• BMB Reports
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• v.39 no.4
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• pp.432-440
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• 2006

A tuber lectin from Arisaema jacquemontii Blume belonging to family Araceae was purified by employing a single step affinity chromatography using column of asialofetuin-linked amino activated silica beads and the bound lectin was eluted with 100 mM glycine-HCl buffer pH 2.5. The purified A. jacquemontii lectin (AJL) showed a single protein band with an apparent molecular mass of 13.4 kDa when submitted to SDS-polyacrylamide gel electrophoresis under reducing as well as non-reducing conditions. The native molecular mass of AJL determined by gel filtration on a Biogel P-200 column was 52 kDa and its carbohydrate content was estimated to be 3.40%. Thus AJL is a tetrameric glycoprotein. The purified lectin agglutinated erythrocytes from rabbit but not from human. Its activity was not inhibited by any of the mono- and disaccharides tested except N-acetyl-D-lactosamine having minimal inhibitory sugar concentration (MIC) 25 mM. Among the glycoproteins tested only asialofetuin was found to be inhibitory (MIC $125\;{\mu}g/mL$). A single band was obtained in native PAGE at pH 4.5 while PAGE at pH 8.3 showed two bands. Isoelectric focusing of AJL gave multiple bands in the pI range of 4.6-5.5. When incorporated in artificial diet AJL significantly affected the development of Bactrocera cucurbitae (Coquillett) larvae indicating the possibility of using this lectin in a biotechnological strategy for insect management of cucurbits. Larvae fed on artificial diet containing sub-lethal dose of AJL showed a significant decrease in acid phosphatase and alkaline phosphatase activity while esterase activity markedly increased as compared to larvae fed on diet without lectin. Out of various human cancer cell lines employed in sulphorhodamine B (SRB) assay, this lectin was found to have appreciable inhibitory effect on the in vitro proliferation of HCT-15, HOP-62, SW-620, HT-29, IMR-32, SKOV-3, Colo-205, PC-3, HEP-2 and A-549 cancer cell lines by 82, 77, 73, 70, 41, 41, 37, 29, 21 and 21% respectively.

• Development and Reproduction
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• v.9 no.2
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• pp.105-114
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• 2005

Embryonic stem(ES) cells have tremendous potential as a cell source for cell-based therapies. Realization of that potential will depend on our ability to understand and manipulate the factors that influence cell fate decision and to develop methods for getting enough cell numbers for clinical applications. Hematopoiesis has been widely studied, and hematopoietic differentiation from ES cells is a good model to study lineage commitment. In this study, we investigated stemness and compared the efficiency of hematopoietic differentiation using two different mouse embryonic stem cell lines TC-1 and B6-1. Although the two cell lines showed known stem cell properties with minor differences, the embryoid body formation efficiency in methylcellulose was much higher in TC-1 than B6-1. When measured potentials of hematopoietic differentiation using functional(colony-forming cell) and phenotypic(specific marker expression) assays, we found that TC-1 can differentiate into hematopoietic cells in methylcellulose culture but B6-1 cannot. These results imply that we can improve the efficiency of hematopoietic cell differentiation by selection of proper cell lines and this may be also applied in the differentiation of human embryonic stem cells.

• Development and Reproduction
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• v.12 no.1
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• pp.107-112
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• 2008

To investigate the $C_{21}$-steroids produced from maturating oocytes in the longchin goby, Chasmichthys dolichognathus, the oocytes ($0.74{\sim}0.97\;mm$) were incubated with radiolabeled $17{\alpha}$-hydroxyprogesterone ($^3H-17{\alpha}OHP$) for 24 hours. The resulting metabolites were analyzed by thin layer chromatography and identified by gas chromatography-mass spectrometry. Two $C_{21}$-steroids, $17{\alpha}$-hydroxy, $20{\alpha}$-dihydroprogesterone ($17{\alpha}20{\alpha}P$) and $17{\alpha}$-hydroxy, $20{\beta}$-dihydroprogesterone ($17{\alpha}20{\beta}P$), were converted from $^3H-17{\alpha}OHP$ in the maturing oocytes. These two main metabolites were detected at 0.80 mm diameter oocytes or greater. In addition, the effects of these metabolites on in vitro germinal vesicle breakdown (GVBD) were tested. The sensitivity of oocytes to the induction of GVBD was greater at $17{\alpha}20{\beta}P$ than $17{\alpha}20{\alpha}P$. This result showed that $17{\alpha}20{\beta}P$ is a major maturation inducing steroid (MIS) in longchin goby, suggesting $17{\alpha}20{\alpha}P$ may play a role in regulating the oocyte maturation process.

