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Plant developmental biology

Name

Mitsuhiro Aida

Affiliation

IROAST
Email: m-aida@kumamoto-u.ac.jp

Research Field

Advanced Green Bio

Title

Professor

1. Research Achievements
Our research aim is to elucidate molecular and genetic mechanisms that regulate the activity ofshoot meristem, a group of dividing cells that is responsible for production of plant aerial organs,such as leaves, stems and floral organs, which are collectively called shoot organs. In
dicotyledonous plants, the shoot meristem is initially formed during embryogenesis in a boundary
region between the two cotyledon primordia, and after germination, it maintains a group of stem
cells at its center and continuously provides new cells that give rise to shoot organs from its
periphery.
We have been focusing on a set of transcription factor encoding genes called CUC1, CUC2
and CUC3, which are expressed in the cotyledon boundary region and required for shoot meristem
formation in Arabidopsis thaliana. We demonstrated that the CUC1 protein, together with thefunctionary redundant transcription factors, promotes expression of key regulatory downstream
genes that are essential for maintaining postembryonic shoot meristem activity, and the mode ofits transcriptional regulation on many of these downstream genes is possibly direct. Of note, apair of the downstream genes, the KNOX gene STM and the cytochrome P450 gene KLU, have
opposite effects on the shoot meristem activity, suggesting that a balancing mechanism for shootmeristem activity upon its initiation (ref. 3).
We also investigated involvement of these transcription factors in reproduction organ
development. The gynoecium, or the female reproductive organ of flowering plants, has anelaborated structure suitable for fertilization and seed dispersal. We have shown that the CUC
genes and its downstream gene BLR interact to ensure proper development of the medial tissues,
which provide physical strength and the ability for efficient seed dispersal to the gynoecium (ref.
5). The medial tissues also undergo post-genital fusion to produce transmitting tract for pollen
tubes and we described changes in the epidermal identity during this event (ref. 4).
Finally, we have also developed a technique to visualize 3D structure oof plant embryo. In
collaboration with the Shinichiro Sawa from FAST and Takashi Ishida from IROAST, we
optimized a tissue clearing method for embryos of the model plant Arabidopsis thaliana, enabling
the collection of high-quality 3D morphological data, which is suitable for quantitative analyses(Figure). Our new method would facilitate analysis of developmental mechanisms that govern
plant embryogenesis, in which a series of coordinated patterning and morphological events
proceed to establish the basic body plan (ref. 2).
2. International Research Collaboration
International collaborative work was made between Michael Lenhard’s group (University of
Potsdam, Germany) and our group, and led to a publication of CUC downstream genes (ref. 3).
Other collaborative work has been carried out with Jose Ireapan Reyes Olalde, a research fellow
from Consejo Nacional de Ciencia y Tecnología (CONACYT) of Mexico, to elucidate hormonal
control of gynoecium development, which led to an oral presentation in the 84th Annual Meetingof the Botanical Society of Japan (September 20th, 2020). A collaboration with Yoshihisa Ikeda
from Palacký University (Czech Republic) has led to a publication of a review article about therole for the plant hormone cytokinin in intracellular signaling (ref 1) and details of this activity
is described elsewhere (Research Unit “Plant Stem Cells and Regeneration” section).
3. Prospect for further research collaboration
The following collaborative research is planned in FY2021.

