• 한국응용곤충학회지
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• 제51권4호
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• pp.349-356
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• 2012

아이코사노이드는 곤충의 다양한 세포성 면역 반응을 중개한다. 본 연구는 면역반응에 따라 혈구세포 밀도 변화에 대한 아이코사노이드의 새로운 중개 기능을 밝히기 위해 수행되었다. 파밤나방(Spodoptera exigua) 5령충은 세균 감염에 따라 2 시간이 지나면 총혈구수의 현격한 증가를 보였다. 이 총혈구수 증가는 주로 부정형혈구와 소구형혈구 밀도의 증가로 해석되었다. 파밤나방 유충에 phospholipase $A_2$ ($PLA_2$) 억제자인 dexamethasone을 처리하면 세균 처리에 의한 총혈구수 변화가 일어나지 않았다. 하지만 dexamethasone을 처리한 유충에 $PLA_2$의 촉매산물인 arachidonic acid를 첨가하면 총혈구수 증가가 회복되었다. 이러한 혈구 밀도 변화에 원인으로서 아이코사노이드 종류를 추적하기 위해 cyclooxygenase (COX)의 억제자인 naproxene을 처리한 결과 총혈구수 증가가 억제되고, lipoxygenase (LOX)의 억제자인 esculetin을 처리하면 총혈구수 증가가 유지되어 COX 산물이 세균 침입에 따른 총혈구수 증가에 관여하는 것으로 나타났다. COX의 생산물인 prostaglandin $E_2$ ($PGE_2$)를 세균 없이 단독으로 처리할 때도 총혈구수의 뚜렷한 증가를 나타냈다. 이러한 결과는 파밤나방의 세포성 면역반응 과정에서 총혈구수 증가를 중개하는 아이코사노이드의 새로운 기능을 제시하고 있다.

• 한국어업기술학회:학술대회논문집
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• 한국어업기술학회 2000년도 춘계수산관련학회 공동학술대회발표요지집
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• pp.449-450
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• 2000

이매패의 hemocytes는 상처 치유, shell repair, 영양물질의 소화, 수송 그리고 분비에 관여한다고 알려져 있으며, 체내 방어 기능에서 필수적이다(Carmen et al., 1997). 패류의 hemocytes에 관해서는 주로 형태, 세포화학적 특징이 연구되어 왔으며(Pipe, 1990 등), 밀도차원심분리법(Bachere et al., 1988)으로 분리된 hemocytes를 면역학적 방법에 의해(Cheng et al., 1980; Yoshino and Granath, 1983) 여러 가지 subpopulatione으로 구분하지만, 현재로서는 같은 종의 패류에서도 분류 방법에 따라 다양하게 분류되고 있다. (중략)

• 한국수산과학회지
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• 제39권spc1호
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• pp.123-131
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• 2006

Immmune parameters of the Manila clam Ruditapes philippinarum collected from four tidal flats, Nudong, Gonam, Hwangdo and Bangpo on Anmyeon-do, Korea were optimized and evaluated at the single cell level using flow-cytometry Hemocytes were withdrawn from the sinus of each clam, and total hemocyte counts (THC), phagocytosis rate, hemocyte mortality (HM) and DNA damage of hemocyte were analyzed. The highest hemocyte counts was recorded from the clams collected from Gonam, followed by Hwangdo, Nudong and Bangpo (P<0.001). Phagocytosis rate and hemocyte mortality of Gonam and Nudong clams were significantly higher than those of clams from Hwangdo and Bangpo (P<0.001). DNA damage in the clams from Nudong was higher twice than that of clams from Gonam (P<0.05). We suggest that the flow-cytometry has a high potential for evaluation of immunity of marine bivalves.

