• Korean Journal of Animal Reproduction
• /
• v.21 no.2
• /
• pp.111-116
• /
• 1997

We determined the effects of follicular fluid fractions in the maturation medium on bovine oocyte maturation, fertilization and subsequent development, as well as on number of cells in blastocysts following culture. Follicular fluid and oocytes from bovine follicles less than 5 mm in diameter were collected from the ovaries of slaughtered cows. Follicular fluid was separated into different molecular weight fractions by untrafiltration through a membrane using a centrifuge at 500$\times$g, for 2h. For the maturation medium, follicular fluid fractions (30%, v/v), whole fluid (30%) or PVP(3mg/ml) were added to TCM 199(0.1$\mu\textrm{g}$/ml estradiol-17$\beta$, 100IU hCG). After maturation for 24h, oocytes were fertilized in vitro with bull frozen-thawed spermatozoa and cultured on a monolayer of granulosa cells for 9 days after fertilization. There were no differences in maturation rates or fertilization rates among any maturation conditions. The rates of development to >2-cell stage of the oocytes were significantly decreased when fraction of follicular fluid below 10,000 MW were added into maturation medium, compared with control and fraction above 10,000 MW(26.0% vs 40.8% to 64.0%, respectveily. p<0.01). Likewise, the rates of development to blastocysts of fertilized oocytes were significantly decreased in maturation medium containing fraction of follicular fluid (<10,000 MW). The average cell number of blastocysts derived from oocytes that matured in the fraction(>10,000 MW) of follicular fluid was 154.7$\pm$13.7. These embryos contained more cells than those matured in whole follicular fluid, or the fraction(<10, 000 MW) of follicular fluid or control(107.0$\pm$8.4, 91.8$\pm$11.8 and 95.8$\pm$6.2, respectively). In conclusion, we found that fractions of follicular fluid contained factors stimulating or inhibiting oocyte cytoplasmic matruation. These suggest that a factor(s) inducing cytoplasmic maturation of oocytes may exist in >10,000 MW fraction of follicular fluid.

• Toxicological Research
• /
• v.27 no.2
• /
• pp.77-83
• /
• 2011

The objective of this study was to evaluate and compare the antitumor activity of different solvent fractions (ethanol, dichloromethane, ethyl acetate, butanol and water) of the Auricularia auricula-judae 70% ethanol extract on the P388D1 macrophage and sarcoma 180 cells. A dose-dependent antitumor activity of each solvent fraction (from 0.01 mg/ml to 0.3 mg/ml) was shown against both cell types. These cytotoxic effects of all the tested fractions were confirmed on the MTT and SRB assays, without statistical differences each other. $IC_{50}$ value of dichloromethane fraction was 94.2 ${\mu}g/ml$ against sarcoma 180 cells lower than any other solvent fractions. The potent antitumor effect of the dichloromethane (DCM) fraction was also found against solid tumor in BALB/c mice. The splenomegaly and higher splenic index were found in tumor-bearing mice, with the DCM fraction returning to the negative control values. Thus, the results indicated the dichloromethane fraction may have potential ingredients as antitumor candidates.

• Korean Journal of Agricultural Science
• /
• v.46 no.2
• /
• pp.369-379
• /
• 2019

Alzheimer's disease (AD) is the most common neurodegenerative disease associated with age, and amyloid beta ($A{\beta}$) is known to cause Alzheimer's disease. In the present study, we investigated the protective effects of Cirsium japonicum var. maackii extract and its fractions against $A{\beta}$-induced neurotoxicity in C6 glial cells. The cells treated with $A{\beta}_{25-35}$ showed a decrease in cell viability and an increase in reactive oxygen species (ROS) production compared with the non-treated cells. However, the cells treated with the C. japonicum var. maackii extract and its fractions increased the cell viability and inhibited the $A{\beta}$-induced ROS production. These results demonstrate the neuroprotective effects of C. japonicum var. maackii against $A{\beta}$. To further examine the protective mechanism, we measured inflammation and apoptosis related protein expressions. The cells treated with extract and fractions from C. japonicum var. maackii down-regulated inflammatory related proteins such as cyclooxygenase-2, interleukin $(IL)-1{\beta}$, and IL-6, and attenuated apoptosis related proteins including B-cell lymphoma-2 (Bcl-2) associated X protein/Bcl-2 ratio. In particular, the ethanol and ethylacetate fraction exhibited higher inhibitory effect against ROS production and apoptosis-related protein expressions among the extract and the other fractions. Therefore, this study demonstrated the protective effects of C. japonicum var. maackii extract and its fractions against $A{\beta}$-induced neurotoxicity in C6 glial cells through the regulation of oxidative stress, inflammation, and apoptosis, suggesting that it might have potential as a therapeutic for AD.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.34 no.10
• /
• pp.1514-1519
• /
• 2005