㻌9 Analysis of transcription factor-target gene interactions during shoot meristem formation
with Simon Scofield in Cardiff University, UK.
9 Mechanisms of fruit development with Stefan de Folter in CINVESTAV, LANGEBIO, Mexico.
9 The role of mechanical signals in the regulation of gene expression with Olivier Hamant in
INRA ENS de Lyon, France.
9 The role of boundary specific signaling peptide in embryogenesis with Keiko U Torii in
Howard Hughes Medical Institute and University of Texas at Austin, USA, and Rüdiger
Simon in Heinrich-Heine University, Germany.
4. Other activities
9 Chairperson of Public Relations Committee, The Japanese Society of Plant Physiologists(March 2020–present)
9 Writing a short article for a scientific exhibition (“Plants: Mainstays of the Planet, National
Museum of Nature and Science)
9 Peer review of three papers
9 Lectures (6 times in Faculty of Science, 4 times in Multidisciplinary Studies, 90 min/time)9 Host of a visiting lecture (Faculty of Science, Prof. Sachihiro Matsunaga from Univ. Tokyo)
9 Supervision of three undergraduate students of Faculty of Science)9 Thesis reviewer for two master course students, Graduate School of Science and Technology
List of Publications (*Corresponding Author)
1. Ikeda Y, Zalabák D, Kubalová I, Králová M, Brenner WG, Aida M (2021). Interpreting
cytokinin action as anterograde signaling and beyond. Front Plant Sci 12, 641257. doi:
10.3389/fpls.2021.641257.
2. Imoto A, Yamada M, Sakamoto T, Okuyama A, Ishida T, Sawa S, *A
Aida M (2021). A
ClearSee-based clearing protocol for 3D visualization of Arabidopsis thaliana embryos.
Plants 10, 190. doi: 10.3390/plants10020190
3. *A
Aida M, Tsubakimoto Y, Shimizu S, Ogisu H, Kamiya M, Iwamoto R, Takeda S, Karim MR,
Mizutani M, Lenhard M, Tasaka M (2020). Establishment of the embryonic shoot meristem
involves activation of two classes of genes with opposing functions for meristem activities.
Int J Mol Sci 21, 5864. doi: 10.3390/ijms21165864.
4. Yokoi A, *A
Aida M (2021). Postgenital fusion and epidermal cell fate control during
gynoecium development. Cytologia 86, 1-2. doi: 10.1508/cytologia.86.1.
5. Yamamoto K, Tasaka M, *A
Aida M. Genetic interactions between the CUP-SHAPEDCOTYLEDON and the BELLRINGER genes indicate their overlapping functions in carpel
boundary development in Arabidopsis thaliana. Plant Morphol, in press.
FigureConfocal image of a cleared Arabidopsis thaliana embryo expressing the auxin response reporter
DR5 (ref 2). An optical section (left) and 3D reconstruction.

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Experiments for anaerobic microbes
For the first one, this year was predominantly spent on preparing reagents and culture media
in order to develop clusters of anaerobic microbes as well as on preparing experimental
apparatus. We have finished preparing almost all culture media planned and broken one rocksample into small pieces under an anaerobic condition. In 2021, we will place the rockfragments on the culture media under an anaerobic condition and aim at observing clusters of
microbes that contribute to the enhancement of carbon precipitation.
Experiments for aerobic microbes (grout technology)
Regarding the second one (application of the grout technology into carbon sequestration), during
the fiscal year of 2020, we have performed the following two experiments: i) creation of a grout
material with urease, ii) image analysis of grouted rock samples with an industrial X-ray CT
scanner. Regarding the grout technology, we have investigated the effect of urease
concentrations on the chemical reaction rate of calcium chloride and urea that results incalcium carbonate. It is found from Fig. 1 that as the urease concentration increases, more
CaCO3 is precipitated, giving an insight into the necessity of optimizing urease concentration
when applying the grout technology to carbon sequestration. This is because extremely highreaction rates would prevent grouts from permeating a fractured rock mass, whilst low reaction
rate does not effectively fill fractures in a rock mass.

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Fig. 1: Effect of urease concentration of the precipitation rate of CaCO3
ii)

Image analysis of grouted rock samples with an industrial X-ray CT scanner

X-ray CT images were taken for rock samples to which grouting was performed using urea,
carbon chloride, and urease. Fig. 2 shows the difference in specific density between an originalrock sample and grouted one. The warm color in the picture corresponds to a region with highspecific density, indicating calcium carbonate precipitated as a result of grouting. As can be seen,
the grouted region is spatially dotted in the rock sample. This indicates that calcium carbonate
was effectively precipitated, thus contributing to the reduction of permeability.

