Natural Toxin의 안전성 평가연구 : Ochratoxin A의 면역독성 Immunotoxicity of ochratoxin A

김주일,한형미,정혜주,김형수,김환묵,정승태,박재현,강선경,김진호 식품의약품안전청 1997 식품의약품안전청 연보 Vol.1 No.-

Ocllratoxin A는 fenicl'flrium과 rtsr☞rgiffus 속 등의 곰팡of가 생성하는 mycotoxin으로서 세포매기성 및 체액성 면역기능에 슨상을 일으키는 것으로 알려져 있다. 본 연구는 OTA가 면역계에 미치는 영향과 OTA의 면역 독성 억제 효과가 있는 물질을 검색하기 위하며 6~7 주령의 Balb/c mice 비장세핀에 T 및 B 세포의 mitogen인 Con A와 LPS로 처리하고 3일간 배양한 후 비장세포의 분화정도를 MTS를 처리하여 관찰하였으며, OTA를 투여한 마우스의 면역장기 형리조직학적 견화를 관찰하였다. 마우스의 비장세포 (2xtD6cel1/rnf)에 OTA(1~6μg/ml)를 상에서 혼합하여 Con A와 LPS로 자극시킨 결과 임파구는 Sfg/mf이상에서 거의 모든 세포가 분화되지 않았다. mice에 OTA (1,5 및 10mg/11g)를 2일 간격으로 6회 복갚 주사한 후 비장세포를 분리하여 Con A와 LPS로 처리한 결과 Smg 투여군에서는 6회 투여 혼에 분화가 억제되었고, 10mg 투여군에서는 3회 및 6회 투여군에서 분화가 억제되었단. Ochratoxin A의 면역독성 억제 효과가 있는 물질을 검색하기 위하여 in vitro 실험에서 마우스의 비장세포 (2×10" cell/mf)에 OTA (3fg/mf)와 L-phenylalanine, indole-3-carbinol, L-ascorbic acid,piroxicaTn, curcumin 및 arethylsalicylic acid를 혼합하고 동일 처리한 결과 L-ascorbic acid, L-phenylala-nine 및 Indole-3-carbinol은 OTA의 면역독성 발현을 운의성 있게 억제하였다. Balt/c mice에 OTA (10mg/kg)와 l'n pifro실험에서 억제효파가 확인된 L-ascorbic acfd, L-phenylalanine 및 indole-3-carbinol을 2일 간격으로 동시에 3회 복강주사하여 관찰한 결과 indote-3-carbine께서 엮제효과를 관찰할수 있었다. Ochratoxin A를 투여할 마우스의 비장 및 홍선에서는 임파구의 빈도가 줄었으며 특히 비장제서는 starry-sky 세포가 증가됨을 관찰할 수 있었다. 이러한 결자로 OTA는 비장세포의 T 및 B 임파구의 분열능을 억제시 키며, 면역장기의 임파구에 손상 야기시키므로서 면역기능을 저하시키고 OTA에 의한 면역독성은 indole-3-carbinol이 경감시킴을 관찰하엿다. Ochratoxin A (OTA) is a Tnyeotoxin produced by several fungal species iBcluded the genera fs4pergiffug and f☞HiciHiHuL and known as one of the major environmental contaminants. Tn thepresent study, the effects of OTA on the imraune system and the prevention of the OTA-inducedimmunosuppression by protective agents were studied in Balb/c rnicf:. Splenocytes were isolated and theproliferative responses to concanavalin A (Con A) and lipopolysaccharide (LPS) were measured in thepreseEce of 1 to 6fg/mf OTA. Sfg/mf OTA completely btocked both Con A and LPS-stimulatedmitogenic responses, causing approximately 50% inhibition at 3fe/mf OTA. This suppression of mitogen-induced proliferative resplonses observed id uifro was reproduced in mice treated with ☞rA in rioafntraperitoneal administration of 5, IDmg/kg OTA 3 to etimes every other day significantly decreasedboth Con A- aud LPS-inttuced mitegenic responses. The effects of possible protective agents on theOTA-induced suppression c)』 mitogenic responses were examined both in uifro and in viua Amoag 6 pos-sible protective agents (1,-phenylalanine, L-ascorbic acid, indole-3- carbinol, curcumin, firoxicam,acethylsaricylic acid) employed in the present study, L-phenylalanine (100ff), L-ascorbic acid (100#M) and iBdote-3-carbinel t 10Df) showed significantly protective effects on the OTA-iBduced suppres-sion of mitogenic responses id uifro. Howeuer, when the protective of·fects of these agents were examinedin rr'ua only indole-3-carbinol showed significantly protective respoases. It has been ebserved that Iyin-phatic cell population in thc spleen and thymus was decreased in raice given OTA by hematoBylin-eosinstaining. These data indicate that OTA suppresses the cell-mediated and humoral immune functioBsand this OTA-iBduced supflression of immune functions can he alleviated by indole-3-carbinol.

