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Mineralized Polysaccharide Transplantation Modules Supporting Human MSC Conversion into Osteogenic Cells and Osteoid Tissue in a Non-Union Defect

  • Ge, Qing;Green, David William;Lee, Dong-Joon;Kim, Hyun-Yi;Piao, Zhengguo;Lee, Jong-Min;Jung, Han-Sung
    • Molecules and Cells
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    • v.41 no.12
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    • pp.1016-1023
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    • 2018
  • Regenerative orthopedics needs significant devices to transplant human stem cells into damaged tissue and encourage automatic growth into replacements suitable for the human skeleton. Soft biomaterials have similarities in mechanical, structural and architectural properties to natural extracellular matrix (ECM), but often lack essential ECM molecules and signals. Here we engineer mineralized polysaccharide beads to transform MSCs into osteogenic cells and osteoid tissue for transplantation. Bone morphogenic proteins (BMP-2) and indispensable ECM proteins both directed differentiation inside alginate beads. Laminin and collagen IV basement membrane matrix proteins fixed and organized MSCs onto the alginate matrix, and BMP-2 drove differentiation, osteoid tissue self-assembly, and small-scale mineralization. Augmentation of alginate is necessary, and we showed that a few rationally selected small proteins from the basement membrane (BM) compartment of the ECM were sufficient to up-regulate cell expression of Runx-2 and osteocalcin for osteoid formation, resulting in Alizarin red-positive mineral nodules. More significantly, nested BMP-2 and BM beads added to a non-union skull defect, self-generated osteoid expressing osteopontin (OPN) and osteocalcin (OCN) in a chain along the defect, at only four weeks, establishing a framework for complete regeneration expected in 6 and 12 weeks. Alginate beads are beneficial surgical devices for transplanting therapeutic cells in programmed (by the ECM components and alginate-chitosan properties) reaction environments ideal for promoting bone tissue.

Effects of hot melt extrusion processed nano-iron on growth performance, blood composition, and iron bioavailability in weanling pigs

  • Lee, JunHyung;Hosseindoust, Abdolreza;Kim, MinJu;Kim, KwangYeol;Choi, YoHan;Moturi, Joseph;Song, ChangHyun;Lee, SongYi;Cho, HyunJong;Chae, ByungJo
    • Journal of Animal Science and Technology
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    • v.61 no.4
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    • pp.216-224
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    • 2019
  • This study was conducted to investigate the effects of hot melt extrusion (HME) nano-iron as an alternative for the common ferrous sulfate on iron (Fe) bioavailability, growth performance, nutrient digestibility, intestinal morphology, and intestinal microbiota of weanling pigs. A total of 200 piglets (Landrace ${\times}$ Yorkshire ${\times}$ Duroc) were randomly allotted to seven treatments on the basis of initial body weight (BW) and sex. Treatments were the INO100 (100 ppm Fe as $FeSO_4$), HME-Fe levels (50, 75, and 100 ppm nano-Fe as $FeSO_4$). ORG100 (100 ppm Fe as iron methionine). In phase 1, the HME50 pigs showed the lowest Fe content in feed and feces. Plasma Fe concentration was increased in HME100 and ORG100 pigs. In phase 2, there were significantly lower concentration of Fe in feed and feces of HME50 pigs (p < 0.01). A lower Fe concentration in the plasma and liver were observed in HME50 pigs compared with HME100 pigs. Concentration of red blood cell (RBC) was the lowest (p < 0.01) for HME50 pigs. During phase 2, the HME100, HME75, and ORG100 pigs showed a higher RBC and hemoglobin values compared with HME50 pigs. Digestibility of gross energy (GE) and crude protein (CP) were significantly higher in HME100 pigs compared with HME50 pigs. There was an increased (p < 0.01) villus height in the duodenum and jejunum of HME100 pigs compared with HME50 pigs. It is concluded that dietary Fe does not improve growth performance of weanling pigs; however, increasing the dietary iron concentration in weanling piglets increased the RBC and hemoglobin. In addition, the potential ability of HME to be used at a lower level (HME75) was observed.