This study was performed to investigate the effects of Porphyra tenera (PT) on cytotoxicity and quinone reductase (QR) activity in the cancer cells. PT was extracted with methanol and further fractionated into five different types: hexane (PTMH), ethyl-ether (PTMEE), ethylacetate (PTMEA) butanol (PTMB) and aquous (PTMA) partition layers. We determined the cytotoxic effect of these layers on C6, HepG2, MCF-7, and HT-29 cell lines by MTT assay. Among the various fractions, hexane (PTMH) of PT showed the strongest cytotoxic effect on C6, HepG2 and MCF-7 cell lines. PTMH displayed very low level of cytotoxicity at the lower concentration levels and at 300 $\mu$g/mL. PTMH resulted in 87.5$\%$ growth inhibition on C6 cell 70 $\%$ on the HepG2 cell and 89$\%$ on the MCF-7 cell, which were significantly high compared to other fractions. A 400 $\mu$g/mL PTMH concentration level, 99$\%$, 94.5$\%$ and 99$\%$ of cell growth inhibition were resulted on the same cell lines. On HT-29 cell line, both hexane (PTMH) and aqueous (PTMA) fraction of PT showed cytotoxic effects, but the Percentage was not as high as previous results tested on other cell lines such as C6 HepG2 and MCF-7 cell lines. Also, we observed quinone reductase (QR) inducing-effects in all fractions of PT on HepG2 cells. The QR inducing effects of the PTMH on HepG2 cells at 150 $\mu$g/mL concentration was 6.6 times higher than the control. Although further studies are needed, the present work suggests that PT was a potential to be used as a chemopreventive.

• Journal of Ginseng Research
• /
• v.14 no.3
• /
• pp.364-372
• /
• 1990

This experiment was performed to investigate the effects of ginseng saponin fractions (total saponin, triol saponin. diol saponin) and lipopolysacrharide (LPS) on the tllmoricidal activity of macrophage. The ginseng saponin fractions had little effect on the tumoricidal activity of macrophage (less than 10%). When the ginseng saponin fractions were treated with LPS, the effects of tumoricidal activation of macrophage increased a relatively high percent, and the total saponin and triol saponin (20-35%) were ulore effectual than diol saponin (15-25%). The effects of ginseng saponin and LPS on the tumoricidal activity of macrophage were mediated by the induction of macrophage-release factor(5) which has(have) the capacity of tumor cell killing. And the quantity of the (actors) was(were) increased by the contact of macrophage with tumor cell.

• Korean Journal of Microbiology
• /
• v.24 no.4
• /
• pp.377-384
• /
• 1986

A study on the production of mycobacterial antogens has been made in order to improve immunological reactivity and specificity, which have long been explored for the better use of immunological diagnosis of mycobacterial infections. Instead of culture filtrate cell extract was used as a starting material for the production of antigens in this study. Cell extract was fractionated though several steps such as salting out, gel filtration and ion exchange column chromatography and reactivity and specificity of the fractions so produced were enaluated by the various serological methods. The result showed that the species-specific antigenic components distributed mostly in the fractions, Tc of M. tuberculosis, Kc of M. kansasii, Sa of M. scrofulaceum, Aa of M. avium and Fa, Fb, Fc (FF1) of M. fortuitum, which were fractionated by ion exchange column prior to concentrating by salting out and molecular sieving.

• Journal of Plant Biology
• /
• v.9 no.1_2
• /
• pp.1-6
• /
• 1966

Using the Chlorella cells which had been uniformly labeled with $^{32}P$, the distribution of phosphorus in various fractions of cell material was investigated. Uniformly $^{32}P$-labeled Chlorella cells were further grown in a P-free medium, and some protions of the cells were taken out at intervals during the culture, and subjected to analyze the contents of $^{32}P$ in various fractins of the cell constituents. 2. Analysis of the $^{32}P$-labeled Chlorella cells showed that the highest in P-content was the fraction of RNA followed by those of lipid, RNA-polyphosphate complex, acid-insoluble polyphosphate, acid-soluble polyphosphate, DNA and protein. 3. During the culture of $^{32}P$-labeled Chlorella cells in a P-free medium, amounts of phosphate in DNA, protein and lipid fractions increased, while the P-contents in the fraction of RNA-polyphosphate complex decreased as well as those of acid-insoluble polyphosphate and acid-soluble polyphosphate fractions. 4. It was inferred that phosphorus used in the syntheses of DNA and protein was taken from polyphosphates of the cells, and RNA-polyphosphate complex would play an important role as a phosphate pool.

