• Korean Journal of Pharmacognosy
• /
• v.19 no.4
• /
• pp.251-255
• /
• 1988

A cytotoxic sesquiterpene against L1210 cell, named pubetalin. was isolated from the herb of Siegesbeckia pubescens Makino. Its structure was identified as $6-formyl-2,\;3,\;3{\alpha},\;4,\;5,\;8,\;9,\;11{\alpha}-octahydro-10-hydroxymethyl-5-methoxy-3-methylene-2-oxocyclodeca[{\beta}]\;furan-4-ylester$ of 2-methyl-2-propenoic acid.

• Asian Pacific Journal of Cancer Prevention
• /
• v.16 no.15
• /
• pp.6581-6589
• /
• 2015

Background: Previously, stingless bee (Trigona spp.) products from East Kalimantan, Indonesia, were successfully screened for in vitro antiproliferative activity against human cancer derived cell lines. It was established that propolis from T. incisa presented the highest in vitro cytotoxicity against the SW620 colon cancer cell line (6% cell survival in $20{\mu}g/mL$). Materials and Methods: Propolis from T. incisa was extracted with methanol and further partitioned with n-hexane, ethyl acetate and methanol. The in vitro cytotoxicity of the extracts was assessed by the MTT assay against human colon (SW620), liver (Hep-G2), gastric (KATO-III), lung (Chago) and breast (BT474) cancer derived cell lines. The active fractions were further enriched by silica gel quick column, absorption and size exclusion chromatography. The purity of each fraction was checked by thin layer chromatography. Cytotoxicity in BT-474 cells induced by cardanol compared to doxorubicin were evaluated by MTT assay, induction of cell cycle arrest and cell death by flow cytometric analysis of propidium iodide and annexin-V stained cells. Results: A cardol isomer was found to be the major compound in one active fraction (F45) of T. incisa propolis, with a cytotoxicity against the SW620 ($IC_{50}$ of $4.51{\pm}0.76{\mu}g/mL$), KATO-III (IC50 of $6.06{\pm}0.39{\mu}g/mL$), Hep-G2 ($IC_{50}$ of $0.71{\pm}0.22{\mu}g/mL$), Chago I ($IC_{50}$ of $0.81{\pm}0.18{\mu}g/mL$) and BT474 (IC50 of $4.28{\pm}0.14{\mu}g/mL$) cell lines. Early apoptosis (programmed cell death) of SW620 cells was induced by the cardol containing F45 fraction at the $IC_{50}$ and $IC_{80}$ concentrations, respectively, within 2-6 h of incubation. In addition, the F45 fraction induced cell cycle arrest at the G1 subphase. Conclusions: Indonesian stingless bee (T. incisa) propolis had moderately potent in vitro anticancer activity on human cancer derived cell lines. Cardol or 5-pentadecyl resorcinol was identified as a major active compound and induced apoptosis in SW620 cells in an early period (${\leq}6h$) and cell cycle arrest at the G1 subphase. Thus, cardol is a potential candidate for cancer chemotherapy.

• Journal of Plant Biotechnology
• /
• v.5 no.2
• /
• pp.121-129
• /
• 2003

Callus cultures of Azadirachta indica flower petals were established on MS medium supplemented with naphthalene acetic acid (1 mg/L), kinetin (0.5 mg/L) and sucrose $(3\%\;w/v)$. Cell cultures of Azadirachta indica were established and studied the growth and production kinetics. Half 85 medium supplemented with dicamba (2 mg/L), kinetin (1 mg/L) and sucrose $(3\%\;w/v)$ was found to be suitable for initiation and maintenance of cell cultures from the calli. MS medium supplemented with naphthalene acetic acid (NAA) (1 mg/L), kinetin (0.5 mg/L) and sucrose $(3\%\;w/v)$ was found to be suitable as production medium. Around $80\%\;(0.05\%\;w/v)$ of azadirachtin was found to be intracellular. The effect of various precursors, elicitors, permeabilizing agents and growth retardants in cell cultures was studied. The addition of precursors sodium acetate (10 mg/L), squalene (10 mg/L), isopentenyl pyrophosphate (1 mg/L) and geranyl pyrophosphate (1 mg/L) to the cell cultures on day 3 has shown significant increase in bioproduction of azadirachtin $(64.94{\pm}4.40\;mg/L,\;72.81{\pm}0.04\;mg/L,\;51.63{\pm}1.26\;mg/L\;and\;30.70{\pm}0.28\;mg/L\;respectively)$ over the control cultures $(4.70{\pm}0.27 mg/L)$. $5\%$ v/v cell extracts of Fusarium solani has shown moderate increase in the content of azadirachtin $(5.71{\pm}0.34\;mg/L)$ when compared to control cultures $(2.40{\pm}0.56\;mg/L)$. The addition of methyl jasmonate $(500\;{\mu}M/L)$ on day 3 has shown $\~4$ fold improvement in bioproduction of azadirachtln $(6.92{\pm}0.11\;mg/L)$ when compared to control cultures $(1.63{\pm}0.02\;mg/L)$. There was no significant effect of the studied growth retardants and permeabilizing agents on bioproduction of azadirachtin. Cells are cultivated in large volumes using the effective precursors.