Transforming Growth Factor-β1에 대한 항체 및 유전자 치료가 피부 반흔 형성에 미치는 영향

김준형,한기환,안종덕,이인규,김은주,황미열,박관규 대한병리학회 2001 Journal of Pathology and Translational Medicine Vol.35 No.4

Regulation of Tumor Neceosis Factor-${\alpha}$ Receptors and Signal Transduction Pathways

Han, Hyung-Mee The Korean Society of Toxicology Korea Environment 1992 Toxicological Research Vol.8 No.2

Tumor necrosis factor-${\alpha}$(TNF), a polypeptide hormone secreted primarily by activated macrophages, was originally identified on the basis of its ability to cause hemorrhagic necrosis and tumor regression in vivo. Subsequently, TNF has been shown to be an important component of the host responses to infection and cancer and may mediate the wasting syndrome known as cachexia. These systemic actions of TNF are reflected in its diverse effects on target cells in vitro. TNF initiates its diverse cellular actions by binding to specific cell surface receptors. Although TNF receptors have been identified on most of animal cells, regulation of these receptors and the mechanisms which transduce TNF receptor binding into cellular responses are not well understood. Therefore, in the present study, the mechanisms how TNF receptors are being regulated and how TNF receptor binding is being transduced into cellular responses were investigated in rat liver plasma membranes (PM) and ME-180 human cervical carcinoma cell lines. $^{125}I$-TNF bound to high ($K_d=1.51{\pm}0.35nM$)affinity receptors in rat liver PM. Solubilization of PM with 1% Triton X-100 increased both high affinity (from $0.33{\pm}0.04\;to\;1.67{\pm}0.05$ pmoles/mg protein) and low affinity (from $1.92{\pm}0.16\;to\;7.57{\pm}0.50$ pmoles/mg protein) TNF binding without affecting the affinities for TNF, suggesting the presence of a large latent pool of TNF receptors. Affinity labeling of receptors whether from PM or solubilized PM resulted in cross-linking of $^{125}I$-TNF into $M_r$ 130 kDa, 90 kDa and 66kDa complexes. Thus, the properties of the latent TNF receptors were similar to those initially accessible to TNF. To determine if exposure of latent receptors is regulated by TNF, $^{125}I$-TNF binding to control and TNF-pretreated membranes were assayed. Specific binding was increased by pretreatment with TNF (P<0.05), demonstrating that hepatic PM contains latent TNF receptors whose exposure is promoted by TNF. Homologous up-regulation of TNF receptors may, in part, be responsible for sustained hepatic responsiveness during chronic exposure to TNF. As a next step, the post-receptor events induced by TNF were examined. Although the signal transduction pathways for TNF have not been delineated clearly, the actions of many other hormones are mediated by the reversible phosphorylation of specific enzymes or target proteins. The present study demonstrated that TNF induces phosphorylation of 28 kDa protein (p28). Two dimensional soidum dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE) resolved the 28kDa phosphoprotein into two isoforms having pIs of 6.2 and 6.1. The pIs and relative molecular weight of p28 were consistent with those of a previously characterized mRNA cap binding protein. mRNA cap binding proteins are a class of translation initiation factors that recognize the 7-methylguanosine cap structure found on the 5' end of eukaryotic mRNAs. In vitro, these proteins are defined by their specific elution from affinity columns composed of 7-methylguanosine 5'-triphosphate($m^7$GTP)-Sepharose. Affinity purification of mRNA cap binding proteins from control and TNF treated ME-180 cells proved that TNF rapidly stimulates phosphorylation of an mRNA cap binding protein. Phosphorylation occurred in several cell types that are important in vitro models of TNF action. The mRNA cap binding protein phosphorylated in response to TNF treatment was purifice, sequenced, and identified as the proto-oncogene product eukaryotic initiation factor-4E(eIF-4E). These data show that phosphorylation of a key component of the cellular translational machinery is a common early event in the diverse cellular actions of TNF.

