• Biotechnology and Bioprocess Engineering:BBE
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• v.3 no.2
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• pp.87-90
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• 1998

Hemolymph is oxidized and darkens visibly during the collection from silkworm due to the activity of tyrosinase in it. Toxic quinones are produced by the oxidation and consequently inhibit the cell growth. Heat treatment can be used to prevent the oxidation ; however, the oxidation may occur during the collection of hemolymph before it is heat-treated. It makes the hemolymph collection difficult especially on a large-scale preparation. Hemolymphs collected from 257 different strains of silkworms were examined to select the slowly oxidized hemolymphs. Hemolymphs collected from mutant strains such as Lemone, TBO, Cre, Y4, and wEb showed relatively slow color changes. Oxidation rates of the hemolymphs were measured by the absorbance change using a spectrophotometer. The hemolymph of wEb showed the slowest oxidation. The absorbance of this mutant hemolymph reached the saturation value at 20$^{\circ}C$ in 450 min, whereas the total oxidation time of the wild-type (Baekokjam) hemolymph at the same temperature was 120 min. We tested if this mutant hemolymph is useful as a medium supplement for insect cell culture. Cell growth rate and final cell concentration in the medium supplemented with the wEb hemolymph were almost same as those in the medium supplemented with the wild-type hemolymph. Hemolymph is collected on a small scale by clipping the abdominal leg; however, this method is not appropriate fro large scale preparation. Centrifugation after chopping the silkworm hemolymph by a blending mixer is a more appropriate procedure for large scale collection. Slowly oxidized wEb hemolymph resulted in higher cell concentration than the wild-type hemolymph when hemolymph was collected by the large scale preparation method.

• 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.

• International Journal of Industrial Entomology and Biomaterials
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• v.40 no.1
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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.

• Biotechnology and Bioprocess Engineering:BBE
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• v.10 no.4
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• pp.353-356
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• 2005

The effect of silkworm hemolymph on the expression of recombinant protein in Escherichia coli was investigated. The addition of silkworm hemolymph to the culture medium in­creased the production of recombinant $\beta$-galactosidase in E. coli. The production was dependent on the concentration of the added silkworm hemolymph, which increased 2-, 5-, and 8-fold in media supplemented with 1, 3, and $5\%$ silkworm hemolymph, respectively. To identify the effective component, the silkworm hemolymph was fractionated by gel filtration column chromatography. A fraction, with a molecular weight of about 30 K was identified as the effective component.

• International Journal of Industrial Entomology and Biomaterials
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• v.47 no.2
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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.

• Korean Journal of Veterinary Research
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• v.39 no.5
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• pp.855-864
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• 1999

Hemal nodes and hemolymph nodes are lymphoid organs that share morphologic and functional characteristics of lymph nodes and spleens. The aim of the present study was to obtain new informations on the distinct morphological structures of hemal nodes and hemolymph nodes according to ages, and to get the basic data for their functions in Korean native goats. Goats were divided into 5 groups, consisting of 3 animals aged 1, 3, 6, 10 and 12 months, respectively. Ultrastructural features of the organs were observed by transmission and scanning electron microscopes. The sinuses of hemal nodes and hemolymph nodes were lined by endothelial-like reticular cells which had euchromatin-rich nuclei and many cytoplasmic processes, surrounding collagen fibrils. Macrophages containing phagocytosed erythrocytes were often noted in the diffuse lymphatic tissues of hemal nodes and hemolymph nodes. Some mast cells were in contact with the plasma cells near the blood vessel. Hemal nodes and hemolymph nodes had venous sinusal-like vessels which were different from the deep sinus. The lymph vessels with valves were observed in the capsule of the hemolymph node. There were no ultrastructural differences of the organs in the age different groups of the animals. These results suggest that hemal nodes and hemolymph nodes may take part in hemopoiesis, blood filtration and immune reaction in Korean native goats.

