• Journal of Sericultural and Entomological Science
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• v.45 no.2
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• pp.103-107
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• 2003

Entomopathogenic nematodes were isolated through the investigation in soils collected from cultivated and non-cultivated fields using silkworms (Bombyx mori) and Galleria mellonella trap. The detectable rate of entomopathogenic nematodes of silkworms trap was higher than the G. mellonella trap. This study indicates the detection of entomopathogenic nematodes from soils that silkworms are sensitive superior to the G. mellonella to entomopathogenic nematodes. The steinernema, rhabditidae, and diplogatroidae strains successfully cultured on the silkworms host as well as on artificial media. Reproductivity in the living silkworm larva and pupa was 1.5 to 3.5${\times}$ 10$\^$5/ nematodes per host However, G. mellonella could be multiplied less than 5${\times}$l0$\^$5/ nematodes. The dried pupa of the silkworm following mositurize was cultured 0.5 to 2${\times}$10$\^$5/) nematodes per host. The culture methods of the steinernema, rhabditidae, and diplogatroidae strains, using silkworm powder, extracted chicken intestine, and food waste fertilizer could be applicative, but rate of reproduction was low.

• Journal of Oral Medicine and Pain
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• v.25 no.1
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• pp.53-62
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• 2000

Bone marrow culture systems are widely used to differentiate osteoclast-like cells in vitro using several osteotropic hormones. In this study, we isolated and cultured the mouse bone marrow cells with or without some osteotropic hormones such as parathyroid hormone(PTH), prostaglandin $E_2(PGE_2)$ and $l,25(OH)_2-vitamin$ $D_3$(Vit. $D_3$). We confirmed the formation of osteoclast-like cells morphologically and functionally by the expression of tartrate-resistant acid phosphatase(TRAP) and by their capability to resorb dentin slices. We also studied the effects of transforming growth $factor-{\beta}(TGF-{\beta})$ and epidermal growth factor(EGF) on the Vit. $D_3-induced$ osteoclast-like cell formation. In control, a few multinucleated cells were formed whereas PTH and $PGE_2$ increased the number of multinucleated cells. PTH, $PGE_2$ and Vit. $D_3$ induced the formation of TRAP-positive multinucleated cells. After culture of mouse bone marrow cells on the dentin slices with or without osteotropic hormones, giant cells with diverse morphology were found on the dentin slices under the scanning electronmicroscopy. After removing the attached cells, resorption pits were identified on the dentin slices, and the shape of resorption pits was variable. EGF increased the osteoclast-like cell formation induced by Vit. $D_3$, however, $TGF-{\beta}$ showed biphasic effect, which at low concentration, increased and at high concentration, decreased the osteoclast-like cell formation induced by Vit. $D_3$.

• Journal of Microbiology and Biotechnology
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• v.2 no.1
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• pp.41-45
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• 1992

Shikonin production was examined in a bubble column bioreactor system with the hairy roots of Lithosphermum erythrorhizon. The volumetric productivity was higher than those obtained from other reactor configurations with free or immobilized cells of the same cell line. The productivities of the bubble column reactor, with and without a product absorption trap, were 7.4 and 4.5 mg of shikonin/l/d, respectively. This indicated the importance of the product removal in the design and operation of the shikonin production system with hairy root culture.

• Journal of Periodontal and Implant Science
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• v.32 no.4
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• pp.733-744
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• 2002

The fibroblasts are the principal cells in the periodontal ligament of peridontium. As the periodontal ligament fibroblasts (PDLF) show similar phenotype with osteoblasts, the PDLF are thought to play an important role in alveolar bone remodeling. Cell-to-cell contacted signaling is crucial for osteoclast formation. Recently it has been reported that PDLJ enhance the bone resorbing activity of osteoclasts differentiated from hematopoietic preosteoclasts. The aims of this study were to $clarify\;^{1)}$ the mechanism of PDLF-induced osteoclastogenesis $and\;^{2)}$ whether we can use preosteoclast cell line instead of primary hematopoietic preosteoclast cells for studying the mechanism of PDLF-induced osteoclastogenesis. Osteoclastic differentiation of mouse macrophage cell line RAW264.7 was compared with that of mouse bone marrow-derived M-CSF dependent cell (MDBM), a well-known hematopoietic preosteoclast model, by examining, 1) osteoclast-specific gene expression such as calcitonin receptor, M-CSF receptor (c-fms), cathepsin K, receptoractivator nuclear factor kappa B (RANK) ,2) generation of TRAP(+) multinucleated cells (MNCs), and 3) generation of resorption pit on the $OAAS^{TM}$ plate. RAW264.7 cultured in the medium containing of soluble osteoclast differentiation Factor (sODF) showed similar phenotype with MDBM-derived osteoclasts, those are mRNA expression pattern of osteoclast-specific genes, TRAP(+) MNCs generation, and bone resorbing abivity. Formation of resorption pits by osteoclastic MNCs differentiated from sODF-treated RAW264.7, was completely blocked by the addition of osteoprotegerin (OPG), a soluble decoy receptor for ODF, to the sODF-containing culture me야um. The effects of PDLF on differentiation of RAW264.7 into　the TRAP(+) multinucleated osteoclast-like cells were examined using coculture system. PDLF were fxed with paraformaldehyde, followed by coculture with RAW264.7, which induced formation of TRAP(+) MNCs in the absence of additional treatment of sODF. When compared with untreated and fixed PDLF (fPDLF), IL-1 ${\beta}$-treated, or lipopolysaccha-ride-treated and then fixed PDLF showed two-folld increase in the supporting activity of osteoclastogenesis from RAW264.7 coculture system. There were no TRAP(+) MNCs formation in coculture system of RAW264.7 with PDLF of no fixation. These findigs suggested that we can replace the primary hematopoietic preosteoclasts for RAW264. 7 cell line for studying the mechanism of PDLF-induced osteoclastogenesis, and we hypothesize that PDLF control osteoclastogenesis through ODF expression which might be enhanced by inflammatory signals.

