쿠싱증후군 환자에서 당 대사 이상 정도에 따른 인슐린 감수성과 인슐린 저항성의 변화

정인경,김성훈,정재훈,민용기,이명식,이문규,유형준,안규정,노정현,김동준,김광원 대한내분비학회 2003 Endocrinology and metabolism Vol.18 No.4

연구배경 당질 코르티코이드는 당 대사에 매우 중요한 호르몬으로 내인성 당질 코르티코이드 과다상태인 쿠싱증후군에서는 말초조직에서 인슐린 저항이 증가하고 이를 보상하고자 인슐린 분비의 증가로 고인슐린혈증이 동반된다고 보고되고 있다. 하지만 생체 내에서와 달리 시험관내에서는 췌도세포에 당질 코르티코이드를 장시간 처리하면, 인슐린 분비 및 생합성이직접적으로 억제됨이 확인된 바 있어 쿠싱증후군 환자에서 당뇨병의 원인으로는 아마도 말초조직에서 증가된 인슐린 저항성 뿐 아니라 이를 충분히 보상하지 못하는 췌장에서의 인슐린 분비 저하가 같이 동반되어있지 않을까 하는 가설을 세우게 되었고, 아직까지 당질코르티코이드가 당대사 이상을 일으키는 기전에 대해 쿠싱증후군을 당대사 정도에 따라 인슐린 감수성과 분비능을 분석한 연구는 없었기에 이를 알아보고자 하였다. 방법: 삼성서울병원에서 쿠싱증후군으로 진단 받은 환자 15명을 대상으로 하였다. 이에 대한 대조군으로는 쿠싱증후군 환자와 같은 성별 그리고 체질량지수를 갖은 15명의 건강한 성인을 대상으로 비교 하였다 쿠싱증후군 환자를 대상으로 경구당부하 검사를 통해 당대사 정도를 정상군, 내당능장애군, 그리고 당뇨병군으로 나눈 후 정맥 당부하 검사를 시행하여 각군의 인슐린 저항성과 인슐린 분비능의 지표를 비교하고, 수술 후 쿠싱증후군이 완치된 상태에서 수술 전후의 당대사 지표의 변화를 조사하였다. 결과: 1) 쿠싱증후군 환자 중 정상인은 20%, 내당능 장애는 27%, 그리고 당뇨병은 53%였다. 체질량지수, 나이, 그리고 발병 기간은 세 군간에 의미 있는 차이가 없었으나, 24시간 소변검사의 코르티솔 농도는 당뇨병군에서 의미있게 높았다. 2) 정맥당부하 검사 결과, 인슐린 감수성 지표인 Sl는쿠싱증추린」서 1.58±0.10[×10^(-4)(min^(-1)(μU/mL)^(-1)]로 정상 대조군의 3.37±0.49[×10^(-4)(min^(-1)(μU/mL)^(-1)]에 비해 의미있게 낮았으나(P=0.024), 쿠싱증후군 환자 중 NGT, IGT, DM 군간에 서로 통계적인 차이는 없었다. 3) SG는 정상 대조군과 쿠싱증후군 환자간에는 의미있는 차이가 없었고, 쿠싱 증후군에 있어서 당대사가 악화될수록 감소하는 경향을 보였으나 의미있는 차이는 없었다. 4) 인슐린 분비능의 지표인 AIRg는 정상인에 비해 전체 쿠싱증후군 환자의 경우 증가하는 경향을 보였으나 의미있는 차이는 없었다. 하지만 쿠싱증후군 환자중에서 당대사 상태에 따라 NGT군은 1299 (1297∼1310)(mu/g/min ×10^(-2))로 정상 대조군(368.9±98.6[mu/g/min ×10^(-2)]) 보다도 의미있게 높았고, DM군{202.2 (91.1~371.4) [mu/g/min ×10^(-2)}은 NGT군에 비해 의미있게 낮았다(P=0.0031). 5) 15명중 현재 완치 상태에 있는 6명에 대해 수술전과 후로 비교하였다. 수술 전 당대사 상태가 1명은정상, 1명은 내당능 장애, 그리고 4명은 당뇨병이었으나 수술 후 시행한 경구 당부하 검사상 모두 정상 당대사 상태를 보였다. 6) 수술 후 완치된 환자 6명에 있어 인슐린 감수성지표인 Sl는 수술전에 중앙값이 1.22[×10^(-4)(min^(-1)(μU/mL)^(-1)]로 대조군에 비해 의미있게 감고』어 있었으나(p.0.05), 수술후 10.95 [×10^(-4)(min^(-1)(μU/mL)^(-1)]로 정상 수준으로 회복되었고(P=0.0022), 인슐린 분비능을 나타내는 AIRg [mu/g/min ×10^(-2)] 값도 정상수준으로 회복되었다. 특히 인슐린 분비능의 회복양상은 혈당농도에 따라 판이하게 나타나서, 정상과 내당능장애 상태에 있던 2명은 수술전에 1201 [mu/g/min ×10^(-2)]로 증가되어 있던 AIRg 값이 수술 후 정상 수준으로 감소하였고, 수술 전에 당뇨병 상태에 있던 4명의 경우 245.9 [mu/g/min ×10^(-2)]로 인슐린 분비능이 감고il어 있었는데 이들은 수술 후 모두 정상 수준으로 증가되었다 (P=0.0286). 결론: 쿠싱증후군 환자에서 당대사 이상은 80%로 높은 유병률을 보였다. 모든 쿠싱증후군환자에서 인슐린 감수성은 정상인에 비해 저하되어 있어 말초조직의 인슐린 저항이 선행됨을 시사하며, 인슐린 분비능은 당대사의 정도에 따라 다르게 나타났는데, 정상 당대사군에서는 인슐린의 저항성을 극복할 만큼 정상 대조군보다 더 많은 양의 인슐린 분비를 하다가 고코르티솔혈증이 심할수록 인슐린 분비능의 감소로 당뇨병으로 진행됨을 확인할 수 있었고, 이런 인슐린 저항성과 인슐린 분비장애는 수술 후 다시 회복되는 가역적인변화를 보였다. Background: Glucocorticoid plays an important role in the control of carbohydrate metabolism. Patients with Cushing's syndrome have been reported to have an increased incidence of carbohydrate intolerance due to peripheral insulin resistance and hyperinsulinemia, although the exact incidence