• Microbiology and Biotechnology Letters
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• v.9 no.1
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• pp.7-19
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• 1981

The production of gluconic acid from glucose by the resting cell system of Aspergillus niger was studied. It was found that the conversion products from glucose by the resting cell system were markedly influenced by the pH, temperature, substrate concentration, aeration, metal ions, cultivation time and storage conditions of the resting cells. Conversion products were identified as gluconic acid by the thin layer chromatography and infrared spectrophotometry. These conversions were greatly stimulated by addition of $Mg^{++}$, and S $n^{++}$, but showed inhibitory effects by C $u^{++}$, H $g^{++}$, C $d^{++}$, A $g^{+}$ and cyanide. For the optimum cell storage, it was effective to be kept at -$25^{\circ}C$ in 0.05M phosphate buffer solution of pH 7.0. The gluconic acid production by the resting cell system was more effective than those of the fermentation with respect to cultivation time, yield, recovery and re-use of the cell.l.l.l.l.l.l.

• THE JOURNAL OF KOREAN ORIENTAL ONCOLOGY
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• v.3 no.1
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• pp.67-84
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• 1997

Bujeonghangamtang(扶正抗癌湯) has been used for cure of tumor as a traditional medicine without any experimental evidence to support the rational basis for its clinical use. This study was carried out to evaluate the possible therapeutic or antitumoral effects of Bujeonghangamtang extract against tumor, and to carry out some mechanisms responsible for its effect. Some kinds of tumor were induced by the typical application of 3-methylcholanthrene. (MCA) or by the implantation(s.c) of malignant tumor cells such as leukemia cells(3LL cells) or sarcoma cells(Sl80 cells). Treatment of the Bujeonghangamtang water-extract (dailly 1mg／mouse, i. p.) was continued for 7 days prior to tumor induction and after that the treatment was lasted for 20 days. Against squamous cell carcinoma induced by MCA, Bujeonghangamtang decreased not only the frequency of tumor production but also the number and the weight of tumors per tumor bearing mice (TBM). Bujeonghangamtang also significantly suppressed the development of 3LL cell and S180 cell-implanted tumors in occurrence-frequency and their size, and some developed tumors were regressed by the continuous treatment of Bujeonghangamtang extract into TBM. In vitro, treatment of Bujeonghangamtang extract had no effect on the growth of some kinds of cell line such as FsaII, A431 strain but significantly inhibited the proliferation of 3LL, S180 cells and augmented the DNA synthesis of mitogen-activated lymphocytes. Bujeonghangamtang also stimulated the migrative ability of leukocyte, the MIF and IL-2 production of T lymphocytes, but not IL 6 production of B cells. Bujeonghangamtang-administration to mice enhanced NK cells activities. These results demonstrated that Bujeonghangamtang extract exhibited a significant prophylactic benefits against tumors and its antitumor activity was manifested depending on the type of tumor cells. And these results also suggested that effect of Bujeonghangamtang might be chiefly due to nonspecific enhancement of NK cell activities and cell-mediated immune responses.

• Biotechnology and Bioprocess Engineering:BBE
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• v.11 no.5
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• pp.442-448
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• 2006

Human lactoferrin (hLf) is an iron-binding glycoprotein that has been considered to play many biological roles in the human, including the stimulation of the immune system, antimicrobial and anti-inflammatory effects, and regulation of iron absorption. We generated transgenic Siberian ginseng (Acanthopanax senticosus) cell cultures producing a functional hLf protein using the signal peptide sequence from the endoplasmic reticulum and driven by an oxidative stress-inducible SWPA2 promoter which is highly expressed in plant cell cultures. The production of hLf increased proportionally to cell growth and showed a maximal level (up to 3.6% of total soluble protein) at the stationary phase in suspension cultures. Full-length hLf protein was identified by immunoblot analysis in transgenic cell cultures of Siberian ginseng. Recombinant hLf (rhLf) was purified from suspension cells of Siberian ginseng by ammonium sulfate precipitation, cation-exchange and gel filtration chromatography. N-terminal sequences of rhLf were identical to native hLf (nhLf). The overall monosaccharide composition of rhLf showed the presence of plant specific xylose while sialic acid is absent. Antibacterial activity of purified rhLf was higher than that of nhLf. Taken together, we anticipate that medicinal Siberian ginseng cultured cells, as demonstrated by this study, will be a biotechnologically useful source for commercial production of functional hLf not requiring further purification.

