• Archives of Plastic Surgery
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• v.37 no.5
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• pp.509-518
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• 2010

Purpose: Major drawbacks of conventional bone marrow stromal cells (BSCs) transplantation method are mainly caused by direct transplanted cell to host cell interactions. We hypothesized that separation of the transplanted cells by a microporous membrane might inhibit most of the potential adverse effects and induce superior effect. The purpose of the study is to determine the optimal condition of the microporous membrane. Methods: First, BSCs were placed in polyethylene terephthalate (PET) transwell inserts with 3, 8, or $12{\mu}m$ pore size, and cultured in 24 well culture plates. After 5 days, bottoms of the plates were observed for presence of attached BSCs in monolayer and cell numbers were evaluated. Second, BSCs were placed PET, polycarbonate (PCT), and mixed cellulose esters (MCE) transwell inserts with 3 and $8{\mu}m$ pore size, and cultured in 24 well culture plates. After 3 days, the supernatants of the media left in culture plate were analyzed for collagen, vascular endothelial growth factor (VEGF), platelet derived growth factor BB (PDGF-BB), and basic fibroblast growth factor (bFGF). Third, BSCs were placed in 15% and 70% of the PET membrane with $3{\mu}m$ pore size. All the experimental conditions and methods were same as the second study. Results: The optimal pore sizes to prevent BSC leakage were $3{\mu}m$ and $8{\mu}m$. The amounts of type I collagen and three growth factors tested did not show significant differences among PET, PCT, and MCE groups. However, the collagen, VEGF, and bFGF levels were much higher in the high (70%) density group than in the low (15%) density group. Conclusion: This study revealed that the optimal pore size of membrane to prevent direct BSC to recipient cell contact is in between $3{\mu}m$ and $8{\mu}m$. Membrane materials and pore sizes do not influence the collagen and growth factor passage through the membrane. The most striking factor for collagen and growth factor transport is pore density of the membrane.

• Journal of Microbiology and Biotechnology
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• v.17 no.8
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• pp.1316-1323
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• 2007

In order to induce high levels of protein secretion, we have constructed a recombinant plasmid, designated pBP244, into which was incorporated key components of the type-II See-dependent secretion system, including LepB (signal peptidase), SecA (ATPase), and SecB (chaperone). The biological activities of the LepB, SecA, and SecB components expressed from genes harbored by pBP244 appeared to play their normal roles. In order to evaluate the protein secretion, a pspA (Streptococcus $\underline{p}neumoniae\;\underline{s}urface\;\underline{p}rotein\;\underline{A}$) gene was cloned into pBP244, resulting in pBP438. S. typhimurium harboring pBP438 grown until the stationary phase, secreted a higher level of PspA into the culture supernatants than did the strain harboring pYA3494. The strain harboring pBP438 secreted a supernatant amount 1.71-fold, a periplasmic space amount 1.47-fold, and an outer membrane amount 1.49-fold higher than that of pYA3494. S. typhimurium ${\chi}8554$ kept the $Asd^+$ plasmid pBP244 and pBP438 for 60 generations in LB broth harboring DAP, thereby indicating that pBP244 and pBP438 were quite stable in the Salmonella strain.

• Food Science and Biotechnology
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• v.14 no.4
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• pp.479-486
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• 2005

Arabinoxylan, a complex polysaccharide in cereal cell walls, has recently received research attention as a biological response modifier. The immunomodulating effect of arabinoxylans from rice bran (AXrb) was studied using a combined process of extrusion and commercial hemicellulase treatment in order to elucidate the augmentation mechanism of cell-mediated immunity in vitro. The cytotoxicity of mouse spleen lymphocytes against YAC-1 tumor cells was significantly enhanced by treatment with AXrb at $10-100\;{\mu}g/mL$. In an attempt to investigate the mechanism by which AXrb enhance NK cytotoxicity, we examined the effect of AXrb on cytokine production by spleen lymphocytes. Culture supernatants of the cells incubated with AXrb were collected and analyzed for IL-2 and IFN-${\gamma}$ synthesis by ELISA. IL-2 and IFN-${\gamma}$ production were increased significantly. These results suggest that AXrb may induce Th1 immune responses. Macrophages play an important role in host defenses against tumors by killing them and producing secretory products, which protect against bacterial, viral infection and malignant cell growth. AXrb were examined for their ability to induce secretory and cellular responses in murine peritoneal macrophages. When macrophages were treated with various concentrations ($10-100\;{\mu}g/mL$) of AXrb, AXrb induced tumoricidal activity, as well as increasing phagocytosis and the production of NO, $H_2O_2$, TNF-${\alpha}$, IL-$1{\beta}$, and IL-6. These results indicate that reactive oxygen species, reactive nitrogen species, and inflammatory cytokines are likely to be the major mediators of tumoricidal activity in AXrb-treated macrophages. Therefore, AXrb may be useful in cancer immunotherapy and it is anticipated that AXrb obtained using extrusion and subsequent enzyme treatment can be used as an ingredient in nutraceuticals and cereal-based functional food.

