• Journal of Sericultural and Entomological Science
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• v.52 no.2
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• pp.159-164
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• 2014

In this study, functionality of products of silkworm hemolymph fermented by Bacillus species was studied such as cell viability, antioxidant effect, and inhibitory effect on tyrosinase activities. A matured silkworm hemolymph was degraded by fermentation with Bacillus subtilis 10854 and Bacillus amyloliquefaciens M27. Especially, proteins of matured silkworm hemolymph were degraded to 3,000 Da by fermentation with B. amyloliquefaciens M27. Cell viability for MTT assay was higher than PBS in hemolymph and fermented hemolymph. A DPPH free radical scavenging activity of fermented hemolymph was as higher as Vitamin C and dependent on sample concentrations. Thus, these results suggest that degraded hemolymph fermented by B. amyloliquefaciens M27 may have antioxidant properties as a material for cosmetics.

• Biotechnology and Bioprocess Engineering:BBE
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• v.11 no.4
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• pp.364-367
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• 2006

In order to enhance the repair of defects in articular cartilage via cell therapy with autologous chondrocytes, as well as with periosteum-derived progenitor cells (PDPCs), silkworm hemolymph (SH) and a variety of cartilage-specific extracellular matrices (ECMs) including type II collagen, proline, chondroitin 4-sulfate, and chondroitin 6-sulfate were assessed with regard to their efficacy as media supplements. SH, a known anti-apoptotic agent, was found to enhance cell growth, as was shown by the results of a 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyltetrazolium bromide (MTT) assay. According to the results of reverse transcriptase polymerase chain reaction (RT-PCR) analyses, the cartilage-specific ECMs were found to stimulate the expression of hyaline cartilage-specific genes, most notably type II collagen and Sox9, in monolayer cultures of PDPCs.

• Journal of Nutrition and Health
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• v.51 no.3
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• pp.201-207
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• 2018

Purpose: Obesity, a worldwide epidemic, is associated with insulin resistance, hyperlipidemia, hypertension, cardiovascular disease, and certain cancers. Many strategies, including natural alternative anti-obesity agents, are used widely to prevent obesity. This study examined the effects of silkworm hemolymph on the weight control of C57BL/6N mice fed with a high-fat diet. Methods: The mice were divided into five groups: normal group (N), high-fat diet group (HFC), high-fat diet and silkworm hemolymph (at dose of 1 mL/kg BW (HFS-1), 5 mL/kg BW (HFS-5) and 10 mL/kg (HFS-10) for 12 weeks. Results: After 12 weeks treatment, the administration of silkworm hemolymph decreased the final body weight significantly along with a decrease in the weights of epididymal fat and total fat. The plasma LDL-cholesterol concentration was significantly lower in the HFS-1, HFS-5, and HFS-10 groups than in the HFC group. In addition, the leptin level of the HFS groups was significantly lower than those of the HFC group without a change in the plasma insulin concentration. Conclusion: These findings suggest that the silkworm hemolymph may have the potential to prevent obesity.

• International Journal of Industrial Entomology and Biomaterials
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• v.24 no.2
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• pp.69-74
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• 2012

Tropical tasar silkworm Antheraea mylitta Drury experiences extreme temperature stress conditions during its life cycle particularly during diapauses and first crop. The present study witnessed the impact of high temperature on some biochemical profiles and egg production (fecundity) of semi-domesticated Daba and Shorea robusta (Sal) based wild ecorace Laria during seed cocoon (pupa) preservation. Cocoons of Daba and Laria were treated with high temperature at $40^{\circ}C$ for 10 days in a BOD incubator. The protein profile and carbohydrate content in the hemolymph and fat body and total haemocyte count (THC) in the hemolymph of pupa were investigated. Further, the fecundity and fertility of egg was assessed. Significant increase in the protein concentration was observed in the hemolymph with reduction in the fat body (p<0.05). The difference in protein concentration was highly significant between the semidomesticated Daba and wild ecorace Laria (p<0.05). High pupal mortality (20%) and reduced fecundity (10-15%) in Daba was noticed compared to wild Laria. Also an increased THC (>28000) was recorded in Laria. The study infers the potentials of wild ecoraces in sustaining the extreme temperature conditions and need of adopting suitable package of practices for the preservation of diapause seed cocoons during extreme summer conditions. There is possibility to introgression thermal stress resistant traits in the semi-domesticated races of tasar silkworm by resorting to conventional breeding plans with wild races and keeping the thermal stress induced response as markers.

• International Journal of Industrial Entomology and Biomaterials
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• v.45 no.1
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• pp.17-21
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• 2022

The present study aimed to determine whether a hemolymph prepared from Antheraea yamamai larvae had the same biological activities using a Bombyx mori hemolymph prior to exposure to lipopolysaccharide (LPS) in order to induce an inflammatory response. The effects of the hemolymph were determined using a reverse transcription-quantitative polymerase chain reaction to assess the expression of pro-inflammatory molecules. The A. yamamai hemolymph exerted anti-inflammatory effects on LPS-activated human monocytic leukemia cells via Toll-like receptor (TLR) 4-mediated suppression, similar to the B. mori hemocyte extract. Treatment with the A. yamamai hemolymph significantly suppressed LPS-induced upregulated inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) mRNA expression at all tested concentrations compared with the control, similar to the B. mori immune-challenged hemolymph. Finally, the A. yamamai hemolymph, like the B. mori immune-challenged hemolymph, suppressed all of these concentrations in a dose-independent manner. These results demonstrate that the hemolymph of A. yamamai exhibited important biologically active substances. Further in-depth functional studies are required to fully understand the mechanisms underlying the biological activities of wild-type silkworm hemolymphs.