• 한국생물공학회:학술대회논문집
• /
• 2002.04a
• /
• pp.321-324
• /
• 2002

In situ recovery of recombinant human granulocyte-macrophage colony stimulating factor (hGM-CSF) was performed in transgenic Nicotiana tabacum cell suspension cultures. Aqueous two-phase systems (ATPS) were used to utilize its biocompatibility. Transgenic plant cells could be grown up to 15.7 g/L in normal medium and 18.6 g/L in ATPS composed of 6% (w/w) polyethylene glycol (PEG) 20,000 and 10% (w/w) dextran 2,000,000. It was proved that the A TPS was not harmful to cell growth. In addition, It is expected to recover hGM-CSF simultaneously with cell growth. Using this method, maximum hGM-CSF concentration at 1.64 ng/mL was obtained on day 3.

• Applied Chemistry for Engineering
• /
• v.19 no.4
• /
• pp.366-369
• /
• 2008

To evaluate the performance of a bipolar packed bed cell (BPBC) filled with granular aluminium, the experiments were carried out in two groups as batch and continuous processes. In a batch process, removal efficiency of total phosphate (T-P) was 88% in case of electrolytic treatment of phosphate solution, T-P 10 mg/L at 6 V during 3 h by BPBC filled with granular aluminium. In a continuous process, residual T-P concentration was about 2 mg/L in case of electrolytic treatment of phosphate solution, 10 mg/L at 6 V, HRT 3 h by BPBC filled with granular aluminium. Break-through point was observed after running for 120 h at hydraulic retention time (HRT) of 3 h.

• Microbiology and Biotechnology Letters
• /
• v.36 no.4
• /
• pp.307-313
• /
• 2008

The yeast surface expression system, pCTXYN (6.8 kb), of Bacillus endoxylanase gene (xynB, 642 bp) was constructed and introduced into Saccharomyces cerevisiae EBY100 cell. The transformed yeast cell showing the highest endoxylanase activity was selected through the active staining of colonies grown on YPDG medium containing xylan. With the yeast transformant, EBY100/pCTXYN, grown on galactose containing medium, it was found that the endoxylanase was successfully displayed on the yeast cell surface and the xylooligosaccharides were efficiently produced from xylan. The most of endoxylanase activity was detected in the cell fraction and reached about 1.9 unit/mL after 48 h cultivation. The optimized conditions for xylooligosaccharides production from xylan were determined as follows: substrate and its concentration, oat spelt xylan 6%; concentration of yeast whole-cell, 5 unit/mL; temperature, $50^{\circ}C$, and reaction time $2{\sim}4\;h$. When the oat spelts xylan and corncob xylan were hydrolyzed by treatment with cell surface-displayed endoxylanase, xylotriose was formed as a main product.

• 한국약용작물학회:학술대회논문집
• /
• 2006.11a
• /
• pp.103-115
• /
• 2006

[ ${\beta}-Glucan$ ] is a glucose polymer that has linkage of ${\beta}-(1,3)$, -(1,4) and -(1,6). As exclusively found in fungal and bacterial cell wall, not in animal, ${\beta}-glucans$ are recognized by innate immune system. Dendritic cells (DC) or macrophages possesses pattern recognition molecule (PRM) for binding ${\beta}-glucans$ as pathogen-associated molecular pattern (PAMP). Recently ${\beta}-glucans$ receptor was cloned from DC and named as dectin-l which belongs to type II C-type lectin family. Human dectin-l is consisted of 7 exons and 6 introns. The polypeptide of dectin-l has 247 amino acids and has cytoplasmic, transmembrane, stalk and carbohydrate recognition domains. Dectin-l could recognize variety of beta-l,3 and/or beta-l,6 glucan linkages, but not alpha-glucans. In our macrophage cell line culture system, dectin-l mRNA was detected in RA W264.7 cells by reverse transcription-polymerase chain reaction (RT-PCR). Dectin-l was also detected in the murine organs of spleen, thymus, lung and intestines. Treatment of RA W264.7 cells with ${\beta}-glucans$ of Ganoderma lucidum (GLG) resulted in increased expression of IL-6 and $TNF-{\alpha}$ in the presence of LPS. However, GLG alone did not increase IL-6 nor $TNF-{\alpha}$ These results suggest that receptor dectin-l cooperate with CD14 to activate signal transduction that is very critical in immunoresponse.

