• Ocean and Polar Research
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• v.40 no.1
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• pp.15-22
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• 2018

In the present study, we investigated temporal variation in the reproductive effort and biochemical contents in adult Manila clam Ruditapes philippinarum at Shi-Heung-Ri beach on the east coast of Jeju Island. Gonad-somatic index (GSI), a ratio of the egg mass to the total biomass determined using ELISA increased dramatically from late May to June (7.1% to 19.5%). In mid-July, GSI dropped to 15.1%, indicating spawning on a small scale during this period. GSI also declined dramatically from late August (15.5%) to September (4.3%), suggesting that massive spawning occurred during this period. A positive correlation was observed between clam size and potential fecundity, while potential fecundity ranged between 1.19-8.40 million eggs/clam. Total proteins and lipids in the tissue increased from late spring to mid-summer, coinciding with an increase in GSI. Protein, lipid and carbohydrate contents in the tissue were lowest in January, indicating that clams were suffering from poor nutritional circumstances, which may be associated with poor food supply from the environment. Monthly increase and/or decrease in the body weight were closely related to gonad maturation and subsequent spawning, which was also linked to temporal changes in the contents of protein, lipid and carbohydrates in the tissue.

• Journal of Marine Life Science
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• v.3 no.2
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• pp.59-66
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• 2018

The assessment of level of health of the tidal flats can be evaluate by health of organisms inhabit the tidal flats. It is possible to evaluate the precise health level of organisms inhabit the tidal flats using analysis of expression of biomarker genes. The purpose of this research is to evaluate the health of the tidal flats on the west coast using biomarker genes such as heat shock protein 70 (Hsp70), heat shock protein 90 (Hsp90), glutathione S-transferases (GST) and thioredoxin (TRX). These genes are stress, immune, and antioxidant related genes that can be used to look at the health of an organism through gene expression. In this study, we collected manila clam (Ruditapes philippinarum) in 8 analysis areas on the west coast. Expression of the genes was analyzed by RT-qPCR method. Results showed that, the expression of Hsp70, Hsp90, GST and TRX genes were differentially expressed in the 8 analysis areas. In particular, the expression of Hsp90 and GST or the expression of Hsp70 and TRX were similar. This means that there is a substance that reacts specifically to each gene. Therefore, I think suggest that the based on the results of physicochemical analysis, it can be selected genes suitable for analysis. These results suggest that Hsp70, Hsp90, GST and TRX were played roles in biomarker for assessment of the health of tidal flats.

• The Korean Journal of Malacology
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• v.24 no.1
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• pp.67-72
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• 2008

This study investigated the proximate composition in the muscle and viscera of five commercially valuable Korean Veneridae, Protothaca jedoensis, Ruditapes philippinarum, Saxidomus purprratus, Cyclina sinensis and Meretrix lusoria. The proximate analysis revealed that the moisture, crude protein and carbohydrate of muscle in higher than viscera. As results, except for the crude lipid and carbohydrate there was little significant differences among the moisture, protein and crude ash. The crude lipid contents was highest in the viscera of Meretrix lusoria, while lowest in the muscle of Ruditapes philippinarum. The carbohydrate content of viscera in the Meretrix lusoria, Ruditapes philippinarum and Protothaca jedoensis were $3.61{\pm}0.45%$, $3.45{\pm}1.4%$ and $3.39{\pm}0.07%$, respectively.

• The Korean Journal of Malacology
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• v.32 no.1
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• pp.45-54
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• 2016

