A Comparison of Lyophilized Amniotic Membrane with Cryopreserved Amniotic Membrane for the Reconstruction of Rabbit Corneal Epithelium

Ahn Jae-Il,Jang In-Keun,Lee Doo-Hoon,Seo Young-Kwon,Yoon Hee-Hoon,Shin Youn-Ho,Kim Jae-Chan,Song Kye-Yong,Lee Hee-Gu,Yang Eun-Kyung,Kim Ki-Ho,Park Jung-Keung The Korean Society for Biotechnology and Bioengine 2005 Biotechnology and Bioprocess Engineering Vol.10 No.3

Many researchers have employed cryopreserved amniotic membrane (CAM) in the treatment of a severely damaged cornea, using corneal epithelial cells cultured on an amniotic membrane (AM). In this study, two Teflon rings were made for culturing the cells on the LAM and CAM, and were then used to support the AM, which is referred to in this paper as an Ahn's AM supporter. The primary corneal epithelial cells were obtained from the limbus, using an ex-plantation method. The corneal epithelium could be reconstructed by culturing the third­passage corneal epithelial cells on the AM. A lyophilized amniotic membrane (LAM) has a higher rate of graft take, a longer shelf life, is easier to store, and safer, due to gamma irradiation, than a (AM. The corneal epithelium reconstructed on the LAM and (AM, supported by the two­Teflon rings, was similar to normal corneal epithelium. However, the advantages of the LAM over that of the (AM make the former more useful. The reconstruction model of the corneal epithelium, using AM, is considered as a good in vitro model for transplantation of cornel epithelium into patients with a severely damaged cornea.

Reconstruction of Rabbit Corneal Epithelium on Lyophilized Amniotic Membrane Using the Tilting Dynamic Culture Method

Ahn, Jae-Il,Lee, Doo-Hoon,Ryu, Yang-Hwan,Jang, In-Keun,Yoon, Mun-Young,Shin, Youn Ho,Seo, Young-Kwon,Yoon, Hee-Hoon,Kim, Jae-Chan,Song, Kye-Yong,Yang, Eun-Kyung,Kim, Ki-Ho,Park, Jung-Keug Blackwell Publishing Inc 2007 Artificial Organs Vol.31 No.9

<P>Abstract: </P><P>Rabbit corneal epithelium was reconstructed using tilting dynamic culture with a self-manufactured, amniotic membrane (AM) supporter and a lyophilized amniotic membrane (LAM). Rabbit corneal epithelial (RCE) cells were cultured and cryopreserved after isolation from the limbus. The second- and third-passage RCE cells were plated onto the epithelial side of the LAM of Ahn's AM supporter. Two days later, the air–liquid interface culture was maintained with third-passage RCE cells for 6 days and second-passage corneal epithelial cells for 9 days. The average viability of thawed RCE cells, assessed using trypan blue dye exclusion, was 77.42%. The reconstructed corneal epithelium was characterized by histological (hematoxylin and eosin) and immunohistochemical staining (proliferating cell nuclear antigen) for light microscopy, and by reverse transcriptase-polymerase chain reaction, glucose assay, and transmission electron microscopy. The basal layer of the reconstructed corneal epithelium was well formed, and the epithelium was tightly constructed due to the increase in cell proliferation and differentiation caused by the tilting dynamic culture, as opposed to static culture. Tilting dynamic culture was useful for the reconstruction of the epithelium using easily damaged epithelial cells and resulted in more stratum cell layers. Moreover, cytokeratin (CK3) mRNA expression in tilting dynamic cultured third-passage RCE cells seeded onto AM was greater than in static cultured third-passage RCE cells. The morphology of the reconstructed corneal epithelium on LAM by tilting dynamic culture for 9 days resembled that of the skin epidermis. This was thought to be because the tilting dynamic culture not only accelerated the proliferation and differentiation of cells by physical or mechanical stimulation, but also ensured that the supply of medium was delivered to the basal cells more efficiently. Thus, the reconstruction of the corneal epithelium using LAM and tilting dynamic culture was considered to be a good in vitro model for autologous or allogeneic transplantation of corneal epithelium and skin epidermis in patients with damaged epithelia. </P>

