• 한국수산과학회지
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• 제45권2호
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• pp.154-160
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• 2012

This study the effects of Cu exposure on biochemical factors in the hemolymph and hepatopancreas of the abalone $Haliotis$ $discus$ $hannai$. Abalone were exposed to 0, 5, 10, 20 and 40 ${\mu}g/L$ Cu for 4 weeks. The calcium concentrations in hemolymph were decreased significantly on exposure to 20 and 40 ${\mu}g/L$ Cu after 2 weeks. The aspartate aminotransferase (AST), alanine aminotransferase (ALT) and alkaline phosphatase (ALP) activities in abalone hemolymph were markedly elevated after exposure to 40 ${\mu}g/L$ Cu for 4 weeks. The hepatopancreas superoxide dismutase (SOD) and catalase (CAT) activities were also significantly increased by exposure to 20 ${\mu}g/L$ Cu for 4 weeks. These biochemical factors may represent a convenient method of monitoring heavy metal pollution in coastal areas. From these results, we conclude that a high copper concentration (40 ${\mu}g/L$) in water may curtail hemolymph homeostasis and anti-oxidative reactions in abalone.

• 한국수산과학회지
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• 제50권4호
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• pp.388-395
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• 2017

We investigated the toxicity of zinc and aluminum hot-dip galvanized sheet steel to abalone Haliotis discus hannai via changes in the gill and hepatopancreas using histological and transmission electron microscopy analysis. Experimental groups were composed of one control and four exposure conditions (direct or indirect exposure to zinc and aluminum hot-dip galvanized sheet steel). In the control group, aluminum exposure groups (direct and indirect), and indirect zinc exposure group, abalone mortality was not observed until the end of the experiment, and no histopathological changes were observed in the gill and hepatopancreas. However, the direct zinc exposure group exhibited 100% mortality. Ultrastructural analysis of the cytoplasm of ciliated and microvilli-bearing epithelial cells from gill filaments revealed electron-dense vesicles near the cell membrane and disruption of the nuclear membrane. We also observed swollen mitochondria and a loss of mitochondrial cristae. The hepatopancreas showed similar changes, and we detected highly electron-dense particles within the vesicles. These results suggest that abalone exposed directly to zinc hot-dip galvanized sheet steel experience acute toxicity, causing damage to cell organelles in the gill and hepatopancreas and, finally, inducing mortality.

• Fisheries and Aquatic Sciences
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• 제3권3_4호
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• pp.188-194
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• 2000

A protease purified from hepatopancreas of shrimp, Penaeus japonicus, had maximum activity at $70^{\circ}C$ and in neutral and alkaline pH ranges. Specific activity at optimum reaction condition of the protease was estimated to be approximately 12 U/mg/min. The protease was stable in neutral and alkaline pH ranges and activity was retained after heat treatment at $50^{\circ}C$ for 30 min. Apparent $K_m$ and $V_{max}$ value against casein substrate were estimated to be $0.29\%$ and $7.8see^{-1}$, respectively, and those against N-CBZ-L-tyrosine p-nitropheny1 ester (CBZ­Tyr-NE) were 0.38 mM and $2,400 see^{-1}$, respectively. The N-termina1 sequence of the protease showed high homology to the trypsin from same species and the proteases from shrimp. Myosin heavy chain (MHC) from shrimp tail meat was the most susceptible to the protease and actin/tropomyosin were degraded progressively during 4 hr incubation, but to a lesser degree than MHC.

• 한국식품영양학회지
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• 제19권4호
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• pp.466-472
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• 2006

붉은 대게 가공공정 중에 얻어지는 자숙액 및 내장의 천연 게향 소재로서의 가능성을 검토하고자 정미성분을 포함한 신품학적인 특성을 분석 비교한 결과는 다음과 같다. 붉은 대게 자숙액을 $40\;^{\circ}Brix$까지 농축시키면서 관능 검사 및 냄새 특성 강도를 수행한 결과 $30\;^{\circ}Brix$가 가장 양호하였다. 그리고 붉은 대게 농축 자숙액$(30^{\circ}Brix)$과 내장의 불휘발성 유기산에서는 lactic acid가 가장 많았고 다음으로 succinic acid이었다. 핵산 관련 물질은 IMP, Hx 및 GMP가 내장에서 대부분이었고, AMP, HxR, IMP 및 GMP 등이 자숙액에서 대부분을 차지하였다. 내장의 유리 아미노산 총량은 농축 자숙액에 비해 5.6배나 많았다. 조성면에서는 methionine, lysine, arginine, valine, histidine, alanine, hydroxy proline 및 glycine 등 8종이 자숙액 전체 아미노산의 72%를 차지하였고, 내장에서는 methyl histidine, leucine, alanine, glutamic acid, glycine, valine, threonine, taurine, isoleucine 및 serine 등 10종이 전체의 74%를 차지하였다. 결론적으로 천연 게향 제조를 위한 소재로서는 $30\;^{\circ}Brix$ 농도의 붉은 대게 농축 자숙액에 0.5%(w/w)의 붉은 대게 내장을 첨가하는 것이 적합하였다