• Journal of Physiology & Pathology in Korean Medicine
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• v.21 no.2
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• pp.414-424
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• 2007

Adenophorae Radix (AR), Liriopis Tuber (LT), Dendrobii Monile (DM), Polygonati Officinalis (PO), Polygonati Rhizoma (PR) have been used to treated a variety condition/diseases in traditional oriental medicine. The present study was conducted to investigate the immunmodulatory effects of the water-extracted AR, LT, DH, PO and PR. In spleen cell proliferation assay, DH was significantly enhanced mitogenic activity compared with control group. In RT-PCR, DH ad PO induced IL-2 and IFNr cytokine gene expression in mouse spleen cells. Methotrexate(MTX), immune supression agent, was significantly inhibited mouse spleen cell proliferation(1600 mg/ml). In spite of MTX treatement, DH and PO sustained the spleen cell proliferation, In the flow cytometry analysis, DH stimulated mouse spleen cells showed an increase in B-cell phenotype (CD45R/B220). The water-extracted DH and PO inhibited NO production and iNOS expression in LPS-stimulated RAW 264.7 macrophage cell. DH induced IL-2 and IFNg gene expression in human peripheral mononuclear cells. The GC-MS analysis show that the main component of water-extracted DH was b-Nitroethyl alcohol. The main components of water-extracted PO were Dipirartril-tropico, Methyl sulfoxide and Demsodrox. These data suggest that among these extracts, DH has a protective effcet of immune suppression caused by MTX. DH may be enhance cellular and humoral immune response by the regulation of cytokine gene expression, NO production and B cell production.

• Journal of Bio-Environment Control
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• v.15 no.1
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• pp.91-99
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• 2006

This study was conducted to determine the influence of postharvest dipping treatment with aminoethoxyvinylglycine (AVG) on ethylene production and composition of non-cellulosic neutral sugars in cell walls of 'Tsugaru' apple fruits during storage. Fruits were harvested on August 20, soaked in AVG 50 and 75 $mg L^{-1}$ solution for 5 minutes, and stored in cold storage chamber at $0{\pm}1^{\circ}C$ for 60 days. Fruit quality factor, ethylene productions, and cell wall component changes were investigated at 20 days interval. As a result, the fruit firmness and acid content were much higher in AVG treated fruits than those of untreated one during 60 days of cold storage. Ethylene production of AVG treated fruits was reduced to the level of 1/10 compared with untreated one. As to the change of non-cellulosic neutral sugars in the cell walls of 'Tsu- garu' fruits, the major sugar was arabinose and galactose in water, CDTA and $Na_2CO_3$ soluble fractions. The content of arabinose and galactose in untreated fruits increased as the softening of fruits was in progress, but the fruits treated with AVG showed a little change during storage, so it is predicted that these two cell wall compositional sugars were not solubilized by the treatment of AVG. Accordingly, the marketability of 'Tsu- garu' fruits could remarkably increase when soaking the fruits in AVG solution after harvest.

• Herbal Formula Science
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• v.5 no.1
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• pp.147-168
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• 1997