• Korean Journal of Veterinary Research
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• v.37 no.4
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• pp.727-734
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• 1997

Hemolymph nodes of the Korean native cattle were observed gross anatomically and light microscopically in this study. Hemolymph nodes of the Korean native cattle were found mainly at the periphery of the thoracic and abdominal aortae although there were some other parts to observe these small organs. These organs were small in size and spherical or ovoid in shape, and color of them was dark red or gray in common with red band. The hemolymph nodes were surrounded by a relatively thick connective tissue capsules composed of dense collagen fibers with many scattered smooth muscle fibers and there was a hilus in each node. The parenchyma was divided into an outer cortex and an inner medulla. A few primary and secondary lymph nodules were observed in the cortex and there were diffuse lymphatic tissues among them. The medullary cords consisted of diffuse lymphatic tissues were separated by the medullary sinuses which filled with erythrocytes. The afferent and efferent lymph vessels were observed at the periphery of the capsule and the hilus area, respectively. The stroma of the hemolymph nodes was composed of reticular cells and fibers, and the meshwork of the stroma was filled with many lymphocytes and erythrocytes, and a few macrophages and megakaryocytes etc. These findings suggest that the hemolymph node of the Korean native cattle is involved in blood filtration and immune reaction.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.45 no.2
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• pp.154-160
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• 2012

This study the effects of Cu exposure on biochemical factors in the hemolymph and hepatopancreas of the abalone $Haliotis$ $discus$ $hannai$. Abalone were exposed to 0, 5, 10, 20 and 40 ${\mu}g/L$ Cu for 4 weeks. The calcium concentrations in hemolymph were decreased significantly on exposure to 20 and 40 ${\mu}g/L$ Cu after 2 weeks. The aspartate aminotransferase (AST), alanine aminotransferase (ALT) and alkaline phosphatase (ALP) activities in abalone hemolymph were markedly elevated after exposure to 40 ${\mu}g/L$ Cu for 4 weeks. The hepatopancreas superoxide dismutase (SOD) and catalase (CAT) activities were also significantly increased by exposure to 20 ${\mu}g/L$ Cu for 4 weeks. These biochemical factors may represent a convenient method of monitoring heavy metal pollution in coastal areas. From these results, we conclude that a high copper concentration (40 ${\mu}g/L$) in water may curtail hemolymph homeostasis and anti-oxidative reactions in abalone.

• Journal of Sericultural and Entomological Science
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• v.41 no.2
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• pp.82-86
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• 1999

Hemolymph proteins and vitellogenin from mulberry longicorn beetle, Apriona germari HOPE, were indentified, and their changed were analyzed during the larval-pupal-adult development and in the newly laid eggs. Thres major hemolymph proteins were observed in the hemolymph during the larval-pupal-adult development and the intensity of their proteins was clearly observed during the pupal stage. From SDS-[olyacrylamide gel electrophoresis analysis, molecular weights of three major hemolymph proteins were approximately 74 kDa, 78kDa and 85kDa. Vitellogenin in A. Germati appeared in the hemolymph of only abult female and is considered to be a product synthersized within 10 days after adult emergence. The molecular weight of vitellogenin was consited of a heavy subunit (165 kDa) and a light subunit (40 kDa).

• Journal of Microbiology and Biotechnology
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• v.11 no.5
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• pp.853-857
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• 2001

The effect of silkworm hemolymph on the inhibition of baculovirus-induced insect cell apoptosis was quantitatively investigated using a flow cytometric analysis. Spodoptera frugiperda (Sf9) cell and Autographa californica nuclear polyhedrous virus (AcNPV) were used as a model for insect cell and baculovirus in this study, respectively. Compared with a mammalian cell cycle, the fraction of G1 cells was relatively small in the Sf9 cell cycle. Silkworm hemolymph did not affect the Sf9 cell-cycle distribution before the baculovirus infection. However, the fraction of cells which are not in the sub-G1 phase remained at a high level for 3 days after the infection in the medium without silkworm hemolymph, while it remained at a high level for 7 days after the infection in the medium supplemented with silkworm hemolymph. The fractions of apoptotic cells in the sub-G1 phase were $4.7\%$, and 4 days after infection, $22.7\%$, in the media with and without silkworm hemolymph, respectively.