Poster Session : PS 1376 ; Nephrology : Delta Neutrophil Index as an Independent Predictor of Mortality in Septic Acute Kidney Injury Patients Under-going Continuous Renal Replacement Therapy

( In Mee Han ),( Dong Ho Shin ),( Youn Kyung Kee ),( Chang Yun Yoon ),( Eunyoung Lee ),( Young Su Joo ),( Seung Gyu Han ),( Hyung Jung Oh ),( Jung Tak Park ),( Seung Hyeok Han ),( Shin Wook Kang ),( T 대한내과학회 2014 대한내과학회 추계학술대회 Vol.2014 No.1

Background: Delta Neutrophil Index (DNI) indicates the fraction of circulating immature granulocytes, which is known to increase in infectious and/or septic conditions. However, the relationship between DNI and mortality in septic acute kidney injury (AKI) patients is not yet fully elucidated. Therefore, we assessed whether DNI is associated with mortality in septic AKI patients requiring continuous renal replacement therapy (CRRT). Methods: We retrospectively enrolled 285 patients with septic AKI who were treated with CRRT at Yonsei University Health System between August 2009 and September 2012. The patients were dichotomized into high and low DNI groups based on the cutoff value from receiver operating characteristics of DNI values at the time of CRRT initiation. Log-rank test and Cox proportional hazards analysis were conducted to evaluate the effect of DNI as a prognostic factor for 28-day all-cause mortality. Results: The mean age of the enrolled patients was 61.0 ± 14.7 years and 180 patients (63.2%) were male. The high DNI group (DNI > 5.6%) was composed of 149 patients (52.3%). During the study period, 192 patients (67.1%) died. Mortality rate during the 28-days was significantly increased in the high DNI group compared to low DNI group (79.9% vs. 53.3%, P < 0.01, log rank test p Conclusions: This study demonstrates that DNI at CRRT initiation could be an useful predictor for mortality in septic AKI patients requiring CRRT.

Decreased Circulating Klotho Levels in Patients Undergoing Dialysis and Relationship to Oxidative Stress and Inflammation

Oh, Hyung Jung,Nam, Bo Young,Lee, Mi Jung,Kim, Chan Ho,Koo, Hyang Mo,Doh, Fa Mee,Han, Jae Hyun,Kim, Eun Jin,Han, Ji Suk,Park, Jung Tak,Yoo, Tae-Hyun,Kang, Shin-Wook,Han, Dae-Suk,Han, Seung Hyeok International Society for Peritoneal Dialysis 2015 Peritoneal dialysis international Vol.35 No.1