LasR Might Act as an Intermediate in Overproduction of Phenazines in the Absence of RpoS in Pseudomonas aeruginosa

  • He, Qiuning;Feng, Zhibin;Wang, Yanhua;Wang, Kewen;Zhang, Kailu;Kai, Le;Hao, Xiuying;Yu, Zhifen;Chen, Lijuan;Ge, Yihe
    • Journal of Microbiology and Biotechnology
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    • v.29 no.8
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    • pp.1299-1309
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    • 2019
  • As an opportunistic bacterial pathogen, Pseudomonas aeruginosa PAO1 contains two phenazine-producing gene operons, phzA1B1C1D1E1F1G1 (phz1) and phzA2B2C2D2E2F2G2 (phz2), each of which is independently capable of encoding all enzymes for biosynthesizing phenazines, including phenazine-1-carboxylic acid and its derivatives. Other previous study reported that the RpoS-deficient mutant SS24 overproduced pyocyanin, a derivative of phenazine-1-carboxylic acid. However, it is not known how RpoS mediates the expression of two phz operons and regulates pyocyanin biosynthesis in detail. In this study, with deletion of the rpoS gene in the $PA{\Delta}phz1$ mutant and the $PA{\Delta}phz2$ mutant respectively, we demonstrated that RpoS exerted opposite regulatory roles on the expression of the phz1and phz2 operons. We also confirmed that the phz1 operon played a critical role and especially biosynthesized much more phenazines than the phz2 operon when the rpoS gene was knocked out in P. aeruginosa. By constructing the translational reporter fusion vector lasR'-'lacZ and the chromosomal fusion mutant $PA{\Delta}lasR::lacZ$, we verified that RpoS deficiency caused increased expression of lasR, a transcription regulator gene in a first quorum sensing system (las) that activates overexpression of the phz1 operon, suggesting that in the absence of RpoS, LasR might act as an intermediate in overproduction of phenazine biosynthesis mediated by the phz1 operon in P. aeruginosa.

Current status on the modification of the scope for GMO regulation on the gene edited plants with no remnants of inserted foreign DNA fragments (외래 DNA단편이 잔존하지 않는 유전자교정식물에 대한 GMO규제 범위의 제외에 관한 국제 동향)

  • Lee, Shin-Woo
    • Journal of Plant Biotechnology
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    • v.46 no.3
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    • pp.137-142
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    • 2019
  • Gene edited crops can be classified as SDN-1, SDN-2 and SDN-3 group depending on their mutation's range and the usage of donor DNA. The SDN-1 and SDN-2 crops, in particular, could be developed as 100% transgene-free, which do not contain any DNA fragment of the vector or guide RNA used for gene editing such as CRISPR Cas9 system. Therefore, there are no scientific methods available for the detection of these crops and differentiation with the one produced by conventional cross breeding techniques. Additionally, it would be impossible to properly implement the existing GMO regulation law, in particular, the national legislation for "GMO labelling". In this regard, Australia has announced that SDN-1 crops will not be subjected to the existing GMO regulation. Furthermore, Argentina and Brazil have established a new policy that GE crops with no transgene (100% transgene-free crops) should be exempted from the scope of the GMO. In addition, Japan has also announced that "an organism that has no remnants of inserted nucleic acid processed extracellularly is not subjected to the Cartagena Act". It means that SDN-2 crops can also be exempted from the scope of GMO. In this trend, in South Korea, I suggested that gene edited crops with no remnants of inserted foreign DNA fragments should be excluded from the existing GMO regulation. Thus, I expect that diverse elite crop lines should be developed by using advanced gene editing technologies

vfr, A Global Regulatory Gene, is Required for Pyrrolnitrin but not for Phenazine-1-carboxylic Acid Biosynthesis in Pseudomonas chlororaphis G05

  • Wu, Xia;Chi, Xiaoyan;Wang, Yanhua;Zhang, Kailu;Kai, Le;He, Qiuning;Tang, Jinxiu;Wang, Kewen;Sun, Longshuo;Hao, Xiuying;Xie, Weihai;Ge, Yihe
    • The Plant Pathology Journal
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    • v.35 no.4
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    • pp.351-361
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    • 2019
  • In our previous study, pyrrolnitrin produced in Pseudomonas chlororaphis G05 plays more critical role in suppression of mycelial growth of some fungal pathogens that cause plant diseases in agriculture. Although some regulators for pyrrolnitrin biosynthesis were identified, the pyrrolnitrin regulation pathway was not fully constructed. During our screening novel regulator candidates, we obtained a white conjugant G05W02 while transposon mutagenesis was carried out between a fusion mutant $G05{\Delta}phz{\Delta}prn::lacZ$ and E. coli S17-1 (pUT/mini-Tn5Kan). By cloning and sequencing of the transposon-flanking DNA fragment, we found that a vfr gene in the conjugant G05W02 was disrupted with mini-Tn5Kan. In one other previous study on P. fluorescens, however, it was reported that the deletion of the vfr caused increased production of pyrrolnitrin and other antifungal metabolites. To confirm its regulatory function, we constructed the vfr-knockout mutant $G05{\Delta}vfr$ and $G05{\Delta}phz{\Delta}prn::lacZ{\Delta}vfr$. By quantifying ${\beta}-galactosidase$ activities, we found that deletion of the vfr decreased the prn operon expression dramatically. Meanwhile, by quantifying pyrrolnitrin production in the mutant $G05{\Delta}vfr$, we found that deficiency of the Vfr caused decreased pyrrolnitrin production. However, production of phenazine-1-carboxylic acid was same to that in the wild-type strain G05. Taken together, Vfr is required for pyrrolnitrin but not for phenazine-1-carboxylic acid biosynthesis in P. chlororaphis G05.

