• Title/Summary/Keyword: Translational Research

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Control of Rattus norvegicus on Uninhabitable Islands - Case of Sasudo Island - (무인도에서 집쥐 개체군의 포획과 제어 - 제주 사수도의 사례 -)

  • Jun-Won Lee;Ga-Ram Kim;Seon-Mi Park;Sung-Hwan Choi;Young-Hun Jung;Hong-Shik Oh
    • Korean Journal of Environment and Ecology
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    • v.36 no.6
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    • pp.575-581
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    • 2022
  • Brown rats (Rattus norvegicusBerkenhout, 1769) were eradicated from Sasudo Island (33°55'13.04" N, 126°38'19.98" E), an uninhabited island designated as Natural Monument No. 333 and the largest breeding site for the streaked shearwater (Calonectris leucomelasTemminck, 1835) in South Korea. Twelve eradication studies were conducted from November 2015 to February 2021. The survey was conducted using a line census method that entailed slow wandering throughout the island and identifying starting and returning points. Capture traps were installed around traces of Rattus norvegicus,such as excrement and identified holes. As a result, 2 to 6 individuals were captured at each instance, except for the last time, when no individuals were captured. It is hypothesized that brown rats arrived at Sasudo Island via vessels arriving at the island for leisure and fishing. After the damage to streaked shearwater caused by brown rats was confirmed in 2006, entry to Sasudo Island was strictly prohibited through security measures, and marine clean-up programs that began in 2013 and continuous capturing since 2015 have been successful in eradicating brown rats. To maintain and manage the condition in Sasudo Island, preservation and management measures, such as strict visitor control, are necessary to prevent the inflow of new brown rats in the future.

Highly efficient genome editing via CRISPR-Cas9 ribonucleoprotein (RNP) delivery in mesenchymal stem cells

  • A Reum Han;Ha Rim Shin;Jiyeon Kweon;Soo Been Lee;Sang Eun Lee;Eun-Young Kim;Jiyeon Kweon;Eun-Ju Chang;Yongsub Kim;Seong Who Kim
    • BMB Reports
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    • v.57 no.1
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    • pp.60-65
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    • 2024
  • The CRISPR-Cas9 system has significantly advanced regenerative medicine research by enabling genome editing in stem cells. Due to their desirable properties, mesenchymal stem cells (MSCs) have recently emerged as highly promising therapeutic agents, which properties include differentiation ability and cytokine production. While CRISPR-Cas9 technology is applied to develop MSC-based therapeutics, MSCs exhibit inefficient genome editing, and susceptibility to plasmid DNA. In this study, we compared and optimized plasmid DNA and RNP approaches for efficient genome engineering in MSCs. The RNP-mediated approach enabled genome editing with high indel frequency and low cytotoxicity in MSCs. By utilizing Cas9 RNPs, we successfully generated B2M-knockout MSCs, which reduced T-cell differentiation, and improved MSC survival. Furthermore, this approach enhanced the immunomodulatory effect of IFN-r priming. These findings indicate that the RNP-mediated engineering of MSC genomes can achieve high efficiency, and engineered MSCs offer potential as a promising therapeutic strategy.

Diurnal Variation in Hydration of the Cervical Intervertebral Disc Assessed Using T2 Mapping of Magnetic Resonance Imaging