<P>♦ <I>Introduction:</I> It has been reported that klotho deficiency is associated with oxidative stress and inflammation in experimental kidney disease models. Patients with endstage renal disease (ESRD) are particularly characterized by increased oxidative stress and inflammation. However, little is known about the relationship between these features and klotho in patients with ESRD.</P><P>♦ <I>Methods:</I> We conducted a single-center, cross-sectional study of 78 patients receiving peritoneal dialysis (PD). Serum concentrations of klotho, high-sensitivity C-reactive protein (hsCRP), interleukin-6 (IL-6), and 8-isoprostane were measured by enzyme-linked immunosorbent assay. To define factors independently associated with klotho, we determined Spearman’s correlation coefficients for between co-variates and conducted multiple linear regression analyses.</P><P>♦ <I>Results:</I> Patients were classified by median concentration of klotho. In patients with klotho levels > 329.6 pg/mL, serum 8-isoprostane and IL-6 levels were significantly higher than in those with klotho levels < 329.6 pg/mL. In correlation analyses, log 8-isoprostane (γ = –0.310, <I>p</I> = 0.006) and log IL-6 (γ = –0.343, <I>p</I> = 0.002) were inversely correlated with log klotho. After adjustment for age, gender, mean arterial pressure, log intact parathyroid hormone, and log IL-6, log 8-isoprostane was independently associated with log klotho (β = –0.158, <I>p</I> = 0.040). However, the significant relationship between klotho and IL-6 was not seen in an adjusted model.</P><P>♦ <I>Conclusions:</I> This study showed that circulating klotho levels were significantly associated with 8-isoprostane levels in patients undergoing PD, suggesting a potential link between klotho deficiency and enhanced oxidative stress in ESRD patients.</P>

종양괴사인자(TNF)가 ME-180 사람 경부 암종세포에서 종양 발생 유전자의 발현에 미치는 영향

한형미(Hyung Mee Han),김형수(Hyung Soo Kim),손경희(Kyung Hee Sohn),최경백(Kyoung Baek Choi),정승태(Seung Tae Chung),김진호(Jin Ho Kim),이병무(Byung Moo Lee),김주일(Joo Il Kim) 大韓藥學會 1997 약학회지 Vol.41 No.5

Tumor necrosis factor-alpha (TNF) induced a cytotoxic response in ME-180 cervical carcinoma cells in vitro. This cytotoxic response was accompanied by a temporal series of mitogenic stimuli : increased c-fos, c-jun and jun-B expression. Depletion of protein kinase C (PKC) by exposure of ME-180 cells to 100ng/ml phorbol myristate acetate (PMA) for 24hours almost completely abolished TNF-mediated increase in these signals, indicating that a PKC-dependent pathway is involved in TNF-mediated increases in the expression of c-fos, c-jun and jun-B. Characteristics of TNF receptors after exposure to 100ng/ml PMA or 24hours were not altered, suggesting that diminished induction of these oncogenes by TNF after PMA treatment is not due to any changes at the receptor level. To examine whether a PKC-dependent pathway is involved in TNF-mediated cytotoxicity in ME-180 cells, cytotoxicity was measured after depletion of PKC. No apparent changes in cytototoxicity after PKC depletion suggest that a PKC-dependent pathway is not involved in TNF-mediated cytotoxicity. Furthermore, results from cytotoxicity tests after exposure to staurosporine (PKC inhibitor) did not show any changes in the TNF-mediated cytotoxicity, confirming that a PKC-dependent pathway is not involved in this process. These data indicate that 1) TNF induces expression of c-fos, c-jun and jun-B oncogenes via a PKC-dependent pathway and 2) PKC-dependent expression of these three oncogenes by TNF may not be involved in TNF-mediated cytotoxicity in ME-180 cells.