and nature of this disorder have remained unclear. Few results have been published about insulin resistance and insulin secretion according to the level of glucose concentration, or about the reversibility of such defects in patients with Cushing's syndrome. Methods: To assess the effect of glucocorticoid on the insulin sensitivity and insulin secretion in Cushing's syndrome, 15 patients with Cushing's syndrome were classified into 3 groups (normal glucose tolerance: NGT, impaired glucose tolerance: IGT, diabetes: DM) according to the degree of glucose tolerance based on the oral glucose tolerance test (OGTT). Insulin modified, frequently sampled, intravenous glucose tolerance test (FSIGT) was performed before and after curative surgery on these patients and on 15 healthy control subjects. Data were evaluated by non-parametric statistical analysis. Results: 1) Among the 15 patients with Cushing's syndrome, 3 (20%) were NGT, 4 (27%) IGT, and 8 (53%) DM, based on OGTT. Twenty-four hour urinary free cortisol (UFC) was significantly higher in the DM group. 2) Insulin sensitivity index (SI) of Cushing's syndrome was significantly lower than that of the control group p=0.0024), but was not significantly different among the three Cushing's syndrome groups of NGT, IGT and DM. 3) Glucose mediated glucose disposal (SG) (Ed- confirm this abbreviation; it does not seem to match the definition) of Cushing's syndrome was not significantly different from that of the control group. 4) Insulin secretion (AIRg) of Cushing's syndrome tended to be high, but it was not significantly different from that of control. However, according to the level of glucose concentration there was significant difference in AlRg among the three Cushing's syndrome groups p=0.0031); AIRg of DM was significantly lower than that of NGT. 5) After surgical treatment, parameters of insulin sensitivity and insulin secretion were normalized in 6 cured patients; 1 with NGT, 1 with IGT, and 4 with DM, preoperatively. Median SI of all 6 patients was significantly improved up to the normal range postoperatively p=0.0022). Median AIRg of these 6 patients was balanced around that of normal control postoperatively p=0.0286). Conclusion: Eighty percent of patients with Cushing's syndrome had abnormality of carbohydrate metabolism. Insulin sensitivity was significantly decreased in Cushing's syndrome. Insulin secretion was significantly higher only in the NGT and IGT groups of Cushing's syndrome. As the hypercortisolemia is exacerbated, insulin secretion is significantly decreased and causes DM, suggesting that glucocorticoid has a direct or indirect toxic effect on the pancreatic beta cell (J Kor SOC Endocrinol 18:392-403, 2003).