  • Chanyuan Liu;Jingyi Wang;Bowen Hou;Yitong Li;John N. Morelli;Peisen Zhang;Jun Ran;Xiaoming Li
    • Korean Journal of Radiology
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    • v.23 no.6
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    • pp.638-648
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    • 2022
  • Objective: The study aimed to investigate the diurnal variation in cervical disc hydration and its relationship with cervical degeneration. Materials and Methods: C3-C7 discs of 86 prospectively enrolled participants (37 males, 49 females; mean age ± standard deviation, 23.5 ± 2.5 years) were assessed using T2 mapping in the morning and evening. All discs were stratified by Miyazaki grade or C2-C7 Cobb angle and T2 values (T2). The degree of diurnal T2 variation (T2-DDV), defined as (morning T2 - evening T2)/morning T2 x 100%, was measured for the entire disc, annulus fibrosus (AF), nucleus pulposus (NP), and endplate zones. Results: T2 of the entire disc decreased significantly after the daytime load (p < 0.001), with a T2-DDV of 13.3% for all discs and 16.0%, 12.2%, and 13.0% for healthy (grade I), mild degenerative (grade II), and advanced degenerative (grade III/IV) discs, respectively. T2 of regional NPs and AFs decreased significantly from morning to evening (p ≤ 0.049) except in the healthy anterior inner AF (p = 0.092). Compared with healthy discs, mild degenerative discs displayed lower T2 and T2-DDV in regional NPs (p < 0.001). Advanced degenerative discs showed higher T2-DDV in the anterior inner AF compared with healthy discs (p = 0.050). Significant diurnal T2 changes in the endplate zones were observed only in healthy discs (p = 0.013). Cervical discs in the low Cobb angle group showed higher T2-DDV in the anterior AFs and anterior NP and lower T2-DDV in the posterior AF than those in the high Cobb angle group (p ≤ 0.041). Conclusion: This study characterized the diurnal variation in hydration of the cervical discs as assessed using T2 mapping and revealed early chemo-mechanical coupling dysfunction in degenerating discs. Cervical sagittal alignment on MRI can affect the diurnal stress patterns of the cervical discs. T2 mapping is sensitive to disc biomechanical dysfunction and offers translational potential from biomechanical research to clinical application.

Adipose Tissue-Derived Mesenchymal Stromal Cells from Ex-Morbidly Obese Individuals Instruct Macrophages towards a M2-Like Profile In Vitro

  • Daiana V. Lopes Alves;Cesar Claudio-da-Silva;Marcelo C. A. Souza;Rosa T. Pinho;Wellington Seguins da Silva;Periela S. Sousa-Vasconcelos;Radovan Borojevic;Carmen M. Nogueira;Helio dos S. Dutra;Christina M. Takiya;Danielle C. Bonfim;Maria Isabel D. Rossi
    • International Journal of Stem Cells
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    • v.16 no.4
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    • pp.425-437
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    • 2023
  • Obesity, which continues to increase worldwide, was shown to irreversibly impair the differentiation potential and angiogenic properties of adipose tissue mesenchymal stromal cells (ADSCs). Because these cells are intended for regenerative medicine, especially for the treatment of inflammatory conditions, and the effects of obesity on the immunomodulatory properties of ADSCs are not yet clear, here we investigated how ADSCs isolated from former obese subjects (Ex-Ob) would influence macrophage differentiation and polarization, since these cells are the main instructors of inflammatory responses. Analysis of the subcutaneous adipose tissue (SAT) of overweight (OW) and Ex-Ob subjects showed the maintenance of approximately twice as many macrophages in Ex-Ob SAT, contained within the CD68+/FXIII-A- inflammatory pool. Despite it, in vitro, coculture experiments revealed that Ex-Ob ADSCs instructed monocyte differentiation into a M2-like profile, and under inflammatory conditions induced by LPS treatment, inhibited HLA-DR upregulation by resting M0 macrophages, originated a similar percentage of TNF-α+ cells, and inhibited IL-10 secretion, similar to OW-ADSCs and BMSCs, which were used for comparison, as these are the main alternative cell types available for therapeutic purposes. Our results showed that Ex-Ob ADSCs mirrored OW-ADSCs in macrophage education, favoring the M2 immunophenotype and a mixed (M1/M2) secretory response. These results have translational potential, since they provide evidence that ADSCs from both Ex-Ob and OW subjects can be used in regenerative medicine in eligible therapies. Further in vivo studies will be fundamental to validate these observations.