To evaluate the applicability of cellular energy allocation (CEA) in the bivalves as a biomarker for the assessment of environmental contamination, the energy contents and energy consumption in several tissues of the Manila clam, Ruditapes philippinarum were analyzed. The contents of lipid, glucose, protein and electron transport system (ETS) activity in the foot, siphons, gills, and body of R. philippinarum exposed to crude oil-spiked sediments were measured at 1, 2, 4, 7, 10 days after exposure. The reserved energy (energy available, EA) in the lipid, glucose and protein decreased as contamination level and exposure time increased. In contrast, the ETS activity (energy consumed, EC) showed the reverse tendency. The order of available energy contents were foot > siphons > gill > body. Significant differences in both EA and EC were found only at the highest contamination level (58.3 mg TPAHs/kg DW). EA decreased significantly in the foot and gill at 1 day, in the body at 2 and 7 days after exposure. EC increased significantly in the body at 4 days after exposure. CEA showed higher sensitivity to the contamination than EA or EC. Especially, CEA in the foot and body decreased significantly at lower ranges of contamination level (as low as 6.5 mg TPAHs/kg DW) during 1 to 7 days after exposure. The CEA is more useful than EA or EC alone for the assessment of sediment contamination at lower level that acute toxicity could not be detected. CEA analyses in the body of R. philippinarum after 4 days' exposure to contaminated sediments seem to be the most sensitive and reliable.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.35 no.5
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• pp.529-533
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• 2002

The peptides inhibiting angiotensin converting enzyme (ACE) were isolated from the hydrolysate of manila clam (Ruditapes philippinamm) proteins prepared with thermolysin. The thermolysin hydrolysate was pretreated with membrane filter (MW cut-off 10,000) to obtain the peptide fraction with ACE inhibition. The crude peptides were applied to a Sephadex LH-20 column and eluted with $30\%$ methanol. The three active fractions (A, B and C) were collected and concentrated, and then applied to a SP-Toyopearl 650S column equilibrated with distilled water and was eluted with a linear gradient of NaCl concentration (0 to 1 M). The four active fractions (A-1, A-2, B-1 and C-1) were collected and concentrated, and then applied to a SuperQ-Toyopearl 650S column equilibrated with distilled water and was eluted with a linear gradient of NaCl concentration (0 to 1 M). The maximum inhibitory activity was observed in the fraction B-1Q showed the IC_{50} values of 0.748 $\mu$g. The abundant amino acids obtained from active fraction B-1Q were leucine, isoleucine, alanine and threonine.

• The Korean Journal of Malacology
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• v.19 no.1
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• pp.41-51
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• 2003

Reproductive cycle with the gonadal development of Ruditapes philippinarum can be classified into five successive stages by histological observations: early active stage (January to March), late active stage (February to May), ripe stage (April to August), partially spawned stage (May to October), and spent/inactive stage (August to February). Changes in total protein contents in the adductor muscle tissues reached the maximum in the early and late active stages (February) and appeared the minimum in the ripe and partially spawned stages (May), while changes in their contents in the visceral mass tissues reached the maximum in the ripe and partially spawned stages (June) and gradually decreased in the partially spawned stage (June to October). On the whole, changes in total protein contents showed a negative correlationship between the adductor muscle and visceral mass tissues (r = -0.292, p < 0.05). Changes in total lipid contents in the adductor muscle tissues reached the maximum in the inactive and early active stages (January) and sharply decreased in the early and late active stages (February), while their contents in the visceral mass tissues reached the maximum in the ripe and partially spawned stages (April) and gradually decreased in the partially spawned stage (to October). On the whole, changes in total lipid contents showed a negative correlationship between the adductor muscle and visceral mass tissues (r = -0.699, p<0.05). Changes in glycogen contents in the adductor muscle tissues reached the maximum in the late active and ripe stages (April) and rapidly decreased in the partially spawned stage (May to October), while their contents in the visceral mass tissues reached the maximum in the early and late active stages (February) and rapidly decreased in the late active stage (March). Thereafter, their levels gradually increased in the ripe and partially spawned stages (April to July). On the whole, changes in glycogen contents appeared no correlationship between the adductor muscle and visceral mass tissues (r = 0.062, p > 0.05). These results indicate that the adductor muscle and visceral mass tissues are an important energy storage and nutrient supply organ in the Manila clams, and the nutrient contents of the adductor muscle and visceral muscle tissues change in response to gonadal energy needs.