6SHFLILFJUDYLW\Fig. 2: Difference in specific density between an original rock and grouted samples㻌1R

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As one of new challenges, we have investigated simultaneous measurement of temperatureand pressure with FLIM-PSP technique. Since we succeeded in obtaining temperature andpressure calibration curves using a photomultiplier tube, we clarified that it is possible tomeasure simultaneously temperature and pressure at the calibration test level. Our next test is
performed by a high speed camera (CMOS) at Kumamoto university. It was not trial by the FLIM
camera in DLR. The calibration curves by some high speed cameras were obtained successfully.
Demonstration tests are underway with simultaneous measurement of temperature and pressure
distribution by this technique using a wall impinging jet model. However, in order to apply it to
the industrial model, it is necessary to investigate to further increase the temperature sensitivity
and temperature sensitivity.
We hope to have collaboration tests using the FLIM camera at DLR in Germany in 2021.

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3UHVHQWDWLRQV 3XEOLFDWLRQV LQFOVXEPLWWHGSDSHUV [Presentations]Ruda LEE
1. Ueno Sho, Takuro Niidome, Ruda Lee. Development of ErB2-targeting Liposomes forEnhancing Delivery to ErbB2- positive Breast cancer. The 15th International Student
Conference on Advanced Science and Technology䠄ICAST 2020 Kumamoto, Dec 3-4 (2020),
Kumamoto, Japan (Online)
2. Seung-Hae Kwon, Ji Hoon Kim, Jung Hoon Choi, Ruda Lee. Non-invasive early detectionof calpain 2-enriched non-small cell lung cancer using a human serum albumin-based
calpain-2 nanosensor. 7th International Conference on Bio-Sensing Technology, Busan,
Korea (Online)
Yong Il PARK
1. Song Yeul Lee, Yoongu Lim, Dohun Kim, Uk Sim, Yong Il Park. The Effect of NaYF4
Upconversion Nanoparticles for the Enhancement of ZnFe2O4/TiO2 Photoanode, KIChE
Fall Meeting and International Symposium, Oct 14-16, Korea (Online).

㻌2. Song Yeul Lee, Yong Il Park. Enhanced Cyclic Performance of Sodium-Ion Capacitor via
Surface Modification, 2020 KSIEC Annual Meeting in Celebration of the 30th Anniversary,
Oct 28-30, Gwangju, Korea.
3. Yoongu Lim, Song Yeul Lee, Dohun Kim, Uk Sim, Yong Il Park. Enhancement of
ZnFe2O4/TiO2 Photoanode Through the Effect of NaYF4 Upconversion Nanoparticles, The
16th International Symposium on Novel and Nano Materials Incorporating theConference of Korean Powder Metallurgy Institute, Nov 3-6, Jeju, Korea.
4. Panju Kim, Eun Young Lee, Byung Joon Moon, Song Yeul Lee, Yong Il Park, Sang Hyun
Lee. Synthesis and Applications of Lead-Free Perovskite Cesium Ytterbium Halide
Nanocrystals. The 30th International Photovoltaic Science and Engineering Conference,
Nov 9-13 (2020), Jeju, Korea.
Jung Hoon CHOI
1. Yoon-Hwan Kim, Min Soo Kang, Tae Hyeong Kim, Yunho Jeong, Jin-Ok Ahn, Jung Hoon
Choi, Jin-Young Chung. Development of Synbio-glucan Functional Spray for Canine
Atopic Dermatitis. 2020 KSVS symposium (Korea, Hongchun 19~20 NOV 2020)
2. Min Soo Kang, Woosuk Kim, Tae Hyeong Kim, Hyo Young Jung, Hyun Jung Kwon, Dae
Won Kim,In Koo Hwang and Jung Hoon Choi. High fat diet accelerates and exacerbates
microgliosis and neuronal damage/death in the somatosensory cortex after transient
forebrain ischemia in gerbils. 2020 KALAS International symposium (Korea,
Pyeongchang 15~18 Jul 2020)
Xiaoxue XU
N/A[Publications]Ruda Lee
1. Yong Il Park, Seung-Hae Kwon, Gibok Lee, Keiichi Motoyama, Min Woo Kim, Min Lin,
Takuro Niidome, Jung Hoon Choi, Ruda Lee*. pH-sensitive multi-drug liposomestargeting folate receptor ǃ for efficient treatment of non-small cell lung cancer. Journal ofControlled Release, 330, 1-14, February 2021.
2. Sho Ueno, Min Woo Kim, Gibok Lee, Yong Il Park, Takuro Niidome, and Ruda Lee*.
Development of ErbB2-Targeting Liposomes for Enhancing Drug Delivery to ErbB2Positive Breast Cancer. Pharmaceutics, 12, 585, June 2020.
3. Xiaoxue Xu, Hongxu Lu and Ruda Lee. Near Infrared Light Triggered Photo/ImmunoTherapy Toward Cancers. Frontiers in Bioengineering and Biotechnology, 8, 488, 2020.
4. Song Yeul Lee*, Ruda Lee*, Eunha Kim, Sanghee Lee and Yong Il Park. Near-Infrared
Light-Triggered Photodynamic Therapy and Apoptosis Using Upconversion Nanoparticles
with Dual Photosensitizers. Frontiers in Bioengineering and Biotechnology, 8, 275. Apr
2020.
Yong Il PARK
1. Yong Il Park, Seung-Hae Kwon, Gibok Lee, Keiichi Motoyama, Minwoo Kim, Min Lin,
Takuro Niidome, Jung Hoon Choi, Ruda Lee. pH-Sensitive Multi-Drug Liposomes
Targeting Folate Receptor ǃ for Efficient Treatment of Non-Small Cell Lung Cancer.
Journal of Controlled Release, 330, 1-14, Feb 2021.
2. Da Hwi Gu, Jungsoo Lee, Hyeong Woo Ban, Gibok Lee, Minju Song, Wooyong Choi,
Seongheon Baek, Hyewon Jeong, Song Yeul Lee, Yonghoon Choi, Jongnam Park, Yong Il
Park,* Jae Sung Son*. Colloidal Suprastructures Self-Organized from Oppositely Charged
All-Inorganic Nanoparticles. Chemistry of Materials, 32, 8662-8671, Oct 2020.
3. Sho Ueno, Min Woo Kim, Gibok Lee, Yong Il Park, Takuro Niidome, Ruda Lee.
Development of ErbB2-Targeting Liposomes for Enhancing Drug Delivery to ErbB2-