• Parasites, Hosts and Diseases
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• v.54 no.2
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• pp.123-132
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• 2016

Trichomonas vaginalis causes the most prevalent sexually transmitted infection worldwide. Trichomonads have been detected in prostatic tissues from prostatitis, benign prostatic hyperplasia (BPH), and prostate cancer. Chronic prostatic inflammation is known as a risk factor for prostate enlargement, benign prostatic hyperplasia symptoms, and acute urinary retention. Our aim was to investigate whether T. vaginalis could induce inflammatory responses in cells of a benign prostatic hyperplasia epithelial cell line (BPH-1). When BPH-1 cells were infected with T. vaginalis, the protein and mRNA of inflammatory cytokines, such as CXCL8, CCL2, IL-$1{\beta}$, and IL-6, were increased. The activities of TLR4, ROS, MAPK, JAK2/STAT3, and NF-${\kappa}B$ were also increased, whereas inhibitors of ROS, MAPK, PI3K, NF-${\kappa}B$, and anti-TLR4 antibody decreased the production of the 4 cytokines although the extent of inhibition differed. However, a JAK2 inhibitor inhibited only IL-6 production. Culture supernatants of the BPH-1 cells that had been incubated with live T. vaginalis (trichomonad-conditioned medium, TCM) contained the 4 cytokines and induced the migration of human monocytes (THP-1 cells) and mast cells (HMC-1 cells). TCM conditioned by BPH-1 cells pretreated with NF-${\kappa}B$ inhibitor showed decreased levels of cytokines and induced less migration. Therefore, it is suggested that these cytokines are involved in migration of inflammatory cells. These results suggest that T. vaginalis infection of BPH patients may cause inflammation, which may induce lower urinary tract symptoms (LUTS).

• Korean Journal of Veterinary Research
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• v.43 no.2
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• pp.247-253
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• 2003

Actinobacillus pleuropneumoniae causes a highly contagious pleuropneumoniae in swine. The bacterium produces several virulence factors such as exotoxin, LPS, capsular polysaccharide, etc. Among them, the exotoxin, called Apx, has been focused as the major virulence factor, and the toxin consists of 4 gene cluster. apx CABD. apxA is the structural gene of toxin and has four different types, I, II, III, and IV. As the first step of development of a new subunit vaccine, the three different types of apxA gene were amplified from A. pleuropneumoniae isolated from Korea by PCR with primer designed based on the N- and C-terminal of the toxin. The sizes of apxIA, IIA and IIIA were 3,073, 2,971 and 3,159bps, respectively. The comparison of whole DNA sequences of apxIA, IIA and IIIA genes with those of the reference strain demonstrated 98%, 99% and 98% homology, respectively. In addition, the phylogenetic analysis was performed based on the amino acid sequences compared with 12 different RTX toxin family using the neighbor-joining method. ApxA proteins of Korean isolates were identical with reference strains in this study. All ApxA proteins were expressed in E. coli with pQE expression vector and identified using Western blot with polyclonal antibodies against culture supernatants of A. pleuropneumoniae serotype 2 or 5. The sizes of each expressed ApxA protein were about 120, 110, 125 kDa (M.W.), respectively. The results obtained in this study could be used for the future study to develop a new vaccine to porcine pleuropneumoniae.

• Microbiology and Biotechnology Letters
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• v.44 no.2
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• pp.201-207
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• 2016

The aim of this study was to investigate the potential of lactic acid bacteria (LAB) strains as probiotics. Two strains were isolated from healthy chicken cecum and their acid and bile tolerance, residual organic acids, antibacterial activity against pathogenic bacteria, and immunomodulation activity were measured. Identification of the isolated strains was performed using the API 50CHL system and phylogenetic analysis using 16S rDNA sequencing. The isolates were determined to be Lactobacillus sakei strains. The acid tolerance of strains L2 and L8 was high enough that 75% of the inoculum survived in pH 2 for 2 h. The bile tolerance of both strains was observed at a 1% Oxgall concentration in MRS broth. The production of organic acids (lactic acid and acetic acid) and pH changes during growth were monitored and the maximum concentrations were obtained after 48 h of incubation. Culture supernatants of the two LAB strains showed strong antibacterial activity against pathogenic bacteria. The heat-killed LAB cells also induced high levels of immune cell proliferation compared with the control, and stimulated IL-6 and TNF-α production in mouse macrophages. Therefore, L. sakei strains L2 and L8 can be considered suitable probiotic bacteria.