• The Korean Journal of Malacology
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• v.22 no.2
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• pp.157-165
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• 2006

The present study was conducted to confirm that a bivalve Ruditapes philippinarum can be used as a biomarker for the monitoring of the heavy metal pollution in the silt of the marine environment. The clams were collected from the silt of Cheonsu-bay, Buheung-ri, and Tan-island of the West Sea, Korea. To observe the normal structures of the target organs (hepatopancreas and gill), they were dissected out for the immunohistochemical study and the electron microscopy with TEM, SEM, and SEM-EDS. The immunohistochemical study showed that the interdiverticular connective tissues of the hepatopancreas, and the outer epithelium of the gill lamellae was strongly reacted to anti-metallothionein (MT), indicating the presence of MT, a metal-binding protein, involved in metal detoxifying process. According to the examinations under the TEM, the epithelial cells of the hepatopancreas of the clams collected from polluted area (Tan-island) showed certain changes such as swollen rER, swollen nuclear envelope and inclusion bodies in the nulcei. In the SEM-EDS analysis, tissue of the hepatopancreas showed relatively higher concentration of S, Zn, and Cd. These elements are supposed to be concerning with the MT-reaction in the hepatopancreas. Considering that the coastal bivalve R. philippinarum showed immediate subcellular responses to heavy metal pollution in the overall experiments conducted, this species might act as one of efficient biomarkers for the heavy metal contamination in the marine environment.

• Preventive Nutrition and Food Science
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• v.8 no.2
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• pp.154-157
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• 2003

A peptide inhibiting HIV-1 pretense was isolated from the hydrolysate of manila clam (Ruditapes philippinarum) proteins digested with thermolysin. The peptide was purified by using membrane filtration, gel permeation chromatography, ion exchange chromatography, and reverse phase HPLC, The amino acid sequence of the peptide was determined to be Ile-Tyr-Glu-Gly. This tetrapeptide sequence exists in some proteins of Physarum polycephalum and Mycobacterium smegmatis. Chemically synthesized Ile-Tyr-Glu-Gly showed the $IC_{50}$/ value of 22.3 $\mu$M.

• The Korean Journal of Malacology
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• v.29 no.2
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• pp.155-161
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• 2013

Cathepsins are lysosomal/cysteine proteases belong to papain family (C1 family) that is involved in intracellular protein degradation, antigen processing, hormone maturation, and immune responses. In this study, member of cathepsin family was identified from Manila clam (Mc-Cathepsin D) and investigated the immune response against brown ring disease (BRD) causing Vibrio tapetis challenge. The identified Mc-Cathepsin D gene encodes characteristic features typical for the cathepsin family including eukaryotic and viral aspartyl protease signature domain and two highly conserved active sites ($^{84}VVFDTGSSNLWV^{95}$ and $^{270}IADTGTSLLAG^{281}$). Moreover, MC-Cathepsin D shows higher identity values (-50-70%) and conserved amino acids with known cathepsin D members. Transcriptional results (by quantitative real-time RT-PCR) showed that Mc-Cathepsin D was expressed at higher levels in gills and hemocytes than mantle, adductor muscle, foot, and siphon. After the V. tapetis challenge under laboratory conditions, Mc-Cathepsin D mRNA was up-regulated in gills and hemocytes. Present study indicates that Mc-Cathepsin D is constitutively expressed in different tissues and potentially inducible when infecting BRD by V. tapetis. It is further suggesting that Mc-Cathepsin D may be involved in multiple role including immune response reactions against BRD.

• Proceedings of the Korean Environmental Health Society Conference
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• 2004.12a
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• pp.63-68
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• 2004

Immunohistochemical and ultrastructural experiments were conducted to find out heavy metal accumulation in some selected organs such as the kidney, the digestive gland, and the gill of the Antarctic clam Laternula elliptica and R. philippinarum. According to the immunohistochemical study the subject organs of the clam showed reactions indicating the presence of MT (metallothionein), a metal-binding protein involved in metal detoxifying process. Examination under the transmission electron microscope also revealed that other ligands may play a role in metal accumulating and detoxifying process in L. elliptica and R. philippinarum. In the artificial exposure of the clam to Cd, the clams showed immediate subcellular responses. The level of the anti-MT reactions became higher in the proportion to the degree of pollution of their habitat and to the period of Cd exposure. These suggest that the two species can be used as efficient biomarkers for Cd exposure in the natural environment.