태연혈(太淵穴)의 유침 시간에 따른 체온 변화 -적외선체열촬영(赤外線體熱撮影)을 중심으로-

이봉효 ( Bong Hyo Lee ),이경민 ( Kyung Min Lee ),박지하 ( Ji Ha Park ),김민서 ( Min Seo Kim ),김산들 ( San Deul Kim ),박병규 ( Byeong Gyu Park ),양현동 ( Hyun Dong Yang ),예성호 ( Sung Ho Yea ),이호정 ( Ho Jung Lee ),최재원 ( Jae 경락경혈학회 2012 Korean Journal of Acupuncture Vol.29 No.2

Objectives: This study was performed to find the desirable remaining time of needle in the acupuncture treatment. Methods: The 21 volunteers were given acupuncture at LU9 vertically and needles were remained for 2 min, 10 min, 15 min, 30 min, respectively. The thermographic change induced by acupuncture was measured with Digital Infrared Thermographic Image at the following acupoints: LU11, LU10, LU9, LU8, LU5, LU1, and PC7. The statistical significance of thermographi change was evaluated using paired t-test and post hoc Wilcoxon test. Results: The most significant changes after acupuncture were produced when needles were remained for 10 min or 15 min. LU11, LU5, LU1, and PC7 were the point at which all of the remaining time produced significant change commonly. At LU11, the biggest change was produced when needle was remained for 15 min, while at LU5, LU1, and PC7, the biggest change was produced when needle was remained for 30 min, and the smallest change was produced when needle was remained for 10 min at all of acupoints of LU11, LU5, LU1, and PC7. The unbalance between left side and right was decreased the most largely in 15 min group. Conclusions: The results of this study suggest that the desirable remaining time of acupuncture needle might be 15 min.

Uncoupling Protein 3의 골격근 세포내 과발현이 OLETF 백서 및 배양된 골격근 세포에서 포도당대사에 미치는 영향

한정희,박혜선,고정민,김하영,강호경,이인규,박중열,홍성관,이재담,이기업 대한당뇨병학회 2002 Diabetes and Metabolism Journal Vol.25 No.6