• International Journal of Industrial Entomology and Biomaterials
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• 제23권1호
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• pp.137-141
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• 2011

A previous data was provided information for tissuespecific expression genes by means of whole-genome oligonucleotide microarray in the silkworm. We analyzed the tissue-specific expression patterns in the hemocyte tissue on 5 days of 5th instar larvae during the development of $B.$ $mori$. Total 5 candidates pick out from the $Bombyx$ $mori$ Microarray Database (BmMDB; http://silkworm.swu.edu.cn/microarray). To verify the hemocyte-specific expression, we analyzed by semi-quantitative and real-time quantitative RT-PCR using the highly expressed endogenous $Actin$ RNA as an intrinsic reference. In this study, we confirmed that one gene-sw17255- out of 5 candidates expressed in the hemocyte tissue, which was consistent with the previous data. Circulating hemocytes in the body fluid of the $B.$ $mori$ are most powerful target organ for producing biomaterials. We need further studies to find hemocyte-specific promoter region from sw17255 gene. Finally, this result can be applied in creating transgenic silkworms as a biomedical insect.

• International Journal of Industrial Entomology and Biomaterials
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• 제5권2호
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• pp.189-194
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• 2002

Silkworm diseases are major constraint in silk cocoon production. Among silkworm diseases, white muscar-dine is highly contagious and most common in winter and rainy seasons. It is suggested that hemocytes involve in defense against invasion of Beauveria bassi-ana and systemic fungicide/chemicals prevent the proliferation of fungi in the hemolymph or preventing the growth of the fungi in the body cavity through enhancing the hemocyte mediated defense response. In the present study the influence of systemic fungicide on hematological changes in silkworms infected with Beauveria bassiana was reported. It is observed that the total hemocyte counts increased in the hemolymph up to 5 days post inoculation in systemic fungicide treated batches while in the inoculated control the increase was up to 3 days indicating the positive hemocyte mediated response in silkworm treated with systemic fungicide. After 2 days in the inoculated control as the multiplication and growth of mycelia increased, defense capacity of the silkworm was decreased. The biochemical changes were also observed in the hemolymph of silkworm infected with B. bassiana. In silkworm infected with Beauveria bassi-ana, the total protein content increased whereas total carbohydrate and total lipids decreased as the infection progresses. In the case of systemic fungicide treated batches the increase in total protein content was comparatively higher and decrease in total carbohydrate and lipids were comparatively lower than the inoculated control.

• International Journal of Industrial Entomology and Biomaterials
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• 제6권1호
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• pp.11-14
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• 2003

Systemic fungicides viz., Bavistin and Beyleton are reported to have curative effect against the infection of Beauveria bassiana in silkworm Bombyx mori L. To understand the influence of the systemic fungicides on the disease suppression/development, hematological studies were carried out. There was an increase in the percent total hemocyte count, granulocyte and plasmatocyte in silkworm treated with the systemic fungicides. It possibly indicates the influence of systemic fungicide on the hemocyte mediated defense system leading to the higher resistance and suppression of disease development.

• Parasites, Hosts and Diseases
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• 제49권3호
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• pp.229-234
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• 2011

In order to assess changes in the activity of immunecompetency present in Crassostrea gigas infected with Marteilioides chungmuensis (Protozoa), the total hemocyte counts (THC), hemocyte populations, hemocyte viability, and phagocytosis rate were measured in oysters using flow cytometry. THC were increased significantly in oysters infected with M. chungmuensis relative to the healthy appearing oysters (HAO) (P<0.05). Among the total hemocyte composition, granulocyte levels were significantly increased in infected oysters as compared with HAO (P<0.05). In addition, the hyalinocyte was reduced significantly (P<0.05). The hemocyte viability did not differ between infected oysters and HAO. However, the phagocytosis rate was significantly higher in infected oysters relative to HAO (P<0.05). The measurement of alterations in the activity of immunecompetency in oysters, which was conducted via flow cytometry in this study, might be a useful biomarker of the defense system for evaluating the effects of ovarian parasites of C. gigas.