LPS로 염증 유도된 RAW 264.7세포에 대한 참콩풍뎅이(Popillia flavosellata) 에탄올 추출물의 항염증 효과

윤영일(Young-Il Yoon),황재삼(Jae-Sam Hwang),김미애(Mi-Ae Kim),안미영(Mi Young Ahn),이영보(Young-Bo Lee),한명세(Myung Sae Han),구태원(Tae-Won Goo),윤은영(Eun-Young Yun) 한국생명과학회 2015 생명과학회지 Vol.25 No.9

본 연구에서는 참콩풍뎅이(Popillia flavosellata) 에탄올 추출물(PFE)의 항염증 효능을 분석하기 위해 PFE를 농도별(500, 1,000, 2,000 ㎍/ml)로 대식세포인 RAW 264.7에 처리 시 최고 처리농도인 2,000 ㎍/ml까지 통계적인 유의성 있는 독성이 없음을 확인하였다. LPS (100 ng/ml)로 염증 유도된 RAW 264.7 세포에 PFE를 농도별(500, 1,000, 2,000 ㎍/ml)로 동시 처리 시 농도 의존적으로 염증성사이토카인인 TNF-α와 IL-6의 단백질 생성을 통계적인 유의성(p<0.001)있게 억제함을 확인하였다. 또한 염증 유도된 RAW 264.7 세포에 PFE 동시 처리 시 NF-κB p65의 핵으로 이동이 차단됨과 iNOS와 COX-2의 단백질 발현을 감소시키는 것을 확인하였다. 이상의 연구결과를 통해 참콩풍뎅이는 염증에 의해 활성화된 TLR-4 신호전달과정을 조절하는 NF-κB p65의 활성과 염증성사이토카인 TNF-α와 IL-6의 생성 및 염증성효소 iNOS와 COX-2의 생성을 억제하는 항염증 효능이 있음을 확인하였다. The beetle Popillia flavosellata has been no reported its functional effects. In this study, we investigated the anti-inflammatory effect of P. flavosellata ethanol extract (PFE) on RAW 264.7 mouse macrophage cells treated with lipopolysaccharide (LPS) for the induction of inflammation. First, we examined the cytotoxicity of PFE in the RAW 264.7 cells at a concentration of 2,000 μg/ml or less. To evaluate the anti-inflammatory effects of PFE, we investigated the expression levels of proinflammatory cytokines, such as tumor necrosis factor (TNF)-α and interleukin (IL)-6, and proinflammatory enzymes, such as inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) in LPS-induced RAW 264.7 cells. In addition, we examined whether PFE inhibited the translocation of nuclear factor kappa B (NF-κB) p65 into the nucleus in the LPS-induced RAW 264.7 cells. We found that the protein levels of TNF-α and IL-6 were decreased in the LPS-induced RAW 264.7 cells after the treatment with PFE in a dose-dependent manner. In addition, we confirmed that PFE inhibited the translocation of NF-κB p65 into the nucleus, as well as the protein expression levels of iNOS and COX-2. Accordingly, we propose that PFE exerts an anti-inflammatory effect through the down-regulation of NF-κB p65, TNF-α, IL-6, iNOS, and COX-2 via the toll like receptor (TLR)-4 inflammatory signaling pathway.

LPS로 유도된 RAW 264.7세포에 대한 벼메뚜기(Oxya chinensis sinuosa) 에탄올 추출물의 항염증 효과

윤영일(Young-Il Yoon),정미연(Mi Yeon Chung),황재삼(Jae-Sam Hwang),구태원(Tae-Won Goo),안미영(Mi-Young Ahn),이영보(Young-Bo Lee),한명세(Myung-Sea Han),윤은영(Eun-Young Yun) 한국생명과학회 2014 생명과학회지 Vol.24 No.4

