• Title/Summary/Keyword: Green tissue engineering

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A Multi-detection Fluorescence Dye with 5-ALA and ICG Using Modified Light Emitting Diodes

  • Yoon, Kicheol;Kim, Eunji;Kim, Kwanggi;Lee, Seunghoon;Yoo, Heon
    • Current Optics and Photonics
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    • v.3 no.3
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    • pp.256-262
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    • 2019
  • Extensive tumor resection accompanied by radiotherapy and chemotherapy is the standard of care for malignant gliomas. However, there is a significant obstacle to the complete resection of the tumor due to the difficulty of distinguishing tumor and normal brain tissue with a conventional surgical microscope. Recently, multiple studies have shown the possibility of fluorescence-guided surgery in malignant gliomas. The most used fluorescence dyes for brain tumor surgery are 5-aminolevulinic acid (5-ALA) and indocyanine green (ICG). In this paper, a new fluorescence guided operation system, which can detect both 5-ALA and ICG fluorescent images simultaneously, is presented. This operation system consists of light emitting diodes (LEDs) which emits 410 nm and 740 nm wavelengths. We have performed experiments on rats in order to verify the operation of the newly developed operation system. Oral administration and imaging were performed to observe the fluorescence of 5-ALA and ICG fluorescence in rats. When LEDs at wavelengths of 410 nm and 740 nm were irradiated on rats, 628 nm wavelength with a violet fluorescence color and 825 nm wavelength with a red fluorescence color were expressed in 5-ALA and ICG fluorescent material, respectively, thus we were able to distinguish the tumor tissues easily. Previously, due to the poor resolution of the conventional surgical microscope and the fact that the color of the vein is similar to that of the tumor, the tumor resection margin was not easy to observe, thus increasing the likelihood for cancer recurrence. However, when the tumor is observed through the fluorescence guided operation system, it is possible to easily distinguish the color with the naked eye and it can be completely removed. Therefore, it is expected that surgical removal of cancerous tumors will be possible and surgical applications and surgical microscopes for cancer tumor removal surgery will be promising in the future.

Histological Changes of Cervical Disc Tissue in Patients with Degenerative Ossification

  • Xiong, Yang;Yang, Ying-Li;Gao, Yu-Shan;Wang, Xiu-Mei;Yu, Xing
    • Journal of Korean Neurosurgical Society
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    • v.65 no.2
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    • pp.186-195
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    • 2022
  • Objective : To explore the histological feature of the cervical disc degeneration in patients with degenerative ossification (DO) and its potential mechanisms. Methods : A total of 96 surgical segments, from cervical disc degenerative disease patients with surgical treatment, were divided into ossification group (group O, n=46) and non-ossification group (group NO, n=50) based on preoperative radiological exams. Samples of disc tissues and osteophytes were harvested during the decompression operation. The hematoxylin-eosin staining, Masson trichrome staining and Safranin O-fast green staining were used to compare the histological differences between the two groups. And the distribution and content of transforming growth factor (TGF)-β1, p-Smad2 and p-Smad3 between the two groups were compared by a semi-quantitative immunohistochemistry (IHC) method. Results : For all the disc tissues, the content of disc cells and collagen fibers decreased gradually from the outer annulus fibrosus (OAF) to the central nucleus pulposus (NP). Compared with group NO, the number of disc cells in group O increased significantly. But for proteoglycan in the inner annulus fibrosus (IAF) and NP, the content in group O decreased significantly. IHC analysis showed that TGF-β1, p-Smad2, and p-Smad3 were detected in all tissues. For group O, the content of TGF-β1 in the OAF and NP was significantly higher than that in group NO. For p-Smad2 in IAF and p-Smad3 in OAF, the content in group O were significantly higher than group NO. Conclusion : Histologically, cervical disc degeneration in patients with DO is more severe than that without DO. Local higher content of TGF-β1, p-Smad2, and p-Smad3 are involved in the disc degeneration with DO. Further studies with multi-approach analyses are needed to better understand the role of TGF-β/Smads signaling pathway in the disc degeneration with DO.