• Current Research on Agriculture and Life Sciences
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• v.6
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• pp.37-41
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• 1988

The present work deals with the effect of agar and auxins concentrations on vitrification in tissue culture of Gypsophila paniculata L. cv. 'Bristol Fairy' in vitro. The results were summarized as follows. 1. Plant growth, that is, plant height, fresh weight and branching were decreased as increasing agar concentration. On the other hand, addition effect of IAA 1.0mg/l+NAA 0.5mg/l and IAA 2.0mg/l+NAA 1.0mg/l on the plant height were increased strikingly. 2. Addition effect of auxins on the days to rooting were little. And the root development showed same tendency as plant growth. 3. The rate of non-vitrified plants were gradually increased as rising agar concentration. But the addition of agar 1.5g/l in the medium resulted in poor growth. 4. From these results, it was found that following media were the most effective for increasing of non-vitrified and good plant growth in Gypsophila paniculata L. tissue culture.

• Journal of Yeungnam Medical Science
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• v.6 no.2
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• pp.195-205
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• 1989

Development of drug resistance in tumors during treatment is a major factor limiting the clinical use of anticancer agents. When tumor cells acquire resistance to anticancer drug, they show cross-resistance to other antitumor agents. In the present study, SNU-1 cell was induced adriamycin $10^{-7}M$ drug resistance, SNU-1/ADR, in vitro culture system. We got the doubling time and number for viability test during 96 hours by MTT assay. To investigate the cross resistance of various anticancer drugs in human stomach cancer cell SNU-1 and SNU-1/ADR. We compared $IC_{50}$ (drug concentration of 50% reduction) and the relative resistance(RR). SNU-1/ADR was expressed multidrug resistant with vinblastine(RR ; 31.62), vincristine(RR ; 29.50), dactinomycin(RR ; 21.37), epirubicin(RR ; 17.78), daunorubicin(RR ; 14.12), adriamycin(RR ; 7.76), and etoposide(RR ; 4.46), and other drugs, 5-fluorouracil, cisplatin, cyclophosphamide, methotrexate, and aclarubicin, have not cross resistant with adriamycin. There was double minute chromosome in SNU-1/ADR by karyotyping although this change was not seen in SNU-1.

• Journal of fish pathology
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• v.27 no.1
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• pp.17-24
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• 2014

To study the possible use of probiotics in fish farming, The in vitro and in vivo antibacterial effects of Pseudomonas aeruginosa MB I-3 (MB I-3) against the fish pathogenic bacterium Listonella anguillarum were evaluated. The inhibitory effects of MB I-3 against vibrios were investigated by the double layer method and the co-culture. The results showed that MB I-3 inhibited the growth of pathogenic vibrios including Listonella anguillarum, Vibrio alginolyticus, Vibrio cholerae, Vibrio fluvialis, Vibrio furnissii, Vibrio harveyi, Vibrio parahaemolyticus and Vibrio vulnificus. Extracellular substances obtained from the cultural supernatant of MB I-3 by ethyl acetate extraction showed inhibitory effects on L. anguillarum. The antibacterial substance of MB I-3 was evaluated to destroy the cell membrane of L. anguillarum in electron micrographs. The probiotic effects of MB I-3 was tested by exposing olive flounder (Paralichthys olivaceus) fry to L. anguillarum with or without MB I-3. The cumulative mortality of olive flounder fry infected with L. anguillarum was 24% in the group with MB I-3, while it was 46% in the control group without MB I-3. These results indicate that MB I-3 has potential applications as a probiotic for the control of fish pathogenic vibrios in fish rearing system.

• Asian Pacific Journal of Cancer Prevention
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• v.14 no.10
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• pp.5825-5831
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• 2013

Background/Objectives: Maintenance of cellular function in culture is vital for transfer and development following adoptive immunotherapy. Dual properties of IL-21 in activating T cells and reducing activation induced cell death led us to explore the mechanism of action of IL-21 enhanced proliferation and cytotoxic potential of CIK cells. Method: CIK cells cultured from PBMCs of healthy subjects were stimulated with IL-21 and cellular viability and cytotoxicity to K562 cells were measured. To elucidate the mechanism of action of IL-21, mRNA expression of cytotoxic factors was assessed by RT-PCR and protein expression of significantly important cytotoxic factors and cytokine secretion were determined through flow cytometry and ELISA. Western blotting was performed to check the involvement of the JAK/STAT pathway following stimulation. Results: We found that IL-21 did not enhance in vitro proliferation of CIK cells, but did increase the number of cells expressing the CD3+/CD56+ phenotype. Cytotoxic potential was increased with corresponding increase in perforin ($0.9831{\pm}0.1265$ to $0.7592{\pm}0.1457$), granzyme B ($0.4084{\pm}0.1589$ to $0.7319{\pm}0.1639$) and FasL ($0.4015{\pm}0.2842$ to $0.7381{\pm}0.2568$). Interferon gamma and TNF-alpha were noted to increase ($25.8{\pm}6.1ng/L$ to $56.0{\pm}2.3ng/L$; and $5.64{\pm}0.61{\mu}g/L$ to $15.14{\pm}0.93{\mu}g/L$, respectively) while no significant differences were observed in the expression of granzyme A, TNF-alpha and NKG2D, and NKG2D. We further affirmed that IL-21 signals through the STAT-3 and STAT-5b signaling pathway in the CIK cell pool. Conclusion: IL-21 enhances cytotoxic potential of CIK cells through increasing expression of perforin, granzyme B, IFN-gamma and TNF-alpha. The effect is brought about by the activation of STAT-3 and STAT-5b proteins.