연구배경:Uncoupling protein(UCP)는 미토콘드리아의 내막에 위치하는 단백질로 세포내의 과다한 에너지를 열로 발산시키는 기능을 가진다. 최근 동물의 갈색지방조직에만 존재하는 UCP와 유사성을 가진 아형들(UCP2,3)이 사람에게도 존재함이 알려져 큰 관심을 끌도 있는데 이중 UCP3는 그 발현이 골격근세포와 갈색지방조직에만 국한된다. 본 연구에서는 UCP3가 체내 인슐린 감수성을 결정하는데 가장 중요한 조직인 골격근에 국한되어 발현되는 점에 착안하여 UCP3를 골격근세포에 과발현시켰을 때 포도당 대사에 어떠한 영향이 나타나는 지를 조사하였다. 방법:25주령의 8마리의 OLETF 백서를 대상으로 하여 4마리는 골격근에 adenovirus 2mL(1×10¹²pfu/mL)를 주사하여 대조군으로 하였고 4마리는 골격근에 재조합법으로 제작된 adenovirus­UCP3 2mL(1×10¹²pfu/mL)를 주사하였다(UCP3 과발현군). UCP3를 투여한 백서에서 먹이섭취가 증가하는 경향이 있어 그 전날 대조군이 먹은 야의 먹이만큼 투여하였다. 골격근에 adenovirus를 주사한 10일 후에 euglycemic hyperinsulinemic clamp를 시행하였다. Adenovirus­UCP를 C2C12 골격근 세포에 transfection시켜 UCP3를 C2C12 골격근 세포에 transfection시켜 UPS3­C2C12를 만들고 C2C12 골격근 세포와 UPS3­C2C12 골격근 세포에서 포도당 수송 및 당원합성을 측정하였다. 결과:UCP3 과발현 OLETF에서 체중이 감소하는 경향을 보였고 인슐린 감수성이 증가하였다. C2C12세포에서 기저상태 포도당 수송은 1.28±0.17μmol/L/min였고 100nM 인슐린으로 2시간 처리한 후 2.67±0.20 μmol/L/min로 증가하였다. UCP3­C2C12 세포에서는 기저상태 포도당 수송이 3.98±0.13μmol/L/min로 증가되었고 인슐린 처리 후 5.74±0.44μmol/L/min로 증가하였다. 인슐린을 처리한 UCP3­C2C12 세포에 P13K 억제제인 wortmannin을 첨가하였을 때 포도당 수송활성이 3.81±0.20μmol/L/min로 감소하였다. 기저상태 당원합성은 C2C12 세포에서 0.25±0.01μmol/L/min였고 인슐린 처리 후 0.45±0.01μmol/L/min로 증가하였다. UCP3­C2C12 세포에서는 기저상태 당원합성이 0.62±0.01μmol/L/min였고 인슐린 처리 후 1.26±454μmol/L/min로 증가하였다. UCP3­C2C12세포에 wortmannin을 첨가하였을 때 당원합성율이 0.80±0.04μmol/L/min로 감소하였다. 결론:UCP3 과발현이 OLETF 백서에서 인슐린 감수성을 증가시켰고 골격근세포에서 포도당 수송 및 당원합성을 증가시켰다. wortmannin을 첨가하였을 때 포도당 수송 및 당원합성이 감소함으로 보아 이 과정이 인슐린 신호전달체계인 P13K에 일부 의존함을 알 수 있었다. Background : UC P3 is a mitochondrial membrane protein expressed selectively in the skeletal muscle and brown adipose tissue. Since the skeletal muscle is the main organ determining insulin sensitivity in the body, it was hypothesized that UCP3 overexpression in skeletal muscle cells would improve glucose metabolism. Methods : An adenovirus-UCP3 was produced by a recombinant DNA method. OLETF rats were divided into 2 groups. Four rats were injected with the adenovirus-UCP3 (UCP3 group) and others were injected with the adenovirus(control group) in the skeletal muscle. The UCP3 group was provided with the same quantity of food as that consumed by the control group on the previous day. Insulin sensitivity was evaluated by the euglycemic hyperinsulinemic clamp method. In a separate experiment, glucose transport and glycogen synthesis we evaluated in C2C212 cells transfected with ether an adenovirus or the adenovirus-UCP3. Results : The insulin sensitivity improved significantly and the body weight decreased in the UCP3 group. The glucose transport and glycogen synthesis were higher in the UCP3-C2C12 skeletal muscle cells at the basal state. After insulin treatment, glucose transport and glycogen synthesis were also higher in the UCP3-C2C12 cells but the increments were reduced after treatment with wortmannin, a PI3K inhibitor. Conclusion : Insulin sensitivity was higher in the UCP3-overexpressed OLETF rats in the in vivo study. UCP3 transfection also increased glucose transport and glycogen synthesis in the cultured skeletal muscle cells by a PI3K dependent mechanism(J Kor Diabetes Asso 25 :460~468, 2001).