• Fisheries and Aquatic Sciences
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• 제19권4호
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• pp.21.1-21.11
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• 2016

Background: The evaluation of suitable reference genes as normalization controls is a prerequisite requirement for launching quantitative reverse transcription-PCR (RT-qPCR)-based expression study. In order to select the stable reference genes in abalone Haliotis discus hannai tissues (gill and hepatopancreas) under heavy metal exposure conditions (Cu, Zn, and Cd), 12 potential candidate housekeeping genes were subjected to expression stability based on the comprehensive ranking while integrating four different statistical algorithms (geNorm, NormFinder, BestKeeper, and ${\Delta}CT$ method). Results: Expression stability in the gill subset was determined as RPL7 > RPL8 > ACTB > RPL3 > PPIB > RPL7A > EF1A > RPL4 > GAPDH > RPL5 > UBE2 > B-TU. On the other hand, the ranking in the subset for hepatopancreas was RPL7 > RPL3 > RPL8 > ACTB > RPL4 > EF1A > RPL5 > RPL7A > B-TU > UBE2 > PPIB > GAPDH. The pairwise variation assessed by the geNorm program indicates that two reference genes could be sufficient for accurate normalization in both gill and hepatopancreas subsets. Overall, both gill and hepatopancreas subsets recommended ribosomal protein genes (particularly RPL7) as stable references, whereas traditional housekeepers such as ${\beta}-tubulin$ (B-TU) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) genes were ranked as unstable genes. The validation of reference gene selection was confirmed with the quantitative assay of MT transcripts. Conclusions: The present analysis showed the importance of validating reference genes with multiple algorithmic approaches to select genes that are truly stable. Our results indicate that expression stability of a given reference gene could not always have consensus across tissue types. The data from this study could be a good guide for the future design of RT-qPCR studies with respect to metal regulation/detoxification and other related physiologies in this abalone species.

• 한국어병학회지
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• 제14권2호
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• pp.81-90
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• 2001

구리에 대한 넙치의 조직학적 반응을 알아보기 위하여 아가미, 간췌장 및 신장조직상의 변화를 조사하였다. 대조구에서는 5주 후 아가미에서 점액세포와 염세포의 증가가 관찰되었다. 간췌장과 신장에서는 혈구의 증가와 일부 흑색 대식세포들이 관찰되었다. 구리 노출구 가운데 조직학적 변화가 처음 관찰된 농도구는 0.05mg/$\ell$/7d 였으며, 아가미에서는 염세포의 증가, 간췌장에서는 간세포의 활성화와 췌장 효소원과립의 감소, 신장에서는 충혈과 흑색 대식세포군이 관찰되었다. 정상적인 기능 수행이 어려울 것으로 판단되는 한계농도는 아가미의 경우 0.18mg/$\ell$/21d 간췌장에서는 0.18mg/$\ell$/14d 그리고 신장에서는 0.08mg/$\ell$/14d 였다.

• Fisheries and Aquatic Sciences
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• 제1권2호
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• pp.201-208
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• 1998