㻌Positive Breast Cancer. Pharmaceutics, 12, 585, Aug 2020.
4. Jinyue Zhang, Mingshi Jin, Yong Il Park, Longyi Jin, Bo Quan. Facile Synthesis of UltraSmall Hollow Manganese Silicate Nanoparticles as pH/GSH-Responsive T1-MRI Contrast
Agents. Ceramics International, 46, 18632–18638, Jun 2020.
5. Song Yeul Lee, Ruda Lee, Eunha Kim, Sanghee Lee,* Yong Il Park*. Near-Infrared LightTriggered Photodynamic Therapy and Apoptosis using Upconversion Nanoparticles with
Dual Photosensitizers. Frontiers in Bioengineering and Biotechnology, 8, 275, Apr 2020.
6. Song Yeul Lee, Gibok Lee, Young-Si Jun, Yong Il Park*. Visible/Near-Infrared Driven
Highly Efficient Photocatalyst Based on Upconversion Nanoparticles/g-C3N4
Nanocomposite. Applied Surface Science, 508, 144839, Apr 2020.
Jung Hoon CHOI
1. Yong Il Park, Seung-Hae Kwon, Gibok Lee, Keiichi Motoyama, Min Woo Kim, Min Lin,
Takuro Niidome, Jung Hoon Choi, Ruda Lee. pH-sensitive multi-drug liposomes targeting
folate receptor ǃ for efficient treatment of non-small cell lung cancer. Journal of Controlled
Release, 330, 1-14, February 2021.
2. Yoon-Hwan Kim, Min Soo Kang, Tae Hyeong Kim, Yunho Jeong, Jin-Ok Ahn, Jung Hoon
Choi*, Jin-Young Chung. Anti-Inflammatory and Immune Modulatory Effects of SynbioGlucan in an Atopic Dermatitis Mouse Model Nutrients. 2021 Apr; 13(4): 1090.
3. Yoon-Hwan Kim, Tae Hyeong Kim, Min Soo Kang, Jin-Ok Ahn, Jung Hoon Choi*, JinYoung Chung. Comparison of the presentation of atopic dermatitis induced bytrinitrochlorobenzene and house dust mite in NC/Nga mice J Vet Sci. 2020 Jul; 21(4): e59.
4. Sumin Yun, Woosuk Kim, Min Soo Kang, Tae Hyeong Kim, Yoonhwan Kim, Jin-Ok Ahn,
Jung Hoon Choi, In Koo Hwang, Jin-Young Chung. Neuropathological changes in dorsal
root ganglia induced by pyridoxine in dogs. BMC Neurosci. 2020; 21: 11.
Xiaoxue XU
1. Deming Liu, Xiaoxue Xu, Yi Du, Jiayan Liao, Shihui Wen, Xiaotong Dong, Yan Jin, Lei
Liu, Dayong Jin, John A. Capobianco, Dezhen Shen. Reconstructing the Surface Structureof NaREF4 Upconversion Nanocrystals with a Novel K+ Treatment. Chem. Mater. 2021,
33, 7, 2548–2556 March 2021.
2. Yongliang Chen, Xiaoxue Xu, Chi Li, Avi Bendavid, Mika Westerhausen, Carlo Bradac,
Milos Toth, Igor Aharonovich, Toan Trong Tran. BottomǦUp Synthesis of Hexagonal Boron
Nitride Nanoparticles with IntensityǦStabilized Quantum Emitters. Small. In press
March 2021.
3. Hongfang Li, Yahui Li, Kai Wang, Liyan Lai, Xiaoxue Xu, Bin Sun, Zhuoqing Yang, Guifu
Ding. Ultra-high sensitive micro-chemo-mechanical hydrogen sensor integrated bypalladium-based driver and high-performance piezoresistor. International Journal of
Hydrogen Energy. 2021, 46, 1, 1434-1445. January 2021.
4. ZX Deng, JW Tao, W Zhang, HJ Mu, HJ Wu, YB Wang, XX Xu, W Zheng. Effect of protein
adsorption on electrospun hemoglobin/gelatin-MWCNTs microbelts modified electrode:
Toward electrochemical measurement of hydrogen peroxide. 2021, 257, 123827. January
2021.
5. Guohua Jia and Bing-Jie Ni Derek Hao, Chuangwei Liu, Xiaoxue Xu, mehran kianinia,
Igor Aharonovich, Xiaojuan Bai, Xiaoqing Liu, Zhijie Chen, Wei Wei. Surface defectabundant one-dimensional graphitic carbon nitride nanorods boost photocatalytic
nitrogen fixation. New J. Chem., 2020, 44, 20651-20658. November 2020.
6. Sarah Meyer, Raquel Gonzalez de Vega, Xiaoxue Xu, Ziqing Du, Philip A Doble, David
Clases. Characterization of Upconversion Nanoparticles by Single-Particle ICP-MS
Employing a Quadrupole Mass Filter with Increased Bandpass. Analytical chemistry 2020,㻌

