• International Journal of Industrial Entomology and Biomaterials
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• 제40권1호
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• pp.16-21
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• 2020

Humans use insects as food and traditional medicine for many years. Hemolymph is the circulating fluid of insects and is a key component of their immune system. However, limited information is available regarding hemolymph identification, development, and differentiation, as well as the related cellular immune responses. In a previous study, hemolymph extracts prepared from Bombyx mori larvae were found to exert anti-inflammatory effects. In this study, we aimed to identify and compare the antioxidant activity of immune-challenged and unchallenged B. mori hemolymph extracts in vitro. For this purpose, human epithelial Caco-2 cells were first exposed to oxidative stress and then treated with various concentrations and incubation times of either immune-challenged or unchallenged B. mori hemolymph extracts. Next, we determined the effect of treatment on the relative expression of GPX-1, SOD-1, and SOD-2 antioxidant marker genes. We found that the expression rates of the three marker genes were markedly higher at a immune-challenged hemolymph extract concentration of 80 ppm compared to those at other concentrations, and the antioxidant effects were enhanced after treatment for 48 hr. Thus, B. mori hemolymph extracts showed antioxidant activity within the limited time and dose. Especially, the immune-challenged B. mori hemolymph extracts showed higher the antioxidant activities than unchallenged one. The activity of silkworm hemolymph extracts could facilitate the development of new types of functional foods, feed additives, and biomaterials with antioxidant properties.

• International Journal of Industrial Entomology and Biomaterials
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• 제47권2호
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• pp.134-139
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• 2023

SIRT5 and PRDx1 play crucial roles in cancer and are involved in the basic mechanisms of reactive oxygen species detoxification. In our previous studies, we showed that hemolymph extracts of immune-challenged Bombyx mori have antioxidant properties. Following H₂O₂ stimulation, immune-challenged B. mori hemolymph extracts elicited SIRT5 downregulation activity, reaching effective activity at the highest concentration of 100 ppm. Additionally, cells treated with immune-challenged B. mori hemolymph extracts demonstrated increased PRDx1 mRNA expression compared to that of PBS-treated cells. Therefore, immune-challenged B. mori hemolymph extracts offer a potential auxiliary means of treating drug-resistant tumors through downregulation of SIRT5 and upregulation of PRDx1 expression. Nevertheless, further studies on the effects of B. mori hemolymph on SIRT5 and PRDx1 regulation are pertinent for using it as a food or pharmaceutical material and understanding its therapeutic effect on tumors, including those that are drug-resistant.

• Animal cells and systems
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• 제2권1호
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• pp.107-111
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• 1998

Two kinds of humoral factors were observed in 2 orders, 7 families, 10 genera, and 15 different species of Korean ascidians. They are the naturally occuring hemagglutinins and/or hemolysins against human erythrocytes A, B, and 0. All but two species showed aggregative activity, although there were considerable variations in titer. The weak agglutinating and lytic activities were increased in the presence of $Ca^{++}$. Much higher activities of agglutination and/or lysis were shown in the hemolymph than extracts from tissues, and a higher response was shown in adults than in juveniles. No distinct differences from collected locations were observed. The hemolymph of Ciona intestinalis showed a strong hemolytic (cytotoxic) and weak agglutinins capacities. In addition, hemolymph of Styela plicata and Styela clava clava also showed hemoagglutining and hemolytic activities. Botryllus tuberatus had hemagglutining and weak lytic activities. Other species showed only hemagglutining activity. These agglutining activities are probably responsible for carbohydrate recognition in solitary or colonial ascidian. The lytic activity is probably responsible for antibacterial defense and nonfusion reactions between allogeneic colonial ascidians, especially the genus of Botryllus. The occurrences of humoral factors in ascidians were independent of their geographic distributions.

• 한국동물학회지
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• 제37권1호
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• pp.76-87
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• 1994

High density lipophorin-L (HDLP-L) was purified from the hemolymph of Bombyx modi using KBr density gradient ultracentrifugation and gel permeation chromatographv (Sephadex G-2001. Lipophorin has native molecular weight of 730 Nd and consists of Apo-Lp I and Apo-Lp II with molecular weights of 250 Kd and 90 Kd, respectively. Lp contains large amounts of glutamine & glutamic acid, threonine, leucine but small amounts of cysteic acid & oxidized cystine, tyrosine, methionine. Lp also contains diacylglycerol, cholesterol, phosphatidylcholine, and phosphatidylethanolamine. Anti-lipophorin showed positive reaction with fat body and ovarial extracts and also revealed immunological identity with lipophorin of Fall webworm, Hyphantria cunea. Lipophorin maintains constant level during larval and pupal stapes but greatlv increases during adult stage in both male and female. Apo-Lp III was purified from adult hemolymph. Hemolymph was subjected to KBr ultracentrifusation and Lp-free fraction was submitted to cation exchange chromatosraphy after ammonium sulfate precipitation. Apo-Lp III has molecular weight of 17 Kd, and similar amino acid composition ar those of other species Lp but contains high amounts of tryptophan which other are tacking in.

• International Journal of Industrial Entomology and Biomaterials
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• 제2권2호
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• pp.167-172
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• 2001

The vitellin of firefly, Pyrocoelia rufa, is composed of three polypeptides, designated Vn1 (175 kDa), Vn2 (160 kDa) and Vn3 (45 kDa) in SDS-polyacrylamide gel electrophoresis. Three subunits of vitellin were presented in the female adult hemolymph, ovary and egg extracts, but not observed in the male. This vitellin was purified from the eggs of P. rufa by the FPLC techniques, anion exchange chromatography and gel permeation chromatography. In nature, vitellin of P. rufa has molecular weight of 400 kDa. Western blot analysis using polyclonal antiserum against purified vitellin showed that the antiserum was reacted with the three polypeptides, Vnl, Vn2 and Vn3 from the female adult hemolymph, ovary and egg extracts. Amino acid residues at N-terminus of three subunits were sequenced. The N-terminal sequences of large subunits, Vnl and Vn2, were similar to each other, But, the N-terminal sequences of small subunits Vn3, did not have any signnificant homology with large subunits.

