• Preventive Nutrition and Food Science
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• 제9권3호
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• pp.276-282
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• 2004

Marginal Zn deficiency is prevalent through the world and yet human zinc status has not been properly assessed due to the lack of a reliable diagnostic indicator. One potential possibility for zinc status assessment using Zn-binding protein, metallothionein (MT)-mRNA, has been proposed. The purpose of the present study was aimed to show whether measurement of mononuclear cell (MNC) MT mRNA, using a competitive-reverse transcriptase-polymerase chain reaction (competitive-RT-PCR) assay, could indicate zinc status in human subjects. In this study, MNC MT-mRNA expression was measured using a competitive-RT-PCR to compare before and after 14 days of zinc supplementation (50 mg Zn/das zinc gluconate). RT-PCR oligonucleotide primers which were designed to amplify both a 278 bp segment of the human MT-2A cDNA and a 198 bp mutant competitor cDNA template from MNCs, were prepared. MT-2A mRNA was normalized by reference to the housekeeping gene, $\beta$-actin, mRNA for which was also measured by competitive-RT-PCR. There was considerable inter-individual variation in MT-mRNA concentration and yet, the mean MT-2A mRNA level increased 4.7-fold after Zn supplementation, as compared to before Zn supplementation. This MT-2A mRNA level was shown as the same pattern and, even more sensitive assay, compared to the conventional plasma and red blood cells (RBCs) Zn assessment in which plasma and RBCs zinc levels increased 2.3- and 1.2-fold, respectively (p<0.05). We suggest that MT competitive-RT-PCR can be a useful assessment tool for evaluating human zinc status.

• 한국동물학회지
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• 제36권4호
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• pp.529-534
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• 1993

흰쥐의 신피질 세포막과 소포체 막을 분리하여 카드를 및 Metallothioneln(MT)을 투여하였을 때 세포막에 존재하는 Ca-ATPase에 미치는 영향을 측정하여 다음과 같은 결과를 얻었다. 전기영동상에서 분리된 MT가 분자량 12KD 정도의 위치에 band가 나타났으며, 분리한 각 세포의 막에 카드윰을 농도별로 처리하였을 때 고농도일수록 Ca-ATPase의 활성도가 감소하였으나, MT를 처리한 경우 신피질 세포막은 거의 대조군과 유사한 결과를 나타냈고, 소포체 막에 MT를 처리한 경우는 20mg/ml의 카드윰을 처리한 경우와 유사하였다. 이와같은 결과로 보아 카드윰은 세포막의 Ca-ATPase의 활성을 저하시켜 세포내 칼슘 항상성에 영향을 미치는 MT는 Ca-ATPase의 활성을 회복시켜 카드뮴에 의한 세포독성의 방어작용에 부분적으로 작용한 것으로 생각된다.

• Ocean and Polar Research
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• 제23권4호
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• pp.337-345
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• 2001

A Cd-binding protein was identified in the renal cytosol of the Antarctic clam Laternula elliptica which naturally contains high concentrations of Cd. The Cd-binding protein showed similar characteristics of metallothionein (MT) in molecular weight (about 10-12 kDa) and low spectral absorbance at 280 nm with relatively high absorbance at 254nm. Results of immuno-histochemical staining suggested that the MT-like Cd-binding protein was mainly located in the epithelial cells of the kidney. The MT-like protein was a major ligand of cytosolic Cd as shown in the elution profiles of chromatography and may play an important role in Cd sequestration and accumulation in L. elliptica kidney. A considerable amount of Cd was also found to be associated with particulate fraction, indicating the sequestration to particulate fraction is as important as binding to the cytosolic MT-like protein in Cd accumulation in the kidney.