92, 22, 15007-15016. November 2020.
7. Christian Clarke, Mandeep Singh, Sherif Abdulkader Tawfik, Xiaoxue Xu, Michelle JS
Spencer, Rajesh Ramanathan, Philipp Reineck, Vipul Bansal, Cuong Ton-That. Mono-to
few-layer non-van der Waals 2D lanthanide-doped NaYF4 nanosheets with upconversion
luminescence. 2D Materials 2020, 8, 1, 015005. October 2020.
8. Laixu Gao, Xuchen Shan, Xiaoxue Xu, Yongtao Liu, Baolei Liu, Sonquan Li, Chenshuo
Ma, Dayong Jin, Fan Wang. Video-rate upconversion display from optimized lanthanide
ion doped upconversion nanoparticles. Nanoscale, 2020,12, 18595-18599. June 2020.
9. ZX Deng, JW Tao, LJ Zhao, W Zhang, YB Wang, HJ Mu, HJ Wu, XX Xu, W Zheng. Effectof protein adsorption on bioelectrochemistry of electrospun core-shell MWCNTs/gelatinHb nanobelts on electrode surface. Process Biochemistry, 2020, 96, 73-79. June 2020.
10. Ziqing Du, Abhishek Gupta, Christian Clarke, Matt Cappadona, David Clases, Deming
Liu, Zhuoqing Yang, Shawan Karan, William S Price, Xiaoxue Xu*. Porous upconversion
nanostructures as bimodal biomedical imaging contrast agents. The Journal of Physical
Chemistry C 124 (22), 12168-12174. May 2020.


