• Ocean and Polar Research
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• v.34 no.2
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• pp.137-149
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• 2012

Hemocyte parameters of the wild Pacific oyster Crassostrea gigas inhabiting intertidal zones in small bays (Gwangyang and Jinhae Bay) on the southern coast of Korea were evaluated using flow cytometry and neutral red retention (NRR) assay. Morphological features, cell count, mortality, DNA damage, phagocytosis, and lysosomal membrane stability of hemocytes were analyzed. Three types of hemocytes were identified in the oyster hemolymph: granulocytes, hyalinocytes, and blast-like cells. Immune related functions of hemocyte including phagocytosis and lysosomal membrane stability were significantly different among the study areas (P<0.05), while cell count, mortality, and DNA damage of hemocytes were not significantly different. In Gwangyang Bay, phagocytosis of granulocytes and lysosomal membrane stability of oyster hemocytes inhabiting inside bay were significantly lower than those of oyster hemocytes in outside bay (P<0.05), indicating that oysters in inside bay of Gwangyang were relatively suppressed the immunological function in hemocytes. Contrary to Gwangyang Bay, immune parameters of oyster hemocytes in Jinhae Bay not showed the difference between sampling sites. In conclusion, flow cytometry and NRR assay using oyster hemocyte has a powerful tool to investigate the cell level in a short time due to no-preprocessing of material.

• Journal of fish pathology
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• v.13 no.2
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• pp.129-136
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• 2000

To understand host-defence mechanism of clam(Ruditapes philippinarum) hemocyte against foreign materials, classification and their seasonal change in the number were performed. clams collected from a farm in Julpo Bay, Gochang, Chollabuk-Do were used in this experiment. Lots of hemocytes were found between the muscle fibers and connetive tissue of posterior adductor muscle. Hemocytes of R. philippinarum were classified into granulocytes and agranulocytes. Granulocytes were composed of three types, basophilic granulocyte, acidophilic granulocyte and fibrocyte in accordance with the staining affinities of their cytoplasmic granules. Fibrocyte has filopodia and vesicle in endoplasm and bigger than other granulocytes in size. Agranulocytes were less in the number and smaller in the size compared to those of granulocytes. Hyalinocytes had no granule in their cytoplasm. The nucleus located in the center of the cell was oval or spherical shaped. In electron microscopic observation, granulocytes and hyalinocytes contained electron-dense vesicles and some small lucid vesicles in their cytoplasm, respectively. Granulocytes phagocytosed more zymosan particles than hyalinocytes. Acidophilic granulocytes showed higher phagocytic ratio than basophilic granulocytes. Total hemocyte numbers showed the highest at April to August and the lowest at October to December. In the composition of each hemocyte, basophilic granulocytes were always more than acidophilic granulocytes and hyalinocytes.

• Proceedings of the Malacological Society of Korea Conference
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• 2003.11a
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• pp.12-12
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• 2003

• BMB Reports
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• v.43 no.4
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• pp.279-283
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• 2010

Polydnavirus genome is segmented and dispersed on host wasp chromosome. After replication, the segments form double- stranded circular DNAs and embedded in viral coat proteins. These viral particles are delivered into a parasitized host along with parasitoid eggs. A serine-rich protein (SRP) is predicted in a polydnavirus, Cotesia plutellae bracovirus (CpBV), genome in its segment no. 33 (CpBV-S33), creating CpBV-SRP1. This study explored its expression and physiological function in the diamondback moth, Plutella xylostella, larvae parasitized by C. plutellae. CpBV-SRP1 encodes 122 amino acids with 26 serines and several predicted phosphorylation sites. It is persistently expressed in all tested tissues of parasitized P. xylostella including hemocyte, fat body, and gut. Its physiological function was analyzed by injecting CpBV-S33 and inducing its expression in nonparasitized P. xylostella by a technique called SERI (segment expression and RNA interference). The expression of CpBV-SRP1 significantly impaired the spreading behavior and total cell count of hemocytes of treated larvae. Subsequent RNA interference of CpBV-SRP1 rescued the immunosuppressive response. This study reports the persistent expression of CpBV-SRP1 in a parasitized host and its parasitic role in suppressing the host immune response by altering hemocyte behavior and survival.

• Applied Microscopy
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• v.24 no.2
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• pp.93-104
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• 1994

Six types of hemocytes were identified in adult grasshopper, Euprepocnemis shirakii Bolivar. The morphological and ultrastructural characteristics of these cells were characterized by phase contrast, scanning and transmission electron microscope. The prohemocytes are small and spherical cells with large nucleus. The plasmatocytes are spindle shaped or polymorphic cells which show numerous cytoplasmic processes on the cell surface and they have lysosomes and vesicles that may be involved in phagocytic function. Especially, multivesicular bodies are observed in the polymorphic cells. The granulocytes I are spherical shaped cells. They are characterized by a number of electron dense granules measuring $0.2-0.3{\mu}m$ in average diameter and marginal band of microtubules which are always in close proximity to the cell membrane. The granulocytes II are oval or spindle-shaped cells. They contain large electron dense granules measuring $0.5-0.8{\mu}m$ in average diameter. Their cytoplasm is filled with numerous granules. The spherulocytes contain large amounts of spherules that most of their cytoplasm. Spherules filled with fine granules or flocculent materials. The oenocytoids are large spherical cells with few cytoplasmic organelles. Their cytoplasm contains peculiar aspect of motochondria and numerous polyrobosome.