Effect of Light and Cadmium on the Activity and Isozyme Pattern of Catalase from Ric(Oryza sativa L.) (빛과 카드뮴이 벼 catalase 활성과 동위효소 발현에 미치는 영향)

  • Kim, Yoon-Kyoung;Lee, Mi-Young
    • Applied Biological Chemistry
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    • v.49 no.4
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    • pp.287-292
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    • 2006
  • The effects of cadmium on the catalase activity and isozyme patterns under light and dark conditions of rice(Oryza sativa L. cv. Dongjin) seedlings were examined. Cadmium treatment resulted in the notable enhancement of $H_2O_2$ contents in the seedling roots and leaves under light and dark conditions. The catalase isozyme patterns in the roots were different from those in the leaves, showing tissue-specific expression of the enzyme. Moreover, the expression patterns of catalase isozymes in the green seedling roots were different from those in the etiolated seedling roots following cadmium treatment. The increase of total catalase activity was about 16 times at 1 mM cadmium and marked inductions of the isozyme CAT1 and CAT2 contributed to this increase in the green seedling roots. On the other hand, in the etiolated seedling roots, total catalase activity was lower than that of control at 0.5 and 1 mM cadmium, even though catalase activity increased about 3 times at 0.1 mM cadmium. The 3 fold increase of total catalase activity was mainly due to the increase of CAT1, CAT3 and CAT4 at 0.1 mM cadmium. However, treatment with higher concentrations of cadmium decreased the activity of CAT2 and CAT4 in the etiolated roots. In the leaves, the catalase existed as three isozymes; one cationic isozyme CATc, one neutral isozyme CATn and one anionic isozyme CAT1 in the control. The isozyme patterns and total activities remained unaffected by cadmium under light and dark conditions in the seedling leaves. Taken together, it seems that cadmium-induced changes of catalase might be regulated by light in the roots, but not in the leaves.

Real-Time PCR for Validation of Minute Virus of Mice Safety during the Manufacture of Mammalian Cell Culture-Derived Biopharmaceuticals (세포배양 유래 생물의약품 생산 공정에서 Minute Virus of Mice 안전성 검증을 위한 Real-Time PCR)

  • Lee, Dong-Hyuck;Cho, Hang-Mee;Kim, Hyun-Mi;Lee, Jung-Suk;Kim, In-Seop
    • Microbiology and Biotechnology Letters
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    • v.36 no.1
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    • pp.12-20
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    • 2008
  • Validation of viral safety is essential in ensuring the safety of mammalian cell culture-derived biopharmaceuticals, because numerous adventitious viruses have been contaminated during the manufacture of the products. Mammalian cells are highly susceptible to minute virus of mice(MVM), and there are several reports of MVM contamination during the manufacture of biopharmaceuticals. In order to establish the validation system for the MVM safety, a real-time PCR method was developed for quantitative detection of MVM in cell lines, raw materials, manufacturing processes, and final products as well as MVM clearance validation. Specific primers for amplification of MVM DNA was selected, and MVM DNA was quantified by use of SYBR Green I. The sensitivity of the assay was calculated to be $6{\times}10^{-2}TCID_{50}/mL$. The real-time PCR method was proven to be reproducible and very specific to MVM. The established real-time PCR assay was successfully applied to the validation of Chinese hamster ovary (CHO) cell artificially infected with MVM. MVM DNA could be Quantified in CHO cell as well as culture supernatant. When the real-time PCR assay was applied to the validation of virus removal during a virus filtration process, the result was similar to that of virus infectivity assay. Therefore, it was concluded that this rapid, specific, sensitive, and robust assay could replace infectivity assay for detection and clearance validation of MVM.