H9c2 심근 세포주에서 외인성 nitric oxide가 허혈에 의한 세포 독성에 미치는 영향

정성구,장현용,김명천,고영관,정주호,배영미,박원서,김대중,유영민,김성수,임성빈 대한응급의학회 2001 대한응급의학회지 Vol.12 No.4

Background: Nitric oxide(NO) is known to have protective effects on an ischemic heart and to exert triggering effects on ischemic preconditioning. However, the effects of NO during the ischemic period have not been investigated. To investigate the role of exogenous nitric oxide in a model of ischemic heart cell death, we studied the effects of ischemic preconditioning and ischemia in a normal and an ischemic buffer. Methods: Rat cardiac myoblast cells(H9c2) were cultured in a normal and an ischemic buffered medium. For the ischemic culture of heart cells, the cells were cultured in a dessicator with GasPak for 5 hrs. In ischemic preconditioning, the cells were pretreated with ischemic buffer for 5 min and then perfused with normal medium for 30 min. For the measurement of the cytotoxicity, a MTT(3-4-Sdimethylthiazol-2-yl-2,5-diphenyl tetrazolium bromide) assay was performed. A DAPI(4',6-diamidino-2-phenylindole dihydrochloride) staining procedure and a flow cytometry analysis were performed to confirm apoptotic cell death by ischemia. Results: Cell viability, as determined by using a MTT assay, showed that the preconditioned group treated with NO showed more cell death than with the not-preconditioned groups in both normal and ischemic buffers. But, In normal medium and not-preconditioned groups, NO showed protective effect according to the concentrations(100,1000μM) . No treatment with NO produced the different results. In normal medium, the protective effect of ischemic preconditioning was demonstrated, but no protective effect of ischemic preconditioning could be seen in the case of the ischemic buffer. The DAPI staining and flow cytometry analysis of heart cells showed characteristic apoptotic features. Conclusion: NO added in the ischemic phase had deterious effects on heart cells. Ischemic preconditioning was more harmful than ischemia alone. The toxicity of the cells was characteristic apoptosis.

Production of Iron-Binding Peptides from Colostral Whey by Enzymatic Hydrolysis

Sang Bum Kim,Myoung Soo Nam,Kwang Seok Ki,Hyeon Shup Kim,Min Jung Ku,Won Mo Cho 한국축산식품학회 2010 한국축산식품학회지 Vol.30 No.6

Colostral whey prepared from colostrum (pooled from first six post-partum milkings) was heated for 10 min at 100oC. Heated colostral whey was incubated with 1% enzymes (protein equivalent basis) for 15, 30, 60, 90, and 120 min at 50oC. Papain, pepsin, trypsin, and alcalase produced different degrees of hydrolysis (DH), 10.66%, 12.42%, 10.83%, and 25.31%, respectively, at an incubation time of 120 min. The SDS-PAGE reveals that significant amounts of bovine serum albumin (BSA), β-lactoglobulin (β-LG), and α-lactalbumin (α-LA) survived papain digestion. In contrast, pepsin completely removed BSA but not β-LG present in heated colostral whey. Alcalase completely eliminated BSA, β-LG, and α-LA. This differential hydrolysis was confirmed by reversed-phase HPLC analysis. Using ion-exchange chromatography, fraction-1 (F-1) was obtained from alcalase hydrolysate at a NaCl gradient concentration of 0.25 M. Reversed-phase HPLC chromatograms of alcalase F-1 showed numerous small peaks, which probably indicate that a variety of new peptides were produced. Iron content of alcalase F-1 was 28.94 ppm, which was the highest among all enzyme fractions, whereas iron content of colostral whey was 36.56 ppm. Main amino acids contained in alcalase F-1 were Thr (15.45%), Glu (14.12%), and Ser (10.39%). Therefore, alcalase can be used to generate good iron-binding peptides in heated colostral whey, and the resulting iron-binding peptides could be suitable as a value-added food ingredient for food supplements.

Genome Wide Expression Profile of Agrimonia pilosa in LPS-stimulated BV-2 Microglial Cells

Sohn, Sung-Hwa,Ko, Eun-Jung,Kim, Sung-Hoon,Kim, Yang-Seok,Shin, Min-Kyu,Hong, Moo-Chang,Bae, Hyun-Su The Korean Society of Toxicogenomics and Toxicopro 2009 Molecular & cellular toxicology Vol.5 No.1

Microglial cells constitute the first line of defense against infection and injury in the brain. This study was conducted to evaluate the protective mechanisms of Agrimonia pilosa (AP) on LPS-induced activation of BV-2 microglial cells. The effects of AP on gene expression profiles in activated BV-2 microglial cells were evaluated using microarray analysis. BV-2 microglial cells were cultured in a 100 mm dish ($1{\times}10^7/mL$) for 24 hr and then pretreated with 1 g/mL AP or left untreated for 30 min. Next, 1 g/mL LPS was added to the samples and the cells were reincubated at $37^{\circ}C$ for 30 min, 3 hr and 6 hr. The gene expression profiles of the BV-2 microglial cells varied depending on the AP. The microarray analysis revealed that MAPK signaling pathway-related genes were down-regulated and IL10 gene was up-regulated in AP-treated BV-2 microglial cells. AP can affect the inflammatory response and MAPK pathway in BV-2 microglial cells.