본 연구에서는 벼메뚜기 에탄올 추출물의 항염증 효능을 분석하기 위해 LPS로 염증 유도된 RAW 264.7 세포를 이용하였다. OCE의 항염증 효능을 확인 하기 위해서, 염증 유도된 RAW 264.7 세포에 대해 OCE 농도 의존적으로 염증성 사이토카인인 TNF-α와 IL-6의 유전자발현 및 단백질 생성을 감소시킴을 real-time PCR과 ELISA로 확인하였다. 또한, NF-κB p65의 핵으로 이동이 차단됨을 면역형광염색으로 확인하였으며, iNOS와 COX-2 단백질 발현을 감소시키는 것을 Western blot 분석으로 확인하였다. 이상의 연구결과를 통해 벼메뚜기는 염증에 의한 NF-κB p65의 활성과 TNF-α와 IL-6의 생성과 iNOS 및 COX-2의 발현을 억제하는 항염증 효능을 갖고 있는 것을 확인하였다. Although the grasshopper Oxya chinensis sinuosa has long been used as food in Korea, there is little data on its functional effects. In this study, we investigated the anti-inflammatory effect of O. c. sinuosa ethanol extract (OCE) in RAW 264.7 mouse macrophage cells treated with lipopolysaccharide (LPS) for induction of inflammation. First, we determined that there is no cytotoxicity at 2,000 μg/ml or less of OCE in RAW 264.7 cells. To evaluate the anti-inflammatory effects of OCE, we investigated expression levels of pro-inflammatory cytokines such as tumor necrosis factor (TNF)-α and interleukin (IL)-6, and pro-inflammatory enzymes such as inducible nitric oxide synthase (iNOS) and cyclo-oxygenase- 2 (COX-2) in LPS-induced RAW 264.7 cells. In addition, we examined whether OCE could inhibit translocation of NF-κB p65 into the nucleus in LPS induced RAW 264.7 cells. As a result, we found that the mRNA and protein levels of TNF-α and IL-6 decreased in LPS-induced RAW 264.7 cells after treatment with OCE in a dose-dependent manner. In addition, we confirmed a 2,000 ug/ml concentration of OCE inhibited translocation of NF-κB p65 by immunnostaining and Western blot analysis, and a decrease in the protein expression levels of iNOS and COX-2. Accordingly, we suppose that OCE has an anti-inflammatory effect through down-regulation of TNF-α, IL-6, iNOS, and COX-2 related to NF-κB p65 inflammatory signaling pathways.

<i>In vivo</i> genotoxicity evaluation of lung cells from Fischer 344 rats following 28 days of inhalation exposure to MWCNTs, plus 28 days and 90 days post-exposure

Kim, Jin Sik,Sung, Jae Hyuck,Choi, Byung Gil,Ryu, Hyeon Yeol,Song, Kyung Seuk,Shin, Jae Hoon,Lee, Jong Seong,Hwang, Joo Hwan,Lee, Ji Hyun,Lee, Gun Ho,Jeon, Kisoo,Ahn, Kang Ho,Yu, Il Je Informa Healthcare 2014 INHALATION TOXICOLOGY Vol.26 No.4

<P>Despite their useful physico-chemical properties, carbon nanotubes (CNTs) continue to cause concern over occupational and human health due to their structural similarity to asbestos. Thus, to evaluate the toxic and genotoxic effect of multi-wall carbon nanotubes (MWCNTs) on lung cells <I>in vivo</I>, eight-week-old rats were divided into four groups (each group = 25 animals), a fresh air control (0 mg/m<SUP>3</SUP>), low (0.17 mg/m<SUP>3</SUP>), middle (0.49 mg/m<SUP>3</SUP>), and high (0.96 mg/m<SUP>3</SUP>) dose group, and exposed to MWCNTs <I>via</I> nose-only inhalation 6 h per day, 5 days per week for 28 days. The count median length and geometric standard deviation for the MWCNTs determined by TEM were 330.18 and 1.72 nm, respectively, and the MWCNT diameters ranged from 10 to 15 nm. Lung cells were isolated from five male and five female rats in each group on day 0, day 28 (only from males) and day 90 following the 28-day exposure. The total number of animals used was 15 male and 10 female rats for each concentration group. To determine the genotoxicity of the MWCNTs, a single cell gel electrophoresis assay (Comet assay) was conducted on the rat lung cells. As a result of the exposure, the olive tail moments were found to be significantly higher (<I>p</I> < 0.05) in the male and female rats from all the exposed groups when compared with the fresh air control. In addition, the high-dose exposed male and middle and high-dose exposed female rats retained DNA damage, even 90 days post-exposure (<I>p</I> < 0.05). To investigate the mode of genotoxicity, the intracellular reactive oxygen species (ROS) levels and inflammatory cytokine levels (TNF-α, TGF- β, IL-1, IL-2, IL-4, IL-5, IL-10, IL-12 and IFN-γ) were also measured. For the male rats, the H<SUB>2</SUB>O<SUB>2</SUB> levels were significantly higher in the middle (0 days post-exposure) and high- (0 days and 28 days post-exposure) dose groups (<I>p</I> < 0.05). Conversely, the female rats showed no changes in the H<SUB>2</SUB>O<SUB>2</SUB> levels. The inflammatory cytokine levels in the bronchoalveolar lavage (BAL) fluid did not show any statistically significant difference. Interestingly, the short-length MWCNTs deposited in the lung cells were persistent at 90 days post-exposure. Thus, exposing lung cells to MWCNTs with a short tube length may induce genotoxicity.</P>