Removal and Inactivation of Human Immunodeficiency Virus(HIV-1) by Cold Ethanol Fractionation and Pasteurization during the Manufacturing of Albumin and Immunoglobulins from Human Plasma

  • Kim, In-Seop;Eo, Ho-Gueon;Park, Chan-Woo;Chong E. Chang;Lee, Soungmin
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.6 no.1
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    • pp.25-30
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    • 2001
  • Viral safety is a prerequisite for manufacturing clinical albumin and immunoglobulins from human plasma pools. This study was designed to evaluate the efficacy of cold ethanol fractionation and pasteurization (60$\^{C}$ heat treatment for 10h) for the removal/inactivation of human immunodeficiency virus type 1 (HIV-1) during the manufacturing of albumin and immunoglobulins. Samples from the relevant stages of the production process were spiked with HIV-1, and the amount of virus in each fraction was quantified by the 50% tissue culture infectious dose(TCID(sub)50). Both fraction IV fractionation and pasteurization steps during albumin processing were robust and effective in inactivating HIV-1, titers of which were reduced from an initial 8.5 log(sub)10 TCID(sub)50 to undetectable levels. The log reduction factors achieved were $\geq$ 4.5 and $\geq$ 6.5, respectively. In addition, fraction III fractionation and pasteurization during immunoglobulins processing were robust and effective in eliminating HIV-1. HIV-1 titers were reduced from an initial 7.3 log(sub)10 TCID(sub)50 to undetectable levels. The log reduction factors achieved in this case were $\geq$ 4.9 and $\geq$ 5.3, respectively. These results indicate that the process investigated for the production of albumin and immunoglobulins have sufficient HIV-1 reducing capacity to achieve a high margin of safety.

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LPS-Induced Modifications in Macrophage Transcript and Secretion Profiles Are Linked to Muscle Wasting and Glucose Intolerance

  • Heeyeon Ryu;Hyeon Hak Jeong;Seungjun Lee;Min-Kyeong Lee;Myeong-Jin Kim;Bonggi Lee
    • Journal of Microbiology and Biotechnology
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    • v.34 no.2
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    • pp.270-279
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    • 2024
  • Macrophages are versatile immune cells that play crucial roles in tissue repair, immune defense, and the regulation of immune responses. In the context of skeletal muscle, they are vital for maintaining muscle homeostasis but macrophage-induced chronic inflammation can lead to muscle dysfunction, resulting in skeletal muscle atrophy characterized by reduced muscle mass and impaired insulin regulation and glucose uptake. Although the involvement of macrophage-secreted factors in inflammation-induced muscle atrophy is well-established, the precise intracellular signaling pathways and secretion factors affecting skeletal muscle homeostasis require further investigation. This study aimed to explore the regulation of macrophage-secreted factors and their impact on muscle atrophy and glucose metabolism. By employing RNA sequencing (RNA-seq) and proteome array, we uncovered that factors secreted by lipopolysaccharide (LPS)-stimulated macrophages upregulated markers of muscle atrophy and pro-inflammatory cytokines, while concurrently reducing glucose uptake in muscle cells. The RNA-seq analysis identified alterations in gene expression patterns associated with immune system pathways and nutrient metabolism. The utilization of gene ontology (GO) analysis and proteome array with macrophage-conditioned media revealed the involvement of macrophage-secreted cytokines and chemokines associated with muscle atrophy. These findings offer valuable insights into the regulatory mechanisms of macrophage-secreted factors and their contributions to muscle-related diseases.

Real-Time RT-PCR for Validation of Reovirus Type 3 Safety During the Manufacture of Mammalian Cell Culture-Derived Biopharmaceuticals (세포배양 유래 생물의약품 생산 공정에서 Reovirus Type 3 안전성 검증을 위한 Real-Time RT-PCR)