Tish study was carried out to evaluate the possible therapeutic or antitumoral effects of Takrisodokyeum extract against tumor, and immunomodulatory effect. Some kinds of tumor were induced by the typical application of 3-methylcholanthrene (MCA) or by the implantation(s.c) of malignant tumor cells such as leukemia cells(3LL cells) or sarcoma cells(S-I80 and Fas II cells). Treatment of the Takrisodokyeum water-extract(daily 1mg mouse, i.p.) was continued for 7 days prior to tumor induction and after that the treatment was lasted for 15 days. Against squamous cell carcinoma induced by MCA, Takrisodokyeum decreased not only the frequency of tumor production but also the number and the weight of tumors per tumor bearing mice (TBM). Takrisodokyeum also significantly suppressed the development of 3LLcell and S-180 cell by frequency and their size, and some developed tumors were regressed by the continuous treatment of Takrisodokyeum extract into TBM. However, when tumor was induced by FsaII cell-implantation, the growth of implanted cells in mice was delayed by the water extract of Takrisodokyeum until day 7 and then rapid growth ensued. In vitro, treatment of Takrisodokyeum extract had no effect on the growth of some kind of cell lines such as FsaII, A-131 strain but significantly inhibited the proliferation of 3LL, S-180 cells. Takrisodokyeum also stimulated the migrative ability of leucocyte, the MIF and IL 2-production of T lymphocytes, but not IL 6 production of B cells. Takrisodokyeum enhanced Arthus reaction and DTH to sheep erythrocytes, and NK cells activities. These results demonstrated that Takrisodokyeum extract different results according to the type of tumor cells. And these results also suggested that antitumor effect of Takrisodokyeum might be chiefly due to nonspecific enhancement of NK cell activities and cell-mediated immune responses.

• Journal of Microbiology and Biotechnology
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• v.22 no.2
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• pp.199-206
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• 2012

Naturally immobilized tannase (tannin acyl hydrolase, E.C. 3.1.1.20) has many advantages, as it avoids the expensive and laborious operation of isolation, purification, and immobilization, plus it is highly stable in adverse pH and temperature. However, in the case of cell-associated enzymes, since the enzyme is associated with the biomass, separation of the pure biomass is necessary. However, tannic acid, a known inducer of tannase, forms insoluble complexes with media proteins, making it difficult to separate pure biomass. Therefore, this study optimizes the production of cell-associated tannase using a "protein-tannin complex" free media. An exploratory study was first conducted in shake-flasks to select the inducer, carbon source, and nitrogen sources. As a result it was found that gallic acid induces tannase synthesis, a tryptose broth gives higher biomass, and lactose supplementation is beneficial. The medium was then optimized using response surface methodology based on the full factorial central composite design in a 3 l bioreactor. A $2^3$ factorial design augmented by 7 axial points (${\alpha}$ = 1.682) and 2 replicates at the center point was implemented in 17 experiments. A mathematical model was also developed to show the effect of each medium component and their interactions on the production of cell-associated tannase. The validity of the proposed model was verified, and the optimized medium was shown to produce maximum cell-associated tannase activity of 9.65 U/l, which is 93.8% higher than the activity in the basal medium, after 12 h at pH 5.0, $30^{\circ}C$. The optimum medium consists of 38 g/l lactose, 50 g/l tryptose, and 2.8 g/l gallic acid.

• KSBB Journal
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• v.16 no.6
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• pp.579-591
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• 2001

The cell growth inhibitor effect of cervical cancer cells was investigated by liposome mediated transfection (pRcCMVp53/lipofectin) and by transfection using adenovirus (AdCMVp57). The papilloma virus cancer cell lines we used in this study were HPV16 positive, having inhibiter gene, wild p53 gene, CaSki, SiHa, HPV18 positive HeLa, HeLaS3 and HPV negative C33A, HT3. LacZ gene of E.coli was used as the marker gene for the transfection efficiency. The effect on the inhibition of tumor cell growth was measured by cell count and cell viability though ELISA analysis and MTT assay. The inhibition of tumor cell growth was confirmed by measuring each assay for six days, comparing with the normal control cell growth. The cell growth of cervical cancer calls by transfection was significantly reduced and showed tittle differences among the cell lines. To eliminate the potential problem of Ad(adenovirus) contamination during rAAV production, rAAV can be produced by a triple transfection of vector plasmic, packaging plasmid, and adenovirus helper plasmid. To examine the helper functions of Ad plasmids on the production of rAAV vector, we carried out cotransfection of three plasmids, AAV vector, packaging construct, and Ad helper plasmids. The optimized transfection condition for calcium phosphate method is 25ug of total DNA per 10-cm-diameter plate of 293 cell. We found that rAAV yields peaked at 48hr after Ad infection. The titer of rAAV was measured by the dot blot analysis to measure the number of particles/ml based on the quantification of viral DNA. Recent1y, Kombucha(fungi) was identified as a very potent antileukefic agent. In the present study, effect of natural toxin(plankton) and Kombucha is PSP(GTXI-3, neoSTX), on various MTT assay cervical cancer cell line. Toxin(GTX 1-3, neoSTX) also inhibited the proliferation in primary cervical cancer calls in a dose-dependent toxin concentration. These results showed that toxin was very potent in inhibiting the proliferation of cervical cancer calls in vitro. Toxins and Kombuoha exhibited a dose dependent inhibition of cellular proliferation in cancer cell line.