Development of Simulation for Estimating Growth Changes of Locally Managed European Beech Forests in the Eifel Region of Germany (독일 아이펠의 지역적 관리에 따른 유럽너도밤나무 숲의 생장변화 추정을 위한 시뮬레이션 개발)

  • Jae-gyun Byun;Martina Ross-Nickoll;Richard Ottermanns
    • Journal of the Korea Society for Simulation
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    • v.33 no.1
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    • pp.1-17
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    • 2024
  • Forest management is known to beneficially influence stand structure and wood production, yet quantitative understanding as well as an illustrative depiction of the effects of different management approaches on tree growth and stand dynamics are still scarce. Long-term management of beech forests must balance public interests with ecological aspects. Efficient forest management requires the reliable prediction of tree growth change. We aimed to develop a novel hybrid simulation approach, which realistically simulates short- as well as long-term effects of different forest management regimes commonly applied, but not limited, to German low mountain ranges, including near-natural forest management based on single-tree selection harvesting. The model basically consists of three modules for (a) natural seedling regeneration, (b) mortality adjustment, and (c) tree growth simulation. In our approach, an existing validated growth model was used to calculate single year tree growth, and expanded on by including in a newly developed simulation process using calibrated modules based on practical experience in forest management and advice from the local forest. We included the following different beech forest-management scenarios that are representative for German low mountain ranges to our simulation tool: (1) plantation, (2) continuous cover forestry, and (3) reserved forest. The simulation results show a robust consistency with expert knowledge as well as a great comparability with mid-term monitoring data, indicating a strong model performance. We successfully developed a hybrid simulation that realistically reflects different management strategies and tree growth in low mountain range. This study represents a basis for a new model calibration method, which has translational potential for further studies to develop reliable tailor-made models adjusted to local situations in beech forest management.

Neuro-Restorative Effect of Nimodipine and Calcitriol in 1-Methyl 4-Phenyl 1,2,3,6 Tetrahydropyridine-Induced Zebrafish Parkinson's Disease Model

  • Myung Ji Kim; Su Hee Cho; Yongbo Seo; Sang-Dae Kim; Hae-Chul Park; Bum-Joon Kim
    • Journal of Korean Neurosurgical Society
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    • v.67 no.5
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    • pp.510-520
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    • 2024
  • Objective : Parkinson's disease (PD) is one of the most prevalent neurodegenerative diseases, characterized by the loss of dopaminergic neurons in the substantia nigra pars compacta. The treatment of PD aims to alleviate motor symptoms by replacing the reduced endogenous dopamine. Currently, there are no disease-modifying agents for the treatment of PD. Zebrafish (Danio rerio) have emerged as an effective tool for new drug discovery and screening in the age of translational research. The neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) is known to cause a similar loss of dopaminergic neurons in the human midbrain, with corresponding Parkinsonian symptoms. L-type calcium channels (LTCCs) have been implicated in the generation of mitochondrial oxidative stress, which underlies the pathogenesis of PD. Therefore, we investigated the neuro-restorative effect of LTCC inhibition in an MPTP-induced zebrafish PD model and suggested a possible drug candidate that might modify the progression of PD. Methods : All experiments were conducted using a line of transgenic zebrafish, Tg(dat:EGFP), in which green fluorescent protein (GFP) is expressed in dopaminergic neurons. The experimental groups were exposed to 500 μmol MPTP from 1 to 3 days post fertilization (dpf). The drug candidates : levodopa 1 mmol, nifedipine 10 μmol, nimodipine 3.5 μmol, diethylstilbestrol 0.3 μmol, luteolin 100 μmol, and calcitriol 0.25 μmol were exposed from 3 to 5 dpf. Locomotor activity was assessed by automated tracking and dopaminergic neurons were visualized in vivo by confocal microscopy. Results : Levodopa, nimodipine, diethylstilbestrol, and calcitriol had significant positive effects on the restoration of motor behavior, which was damaged by MPTP. Nimodipine and calcitriol have significant positive effects on the restoration of dopaminergic neurons, which were reduced by MPTP. Through locomotor analysis and dopaminergic neuron quantification, we identified the neuro-restorative effects of nimodipine and calcitriol in zebrafish MPTP-induced PD model. Conclusion : The present study identified the neuro-restorative effects of nimodipine and calcitriol in an MPTP-induced zebrafish model of PD. They restored dopaminergic neurons which were damaged due to the effects of MPTP and normalized the locomotor activity. LTCCs have potential pathological roles in neurodevelopmental and neurodegenerative disorders. Zebrafish are highly amenable to high-throughput drug screening and might, therefore, be a useful tool to work towards the identification of disease-modifying treatment for PD. Further studies including zebrafish genetic models to elucidate the mechanism of action of the disease-modifying candidate by investigating Ca2+ influx and mitochondrial function in dopaminergic neurons, are needed to reveal the pathogenesis of PD and develop disease-modifying treatments for PD.