  • Lee, Dong-Hyuck;Jeong, Hyo-Sun;Kim, Tae-Eun;Oh, Seon-Hwan;Lee, Jung-Suk;Kim, In-Seop
    • Korean Journal of Microbiology
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    • v.44 no.3
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    • pp.228-236
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    • 2008
  • Validation of viral safety is essential in ensuring the safety of mammalian cell culture-derived biopharmaceuticals, because numerous adventitious viruses have been contaminated during the manufacture of the products. Mammalian cells are highly susceptible to Reovirus type 3 (Reo-3), and there are several reports of Reo-3 contamination during the manufacture of biopharmaceuticals. In order to establish the validation system for the Reo-3 safety, a real-time RT-PCR method was developed for quantitative detection of Reo-3 in cell lines, raw materials, manufacturing processes, and final products as well as Reo-3 clearance validation. Specific primers for amplification of Reo-3 RNA was selected, and Reo-3 RNA was quantified by use of SYBR Green I. The sensitivity of the assay was calculated to be $3.2{\times}10^0\;TCID_{50}/ml$. The real-time RT-PCR method was proven to be reproducible and very specific to Reo-3. The established real-time RT-PCR assay was successfully applied to the validation of Chinese hamster ovary (CHO) cell artificially infected with Reo-3. Reo-3 RNA could be quantified in CHO cell as well as culture supernatant. When the real-time RT-PCR assay was applied to the validation of virus removal during a virus filtration process, the result was similar to that of virus infectivity assay. Therefore, it was concluded that this rapid, specific, sensitive, and robust assay could replace infectivity assay for detection and clearance validation of Reo-3.

Occurrence of White Smut Caused by Entyloma cosmi on Cosmos bipinnatus in Korea (Entyloma cosmi에 의한 코스모스 흰깜부기병 발생)

  • Park, Ji-Hyun;Cho, Sung-Eun;Park, Mi-Jeong;Hong, Sun-Hee;Shin, Hyeon-Dong
    • Research in Plant Disease
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    • v.18 no.3
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    • pp.250-254
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    • 2012
  • In July 2011, hundreds of garden cosmos (Cosmos bipinnatus) were found to be infected with a white smut in Namyangju. Further surveys in central Korea showed that the disease occurred in all the nine localities visited, with disease incidence rate of >50%. Symptoms manifested as pale-green to chlorotic areas on the leaves, followed by white mildew development on lesions. The lesions usually coalesced and became necrotic, considerably reducing the aesthetic value. Caespituli were initially hypophyllous but later epiphyllous as well. Conidiophores were 10-40 in moderately dense fascicles, hyaline throughout, mostly guttulate, $20-45{\times}2-4{\mu}m$, obtuse at the apex, and had inconspicuous conidial scars. Conidia were solitary, botuliform, hyaline, aseptate, mostly guttulate, apex rounded, base rounded to truncate, $17-24{\times}3.5-5{\mu}m$, hilum inconspicuous, unthickened, and not darkened. Sori were rounded to elongated, resulting in first white and later brown leaf spots. Teliospores embedded in the leaf tissue were subglobose to ellipsoidal, subhyaline to pale yellowish-brown, $12.5-20{\times}10-15{\mu}m$. These morphological characteristics corresponded with those of Entyloma cosmi V$\acute{a}$nky, Horita & Jage. This is the first report of white smut disease of garden cosmos due to infection of E. cosmi in Korea.

Hepatoprotective Effects of Semisulcospira libertine Hydrolysate on Alcohol-induced Fatty Liver in Mice (알코올성 지방간 유발 마우스에서 다슬기 유래 가수분해물의 간 보호 효과)