• 생태와환경
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• 제40권4호
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• pp.489-494
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• 2007

Transcriptional modulation of metallothionein (MT) during acute metal exposures (cadmium, copper or zinc) was examined in fry of Hemibarbus mylodon, a threatened fish species in Korean peninsula. Viability of H. mylodon fry was most affected by copper exposure (up to 79% of mortality at 1 ppm for 48 hours) and considerably by cadmium exposure (21 to 54% of mortality). On the other hand, Zn showed the least adverse effect on the viability (0 to 13% of mortality) of this species. Based on the semi-quantitative RT-PCR analysis, the stimulation of MT mRNA in response to metal exposures followed generally in a dose-dependent fashion where cadmium was the most potent inducer for the induction of MT transcripts in fry (up to more than 5-fold) while the lowest response was observed in zinc-exposed group (2-fold at maximum). From the exposure using environmentally realistic doses of cadmium (0 to 0.05 ppm for 24 hours), MT expression at mRNA level was also sensitively modulated toward upregulation up to more than 3-fold as relative to non-exposed control. Results from the present study would be a good basis for understanding the adaptive capacity and stress physiology of this endangered fish species during metal pollution.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2000년도 춘계학술발표대회
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• pp.426-429
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• 2000

Recombinant E. coli JM109(pZH3-5/pMT) harboring manganese transport gene(mntA) and metal sequestering protein, metallothionein(MT), was cultivated to accumulate cadmium in aqueous phase. Bioaccumulation followed Michaelis-Menten type kinetics. Equilibrium isotherm showed Langmuir type isotherm. The optimum pH for $Cd^{2+}$ uptake was 7-7.5.

• 한국식품영양과학회지
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• 제24권2호
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• pp.183-194
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• 1995

식이내 비타민 E 함량을 0mg vit. E/kg diet(STZ-OE군), 40mg vit. E/kg diet(STZ-40E군) 그리고 40mg vit. E/kg diet(STZ-400E군)로 달리하여 4주간 사육한 후 streptozotocin으로 당뇨병을 유발한 후 4일째에 희생시켜, 당뇨병 쥐에 있어서 MT합성에 미치는 비타민 E의 영향을 관찰한 결과는 다음과 같다. 체중증가는 STZ을 투여하기 전까지는 대조군에 비해 STZ-0E군만 낮았으나 STZ투여한 후에는 STZ투여군들 모두가 현저하게 감소하였다. 실험군간을 비교해 보면 STZ-400E군이 STZ-0E, STZ-40E군에 비해 STZ을 투여한 후 감소율이 낮았다. 혈당량은 당뇨군들이 대조군에 비해 약 3배 정도 높았으나, 당뇨군들 중에서 비타민 E의 영향은 없었다. 간조직 중의 비타민 E 함량은 STZ-0E 및 STZ-40E군에서는 대조군 보다 각각 50, 36%씩 크게 감소되었으나 STZ-400E군은 차이가 없었다. Glutathione(GSH) 함량은 비타민 E의 다량 첨가군(STZ-400E)에서 GSH 함량과 GSH/GSSG비가 증가되었다. 간조직 중의 지질과산화가는 STZ-0E군과 STZ-40E군이 대조군 보다 5.6, 2.5배씩 높았으나 STZ-400E군은 유의적인 차이가 없었다. Piasma insulin level은 대조군에 비해 당뇨유발군 모두가 현저하게 감소하였고, 비타민 E의 효과는 관찰되지 않았다. Piasma cortisol level은 대조군에 비해 STZ-0E, STZ-40E군이 증가하였으나 STZ-400E군은 이들 두군에 비해 유의적으로 감소하여 대조군 수준이었다. 간장, 신장, 소장조직의 MT 함량은 대조군에 비해 당뇨유발군이 각각 5.3, 2.8, 16배 정도 현저히 증가되었는데 STZ-400E군은 STZ-0E, STZ-40E 이들 두군에 비해 현저하게 감소되었다. Sephadex G-75를 사용하여 간조직 중의 Zn-MT를 분리한 결과 당뇨 유발군에서 대조군 보다 Zn-MT peak가 증가되었으나 Cu-MT는 변화가 없었다. Zn-MT는 DEAE A-25를 사용하여 MTI, MTII peak를 확인할 수 있었다. 이상의 결과에서 STZ유발 당뇨쥐는 산화적 stress에 더욱 감수성이 높으므로 지질과 산화 반응이 정상군에 비해 촉진되었으며, 식이 중 공급되는 비타민 E 함량이 부족시는 더욱 가속화되었다. 더불어 간조직 중의 MT합성도 같은 양상으로 증가되어 당뇨병이 free radical 생성으로 인해 MT를 유도하는 하나의 새로운 인자인 것으로 확인하였다. 그러나 식이 중 비타민 E의 다량 급여시는 생체내 항산화계 효소들이 정상군에 가깝게 활성화되고, 생리적 항산화물질이 증가되었다. 따라서 식이비타민 E가 당뇨병으로 인한 지질과산화에 대한 항산화적 방어기구를 강화시키므로써 조직의 과산화적 손상을 환화시키는데 크게 기여함을 알 수 있었다. 또한 비타민 E 다량 투여로 간조직과 신장, 소장조직내의 total MT 함량이 감소되고 분리정제한 Zn-MT의 peak가 감소하는 것으로 보아 MT가 free radical scavenger로서 작용한 것이라고 생각된다.

