, respectively. The proliferation rate of human colon cancer cells was measured by using a 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. Results: Expression of ${\gamma}$-aminobutyric acid B receptor 1 (GABABR1) showed a positive correlation with galectin-3 at both the transcriptional and the translational levels. Down-regulation of galectin-3 decreased not only GABABR1 expression but also the proliferation rate of human colon cancer cells. However, Korean herbal extract, HangAmDan-B (HAD-B), decreased expression of GABABR1 without any expressional change of galectin-3, and offset ${\gamma}$-aminobutyric acid (GABA)-enhanced human colon cancer cell proliferation. Conclusions: Our present study confirmed that GABABR1 expression was regulated by galectin-3. HAD-B induced galectin-3-independent down-regulation of GABABR1, which resulted in a decreased proliferation of human colon cancer cells. The therapeutic effect of HAD-B for the treatment of human colon cancer needs to be further validated.

Development of Master-Slave Type Tele-Operation Control Robotic System for Arrhythmia Ablation (부정맥 시술을 위한 마스터-슬레이브 원격제어·로봇 시스템 개발)

  • Moon, Youngjin;Park, Sang Hoon;Hu, Zhenkai;Choi, Jaesoon
    • Journal of Institute of Control, Robotics and Systems
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    • v.22 no.8
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    • pp.585-589
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    • 2016
  • Recently, the robotic assist system for cardiovascular intervention gets continuously growing interest. The robotic cardiovascular intervention systems are largely two folds, systems for cardiac ablation procedure assist and systems for vascular intervention assist. For the systems, the clinician controls the catheter inserted through blood vessel to the heart via a master console or master manipulator. Most of the current master manipulators have structure of joystick-like pivoting 2 degree of freedom (DOF) handle in the core, which is used in parallel with other sliding switches and input devices. It however is desirable to have customized and optimized design manipulator that can provide clinician with intuitive control of the catheter motion fully utilizing the advantage of the use of robotic structure. A 6 DOF kinematic mechanism that can capture the motion control intention of the clinician in translational 3 DOF and rotational 3 DOF is proposed in this paper. Also, a master-slave motion relationship specially designed for the cardiac catheter manipulation motion is proposed and implemented in an experimental prototype. Design revision for implementation of more efficient motion and experiment in combination with an experimental slave robot system for catheter manipulation are underway.

Novel Biomarkers for Prediction of Response to Preoperative Systemic Therapies in Gastric Cancer

  • Cavaliere, Alessandro;Merz, Valeria;Casalino, Simona;Zecchetto, Camilla;Simionato, Francesca;Salt, Hayley Louise;Contarelli, Serena;Santoro, Raffaela;Melisi, Davide
    • Journal of Gastric Cancer
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    • v.19 no.4
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    • pp.375-392
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    • 2019
  • Preoperative chemo- and radiotherapeutic strategies followed by surgery are currently a standard approach for treating locally advanced gastric and esophagogastric junction cancer in Western countries. However, in a large number of cases, the tumor is extremely resistant to these treatments and the patients are exposed to unnecessary toxicity and delayed surgical therapy. The current clinical trials evaluating the combination of preoperative systemic therapies with modern targeted and immunotherapeutic agents represent a unique opportunity for identifying predictive biomarkers of response to select patients that would benefit the most from these treatments. However, it is of utmost importance that these potential biomarkers are corroborated by extensive preclinical and translational research. The aim of this review article is to present the most promising biomarkers of response to classic chemotherapeutic, anti-HER2, antiangiogenic, and immunotherapeutic agents that can be potentially useful for personalized preoperative systemic therapies in gastric cancer patients.