• 한국응용곤충학회지
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• 제15권4호
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• pp.169-178
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• 1976

솔잎혹파리 (Thecodiplosis japonensis Uchi. et Inouye)의 충영속 유충과 충영을 탈출하여 낙하한 토중의 유충을 대상으로 체액을 분석, 측정코저 종이 크로마토그라피법, micro-Kjeldahl법, 박층 크로카토그라피 법, 왈부르그 검압계법, Bessey-Lowry법과 Reitman-Frankel법으로 측정 또는 분석하였다. 충영과 유충과 토중의 유충에 따라 유리 아미노산, 총 질소함량의 변화, 탄수화물의 변화, 지질의 종류와 변화, 호흡능, phosphatase, GOT, ?의 활성도가 측정되었다. 유리 아미노산, 총 질소함량, 지질함량, 호흡능, acid phosphatase의 황성, GPT의 활성은 충영속의 유충에서 토중의 유충으로 진행함에 따라 감소하는 경향을 나타냈다. 그러나 trehalose의 함량과 alkaline phosphatase의 활성은 증가하는 경향을 나타냈다.

• International Journal of Industrial Entomology and Biomaterials
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• 제1권2호
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• pp.131-135
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• 2000

The vitellin of the fireflies, Luciola unmunsana and L. lateralis was characterized. The vitellin of L. unmunfon is composed of two subunits, designated Vnl (195 kDa) and Vn2 (185 kDa) in SDS-polyacryamide gel electrophoresis. These two subunits of vitellin of L. unmunsana gradually decreased during embryogenesis. As expected, these protein bands were presented in female adult hemolymph and egg extracts, but not in male. The vitellin of L. lateralis is also composed of two subunits, designated Vnl (195 kDa) and Vn2 (180 kDa) in SDS-PAGEi and these two protein bands gradually decreased during embryogenesis. Western blot analysis using each of polyclonal antiserum against vitellins of L. unmunsana and L. lateralis showed that two antisera strongly crossereacted with vitellin subunits of L. unmunsana and L. lateralis, suggesting that vitellins of L. unmunsana and L. lateralis have similarity with each other.

• International Journal of Industrial Entomology and Biomaterials
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• 제2권1호
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• pp.55-59
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• 2001

The vitellin of the red flour beetled Tribolium castaneum Herbst was purified and characterized. The vitellin of T. castaneum was purified by the FPLC techniques, anion exchange chromatography and gel permeation chromatography. In native-polyacrylamide gel electrophoresis, vitellin of T. castaneum was detected as a single band. This native vitellin has molecular weight of 440 kDa. The vitellin of T. castaneum is composed of three polypeptides, designated Vnl (178 kDa), Vn2 (168 kDa) and Vn3 (52 kDa) in SDS-polyacrylamide gel electrophoresis. Three subunits of vitellin were presented in the female adult hemolymph and egg extracts, but not observed in the male. These three polypeptides gradually decreased during embryogenesis. Polyclonal antiserum raised against purified vitellin reacted with the three polypeptides, Vnl, Vn2 and Vn3. Antisera raised against Vn1 and Vn2 cross-reacted with the two large subunits, Vnl and Vn2, respectively. Another subunits Vn3, however, was not cross-reacted with these two antisera. Also, antiserum raised against Vn3 did not cross-react with the Vn1 and Vn2.

• International Journal of Industrial Entomology and Biomaterials
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• 제17권2호
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• pp.217-221
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• 2008

Dung beetle larvae at the $3^{rd}$ instar were injected with lipopolysaccaride and inducible proteins were examined within a pI level of 3-10 and a size level by proteomics, including 1-D SDS PAGE analysis and antibacterial assay. The immune infected larvae extracts provided seven protein bands in one-dimensional electrophoresis and its antibacterial activity also checked. Hemolymph protein from immune infected larvae of the dung beetle were separated by twodimensional gel electrophoresis and compared with those from native larvae. In 2-D gel electrophoresis, we detected 63 immune infected unique and 32 up-regulated proteins, and 36 proteins that were down-regulated or not present in treated gel. Ten protein spots from unique proteins and those presented as different level of abundance in infected and native larvae were specially expressed. These differentially expressed proteins were proposed to be involved in the defense mechanism against microorganism.

• International Journal of Industrial Entomology and Biomaterials
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• 제37권2호
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• pp.95-99
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• 2018

In insect defense system, antimicrobial peptides (AMPs) are one of important biological molecules to survive in a variety of environments. Insect can synthesize AMPs to protect against invading pathogens in humoral immune response. Taking more advantage of biological antimicrobial molecules, we report antibacterial activity of two cecropin type peptides, cecropin and moricin, isolated from the silkworm against four salmonella species. In this work, we purified antimicrobial candidate peptides (AMCP) from the extracts of immune challenged silkworm larval hemolymph by two-step chromatographic purification procedure, cation exchange and gel permeation chromatography. The molecular weights of purified peptides were estimated to be about 4 ~ 5 kDa by Tricin SDS-PAGE analysis, and identified as silkworm cecropin and moricin by NCBI BLAST homology search with their N-terminal amino acid sequences. As antibacterial activity assay, the purified peptides showed stronger antibacterial activity against Salmonella pathogens with an MIC value of $1{\sim}4{\mu}g/mL$. Therefore two cecropin type peptides purified from the silkworm will be valuable potential materials for development of new natural antibiotics.