제왕절개술 환자에서 수술전 투여한 ketamine의 혈청 interleukin-6 및 수술후 통증에 대한 효과

김진수,김유재,안기량,김천숙,김일호,한찬수 순천향의학연구소 1997 Journal of Soonchunhyang Medical Science Vol.3 No.2

The inflammatory reaction to tissue damage during surgery may induce central sensitization followed by hyperalgesia. Previous studies suggest that central sensitization is related to N-methyl-D-aspartate (NMDA) receptor activation, which can be blocked with NMDA antagonist, ketamine. Thus, we compared the effect of preoperative intravenous and epidural low doses of ketamine with placebo on serum interleukin-6 (IL-6) level and postoperative pain. ASA class I and II women scheduled for C-sections received intravenous ketamine 0.15mg/kg(group 2) or placebo(group 1), or epidural ketamine(0.15 mg/kg) before the operation. IL-6 levels were measured before and during the operation, and 8, 24, and 48 hrs after the operation. Visual Analogue Scales(VAS) and Verbal Ration Scales(VRS) were measured at 8, 24, and 48 hrs after the operation. Serum IL-6 levels at 8, 24, and 48 hrs after the operation were significantly lower in the intravenous ketamine and epidural ketamine groups than in the control group. VAS at 8 hrs and 48 hrs after the operation were significantly lower in the epidural ketamine group. VAS at 8 hrs and 48 hrs after the operation were significantly lower in the epidural ketamine group than in the control and intravenous ketamine groups. In conclusion, in the preoperative intravenous and epidural administration of low doses(0.15mg/kg) of ketamine, both are effective in reducing postoperative IL-6 levels. Epidural Ketamine is more effective than intravenous ketamine in postoperative pain control.

Effects of Moxibustion to Zusanli(ST36) on Alteration of Natural Killer Cell Activity in Rats

Choi, Gi Soon,Han, Jae Bok,Park, Joon Ha,Oh, Sang Deog,Lee, Gi Seog,Bae, Hyun Su,Jung, Sung Ki,Cho, Young Wuk,Ahn, Hyun Jong,Min, Byung Il WHO COLLABORATING CENTRE FOR TRADITIONAL MEDICINE 2004 東西醫學硏究所 論文集 Vol.2004 No.-

Moxibustion is one of the major healing techniques in Oriental medicine. It has been widely used in many diseases such as rheumatoid arthritis, Hashimoto disease, breech presentation, etc. However, till now, effects of moxibustion on natural killer (NK) cell activity and relations between sympathetic nerve system (SNS) and the immune alteration induced by moxibustion wert not well studied. This study was designed to evaluate effects of moxibustion on NK cell activity and the Intervention of SNS in the alteration of NK cell activity induced by moxibustion. Splenic NK cell cytotoxicity was measured in a standard 4-hour ^(51)Cr release assay. We measured the NK cell cytotoxicity after moxibustion stimulation for 1, 3, 5 and 7 days, and also measured the NK cell cytotoxicity after 3 and 7 days burn stimulation with similar temperature. Interleukin (IL)-2, -4 and interferon (INF)-γ in serum were measured by rat IL-2, -4 and INF-γ ELISA test kit. To evaluate the effects of sympathectomy on alteration of NK cell cytotoxicity, 6-hydroxydopamine (6-OHDA: 50 mg/kg) was used. We showed that NK cell activity of moxibustion stimulation group incrcased at the 3rd day, and declined at the 7th day in comparison with that of the control group. In thc moxibustion stimulation group, NK cell activity was significantly higher than the sham group at the 3rd day. On the contrary, in the burn stimulation group, NK cell activity was significantly higher than that of the sham groups at 3rd and 7th days. INF-γ level after 3 days in the moxibustion stimulation group was significantly higher than that of the sham group. IL-2 Ievel among groups were not different. IL-4 was not detected in serum with this method. Sympathectomy abolished the NK cell activity alteration induced by moxibustion. The results suggest that moxibustion modulates NK cell activity, along with INF-γ, and SNS is mediating these effects.