Effect of Postmortem Phases on Lamb Meat Quality: A Physicochemical, Microstructural and Water Mobility Approach

  • Ge, Yue;Zhang, Dequan;Zhang, Huimin;Li, Xin;Fang, Fei;Liang, Ce;Wang, Zhenyu
    • Food Science of Animal Resources
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    • v.41 no.5
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    • pp.802-815
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    • 2021
  • To investigate the effect of postmortem phases on lamb meat quality, the physicochemical quality, microstructure and water mobility of oyster cut, short loin, knuckle and silverside muscles from Small-Tail Han sheep were evaluated in the pre-rigor, rigor mortis and post-rigor phases. Pre-rigor lamb meat had higher pH and water holding capacity (WHC), whereas lower CIE L*, b*, hue angle values than rigor mortis and post-rigor meat (p<0.05). The Warner-Bratzler shear force (WBSF) values were higher in rigor mortis short loin and silverside than their pre-rigor and post-rigor counterparts, pre-rigor short loin had lower WBSF value than its post-rigor counterpart (p<0.05). Muscle fibers shrank laterally and longitudinally during the onset of rigor mortis. Rigor mortis and postrigor lamb meat exhibited wide I-bands, dark A-bands, short sarcomeres and large inter-myofibrillar spaces. The shift of immobilized water to free water and repulsion from the intra-myofibrillar space to the extracellular space result in the increase of water loss in rigor mortis and post-rigor lamb meat. The results of the principal component analysis (PCA) indicated that rigor mortis and post-rigor lamb meat had similar quality properties but different from pre-rigor lamb meat. In conclusion, the lamb meat in the pre-rigor phase had good tenderness, color and WHC. The results of this research could provide some theoretical references for lamb meat production and processing.

A Scalable Montgomery Modular Multiplier (확장 가능형 몽고메리 모듈러 곱셈기)

  • Choi, Jun-Baek;Shin, Kyung-Wook
    • Journal of IKEEE
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    • v.25 no.4
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    • pp.625-633
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    • 2021
  • This paper describes a scalable architecture for flexible hardware implementation of Montgomery modular multiplication. Our scalable modular multiplier architecture, which is based on a one-dimensional array of processing elements (PEs), performs word parallel operation and allows us to adjust computational performance and hardware complexity depending on the number of PEs used, NPE. Based on the proposed architecture, we designed a scalable Montgomery modular multiplier (sMM) core supporting eight field sizes defined in SEC2. Synthesized with 180-nm CMOS cell library, our sMM core was implemented with 38,317 gate equivalents (GEs) and 139,390 GEs for NPE=1 and NPE=8, respectively. When operating with a 100 MHz clock, it was evaluated that 256-bit modular multiplications of 0.57 million times/sec for NPE=1 and 3.5 million times/sec for NPE=8 can be computed. Our sMM core has the advantage of enabling an optimized implementation by determining the number of PEs to be used in consideration of computational performance and hardware resources required in application fields, and it can be used as an IP (intellectual property) in scalable hardware design of elliptic curve cryptography (ECC).

PPIA, HPRT1, and YWHAZ are suitable reference genes for quantitative polymerase chain reaction assay of the hypothalamic-pituitary-gonadal axis in sows

  • Kim, Hwan-Deuk;Jo, Chan-Hee;Choe, Yong-Ho;Lee, Hyeon-Jeong;Jang, Min;Bae, Seul-Gi;Yun, Sung-Ho;Lee, Sung-Lim;Rho, Gyu-Jin;Kim, Seung-Joon;Lee, Won-Jae
    • Animal Bioscience
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    • v.35 no.12
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    • pp.1850-1859
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    • 2022
  • Objective: The quantitative reverse transcription polymerase chain reaction (qPCR) is the most accurate and reliable technique for analysis of gene expression. Endogenous reference genes (RGs) have been used to normalize qPCR data, although their expression may vary in different tissues and experimental conditions. Verification of the stability of RGs in selected samples is a prerequisite for reliable results. Therefore, we attempted to identify the most stable RGs in the hypothalamic-pituitary-gonadal (HPG) axis in sows. Methods: The cycle threshold values of nine commonly used RGs (18S, HPRT1, GAPDH, RPL4, PPIA, B2M, YWHAZ, ACTB, and SDHA) from HPG axis-related tissues in the domestic sows in the different stages of estrus cycle were analyzed using two RG-finding programs, geNorm and Normfinder, to rank the stability of the pool of RGs. In addition, the effect of the most and least stable RGs was examined by normalization of the target gene, gonadotropin-releasing hormone (GnRH), in the hypothalamus. Results: PPIA, HPRT1, and YWHAZ were the most stable RGs in the HPG axis-related tissues in sows regardless of the stages of estrus cycle. In contrast, traditional RGs, including 18S and ACTB, were found to be the least stable under these experimental conditions. In particular, in the normalization of GnRH expression in the hypothalamus against several stable RGs, PPIA, HPRT1, and YWHAZ, could generate significant (p<0.05) elevation of GnRH in the preovulatory phase compared to the luteal phase, but the traditional RGs with the least stability (18S and ACTB) did not show a significant difference between groups. Conclusion: These results indicate the importance of verifying RG stability prior to commencing research and may contribute to experimental design in the field of animal reproductive physiology as reference data.