Dry etching of polydimethylsiloxane using microwave plasma

Hwang, Sung Jin,Oh, Dong Joon,Jung, Phill Gu,Lee, Sang Min,Go, Jeung Sang,Kim, Joon-Ho,Hwang, Kyu-Youn,Ko, Jong Soo IOP 2009 JOURNAL OF MICROMECHANICS AND MICROENGINEERING - Vol.19 No.9

<P>This paper presents a new polydimethylsiloxane (PDMS) dry-etching method that uses microwave plasma. The applicability of the method for fabricating microstructures and removing residual PDMS is also verified. The etch rate of PDMS was dominantly influenced by the gas flux ratio of CF<SUB>4</SUB>/O<SUB>2</SUB> and the microwave power. While the PDMS etch rate increased as the flux ratio of CF<SUB>4</SUB> was increased, the etch rate decreased as the flux ratio of O<SUB>2</SUB> was increased. The maximum etch rate of 4.31 µm min<SUP>−1</SUP> was achieved when mixing oxygen (O<SUB>2</SUB>) and tetrafluoromethane (CF<SUB>4</SUB>) at a 1:2 ratio at 800 W power. The PDMS etch rate almost linearly increased with the microwave power. The ratio of the vertical etch rate to the lateral etch rate was in a range of 1.14–1.64 and varied with the gas fluxes. In consideration of potential applications of the proposed PDMS etching method, array-type PDMS microwells and network-type microprotrusion structures were fabricated. The contact angle was dramatically increased from 104° (non-etched PDMS surface) to 148° (etched PDMS surface) and the surface was thereby modified to be superhydrophobic. In addition, a thin PDMS skin that blocked holes and PDMS residues affixed in nickel microstructures was successively removed.</P>

한국인에서 혈소판 당단백 Ⅱb/Ⅲa 유전자 다형성과 관동맥 성형술 후 재 협착과의 관계

이민수,이정우,김보영,임대승,강정아,김정희,김윤철,성보영,최성준,성인환,전은석 충남대학교 의과대학 지역사회의학연구소 2000 충남의대잡지 Vol.27 No.2

Platelet aggregation is the final pathway of acute coronary syndrome such as acute myocardial infarction and unstable angina. Platelet glycoprotein IIb/IIIa is a membrane receptor for fibrinogen and yon Willebrand factor and it plays an important role in platelet aggregation and in the pathogenesis of acute coronary syndrome. It is known that polymorphism of the gene that encoding platelet glycoprotein IIb/IIIa(PI^A1/A2) is strongly related to acute coronary syndrome in Caucasian, but not in Koreans. We investigated relationship between platelet glycoprotein llb/Illa gene polymorphism and restenosis of coronary artery after angioplasty in Koreans. Total 371 patients(M=251. F=120) were enrolled. Angioplasty group comprised 143 patients who underwent coronary angioplasty, and in the angioplasty group, restenosis group comprised with the 65 patients who had restenotic lesion over 50% of luminal diameter in follow-up coronary angiography. Normal group comprised 153 patients who had no significant angiographic lesion and variant angina group comprised 75 patients who were positive in ergonovine test. Genomic DNA was extracted from peripheral arterial blood. To determine the frequency of P1^A1/A2 genotype, polymerase chain reaction(PCR) was done and the product was restricted with Mspl. 3%. agarrose gel electrophoresis showed restriction fragment length polymorphism. Clinical profile and risk factor were also reviewed. Among all 371 patients of study group, genotype of only one patients in restenosis group if is proven to be PI^A1/A2 heterozygote. All patients of normal study group, no restenosis group, and the other patients in restenosis group have an PI^A1 homozygote genotype. In our study, platelet glycoprotein IIb/Illa polymorphism has no relationship with restenosis of the coronary artery after angioplasty in Koreans. But the genotypic frequency of platelet glycoprotein IIb/IIIa gene polymorphism in Koreans is concordant with that of previous studies.