Antiviral and anti-inflammatory activity of budesonide against human rhinovirus infection mediated via autophagy activation

Kim, Seong-Ryeol,Song, Jae-Hyoung,Ahn, Jae-Hee,Lee, Geun-Shik,Ahn, Huijeong,Yoon, Sung-il,Kang, Seung Goo,Kim, Pyeung-Hyeun,Jeon, Sang-Min,Choi, Eun-Ji,Shin, Sooyoung,Cha, Younggil,Cho, Sungchan,Kim, Elsevier 2018 Antiviral research Vol.151 No.-

<P>Human rhinovirus (HRV) infection causes more than 80% of all common colds and is associated with severe complications in patients with asthma and chronic obstructive pulmonary disease. To identify antiviral drug against HRV infection, we screened 800 FDA-approved drugs and found budesonide as one of the possible drug candidates. Budesonide is a corticosteroid, which is commonly used to prevent exacerbation of asthma and symptoms of common cold. Budesonide specifically protects host cells from cytotoxicity following HRV infection, which depend on the expression of glucocorticoid receptor. Intranasal administration of budesonide lowered the pulmonary HRV load and the levels of IL-1 beta cytokine leading to decreased lung inflammation. Budesonide regulates IL-1 beta production following HRV infection independent of inflammasome activation. Instead, budesonide induces mitochondrial reactive oxygen species followed by activation of autophagy. Further, the inhibition of autophagy following chloroquine or bafilomycin Al treatment reduced the anti-viral effect of budesonide against HRV, suggesting that the antiviral activity of budesonide was mediated via autophagy. The results suggest that budesonide represents a promising antiviral and anti-inflammatory drug candidate for the treatment of human rhinovirus infection.</P>

Graphene–Ionic Liquid Based Hybrid Nanomaterials as Novel Lubricant for Low Friction and Wear

Khare, Varsha,Pham, Minh-Quan,Kumari, Nitee,Yoon, Hae-Sung,Kim, Chung-Soo,Park, Jae-IL,Ahn, Sung-Hoon American Chemical Society 2013 ACS APPLIED MATERIALS & INTERFACES Vol.5 No.10

<P>Hybrid nanomaterials offer potential scope for an increasing number of novel applications when engineered to deliver usefully functional properties. Recent advancements in the design of new material products that result from interactions among different compositions at the nanoscale and microscale has led to innovative ways to fabricate and process hybrids with altered structural physicochemical properties. An example is the development of novel “lubricants” that make use of ionic liquids (ILs) and their ability to induce exploitable molecular assemblies at the IL–graphene interface. In the present study, we report the potential of graphene–IL hybrid nanomaterials for engineering applications with a focus on “lubricant” properties to reduce frictional forces to enhance tribological performance. The present contribution outlines the wear and tribological properties (friction and lubrication) of a highly viscous IL [BMIM][I] and its comparison with its nanohybrid material counterpart. Detailed structural–microstructural investigations of the nanohybrid materials were performed using X-ray diffraction and microscopic techniques employing scanning electron (SEM), transmission electron (TEM), and high resolution transmission electron (HRTEM) microscopies. A comparative study of the morphology of friction track and wear behavior was assessed by SEM and TEM. These characteristic properties within and outside the friction track were further correlated with physical and chemical interactions obtained by contact angle measurements and Raman spectroscopy and energy dispersive analysis by X-ray (EDAX).</P><P><B>Graphic Abstract</B> <IMG SRC='http://pubs.acs.org/appl/literatum/publisher/achs/journals/content/aamick/2013/aamick.2013.5.issue-10/am302761c/production/images/medium/am-2012-02761c_0016.gif'></P><P><A href='http://pubs.acs.org/doi/suppl/10.1021/am302761c'>ACS Electronic Supporting Info</A></P>

고속 유리하알 변환기와 역변환기의 설계

안두수,이재춘,김재일 成均館大學校 科學技術硏究所 1995 論文集 Vol.46 No.2

This paper presents the transformers design to implement a rationalized Haar transform and a inverse transform algorithm. The transformer consists of a FRHTer to take a RHaar coefficients and a FIRHTer to synthesize signals. The FRHTer and FIRHTer are designed for reduced adder stages compared with other orthogonal transforms, and rationalized coefficients are designed for bit shifter In the FRHTer, the input signal is analyzed into the RHaar spectra which are drawn to the output. In the FIRHTer, the signal is synthesized and produced as the waveform.