• Journal of Life Science
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• v.16 no.7 s.80
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• pp.1087-1089
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• 2006

Shrimp has efficient non-specific defense mechanisms based on activities of the hemocytes against pathogens. Up to now, it has been known that one of the non-specific immune reactions is related to mutual association among types of hemocyte, granular cell, semi-granular cell, and hyaline cell. In this study, we tried to know the effectiveness of feed-additives such as 5-aminolevulinic acid(ALA), chitosan, and hot-water extract of herb on immunity of shrimp (Penaeus chinensis) by hemocytic observation and SDS-PAGE analysis. Finally, we suggest a principle of the examination system for effects of various feed-additives.

• Journal of Life Science
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• v.27 no.11
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• pp.1276-1289
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• 2017

This study isolated and purified the antimicrobial peptide McSSP-31 from an acidified hemocyte extract of a Mytilus coruscus. The antimicrobial peptide was purified by using a $C_{18}$ reversed-phase high-performance liquid chromatography (HPLC). The peptide was determined to be 3330.549 Da by matrix assisted-laser desorption ionization time-of-flight mass spectrophotometer (MALDI-TOF/MS). The N-terminus of a 14 amino-acid sequence was identified as P-S-P-T-R-R-S-T-S-R-S-K-S-R by Edman degradation method. The acquired sequence showed a 93% similarity with the sperm-specific protein Phi-1, which is from M. californianus. The identified open-reading frame (ORF) of peptide was 306 bp encoding 101 amino acids, which was analyzed by rapid amplification of cDNA ends (RACE), cloning and sequencing analysis. We compared the full sequence with other known proteins that reveal the sperm-specific protein Phi-1 (93.5%) of M. californianus. Synthesized antimicrobial peptide (McSSP-31) showed antibacterial activity against gram-positive bacteria including B. subtilis, S. mutans, S. aureus and gram-negative bacteria including E. coli, K. pneumoniae, P. mirabilis, P. aeruginosa and fungi, C. albicans. Also, synthesized peptide showed strong antibacterial activity against antibiotic-resistant strains, including S. aureus. The cytotoxicity of the peptide was determined by using the HUVEC human cell line. The peptide did not exhibit any significant cytotoxic effects on the normal human cell line, and it had very low hemolytic activity with flounder hemoglobin. The results demonstrated that peptide purified from the hemocyte of a M. coruscus exhibits antibacterial activity against various bacteria and has the potential to be an alternative antibiotic agent.

• Journal of fish pathology
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• v.18 no.3
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• pp.247-258
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• 2005

To understand responses of bivalve tissues to various injurious agents, histopathological observation was done in the Manila clam, Ruditapes hilippinarum, during 240 hours after an intramuscular injection of N-methyl-N-nitro-N-nitrosoguanidine (MNNG). Some histopathological changes were observed with the epithelia of stomach, midgut, hindgut, digestive diverticula and gonads after injecting MNNG. The earliest degenerative changes were recognized in the epithelium of midgut 12 hours after the injection. Partial or complete, epithelial destruction was constantly accompanied by the massive infiltration of a mononuclear, necrotic cell group including hemocytes. At 144 hours later, gonads showed the necrotic desquamation of their germinal epithelia. Nearly all of the infiltrated cells within the destructive epithelium were suggested to be hemocytes, from the binding property with a hemocyte-specific lectin, Ricinus communis (RCA-1). From these results it was concluded that the epithelium of digestive system is vulnerable to parenteral MNNG and hemocytes might be deeply involved in this MNNG-induced destruction of the digestive epithelium.

• Proceedings of the Korean Society of Fisheries Technology Conference
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• 2001.05a
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• pp.230-231
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• 2001

지금까지 많은 항균성 펩타이드가 포유류 (1), 양서류 (2), 곤충 (3) 및 식물 (4)로 부터 단리ㆍ정제되었고, 구조-활성간의 연구가 활발히 진행되고 있지만, 이러한 연구는 육상생물에 제한적으로 수행되어 겼으며 해양생물을 대상으로 한 연구는 극히 제한되어 있는 상태이다. 해양생물로부터 정제된 항균성 펩타이드로서는 Flounder skin 분비물에서 pleurocidin (5), Moses sole fish에서 pardaxins (6), Horseshoe crab에서 Tachyplesin I(7), Tunicates hemocytes에서 Clavanins (8), Mollusc에서 Mytilins (9) 등이 보고되고 있지만 불가사리를 이용한 항균성물질의 연구는 희박한 실정이다. (중략)

• Fisheries and Aquatic Sciences
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• v.5 no.4
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• pp.263-270
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• 2002

The Pacific oysters, Crassostrea gigas, were stressed with different concentrations of benzo(a) pyrene and depurated to determine the hemocyte lysosomal membrane stability and hydrolytic enzymatic activity as a biomarker candidate to the chemical, using NRR (neutral red retention) and API ZYM System, respectively. The membrane damage measured as NRR decrease was significant with the increase of chemical concentration and exposure time (P<0.05), providing a possible tool for biomarker. Interestingly, the control showed intrinsic stress probably due to captive life in the laboratory, and a recovering trend was also found during the depuration. The benzo(a)pyrene-exposed oysters showed increased enzyme activities in alkaline phosphatase, esterase (C4), acid phosphatase, naphthol-AS-BI-phospho­hydrolase, $\beta$-galactosidase, $\beta$-glucuronidase, and N-acetyl- $\beta$-glucosaminidase. Of them, only two enzymes, acid phosphatase and alkaline phosphatase, showed some potential available for the generation of enzymatic biomarker in the oyster. The results are suggestive of the potential availability of the cellular and enzymatic properties as a biomarker. However, considering that a robust biomarker should be insensitive to natural stress coming from normal physiological variation, but sensitive to pollutants, a concept of intrinsic stress the animal possesses should be taken into consideration. This reflects the necessity of further research on the intrinsic stress affecting the cellular and enzymatic properties of the chemical­stressed oysters prior to using the data as a biomarker.