Knockdown of LKB1 Sensitizes Endometrial Cancer Cells via AMPK Activation

  • Rho, Seung Bae;Byun, Hyun Jung;Kim, Boh-Ram;Lee, Chang Hoon
    • Biomolecules & Therapeutics
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    • v.29 no.6
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    • pp.650-657
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    • 2021
  • Metformin is an anti-diabetic drug and has anticancer effects on various cancers. Several studies have suggested that metformin reduces cell proliferation and stimulates cell-cycle arrest and apoptosis. However, the definitive molecular mechanism of metformin in the pathophysiological signaling in endometrial tumorigenesis and metastasis is not clearly understood. In this study, we examined the effects of metformin on the cell viability and apoptosis of human cervical HeLa and endometrial HEC-1-A and KLE cancer cells. Metformin suppressed cell growth in a dose-dependent manner and dramatically evoked apoptosis in HeLa cervical cancer cells, while apoptotic cell death and growth inhibition were not observed in endometrial (HEC-1-A, KLE) cell lines. Accordingly, the p27 and p21 promoter activities were enhanced while Bcl-2 and IL-6 activities were significantly reduced by metformin treatment. Metformin diminished the phosphorylation of mTOR, p70S6K and 4E-BP1 by accelerating adenosine monophosphate-activated kinase (AMPK) in HeLa cancer cells, but it did not affect other cell lines. To determine why the anti-proliferative effects are observed only in HeLa cells, we examined the expression level of liver kinase B1 (LKB1) since metformin and LKB1 share the same signalling system, and we found that the LKB1 gene is not expressed only in HeLa cancer cells. Consistently, the overexpression of LKB1 in HeLa cancer cells prevented metformin-triggered apoptosis while LKB1 knockdown significantly increased apoptosis in HEC-1-A and KLE cancer cells. Taken together, these findings indicate an underlying biological/physiological molecular function specifically for metformin-triggered apoptosis dependent on the presence of the LKB1 gene in tumorigenesis.

Optimized phos-tag mobility shift assay for the detection of protein phosphorylation in planta

  • Hussain, Shah;Nguyen, Nhan Thi;Nguyen, Xuan Canh;Lim, Chae Oh;Chung, Woo Sik
    • Journal of Plant Biotechnology
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    • v.45 no.4
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    • pp.322-327
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    • 2018
  • Post-translational modification of proteins regulates signaling cascades in eukaryotic system, including plants. Among these modifications, phosphorylation plays an important role in modulating the functional properties of proteins. Plants perceive environmental cues that directly affect the phosphorylation status of many target proteins. To determine the effect of environmentally induced phosphorylation in plants, in vivo methods must be developed. Various in vitro methods are available but, unlike in animals, there is no optimized methodology for detecting protein phosphorylation in planta. Therefore, in this study, a robust, and easy to handle Phos-Tag Mobility Shift Assay (PTMSA) is developed for the in vivo detection of protein phosphorylation in plants by empirical optimization of methods previously developed for animals. Initially, the detection of the phosphorylation status of target proteins using protocols directly adapted from animals failed. Therefore, we optimized the steps in the protocol, from protein migration to the transfer of proteins to PVDF membrane. Supplementing the electrophoresis running buffer with 5mM $NaHSO_3$ solved most of the problems in protein migration and transfer. The optimization of a fast and robust protocol that efficiently detects the phosphorylation status of plant proteins was successful. This protocol will be a valuable tool for plant scientists interested in the study of protein phosphorylation.

Metabolic Signatures of Adrenal Steroids in Preeclamptic Serum and Placenta Using Weighting Factor-Dependent Acquisitions

  • Lee, Chaelin;Oh, Min-Jeong;Cho, Geum Joon;Byun, Dong Jun;Seo, Hong Seog;Choi, Man Ho
    • Mass Spectrometry Letters
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    • v.13 no.1
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    • pp.11-19
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    • 2022
  • Although translational research is referred to clinical chemistry measures, correct weighting factors for linear and quadratic calibration curves with least-squares regression algorithm have not been carefully considered in bioanalytical assays yet. The objective of this study was to identify steroidogenic roles in preeclampsia and verify accuracy of quantitative results by comparing two different linear regression models with weighting factor of 1 and 1/x2. A liquid chromatography-mass spectrometry (LC-MS)-based adrenal steroid assay was conducted to reveal metabolic signatures of preeclampsia in both serum and placenta samples obtained 15 preeclamptic patients and 17 age-matched control pregnant women (33.9 ± 4.2 vs. 32.8 ± 5.6 yr, respectively) at 34~36 gestational weeks. Percent biases in the unweighted model (wi = 1) were inversely proportional to concentrations (-739.4 ~ 852.9%) while those of weighted regression (wi = 1/x2) were < 18% for all variables. The optimized LC-MS combined with the weighted linear regression resulted in significantly increased maternal serum levels of pregnenolone, 21-deoxycortisol, and tetrahydrocortisone (P < 0.05 for all) in preeclampsia. Serum metabolic ratio of (tetrahydrocortisol + allo-tetrahydrocortisol) / tetrahydrocortisone indicating 11β-hydroxysteroid dehydrogenase type 2 was decreased (P < 0.005) in patients. In placenta, local concentrations of androstenedione were changed while its metabolic ratio to 17α-hydroxyprogesterone responsible for 17,20-lyase activity was significantly decreased in patients (P = 0.002). The current bioanalytical LC-MS assay with corrected weighting factor of 1/x2 may provide reliable and accurate quantitative outcomes, suggesting altered steroidogenesis in preeclampsia patients at late gestational weeks in the third trimester.