Influence of crown-to-implant ratio of short vs long implants on implant stability and marginal bone loss in the mandibular single molar implant (하악 구치 단일임플란트 수복에서 임플란트 길이에 따른 치관-임플란트 비율이 임플란트안정성 및 변연골소실에 주는 영향)

  • Baek, Yeon-Wha;Kim, Bongju;Kim, Myung-Joo;Kwon, Ho-Beom;Lim, Young-Jun
    • Journal of Dental Rehabilitation and Applied Science
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    • v.34 no.4
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    • pp.280-289
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    • 2018
  • Purpose: The purpose of this randomized clinical trial is to determine whether implant length and the crown-to-implant (C/I) ratio influence implant stability and peri-implant marginal bone loss (MBL). Materials and Methods: 46 patients with single tooth missing in the posterior molar region of the mandible were included in this study. 19 implants (CMI IS-III $active^{(R)}$ long implant) of 5.0 mm diameter and 10 mm length were installed for the control group, while 27 implants (CMI IS-III $active^{(R)}$ short implant) of 5.5 mm diameter and 6.6, 7.3 or 8.5 mm length were placed for the experimental group. Each implant was inserted and immediately loaded using the digitally pre-fabricated surgical template and provisional restoration. The CAD-CAM monolithic zirconia crown was fabricated at 3 months after the surgery as a definitive restoration. The ISQ value and the MBL was measured at 48 weeks after the surgery. The correlation between the C/I ratio, MBL, and secondary implant stability was analyzed. Results: Successful results in terms of ISQ and MBL were achieved with both groups. There was no significant difference between the groups in terms of ISQ values and MBL at 48 weeks after the surgery (P > 0.05). No significant correlation was found between the C/I ratio and secondary stability as well as the C/I ratio and the MBL (P > 0.05). Conclusion: The influence of C/I ratio in both groups was not shown on the stability nor the marginal bone loss in implants supporting single crown of the mandible. Short implant could be a preferable alternative option in the reduced bone height mandible under the limited condition despite its higher C/I ratio.

Questionnaire survey for the clinical trial participants who experienced both digital and conventional impression (디지털 인상법과 종래인상법을 동시에 경험한 임상시험자를 대상으로 한 설문지분석)

  • Yang, Eunbee;Kim, Bongju;Lee, Jun Jae;Lee, Seung-Pyo;Lim, Young-Jun
    • Journal of Dental Rehabilitation and Applied Science
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    • v.34 no.4
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    • pp.270-279
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    • 2018
  • Purpose: The aim of this study was to assess the patients' perception, acceptance, and preference of the difference between a conventional impression and digital impression through questionnaire survey. Materials and Methods: Thirteen (6 male, 7 female) subjects who experienced both digital and conventional impression at the same day were enrolled in this study. Conventional impression were taken with polyvinylsiloxane and digital impression were performed using a newly developed intra-oral scanner. Immediately after the two impressions were made, a survey was conducted with the standardized questionnaires consisting of the following three categories; 1) general dental treatment 2) satisfaction of conventional impression 3) satisfaction of digital impression. The perceived source of satisfaction was evaluated using Likert scale. The distribution of the answers was assessed by percentages and statistical analyses were performed with the paired t-test, and P < 0.05 was considered significant. Results: There were significant differences of the overall satisfaction between two impression methods (P < 0.05). Digital impression showed high satisfaction in less shortness of breath and odor to participants compared to conventional impression. The use of an oral scanner resulted in a discomfort of TMJ due to prolonged mouth opening and in lower score of the scanner tip size. Conclusion: It was confirmed that the preference for the digital impression using intraoral scanner is higher than the conventional impression. Most survey participants said they would recommend the digital impression to others and said they preferred it for future prosthetic treatment.