• KSBB Journal
• /
• v.4 no.1
• /
• pp.18-20
• /
• 1989

Solid gelatin microcarrier with the size distribution between $100{\mu}{\textrm}{m}$ and $400{\mu}{\textrm}{m}$ was prepared for the attachment and growth experiment for anchorage-dependent animal cell lines, i.e., L 929 and BHK 21. The growth and the maximum cell densities on this gelatin based and polyacrylamide (PAA) microcarriers were compared with those on the commercial dextran based Cytodex 3 microcarrier. Both cell lines showed good comparable attachment and growth patterns on the above three microcarriers. The mouse fibroblast, L 929 showed about the same maximum cell density on PAA, gelatin and Cytodex 3 MC'S BUT BHK 21, the baby hamster kidney cell line, showed the best result on Cytodex 3, which was about $4\times10^6$ cells/ml with the microcarrier concentration of 10 g/1.

• Journal of Animal Science and Technology
• /
• v.55 no.1
• /
• pp.19-23
• /
• 2013

The current study was carried out to determine the effects of Kohlrabi (Brassica oleracea var. gongylodes) on proliferation and differentiation of pig preadipocytes and $_3T_3-L_1$ cells. Pig preadipocytes were isolated from the backfat of the new-born pigs. Twenty-four hours after seeding, the cells were washed with DMEM/F-12 (designated day 0). To measure the cell proliferation, the cells were treated with 25 ng/ml and 100 ng/ml ethanol extracts of Kohlrabi (peel and flesh) for two days (day 0 ~ 2). To measure differentiation, the cells were treated with Kohlrabi for two days (day 0 ~ 2) and cell differentiation was measured on day 6. Twenty-five ng/ml and 100 ng/ml of Kohlrabi peel decreased proliferation of pig preadipocytes by 4.59% and 17.7%, respectively, compared with the control and Kohlrabi flesh by 11.4% and 19.2%, respectively. However, Kohlrabi did not inhibit cell differentiation. To measure the effects of Kohlrabi on proliferation and differentiation of $_3T_3-L_1$ cells, the cells were treated with Kohlrabi for two days in culture, like pig preadipocytes. Kohlrabi (both peel and flesh) did not show any effects on cell proliferation and differentiation. In summary, the results of the current study showed that Kohlrabi decreased proliferation of pig preadipocytes, but no inhibitory effects on differentiation of the cells. Kohlrabi had no effects on proliferation and differentiation of $_3T_3-L_1$ cells.

• Journal of the East Asian Society of Dietary Life
• /
• v.25 no.6
• /
• pp.990-998
• /
• 2015

The aim of this study was to examine the cytotoxicity and potential inhibitory effects from four types of edible domestic brown seaweeds, Undaria pinnatifida (UP), Laminaria japonica (LJ), Sargassum fulvellum (SF), and Hizikia fusiforme (HF), on preadipocyte differentiation in 3T3-L1 cell line. Water and ethanol extracts from the four types of seaweeds were prepared and tested for cell viability in the 3T3-L1 cell line by using MTT assay. In addition, various doses of the water extract of seaweeds (WES) and ethanol extract of seaweeds (EES) were treated at the beginning of 3T3-L1 differentiation and continued until the cells were fully differentiated to adipocytes. Oil Red-O staining was performed to determine the potential cell differentiation inhibitory effects of the WES and EES by measuring the levels of lipid droplet accumulation in 3T3-L1 adipocytes. $PPAR{\gamma}$ mRNA expression levels were significantly reduced by WESs of UP, LJ, and HF as well as EESs of LJ and HF. As a result, we observed the superior cell differentiation inhibitory effects of WES compared to that of EES in a dose-dependent manner without any significant cytotoxicity in mouse adipocytes.