• Journal of Nutrition and Health
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• 제29권6호
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• pp.597-607
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• 1996

This study has been investigated the potenial of increased dietary cysteine to alter the effects of cadmium and lead on tissure and bone metal concentrations, excretion and tissue metallothionein(MT) concentrations. Fifty-four male rats of Sprgue-Dawley strain weighing 149$\pm$17g were divided into 9 groups according to body weight. Nine experimental diets with different cadmium (0ppm, 400ppm), lead(0ppm, 710ppm) and cysteine (0.06%, 0.45%, 0.90%) levels were given to rats for 30 days ; Food intake, weight gain, F.E.R, and weights of liver, kidney and femur were decreased in cadmium supplied groups than in cadmium free groups. Urinary and fecal cadmium excretions were increased and MT synthesis we induced in liver, kidney and small intestine in cadmium supplied groups. In lead supplied groups, weight gain and F.E.R were decreased. With cysteine supplementation in cadmium supplied groups, weight gain and F.E.R, and weights of liver, kidney and femur were increased. Cadmium excretion in feces and MT concentrations in liver and kidney were also increased with cysteine supplementation. In lead supplied groups, there was no significant increase in food intake, weight gain and F.E.R with cysteine supplementation. Lead excretion in feces was increased in cysteine supplemented groups. In conclusion, effect of cadmium administration was more toxic than lead adminstration. Cysteine alleviated cadmium and lead toxicity by increasing metallothionein concentration and fecal excretions of heavy metals. Especially, effect of cysteine supplementation was more effective in cadmium groups than in lead groups. Effect of cysteine supplementation was not different with level of cysteine supplementation in both cadmium and lead groups.

• Journal of Microbiology and Biotechnology
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• 제21권5호
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• pp.455-463
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• 2011

Metallothioneins are a class of small cysteine-rich proteins that have been associated with increased tolerance to metal and oxidative stresses in animals, plants, and fungi. We investigated a metallothionein-like (mt-like) gene shown previously to be upregulated in fruiting bodies of the fungus Agaricus bisporus in response to post-harvest storage. Analysis of an A. bisporus genomic DNA cosmid library identified two similar mt-like genes (met1 and met2) arranged as a bidirectional gene pair transcribed from the same promoter region. The promoter contained regulatory elements including 9 metal responsive elements and a CAAT box region 220 bp downstream of met1 that showed striking similarity to a feature in Coprinopsis cinerea mt-like gene promoters. Transcriptional analysis showed that both met genes are significantly and rapidly (within 3 hours) upregulated during post-harvest storage and expression is significantly greater in stipe and cap tissues compared with the gills. However, a strong directionality of the promoter was demonstrated, as transcript levels of met1 were at least two orders of magnitude greater than those of met2 in all samples tested.