A protease, which had no tryptic and chymotryptic activity, was purified from the hepatopancreas of shrimp, P. japonicus, through ammonium sulfate fractionation, Q­Sepharose ionic exchange, benzamidine Sepharose 6B affinity, and Sephacryl S-100 gel chromatography. Molecular weight (M.W.) of the protease was estimated to be 24 kDa by gel filtration and showed a single peptide band by sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE). The protease had a low ratio of acidic to basic amino acids, which is different with pro teases from marine animals. The enzyme was partially inhibited by benzamidine, tosyl-L-lysine chioromethyl ketone (TLCK), phenylmethylsulfonyl fluoride (PMSF), soybean trypsin inhibitor (SBTI), and pepstatin. The enzyme did not have any activity against benzoyl-D,L-arginine p-nitroanilide (BAPNA) or benzoyl-L-tyrosine ethyl ester (BTEE) which is a specific substrate of trypsin and chymotrypsin, respectively. However, the enzyme showed activity forward N-CBZ-L-tyrosine p-nitrophenyl ester (CBZ-Tyr-pNE), N­CBZ-L-tryptophan p-nitrophenyl ester (CBZ-Trp-pNE), and N-CBZ-L-proline p-nitrophenyl ester (CBZ-Pro-pNE). The protease did not showed tryptic and chymotryptic activity, which was not reported in shrimp hepatopancreas.

• Fisheries and Aquatic Sciences
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• 제2권2호
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• pp.218-225
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• 1999

A protease without tryptic and chymotryptic activities was purified from the hepatopancreas of shrimp, Penaeus orientalis, using Q-Sepharose ionic exchange, benzamidine Sepharose-6B affinity, Mono-Q, and gel chromatography. Molecular weight (M.W.) of the protease was estimated to be 27kDa by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS­PAGE). The amino acid composition of the protease was different from that of protease from P. japonicus or trypsin from P. orientalis. The protease was completely inhibited by benzamidine, $N\alpha-p-tosyl-L-lysine$ chloromethyl ketone (TLCK), and phenylmethylsulfonyl fluoride (PMSF) and was not affected by leupeptin, pepstatin, N-tosyl-L-phenylalanine chloromethyl ketone (TPCK), iodoacetate, and ethylenediamine tetra acetate (EDTA). The enzyme did not have any activity against Na-benzoyl-DL-arginine p-nitroanilide (BAPNA) or N-benzoyl-L-tyrosine ethyl ester (BTEE) which are specific substrates of trypsin and chymotrypsin, respectively. However, the protease showed hydrolytic activity for a carboxyl terminal of Tyr, Trp, Phe, Glu, and Cys.

• Fisheries and Aquatic Sciences
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• 제17권2호
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• pp.181-187
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• 2014

This study was performed to identify the optimum fractionation method and conditions to obtain exopeptidase-active fractions from octopus hepatopancreas (HP) crude extracts (CEs) using four techniques: solid ammonium sulfate fractionation, polyethylene glycol (PEG) fractionation, anion exchange chromatography, and gel filtration chromatography. The fractions with the highest total activity toward L-leucine-p-nitroanilide (Leu-pNA) were fraction IV from the ammonium sulfate and PEG fractionation, and fraction II in ion exchange and gel filtration chromatography. The total exoprotease activity of these fractions was highest in fraction IV (4,050.20 U) of ammonium sulfate fractionation, followed by fraction II (3,600.28 U) from gel filtration chromatography, fraction IV (2,861.30 U) from PEG fractionation, and fraction II (2,576.28 U) from ion exchange chromatography. These results suggest that ammonium sulfate fractionation using 60-80% ammonium sulfate was the most efficient method for separating the exoprotease active fractions from CEs of octopus HP.

• 한국어병학회지
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• 제25권2호
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• pp.103-109
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• 2012

The objective of this study was to investigate the effects of exposured $Cu^{2+}$ on growth and bioaccumulation of abalone Haliotis discus hannai. Abalone were exposed to various concentration of $Cu^{2+}$ (0, 5, 10, 20, $40{\mu}g/L$). Bioaccumulation of tissues, hepatopancreas, muscles and gills were measured. hepatopancreas and gills $Cu^{2+}$ concentration of abalone increases to extent during the 4 weeks accumulation time. But muscles showed no significant changes, with respect to control. These data indicate that abalone Haliotis discus hannai hepatopancreas and gills can be considered adequate target tissues for waterborne exposured of $Cu^{2+}$. Weight growth rate of abalone exposed to $Cu^{2+}$ was significantly decreased in 20 and $40{\mu}g/L$ groups compared to control. This study revealed that high $Cu^{2+}$ concentration (${\geq}20{\mu}g/{\ell}$) reduced growth of abalone. These data indicate that excessive waterborne $Cu^{2+}$ can affect the toxicity of xenbiotics to abalone through alterations in growth rate. Thus, environmental standard of $Cu^{2+}$ $20{\mu}g/L$, should be considered a potential source of variation in toxicological studies with abalone.