Production of Iron-Binding Peptides from Colostral Whey by Enzymatic Hydrolysis

Kim, Sang-Bum,Ku, Min-Jung,Cho, Won-Mo,Ki, Kwang-Seok,Kim, Hyeon-Shup,Nam, Myoung-Soo Korean Society for Food Science of Animal Resource 2010 한국축산식품학회지 Vol.30 No.6

Colostral whey prepared from colostrum (pooled from first six post-partum milkings) was heated for 10 min at $100^{\circ}C$ Heated colostral whey was incubated with 1% enzymes (protein equivalent basis) for 15, 30, 60, 90, and 120 min at $50^{\circ}C$. Papain, pepsin, trypsin, and alcalase produced different degrees of hydrolysis (DH), 10.66%, 12.42%, 10.83%, and 25.31%, respectively, at an incubation time of 120 min. The SDS-PAGE reveals that significant amounts of bovine serum albumin (BSA), ${\beta}$-lactoglobulin (${\beta}$-LG), and ${\alpha}$-lactalbumin (${\alpha}$-LA) survived papain digestion. In contrast, pepsin completely removed BSA but not ${\beta}$-LG present in heated colostral whey. Alcalase completely eliminated BSA, ${\beta}$-LG, and ${\alpha}$-LA. This differential hydrolysis was confirmed by reversed-phase HPLC analysis. Using ion-exchange chromatography, fraction-1 (F-1) was obtained from alcalase hydrolysate at a NaCl gradient concentration of 0.25 M. Reversed-phase HPLC chromatograms of alcalase F-1 showed numerous small peaks, which probably indicate that a variety of new peptides were produced. Iron content of alcalase F-1 was 28.94 ppm, which was the highest among all enzyme fractions, whereas iron content of colostral whey was 36.56 ppm. Main amino acids contained in alcalase F-1 were Thr (15.45%), Glu (14.12%), and Ser (10.39%). Therefore, alcalase can be used to generate good iron-binding peptides in heated colostral whey, and the resulting iron-binding peptides could be suitable as a value-added food ingredient for food supplements.

난치성 백반증에 대한 흡입물집을 이용한 비배양 표피세포이식술 20례

배정민 ( Jung Min Bae ),정한미 ( Han Mi Jung ),이한나 ( Han Na Lee ),이로우 ( Ro Woo Lee ),은성혜 ( Sung Hye Eun ),권혁선 ( Hyuck Sun Kwon ),이지혜 ( Ji Hae Lee ),김경문 ( Gyong Moon Kim ) 대한피부과학회 2018 대한피부과학회지 Vol.56 No.7

Background: As nonsurgical interventions for vitiligo are not always successful, various surgical modalities have been used in patients with refractory vitiligo. Of these, non-cultured epidermal suspension transplantation (NCES) was recently introduced to treat large recipient sites using cells from small donor tissue. Objective: We assessed the effectiveness and safety of NCES as a surgical treatment for patients with refractory vitiligo. Methods: We retrospectively reviewed 20 cases in 17 patients (11 females; median age 25 years) who underwent NCES from July 2015 through March 2018. Suction blisters (20 mm in diameter) were collected from the patient’s inner thigh at a donor-to-recipient area ratio of 1:5. After the addition of 5 mL recombinant trypsin solution to the suction blisters, followed by incubation at 37°C for 60 min, epidermal cells were manually scraped off the blister surface, and epidermal cell suspension was obtained by centrifugation at 1,500 RPM for 5 min. The suspension was applied to the vitiligo regions after epidermal ablation of those regions. Phototherapy resumed 1 month later. Treatment success was defined as ≥75% repigmentation of the surgical site, and all adverse events were noted. Results: Overall, 85.0% of cases (17/20) exhibited treatment success. Adverse events included hyperpigmentation (20%) and surgical site infection (5%), but the treatment was tolerable in all cases. Conclusion: NCES is a reliable surgical option for patients with vitiligo refractory to nonsurgical treatment. Large areas of vitiligo can be treated by NCES, and use of this technique should be encouraged in Korea. (Korean J Dermatol 2018;56(7):426∼432)