• Journal of Sericultural and Entomological Science
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• v.49 no.1
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• pp.28-32
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• 2007

This study was carried out to investigate the utilization of hemolymph of silkworm, Bombyx mori, as a substitute for fetal bovine serum(FBS) in the insect cell culture. Hemolymph is collected on a small scale by clipping the abdominal leg; however, this method is not appropriate for large scale collection. The hemolymph was collected from 5 th instar larva by centrifugation after cutting of the abdominal legs was more appropriate procedure for large scale collection. The cell growth in the medium supplemented with hemolymph(Baekokjam) collected in large scale was almost same as that in the medium hemolymph supplemented with hemolymph collected in small scale. However, the mutant($wE^b$) hemolymph collected in large scale was still less effective in the cell growth, as compared to the Baekokjam hemolymph collected in large scale. The optimum centrifugation condition for large-scale bleeding was 500 rpm and 15 min.

• Journal of Sericultural and Entomological Science
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• v.37 no.1
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• pp.52-56
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• 1995

To investigate the effect of hemolymph of silkworm larvae on the multiplication of Bombyx mori nuclear polyhedrosis virus (BmNPV) in BmN-4 cells, BmN-4 cells were infected with BmNPV, which were sequentially Heated with the hemolymph exracted from B. mori larvae. When the culture media TC-100 containing 3% fetal bovine serum was mixed with 10% hemolymph heated at 65$^{\circ}C$ for 30 minutes, the released polyhedra by multiplication of BmNPV in BmN-4 cells were increased more than those of non-treated. However, multiplication of BmNPV in BmN-4 cells treated with non-heated hemolymph was not effective, since non-heated hemolymph was toxic for the cell growth. The result of plaque assay showed that plaque forming units in BmN-4 cells treated with heated hemolymph are significantly increased, suggesting that efficiency of multiplication of BmNPV in BmN-4 cells is due to increase not of cell growth but of infectivity of BmNPV.

• 한국생물공학회:학술대회논문집
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• 2002.04a
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• pp.62-65
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• 2002

We have previously shown that the addition of silkworm hemolymph to a culture medium increases the longevity of insect and mammalian cells by inhibiting apoptosis. This indicates that the component which inhibits apoptosis is contained in the silkworm hemolymph, The apoptosis-inhibiting component was isolated from silkwonn hemolymph and characterized in our previous study. A database search using the N-terminal amino acid sequence of this component as a template resulted in a 95% homology with a low molecular weight lipoprotein, the so called ’30K protein' of unknown function. In this study, the 30K protein gene was expressed in mammalian and insect cells to confirm the apoptosis-inhibiting effect. The overexpression of 30K protein in mammalian cell inhibited the staurosporin-induced apoptosis by the prevention of the activation of caspase 3. Using an Autographa californicanuclear polyhedrosis virus (AcNPV) system, the 30K protein was overexpressed also in insect cells. The expression of the 30K protein increased the longevity of baculovirus-infected insect cells by inhibiting apoptosis. These results suggest that the 30K protein is a novel anti-apoptotic protein.

• Journal of Microbiology and Biotechnology
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• v.17 no.12
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• pp.1944-1948
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• 2007

Silkworm hemolymph (SH), prepared from fifth-instar larvae of Bombyx mori and heat-treated at $60^{\circ}C$ for 30 min, was used to improve cell viability and the production of human cytotoxic T-lymphocyte antigen 4-immunoglobulin (hCTLA4Ig) in transgenic Oryza sativa L. cell suspension cultures. Even though SH could not elevate cell viability at the concentrations up to 3% (v/v), addition of 0.3% (v/v) SH to a culture medium enhanced the production of hCTLA4Ig by 36.8% over an SH-free medium. Moreover, the production period of hCTLA4Ig could be shortened in a 0.3% (v/v) SH-added medium compared with that in an SH-free culture. As a result, addition of 0.3% (v/v) SH improved the productivity of hCTLA4Ig significantly in transgenic rice cell cultures.

• Journal of Sericultural and Entomological Science
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• v.30 no.1
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• pp.20-24
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• 1988

Adult specific proteins, referred to as ASP-I(adult specific protein of slow mobility) and ASP-II(adult specific protein of slow mobility) at the pharate adult stage of the silkworm, Bombyx mori, were detected by polyacrylamide gel electrophoresis. The adult specific proteins (ASP-I and ASP-II) were defined as a kind of adult hemolymph proteins without sex specificity, and the was no variation in the respective electrophoretic mobility of ASP-I and of ASP-II among the forty-one silkworm varieties. tested.