Disease vector occurrence and ecological characteristics of chiggers on the chestnut white-bellied rat Niviventer fulvescens in Southwest China between 2001 and 2019

  • Yan-Ling Chen;Xian-Guo Guo;Wen-Yu Song;Tian-Guang Ren;Lei Zhang;Rong Fan;Cheng-Fu Zhao;Zhi-Wei Zhang;Wen-Ge Dong;Xiao-Bin Huang;Dao-Chao Jin
    • Parasites, Hosts and Diseases
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    • v.61 no.3
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    • pp.272-281
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    • 2023
  • Chigger mites are the vector of scrub typhus. This study estimates the infestation status and ecological characteristics of chiggers on the chestnut white-bellied rat Niviventer fulvescens in Southwest China between 2001 and 2019. Chiggers were identified under the microscope, and infestation indices were calculated. The Preston's log-normal model was used to fit the curve of species abundance distribution. A total of 6,557 chiggers were collected in 136 of 342 N. fulvescens rats, showing high overall infestation indices (prevalence=39.8%, mean abundance=19.2, mean intensity=48.2) and high species diversity (S=100, H'=3.0). Leptotrombidium cangjiangense, Neotrombicula japonica, and Ascoschoengastia sifanga were the three dominant chigger species (constituent ratio=42.9%; 2,736/6,384) and exhibited an aggregated distribution among different rat individuals. We identified 100 chigger species, with 3 of them (Leptotrombidium scutellare, Leptotrombidium wenense, and Leptotrombidium deliense) as the main vectors of scrub typhus in China and nine species as potential vectors of this disease. Disease vector occurrence on N. fulvescens may increase the risk of spreading scrub typhus from rats to humans. Chigger infestation on N. fulvescens varied significantly in different environments. The species abundance distribution showed a log-normal distribution pattern. The estimated number of chigger species on N. fulvescens was 126 species.

A Study of the Metal Artifact Reduction using Dual Energy CT : Clinical Applications of Dual Energy and MAR Algorithm (Dual Energy CT를 이용한 금속물질 인공물 감소방법 : Dual Energy와 MAR알고리즘의 임상적 응용)

  • Park, Ki Seok;Choi, Woo Jeon;Kim, Dong Hyun
    • Journal of the Korean Society of Radiology
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    • v.15 no.3
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    • pp.273-279
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    • 2021
  • Metal material inserted into the body have a large difference in density from human tissues or bones around the Metal during CT scans.. Therefore, the Metal material inserted into the body produces Artifact. Metal Artifact, which occurs around Metals, can degrade the quality of CT images, causing confusion when medical team diagnose lesions. Through this experiment, we confirm that the occurrence of Artifacts decrease by using Dual energy CT and MAR algorithm in Single source Dual energy CT. We also want to present basic data on clinical application methods by comparing and analyzing the characteristics of images obtained by each method. Using GE 750HD CT, artificial implants were scanned using general method and Dual energy. Then we apply the MAR algorithm to each image obtained. And all previously acquired images were compared and analyzed the characteristics of the examination, such as image quality evaluation and dose evaluation. Images with MAR algorithm and Dual Energy confirmed a decrease in Metal Artifact. Images with MAR algorithm have reduced Metal Artifact, but have the disadvantage of distorting the details of artificial joint implants. On the other hand images teseted with Dual Energy have the advantage of being able to implement details than those applied with MAR algorithms, it takes longer to reconstruct the image and the exposure dose was about four times higher than those applied with MAR algorithm. In order to locate Metals, such as the post-operative follow-up period, it is useful to apply MAR algorithm to obtain images. And it is more useful to examine with Dual Energy when micro lesion identification, such as cardiac examination, and surgical planning or when tests are performed in diagnostic way.