  • Song, Eun Jin;Cho, Kyoung Hwan;Choo, Ho Jin;Yang, Eun Young;Jung, Yoon Kyoung;Seo, Min Gyun;Kim, Jong Cheol;Kang, Eun Ju;Ryu, Gi Hyung;Park, Beom Yong;Hah, Young-Sool
    • Food Engineering Progress
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    • v.21 no.4
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    • pp.318-325
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    • 2017
  • Alcoholic steatosis is a fundamental metabolic disorder and may precede the onset of more severe forms of alcoholic liver disease. In this study, we isolated enzymatichydrolysate from Semisulcospira libertine by alcalase hydrolysis and investigated the protective effect of Semisulcospira libertine hydrolysate on liver injury induced by alcohol in the mouse model of chronic and binge ethanol feeding (NIAAA). In an in vitro study, the hydrolysate protects HepG2 cells from ethanol toxicity. Liver damage was assessed by histopathological examination, as well as by quantitating activities of serum aspartate aminotransferase (AST), alanine aminotransferase (ALT), and alkaline phosphatase (ALP). After the administration of S. libertina hydrolysate, fat accumulation and infiltration of inflammatory cells in liver tissues were significantly decreased in the NIAAA mouse model. The elevated levels of serum AST, ALT, and ALP activities, along with the lipid contents of a damaged liver, were recovered in experimental mice administrated with S. libertina hydrolysate, suggesting its role in blood enzyme activation and lipid content restoration within damaged liver tissues. Moreover, treatment with S. libertine hydrolysate reduced the expression rate of cyclooxygenase (COX-2), interleukin $(IL)-1{\beta}$, and IL-6, which accelerate inflammation and induces tissue damage. All data showed that S. libertine hydrolysate has a preventive role against alcohol-induced liver damages by improving the activities of blood enzymes and modulating the expression of inflammation factor, suggesting S. libertine hydrolysate could be a commercially potential material for the restoration of hepatotoxicity.

FABRICATION OF MYOMUCOSAL FLAP USING CULTURED ORAL EPITHELIUM IN RABBIT MODEL (가토모델에서 배양 구강상피를 이용한 근-점막 피판의 형성에 관한 연구)

  • Shin, Young-Min;Chung, Hun-Jong;Ahn, Kang-Min;Park, Hee-Jung;Sung, Mi-Ae;Kim, Soung-Min;Hwang, Soon-Jung;Kim, Myung-Jin;Jahng, Jeong-Won;Kim, Sung-Po;Yang, Eun-Kyung;Song, Kye-Yong;Lee, Jong-Ho
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.27 no.3
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    • pp.226-237
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    • 2005
  • Purpose : Extensive defect of oral and maxillofacial area is usually reconstructed with composite flap including skin paddle. However, if the defects are lined with only skin components, the mucosa's role in mastication and texture are not restored. Furthermore, stiffness and hair-growing prevent denture rehabilitation and good oral hygiene. This study was performed to overcome the disadvantages of composite soft tissue flaps including the skin and to make a model for myo-mucosal flaps. Materials and methods : Buccal mucosa sized $0.5\times1.0\;cm^2$ from New Zealand rabbit (around 1.5kg) was harvested and cultivated by the modification of Rheinwald and Green's keratinocyte culture method. Cultured mucosa was grafted on the fascia of latismus dorsi as form of mucosal sheet. After 7, 10, 14 days, the myomucosal flap was excised and evaluated under light microscope with H & E and immunohistochemical staining. As control group, harvested buccal mucosa from rabbit was transplanted to gracilis muscle(n=6). Results : From 7 days after prelamination, the basal layer of the grafted mucosa resembled that of normal mucosa. As control group, transplanted mucosa had original shape but there's slight inflammatory reaction. Prelaminated mucosa has 19.8$\pm$4.59 cell layers and some samples have more than 20 layers. The expression rate of PCNA was relatively strong (42.9%$\pm$14.1) at the basal layer of grafted mucosa and the laminin was found at the basal layer. On the contrary, prelaminated mucosa at 10 days showed moderate expression rate of PCNA(32.4%$\pm$4.62). We found the mucosal layer was somehow disappeared and there is strong inflammatory reaction. After 14 days prelamination, the grafted oral keratinocytes were almost disappeared and expression of PCNA was not observed. Conclusion : We can make 75 fold large mucosal($3850mm^2$) sheet from small samples of mucosa $(50mm^2)$. Epithelial sheet that grafted on the fascia of muscle underwent differentiation and proliferation. But after 10, 14 days, there was strong inflammatory reaction and the grafted mucosa was destroyed from surface layer. In rabbit model, transfer of fascio-mucosal flap should be done from 7 to 10 days after prelamination.