• Journal of Microbiology and Biotechnology
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• 제19권6호
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• pp.610-615
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• 2009

As a provirus, polydnavirus has a segmented DNA genome on chromosome(s) of host wasp. It contains several genes in each segment that presumably play critical roles in regulating physiological processes of target insect parasitized by the wasp. A cysteine-rich protein 1 (CRP1) is present in the polydnavirus Cotesia plutellae bracovirus (CpBV) genome, but its expression and physiological function in Plutella xylostella parasitized by the viral host C. plutellae is not known. This CpBV-CRP1 encoding 189 amino acids with a putative signal peptide (20 residues) was persistently expressed in parasitized P. xylostella with gradual decrease at the late parasitization period. Expression of CpBV-CRP1 was tissue-specific in the fat body/epidermis and hemocyte, but not in the gut. Its physiological function was analyzed by inducing transient expression of a CpBV segment containing CpBV-CRP1 and its promoter, which caused significant reduction in hemocyte -spreading and delayed larval development. When the treated larvae were co-injected with double-stranded RNA of CpBV-CRP1, the expression of CpBV-CRP1 disappeared, whereas other genes encoded in the CpBV segment was expressed. These co-injected larvae significantly recovered the hemocyte-spreading capacity and larval development rate. This study reports that CpBV-CRP1 is expressed in P. xylostella parasitized by C. plutellae and its physiological function is to alter the host immune and developmental processes.

• BMB Reports
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• 제31권4호
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• pp.364-369
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• 1998

Insect plasma protein is abundant in the hemolymph of holometabolous insect larvae and is used as a source of amino acids and energy for construction of adult structures during metamorphosis. In order to understand the mechanism of decomposition of larval plasma proteins by hemocyte protease, we tried to purify a cysteine protease from the hemocyte lysate by using Carbobenzoxy-L-Phenylalanyl-L-Arginine-4-Methyl-Coumaryl-7-Amide (Z-Phe-Arg-MCA) as substrate and to identify plasma proteins that are selectively susceptible to the purified protease. Here, we describe the purification and characterization of a cysteine protease that specifically hydrolyzes the plasma protein of the coleopteran insect, Tenebrio molitor, larvae. The molecular mass of this enzyme was 25 kDa, as determined by SDS-PAGE under reducing conditions. The amino acids sequence of its $NH_{2}-terminus$ was determined to be Leu-Pro-Gly-Gln-Ile-Asp-Trp-Arg-Asp-Lys-Gly. This sequence contained Pro, Asp, and Arg residues, conserved in many papain superfamily enzymes. The specific cysteine protease inhibitors, such as E-64 and leupetin, inhibited its hydrolytic activity. One plasma protein with a molecular mass of 48 kDa was selectively hydrolyzed within 3 h when the purified enzyme and plasma proteins were incubated in vitro. However, the 48 kDa protein was not hydrolyzed by the purified 25 kDa protease in the presence of E-64. Western blotting analysis at various developmental stages showed that the purified enzyme was detected at larvae, pupae, and adult stages, but not the embryo stage.

• Fisheries and Aquatic Sciences
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• 제6권2호
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• pp.74-80
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• 2003

Two cellular biomarker parameters of the farmed Pacific oyster Crassostrea gigas were studied in vivo and in vitro after exposure to concentrations of polychlorinated biphenyls in terms of neural red uptake (NRU) and lysozyme activity. The oysters exposed in vivo to the xenobiotic concentrations, 0, 30, 90, and 180 ng/g for 14 days, enhanced hemocyte NRU with occasional significant differences (P<0.05), depending on the chemical concentration and duration. An adverse tendency was manifest in the lysozyme activities both in the hemocyte and serum of the oyster treated with the chemical in a same manner, rendering these two cellular parameters as biomarker candidates against the chemical. The oysters exposed in vitro to the chemical concentrations, 0, 1, 5, 10, 100, 1,000, and 10,000 ng/g for 24 hrs at $10^{\circ}C$ showed a similar tendancy as those exposed in vivo to the chemical. Unlike in vivo response, however, the in vitro NRU was first influenced by very low concentration of the chemical. In in vitro results, marked but not significant increase of hemocyte NRU was noticed at the chemical concentration of 5 ng/g, where the value was almost as high as those exposed to higher chemical concentrations, up to 10,000 ng/g. An unusual result was observed in the in vitro lysozyme activity of hemocyte in which significant decrease was first noticed at the chemical concentration of 100 ng/g.