• 한국환경보건학회지
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• 제18권1호
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• pp.95-104
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• 1992

The influence of selenium to several toxic effects of cadmum, including lethality has been shown following pretreatment with cadmium, zinc and seleniurm Five groups of rats, each consisting of 16 rats, were studied and each group was divided into four subgroups, 4 rats for each subgroup. After subcutaneous pretreatment during 5 days with saline, CdCl$_{2}$ (0.5mg/kg, ZnCl$_{2}$ (13.0mg/kg) and $Na_{2}SeO_{3}$(1.0mg/kg), rats were given intraperitioneal administration of various dosage of or cadium of cadmium and selenium. After giving the challenge dose, cadmium and metallothionein(MT) concentrations were determined in liver and kidney The concentration of cadmium in liver and kideny increased proportionally to the increase of challenge dosage. The simultaneous administration of cadmium and selenium significantly more decrease cadmium concentrations in liver and kidney than those of the administration of cadmium only. However, MT concentrations in liver and kideny were increased by the pretreatment of cacmium, zinc and selenium. Our results suggest that increasing cadmium concentrations, gradully accumulating in the tissues of liver and kidney as a result of the pretreatment, served to induced the synthesis of MT, thus making them resistant to the challenge from cadmium.

• Applied Microscopy
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• 제34권3호
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• pp.171-178
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• 2004

메탈로사이오닌 (metallothionein: MT)은 시스테인(cysteine)이 다수 포함되어 있고, 저분자량인 단백질로서 중금속이온이나 다양한 세포 독성 인자에 의해 유도되며, 최근 발암과정 (carcinogenesis)이나 혹은 세포분화와 연관하여 활발하게 연구가 이루어지고 있다. 본 연구는 흰쥐를 수정시킨 후 발생과정 또는 출생 후 성장과정 단계에서 태아와 신생 흰쥐의 간 조직 및 세포내 MT의 분포 양상을 조사하고자 하였다. 흰쥐 간 조직내 MT의 분포는 면역조직화학적 방법을 적용하여 광학현미경으로 관찰하였고, 세포내 미세구조적 위치 (localization)는 금 입자 표지를 이용한 면역세포화학적 방법을 이용하여 투과전자현미경으로 관찰하였다. MT는 발생 13일이 경과한 흰쥐 태아의 간에서 처음으로 관찰되기 시작하여, 출생 당일의 신생 흰쥐에서 가장 높게 관찰되었다. 면역 조직화학 및 세포화학적 방법으로 관찰한 결과, MT 염색성은 발생 후기에서 출생후 10일째까지 비교적 높게 나타났다. MT에 대한 양성반응은 난형 세포 (oval cell)들과 간 실질세포 (liver parenchymal cell)들에서 뚜렷하게 존재하였다. 발생과정과 신생 흰쥐의 간 조직에서는 핵과 세포질 양쪽에서 공통적으로 양성 반응을 보였으나, 생후 30일 이후부터 성체에 이르는 시기까지는 세포질에 한정되어 있는 양상을 보였다. 금 입자 표지법에 의한 MT의 세포질내 분포를 보면, 발생 중인 흰쥐 간 세포에서는 핵과 세포질내에 고르게 산재되어 있었다. 즉, 핵내 염색질, 인, 조면소포체 및 cytosol에서도 MT에 대한 면역 금 입자들이 관찰되었으나, 미토콘드리아와 지질 소포 등에서는 거의 관찰되지 않았다. 상기의 결과들은 발생중 혹은 성장중인 흰쥐의 간에서 MT가 점차 증가하는 것으로 요약될 수 있는 것으로서, 이는 MT가 세포들의 증식과 분화에 금속 이온의 저장과 방출 혹은 전사인자 (transcription factor)로서 작용할 수 있음을 암시하는 것으로 판단된다.