• International Journal of Oral Biology
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• v.30 no.2
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• pp.39-45
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• 2005

The present study was performed to investigate whether Achyranthes Radix extracts play roles in the bone metabolism. Three kinds of Achyranthes Radix extracts (methylene chloride (MC), ethylacetate (Ea), and water (W)) were used for bioassay. We examined cellular activities of osteoblasts by measurement of cell proliferation rate, alkaline phosphatase (ALP) activity, and calcified nodule formation. Osteoclast generation was assayed by measuring the number of tartrate-resistant acid phosphatase (TRAP) (+) multinucleated cells after culture of osteoclast precursor cells. There was a maximum 20% increase in proliferation rate of osteoblastic cells after treatment with MC. First and second subfraction of MC layer increased proliferation of osteoblast. Ea layer and second subfraction of MC layer increased ALP activity. Also MC layer and second subfraction of MC layer from Achyranthes Radix extracts increased the calcified nodule. MC layer and second subfraction of MC layer from Achyranthes Radix extracts significantly decreased in the number of TRAP (+) multinucleated cells. Taken together, Achyranthes Radix stimulates the proliferation and bioactivities of bone-forming osteoblasts, and inhibits activities of bone-resorbing osteoclasts.

• Interdisciplinary Bio Central
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• v.2 no.4
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• pp.15.1-15.5
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• 2010

RIKEN BioResource Center (BRC) has collected, preserved, conducted quality control of, and distributed mouse resources since 2002 as the core facility of the National BioResource Project by the Ministry of Education, Culture, Sports, Science and Technology (MEXT), Japan. Our mouse resources include over 5,000 strains such as humanized disease models, fluorescent reporters, and knockout mice. We have developed novel mouse strains such as tissue-specific Cre-drivers and optogenetic strains that are in high demand by the research community. We have removed all our specified pathogens from the deposited mice and used our quality control tests to examine their genetic modifications and backgrounds. RIKEN BRC is a founding member of the Federation of International Mouse Resources and the Asian Mouse Mutagenesis and Resource Association, and provides mouse resources to the one-stop International Mouse Strain Resource database. RIKEN BRC also participates in the International Gene Trap Consortium, having registered 713 gene-trap clones and their sequences in a public library, and is an advisory member of the CREATE (Coordination of resources for conditional expression of mutated mouse alleles) consortium which represents major European and international mouse database holders for the integration and dissemination of Cre-driver strains. RIKEN BRC provides training courses in the use of advanced technologies for the quality control and cryopreservation of mouse strains to promote the effective use of mouse resources worldwide.

• International Journal of Oral Biology
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• v.34 no.2
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• pp.81-86
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• 2009

Baicalin is a flavonoid purified from the medicinal plant Scutellaria baicalensis. It has been reported that baicalin exhibits antibacterial, anti-inflammatory and analgesic effects. The present study was undertaken to determine the underlying cellular mechanisms of baicalin action in preosteoclasts. The effects of this flavonoid on preosteoclasts were determined by measuring osteoclast generation and osteoclast activity in macrophage-colony stimulating factor (M-CSF)-dependent bone marrow cells (MDBMCs) and in co-cultures of MDBMCs and osteoblasts. Osteoclast generation was assayed by measuring the number of tartrateresistant acid phosphatase (TRAP) (+) multinucleated cells after culture. Osteoclast activity was assayed by measuring the area of the resorption pit after culture. We found that osteoclast generation was induced by M-CSF and receptor activator of NF-kB ligand (RANKL), and by the 1.25-dihydroxycholecalciferol in our cultures. Baicalin decreased both osteoclast generation and activity in MDBM cultures and co-cultures indicating that it may inhibit bone resorption.