• Tuberculosis and Respiratory Diseases
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• v.80 no.1
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• pp.60-68
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• 2017

Background: Mucus hypersecretion from airway epithelium is a characteristic feature of airway inflammatory diseases. Tumor necrosis factor ${\alpha}$ (TNF-${\alpha}$) regulates mucin synthesis. Glucocorticoids including mometasone fuorate (MF) have been used to attenuate airway inflammation. However, effects of MF on mucin production have not been reported. Methods: Effects of MF and budesonide (BUD) on the phorbol-12-myristate-13-acetate (PMA)-induction of mucin and TNF-${\alpha}$ in human airway epithelial cells (NCI-H292) were investigated in the present study. Confluent NCI-H292 cells were pretreated with PMA (200 nM) for 2 hours. Subsequently, the cells were stimulated with MF (1-500 ng/mL) or BUD (21.5 ng/mL) for 8 hours. Dexamethasone ($1{\mu}g/mL$) was used as the positive control. Real-time polymerase chain reaction was used to determine MUC2 and MUC5AC mRNA levels. The level of total mucin, MUC2, MUC5AC, and TNF-${\alpha}$ in culture supernatants were measured using enzyme-linked immunosorbent assay. Results: MF and BUD significantly suppressed MUC2 and MUC5AC gene expression in PMA-stimulated NCI-H292 cells. The inhibitory effects of the two steroid drugs were also observed in the production of total mucin, MUC2 and MUC5AC proteins, and TNF-${\alpha}$. Conclusion: Our findings demonstrated that MF and BUD attenuated mucin and TNF-${\alpha}$ production in PMA-induced human airway epithelial cells.

• Food Science and Biotechnology
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• v.17 no.5
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• pp.1003-1009
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• 2008

This study was performed to determine the effects of $\gamma$-irradiation on the antiallergic activity of alginate. An alginate aqueous solution was $\gamma$-irradiated at 3, 5, 7, 10, 20, and 100 kGy. First, the molecular weight (Mw) of alginate rapidly decreased as the $\gamma$-irradiation dose increased up to 20 kGy. Then, the antiallergic activity of the $\gamma$-irradiated alginate was measured. Interlukin (IL)-4 cytokine and IgE level were significantly decreased in the $\gamma$-irradiated alginate groups as compared to the control group in vitro. The mice intraperitoneally administered with the $\gamma$-irradiated alginate exhibited lower ovalbumin-specific IgE and IgG1 level in serum than the control mice. Furthermore, the $\gamma$-irradiated alginate suppressed total and ovalbumin-specific IgE secretions in the splenocytes. Increased IL-2 level was observed in the culture supernatants of the splenocytes that were obtained from the mice administerd with the $\gamma$-irradiated alginate, while IL-4 level decreased. The present study indicates that $\gamma$-irradiated alginate can suppress allergy in a mouse allergy model.

• Korean Journal of Pharmacognosy
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• v.41 no.2
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• pp.122-129
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• 2010

Mistletoe (Viscum album) is a common name for many species of semi-parasitic plants which grow on deciduous trees all over the world. In this study, the immunomodulatory activity of the water-extract of Korean mistletoe fruits (KMFWE), was investigated on murine peritoneal macrophages. The culture supernatants of KMF-WE-stimulated murine peritoneal macrophages showed the increased production of IFN-$\gamma$, IL-$1{\beta}$ and TNF-$\alpha$, in a dose-dependent manner. KMF-WE also induced chemokine production from murine peritoneal macrophages such as RANTES, MCP-1, MIP-$1{\alpha}$ and MIP-$1{\beta}$, as well as nitric oxide (NO) production, in a dose-dependent manner. The gel filtration fraction revealed F-1, which is the early-eluted and high molecular weight product, is the major fraction of KMF-WE to activate the murine peritoneal macrophage to induce cytokines, chemokines and NO. The nature of F-1 fraction needs to be examined in detail in further studies to define the regulatory mechanisms of cytokine or chemokine induction by KMF-WE on macrophages. These results suggest that KMF-WE possess a potent immunostimulant activity and can be a promising candidate available for development of immunomodulators.

• Korean Journal of Pharmacognosy
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• v.41 no.4
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• pp.275-281
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• 2010

To evaluate the immunomodulatory activity of a water extract (KMF-WE) of Korean mistletoe (Viscum album var. coloratum) fruit, we examined its ability to induce humoral and cellular immune response against keyhole limpet hemocyanine (KLH). Immunized mice with KLH admixed with KMF-WE (KLH/KMF-WE) showed significant induction of KLH-specific antibodies compared to mice immunized with KLH alone. The assay for determining isotypes of antibodies revealed that KMFWE augmented KLH-specific-IgG1 and -IgG2a production. In vitro T lymphocyte proliferation analysis against KLH revealed that the splenocytes of mice immunized with KLH/KMF-WE showed a significantly higher proliferative ability than those from mice immunized with KLH alone. The culture supernatants of splenocytes, which were harvested from mice immunized with KLH/KMF-WE, showed higher levels of both Th-1 type (IL-2, IFN-${\gamma}$) and Th-2 type (IL-4) cytokines in response to KLH stimulation compared to those from mice immunized with KLH alone. Also, in delayed-type hypersensitivity (DTH) assay, mice immunized with KLH/KMF-WE showed a significantly higher reaction to KLH than mice treated with KLH alone. These results suggest that KMF-WE possess immunoadjuvant activity to enhance both antigen-specific humoral and cellular immune responses against protein antigens (KLH).