• Asian-Australasian Journal of Animal Sciences
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• v.21 no.9
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• pp.1330-1338
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• 2008

This study was conducted to investigate the effects of Acanthopanax senticosus extract (ASE) as a dietary additive on gut microflora in weaned piglets. A total of sixty pigs were weaned at 21 d of age (BW = $5.64{\pm}0.23kg$) and allocated on the basis of BW and litter to three dietary treatments in a randomized complete block design. The dietary treatments were: control group (basal diet), antibiotics group (basal diet+0.02% colistin), and ASE group (basal diet+0.1% ASE). On d 7, 14 and 28 after consuming the experimental diets, five piglets per group were sacrificed and then the contents from the jejunum, ileum and cecum were collected to determine changes in the microbial community by using a polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) technique and estimating the contents of Lactobacillus and E. coli by in vitro culturing methods. The results showed that the ASE promoted the microflora diversity in the cecum. Enumeration of bacteria in the gut contents showed that the number of Lactobacillus increased (p<0.05), while that of E. coli decreased (p<0.05) when compared with the other 2 groups as the days of age progressed post-weaning. These findings suggested that the ASE, as a substitute for dietary antimicrobial products, could improve the development of the normal gut microflora and suppress bacterial pathogens, and effectively promote a healthy intestinal environment.

• Journal of Plant Biotechnology
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• v.33 no.4
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• pp.297-302
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• 2006

In general, the proliferation of orchids via somatic embryos has been used for mass production of somatic clones because of high propagation efficiency. In spite of high propagation rate, this method often brings somaclonal variation, especially polyploid frequency. Therefor we here concentrated to investigate the relationship between endopolyploidization patterns of explants and the occurrence of tetraploid variant in clonally proliferated Doritaenopsis via somatic embryo regeneration system. In the fully developed somatic embryo, upper part contained 2C to 16C while middle and lower parts showed 2C to 32C DNA content. Two-week-old embryo contained 2C to 16C, whereas those regenerated after 4 to 10-week-old contained 2C to 64C nuclei. Results showed that endoreduplication was variable depending upon tissue types, ages, and parts in one species. lower part of somatic embryo having high endoreduplication degree increased the regeneration of tetraploid variants by about 3-fold comparing to upper part of somatic embryo culture. polyploid frequency occurrence might be closely related to the high levels of endoreduplication of somatic embryos used as explant. It suggested that the upper part of somatic embryo having comparatively low endoreduplication degree is suitable for the stable in vitro propagation system.

• Korean Journal of Microbiology
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• v.35 no.1
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• pp.82-88
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• 1999

In this study, to evaluate newly developed Japanese encephalitis (JE) vaccine candidate CJ50003, we assessed its immunogenicity along with a previously commercialized inactivated JE Biken vaccine. The CR0003 viral antigens produced in Vero cells were administered suhcutaneouly to mice either with alum-adjuvanled or free form. The ELISA titers and neutralizing (NEUV antibody titers accounting for major protective immunity in JE were determined. Mice given alum-adjuvanted vaccine had a 10 times higher antigen-specific NEUT antibody response than did those which {lad received free antigens. This NEUT antibody response was maintained until day 168 with NEUT titer more than 1:160. Even with the 0.5 ng of alum-adjuvanted antigen dose, NEUT titer was induced more than 1:10 which is considered as an evidence for seroconversion and protection. Thc mice immune sera had a similar rate of cross-reactivity against three different viral antigens, Nakayama-NlH, P3 and SA14; as determined by ELISA assay. In a mice challenge model, vaccination with the GI50003 conferred more protection than with commercialized Biken vaccine against Nakayama virus. These data demonstrated that CJ50003 vaccine candidate has an excellent prophylactic efficacy and implicated it has a strong potential for further development and commercialization.