3. Application & acquisition status of KAKENHI and other external grants
Ruda LEE
1. CRDF Global female young researcher grant, Applied (Principal Investigator)
2. Mitsubishi young researcher grant, Applied (Principal Investigator)
Yong Il PARK
1. National Research Foundation of Korea (NRF), Basic Science Research Program,2020.06.01~2021.05.31 (Principal Investigator)
2. National Research Foundation of Korea (NRF), Korea Research Fellowship (KRF) for
Postdoctoral Researchers, 2020.08.01~2022.12.31 (Principal Investigator)
3. Korea Basic Science Institute, 2021 Research Facility Council Development Support
Project, 2021.04.01~2021.11.30 (Principal Investigator)
4. National Research Foundation of Korea (NRF), Basic Science Research Program, Applied(Principal Investigator)
Jung Hoon CHOI
1. National Research Foundation of Korea funded by the Ministry of Education (NRF2019R1I1A3A01061857) (Principal Investigator)
Xiaoxue XUN/A

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This project entitled “development of microbially-aided carbon sequestration technology” can bedivided into two parts: enhancement of carbon precipitation using anaerobic microbes andclosure of rock fractures in an aquifer with a grout technology using aerobic microbes.
Experiments for anaerobic microbes
For the first one, this year was predominantly spent on preparing reagents and culture media
in order to develop clusters of anaerobic microbes as well as on preparing experimental
apparatus. We have finished preparing almost all culture media planned and broken one rocksample into small pieces under an anaerobic condition. In 2021, we will place the rockfragments on the culture media under an anaerobic condition and aim at observing clusters of
microbes that contribute to the enhancement of carbon precipitation.
Experiments for aerobic microbes (grout technology)
Regarding the second one (application of the grout technology into carbon sequestration), during
the fiscal year of 2020, we have performed the following two experiments: i) creation of a grout
material with urease, ii) image analysis of grouted rock samples with an industrial X-ray CT
scanner.
i) Creation of a grout material with urease
We have investigated the effect of urease concentrations on the chemical reaction rate of
calcium chloride and urea that results in calcium carbonate. It is found from Fig. 1 that as theurease concentration increases, more CaCO3 is precipitated, giving an insight into the necessityof optimizing urease concentration when applying the grout technology to carbon sequestration.
This is because extremely high reaction rates would prevent grouts from permeating a fractured

㻌rock mass, whilst low reaction rate does not effectively fill fractures in a rock mass.

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Fig. 1: Effect of urease concentration of the precipitation rate of CaCO3ii) Image analysis of grouted rock samples with an industrial X-ray CT scanner
X-ray CT images were taken for rock samples to which grouting was performed using urea,
carbon chloride, and urease. Fig. 2 shows the difference in specific density between an originalrock sample and grouted one. The warm color in the picture corresponds to a region with highspecific density, indicating calcium carbonate precipitated as a result of grouting. As can be seen,
the grouted region is spatially dotted in the rock sample. This indicates that calcium carbonate
was effectively precipitated, thus contributing to the reduction of permeability.

6SHFLILFJUDYLW\㻌Fig. 2: Difference in specific density between an original rock and grouted samples
Plan to apply for Kakenhi
As shown, preliminary experiments were conducted for this research project in 2020. In thefiscal year of 2021, we will continue to conduct preliminary experiments. Specifically, regarding
the experiment for anaerobic microbes, we will obtain further rock samples from a deep
underground coal mine located in Hokkaido, with which we will try to find a new type of
microbes that contributes to the generation of calcium carbonate under an anaerobic condition.
In addition, rock samples will be also collected from mountain tunnels. As for the experiment ofgrouting, new types of urease will be tested in 2021 because the urease used in 2020 was foundto have a very high enzyme activity that may not be suitable for filling rock fractures in an
aquifer to which CO2 is injected. As such, we have purchased several types of urease withvarious enzyme activities. Using the different ureases, we aim to select a urease with the mostappropriate enzyme activity. Based on the result, we aim to apply for Kakenhi in 2022.

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