Comparison of shear bond strength according to various surface treatment methods of zirconia and resin cement types (지르코니아의 다양한 표면처리 방법과 레진시멘트 종류에 따른 전단결합강도 비교)

  • Bae, Ji-Hyeon;Bae, Gang-Ho;Park, Taeseok;Huh, Jung-Bo;Choi, Jae-Won
    • The Journal of Korean Academy of Prosthodontics
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    • v.59 no.2
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    • pp.153-163
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    • 2021
  • Purpose: The aim of this study was to evaluate the effects of four surface treatment methods to improve zirconia roughness and three types of resin cement on the shear bond strength (SBS). Materials and methods: A total of 120 zirconia blocks were randomly divided into four surface treatments: non-treatment (Control), airborne-particle abrasion (APA) with 50 ㎛ Al2O3 (APA50), APA with 125 ㎛ Al2O3 (APA125), and ZrO2 slurry (ZA). Three resin cements (Panavia F 2.0, Superbond C&B, and Variolink N) were applied to the surface-treated zirconia specimens. All specimens were subjected to SBS testing using a universal testing machine. The surface of the representative specimens of each group was observed by scanning electron microscope (SEM). SBS data were analyzed with oneway ANOVA, two-way ANOVA test and post-hoc Tukey HSD Test (α=.05). Results: In the surface treatment method, APA125, APA50, ZA, and Control showed high shear bond strength in order, but there was no significant difference between APA125 and APA50 (P>.05). Also, ZA showed significantly higher shear bond strength than Control (P<.05). In the resin cement type, Panavia F 2.0, Superbond C&B, and Variolink N showed significantly higher shear bond strength in order (P<.05). In SEM images, the zirconia surfaces of the APA50 and APA125 showed quite rough and irregular shapes, and the zirconia surface of the ZA was observed small irregular porosity and rough surfaces. Conclusion: APA and ZrO2 slurry were enhanced the surface roughness of zirconia, and Panavia F 2.0 containing MDP showed the highest shear bond strength with zirconia.

Role of CopA to Regulate repABC Gene Expression on the Transcriptional Level (전사 수준에서 repABC 유전자 발현을 조절하는 CopA 단백질의 역할)

  • Sam Woong Kim;Sang Wan Gal;Won-Jae Chi;Woo Young Bang;Tae Wan Kim;In Gyu Baek;Kyu Ho Bang
    • Journal of Life Science
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    • v.34 no.2
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    • pp.86-93
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    • 2024
  • Since replication of plasmids must be strictly controlled, plasmids that generally perform rolling circle replication generally maintain a constant copy number by strictly controlling the replication initiator Rep at the transcriptional and translational levels. Plasmid pJB01 contains three orfs (copA, repB, repC or repABC) consisting of a single operon. From analysis of amino acid sequence, pJB01 CopA was homologous to the Cops, as a copy number control protein, of other plasmids. When compared with a CopG of pMV158, CopA seems to form the RHH (ribbon-helix-helix) known as a motif of generalized repressor of plasmids. The result of gel mobility shift assay (EMSA) revealed that the purified fusion CopA protein binds to the operator region of the repABC operon. To examine the functional role of CopA on transcriptional level, 3 point mutants were constructed in coding frame of copA such as CopA R16M, K26R and E50V. The repABC mRNA levels of CopA R16M, K26R and E50V mutants increased 1.84, 1.78 and 2.86 folds more than that of CopA wt, respectively. Furthermore, copy numbers owing to mutations in three copA genes also increased 1.86, 1.68 and 2.89 folds more than that of copA wt, respectively. These results suggest that CopA is the transcriptional repressor, and lowers the copy number of pJB01 by reducing repABC mRNA and then RepB, as a replication initiator.