• The korean journal of orthodontics
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• v.25 no.6 s.53
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• pp.715-722
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• 1995

Orthodontic tooth movement in response to orthodontic force results from actions of osteoclasts and osteeoblasts in the cell level. Convincing evidence has now been provided to support the view that osteoclasts are derived from mononuclear cells that originate in the bone marrow or other hematopoietic organs and they migrate to the bones via vascular routes. Nitric oxide(NO), which accounts for the biological properties of endothelium-derived relaxing factor(EDRF), is the endogenous stimulator of soluble guanylate cylase. The discovery of the formation of nitric oxide(NO) from L-arginine in mammalian tissues and its biological roles has, in the last 7 years, thrown new light onto many areas of research. Data from experiments in vitro showed that N-metyl-L-arginine(L-NMA) and L-nitro-L- arginine(L-NAME) are competitive inhibitors of nitric oxide synthase. This study suggest that the multinucleated cells in our culture have characteristics of osteoclasts and that the potential bone cell activity of nitric oxide in vitro may be mediated in part by stimulation of marrow mononuclear cells to form osteoclast-like cells. Bone marrow cells were obtaineed from tibia of 19-days old chick embryo. After sacrifice, tibia was quickly dissected and the bone were then split to expose the medullary bone. The cells were attached for 4 hours and the nonadherent cells were collected. Marrow cells weere cultured in 96-well plate in medium 199. To examine the number of TRAP-positive multinucleated cells(MNCs), $10^{-8}\;M\;Vit=D_3$ and various concentration of L-NMA and L-NAME weere added at the beginning of cultures and with each medium change. After 7 days of culture. tartrate-resistant acid phosphatase(TRAP) staining was performed for microscopic evaluation. Cells haying more than three nuclei per cell were counted as MNCs. The obsrved results were as follows;1. 1,25-dihydroxyvitamine $D_3$ stimulated the osteoclast-like multinucleated cells in cultures of chick embryo bone marrow. 2. Nitric oxide synthase inhibitors(NOSI ; N-NMA, N-NAME) stimulated the osteoclast-like cells in cultures of chick embry bone marrow. 3. 1,25-dihydroxyvitamine$D_3$ and nitric oxide synthase inhibitors did not appear to have additive effect on the generation of TRAP-positive MNCs. These results suggest that nitric oxide synthase inhibitors may stimulate the osteoclast-like multinucleated cell formation and fusion in cultures of chick bone marrow.

• The Korean Journal of Pesticide Science
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• v.4 no.2
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• pp.50-55
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• 2000

Occurrence, Injury aspects and control effect of some chemicals were investigated on tomato in plastic house from 1997 to 1998. Adults trapped by yellow sticky trap and sweeping net were increased from late May abruptly. Survey of population densities by yellow sticky trap was more clear than sweeping net. Damaged leaves were found out all the year round in continuous cropping fields and rate of damaged leaves was below 20% in early April and increased gradually up to 80% level after late May. Otherwise, damaged leaves were appeared from 6 weeks after transplanting in first growing field and then damaged leaves was 80% level in spring culture. In autumn culture, rate of damaged leaves was maintained 80% level from transplanting to harvesting date regardless of cultivation years. Number of adults was trapped from late May and peak was late June but mature larva was $6{\sim}8$ individuals in spring culture. In autumn culture, adult and larval densities was maintained high for 3 weeks after transplanting but begun to decrease from 4 weeks(early September). Chlorfenapyr EC, cyromazine WP and abamectin EC could suppress larval populations of Liriomyza trifolii effectively.

• Microbiology and Biotechnology Letters
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• v.27 no.3
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• pp.236-245
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• 1999

Andong Soju is a Korean traditional distilled liquor brewed with Nuruk which is cultured with wild microorganisms. To provide useful information for scientific production and systematic quality control of traditional distilled liquor, the effects of mixed culture of the alcoholic yeasts and saccharifying molds isolated from the Nuruk, and mashes on the flavor and sensory characteristics were investigated. Distillate from mashes cocultured with Saccharomyces cerevisiae and Hansenula anomala using Mucor Nuruk was compared with distillate from mashes brewed with Andong Nuruk and with distillate from plant fermented mashes to analyze their flavor characteristics. The volatile flavor compounds in distillates were analyzed by GC and GC-MS using direct injection, solvent extraction, and purge & trap methods. Alcohols such as 3-methyl-1-butanol, 2-methyl-1-propanol, 1-propanol, and 2-phenyl ethanol; aldehydes such as acetaldehyde and 2-furancar-boxaldehyde; esters such as ethyl ester of acetic acid, hexanoic acid, octanoic acid, decanoic acid; alkanes, alkenes, ketone, sulfur, and pyrone compounds were detected. Alcohols were chief components of flavor compounds. No significant difference in overall acceptability test was shown among three experimental groups(p<0.05), but Nuruk-like aroma, Kaoliangchiew-like aroma, sweet taste, and well rounded mouthfeel showed significant differences among them(p<0.05).