• Molecules and Cells
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• v.26 no.1
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• pp.67-73
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• 2008

Herpetic stromal keratitis (HSK) is an inflammatory disorder induced by HSV-1 infection and characterized by T cell-dependent destruction of corneal tissues. It is not known what triggers $CD4^+$ T cell migration into the stroma of HSV-1-infected corneas. The keratocyte is a fibroblast-like cell that can function as an antigen-presenting cell in the mouse cornea by expressing MHC class II and costimulatory molecules after HSV-1 infection. We hypothesized that chemokines produced by stromal keratocytes are involved in $CD4^+$ T cell infiltration into the cornea. We found that keratocytes produce several cytokines and chemokines, including MCP-1, RANTES, and T cell activation (TCA)-3. HSV-1 infection increased the production of MCP-1 and RANTES by keratocytes, and these acted as chemoattractants for HSV-1-primed $CD4^+$ T cells expressing CCR2 and CCR5. ExpreHerpetic stromal keratitis (HSK) is an inflammatory disorder induced by HSV-1 infection and characterized by T cell-dependent destruction of corneal tissues. It is not known what triggers $CD4^+$ T cell migration into the stroma of HSV-1-infected corneas. The keratocyte is a fibroblast-like cell that can function as an antigen-presenting cell in the mouse cornea by expressing MHC class II and costimulatory molecules after HSV-1 infection. We hypothesized that chemokines produced by stromal keratocytes are involved in $CD4^+$ T cell infiltration into the cornea. We found that keratocytes produce several cytokines and chemokines, including MCP-1, RANTES, and T cell activation (TCA)-3. HSV-1 infection increased the production of MCP-1 and RANTES by keratocytes, and these acted as chemoattractants for HSV-1-primed $CD4^+$ T cells expressing CCR2 and CCR5. Expression of MCP-1 in the corneal stroma was confirmed in vivo. Finally, when HSV-1-primed $CD4^+$ T cells were adoptively transferred into wild type and MCP-1-deficient mice that had been sublethally irradiated to minimize chemokine production from immune cells, infiltration of $CD4^+$ T cells was markedly reduced in the MCP-1-deficient mice, suggesting that it is the MCP-1 from HSV-1-infected keratocytes that attracts $CD4^+$ T cells into the cornea.

• KSBB Journal
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• v.19 no.1
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• pp.17-26
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• 2004

In this study the extracellular production of 5-aminolevulinic aicd (ALA) by recombinant E. coli BL2l (DE3) pLysS harboring the plasmid pFLS45 are investigated. Optimum concentrations of succinic acid and glycine for cell growth and ALA production were found to be 30 mM and 15 mM, respectively. Levulinic acid (LA) as an inhibitor of ALAD was added to the culture medium in the end of exponential cell growth phase and its optimum concentration was 30 mM. Growth of recombinant E. coli BL2l (DE3) pLysS (pFLS45) was largely dependent upon the pH value of culture medium. When the pH of culture medium was in the range of 6.0 and 6.5, high cell mass and ALA production were obtained. IPTG induction for the expression of the fusion gene did not enhance the production of ALA. Recombinant cell grew at 30't faster than at 37$^{\circ}C$, but ALA productivity was lower than at 37$^{\circ}C$. Repeated addition of glycine, succinic acid, and LA increased the production of ALA and the inhibition of intracellular ALA dehydratase activity, with up to 1.3 g/L ALA having been produced in the cultivation.