The Complete Nucleotide Sequence and Gene Organization of the Mitochondrial Genome of the Bumblebee, ignitus (Hymenoptera: Apidae)

( Hyung Joo Yoon ),( Eun Mee Lee ),( Myung Hee Yoon ),( Jae Sam Hwang ),( Byung Rae Jin ),( Yeon Soo Han ),( Ik Soo Kim ),( So Young Cha ) 한국응용곤충학회 2006 한국응용곤충학회 학술대회논문집 Vol.2006 No.9

Presence of Several tRNA-Like Sequences in the Mitochondrial Genome of the Bumblebee, Bombus hypocrita sapporoensis (Hymenoptera: Apidae)

Mee Yeon Hong,So Young Cha,Dong Young Kim,Hyung Joo Yoon,Seong Ryul Kim,Jae Sam Hwang,Ki Gyoung Kim,Yeon Soo Han,Ik Soo Kim 한국유전학회 2008 Genes & Genomics Vol.30 No.4

We have sequenced the mitochondrial genome (mitogenome) of the bumblebee, Bombus hypocrita sapporoensis (Hymenoptera: Apidae). The nearly completed circular genome is 15,468-bp long, excluding a region spanning from srRNA, tRNA(Gln) to the A+T-rich region, with regard to the previously published mitogenome of B. ignitus. The gene arrangement of B. hypocrita mitogenome was identical to the within-generic species, B. ignitus, the tRNAs of which are highly rearranged compared with the positions observed most frequently in insect mitogenome structures. The most unusual feature of the genome is the presence of a number of intergenic spacer sequences which harbor partial tRNA-like sequences capable of forming a clover-leaf secondary structure with the proper anticodons, although the stem regions are not paired off well. Such features appear to be prevalent in hymenopteran species. The ATPase8, ATPase6, ND4L, and ND6 genes, which are adjacent to another PCG at their 3` end, all harbored potential sequences for the formation of a hairpin structure. This may indicate the importance of such structures for the precise cleavage of the mRNA of mature PCGs.

Complete Mitogenome Sequence of the Endangered Jewel Beetle, Chrysochroa fulgidissima (Coleoptera: Buprestidae)

Mee Yeon Hong,Heon Cheon Jeong,Dong Young Kim,Hyung Wook Jeong,Ki-Gyoung Kim,Yeon Soo Han,Byung Rae Jin,Iksoo Kim 한국응용곤충학회 2008 한국응용곤충학회 학술대회논문집 Vol.2008 No.05

We determined the complete mitochondrial genome (mitogenome) of the jewel beetle, Chrysochroa fulgidissima (Coleoptera: Buprestidae) from two overlapping fragments and subsequent sub fragments. The 15,592-bp long C. fulgidissima mitogenome contains gene arrangement and content identical to the most common arrangement found in insects. Most individual C. fulgidissima mitochondrial (mt) genes were well within the range found in the respective genes of other insects. The 875-bp A+T-rich region is shortest among the coleopteran mitogenomes sequenced in their entirety. The region is interesting in that it contains several stem-and-loop structures and tRNA-like structure found in the A+T-rich regions of other insect mitogenomes. As seen in other insect motogenomes the start codon of C. fulgidissima COI gene also is unusual because no typical start codon is available. Three of the 13 protein-coding genes have incomplete termination codon T or TA. All tRNA formed stable stem-and-loop structure, except for tRNASer(AGN), the DHU arm of which formed a simple loop as seen in many other metazoan mt tRNASer(AGN).

Artificial tooth selection and occlusion in complete dentures

Han-Eol Choe,Hyung-Seok Kim,Mee-kyoung Son,허유리 조선대학교 치의학연구원 2019 Oral Biology Research (Oral Biol Res) Vol.43 No.3

It is important to select artificial teeth and arrange them in a proper way especially when patients have severe alveolar ridge resorption or abnormal intermaxillary relation. The purpose of this study is to suggest the criteria for decision making in several clinical cases regarding the type of artificial teeth and concept of occlusion. Several types of artificial teeth are nowadays available. Anatomic tooth, which mimics the cusps and grooves of the nature tooth, is widely used in most cases. But in cases of severe alveolar bone loss or in cases where it is difficult to induce centric occlusion, artificial tooth with nonanatomic and semianatomic cusps could be a better choice. The occlusion scheme is also determined regarding whether to arrange teeth in lingual occlusion or crossbite considering the intermaxillary relation and severity of alveolar ridge resorption.