• Proceedings of the Korean Society of Crop Science Conference
• /
• 2017.06a
• /
• pp.292-292
• /
• 2017

This study was conducted to evaluate the effect of anti-obesity and antioxidant enzyme activities in vitro by different solvent fractions from the roots of Codonopsis lanceolata. The cytotoxicity of different solvent fractions of C. lanceolata on 3T3-L1 preadipocytes were evaluated using the MTT assay, the rate of cell survival progressively decreased in a dose-dependent manner. Butyl alcohol fraction at $200{\mu}g/mL$ exhibited a pronounced cytotoxic effect (75.73%) on 3T3-L1 cell comparable to that of the hexane fraction (79.82%), methylene chloride fraction (84.02%), ethyl acetate fraction (87.62%) and DW fraction (86.30%) at the same concentration. The Oil Red O solution was used to determine whether different solvent fractions of C. lanceolata induce adipocyte differentiation in 3T3-L1 preadipocytes. Confluent 3T3-L1 cells were treated with $50{\mu}g/mL$ concentration of solvent fraction extracts from C. lanceolata. Inhibitory degree of lipid accumulation against solvent fraction extracts showed a significant level compared with the control. Both lipid accumulation and adipocyte differentiation showed relatively high effect on methyl chloride fraction. The root extract of C. lanceolata had the highest SOD enzyme activity of 84.5% in ethyl acetate partition layer and while water partition layer of diploid showed the lowest SOD enzyme activity of 57.9%. The activity of CAT, APX and POD showed a significantly higher activity in ethyl acetate partition layer compared with the other fraction. These results suggested that the roots of C. lanceolata may assist in the potential biological activity on anti-obesity and antioxidant capacity.

• Asian Pacific Journal of Cancer Prevention
• /
• v.17 no.12
• /
• pp.5139-5145
• /
• 2016

There has been an increment in the number of studies focused on marine bioactive materials. Many peptides and other biomaterials with anticancer potential have been extracted from various marine animals. Artemia extracts have found uses in sun-light protection cosmetics and anti-aging products. However, contents of biochemical compounds in Artemia spp. and molecular mechanisms of have not been clearly studied in leukemic cells in vitro. In this work, we isolated and purified proteins of Artemia Urmiana. Six clear fractions (A-F) observed on DEAE-cellulose chromatography were assayed for effects on cell growth, differentiation and apoptosis using the human leukemic HL-60 cell line. Cell proliferation analysis by MTT and BrdU assays indicated that did not affect cells, growth. Cells treated with crude extract and fractions A, B and C, but not E and F (up to $100{\mu}g/mL$), exhibited increase of cell growth in a dose dependent manner. Stimulatory effects of fraction D were observed at concentrations of $10{\mu}g/mL$ and above. In nitro blue tetrazolium (NBT) reduction assays, treatment with $100{\mu}g/mL$ of fraction E or F for 96 hr increased the fraction of differentiated cells up to $14.8{\pm}3.56%$ and $16.5{\pm}2.08%$ respectively. Combination of those fractions with retinoic acid had significant synergistic effects on the differentiation of cells ($56.8{\pm}3.7%$ and $67.4{\pm}4.2%$, $p{\leq}0.01$). Annexin-V FITC staining for apoptosis and flow cytometric assays indicated induction of apoptosis by fractions E and F up to 23.8 and 31.8% of cells.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.30 no.6
• /
• pp.1266-1271
• /
• 2001

The cytotoxic effects of lipid extracts from soybean products were studied using K562 human leukemia cell, Yac1 mouse leukemia cell and S 180 mouse sarcoma cell. Total lipids from soybean powder, soybean curd residue and doenjang were extracted with chloroform/methanol (2 : 1) and water saturated butanol, consecutively, and fractionated into acetone supernatants (AS fraction) and acetone precipitates (AP fraction) by adding excess acetone. AS fraction of doenjang lipids showed the strongest cytotoxic effects on K562, Yac1 and S180 cancer cells, whereas each lipid fraction of soybean curd residue also showed relatively weak cytotoxic effects on cancer cells but soybean powder did not. AS and AP fractions of doenjang contained more free fatty acids than those of soybean curd residue and soybean. And when lipid fractions were digested with 0.4 N KOH/methanol, doenjang lipid fractions showed to contain some alkali-stable substances which showed positive reaction with ninhydrin solution on silica TLC separation.