• THE JOURNAL OF KOREAN ORIENTAL ONCOLOGY
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• v.6 no.1
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• pp.81-97
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• 2000

Objectives: This experimental study was carried out to evaluate the effects of aqueous and methanol extracts of Hedyotis diffusa which has long been used for cancer treatment in oriental medicines on the induction of apoptotic cell death in human lymphoid leukemia cell line, HL-60. Methods: Cells were treated with various concentrations (200 to $0.4{\mu}g$) and periods (6 to 30 hr) of $H_2O$ and methanol extracts of Hedyotis diffusa. Then, cells were tested for viability by MTT assay. Cells wrere treated with $200{\mu}g/ml$ of methanol extract fork various periods. Genomic DNA was isolated, separated, on 1.5% agarose gels, stained with ethidium bromide and visualized under UV light. Cells were treated with $200{\mu}g/ml$ of each extract for 16 hr. Then, cells were treated with Hoechst dye 33342 and observed by fluorescence microscopy. Cells were treated with various doses of each for 12 hr and $100{\mu}g/ml$ of methanol extract for various periods. Lysate from the cells used to measure the activity of Caspase-1 and-3 proteases by using fluorogenic peptide substrates including acetyl-YVAD-AMC and acetyl-DEVD-AMC, respectively. Cells were treated with $200{\mu}g/ml$ of each extract for various periods. Cell lysates were immunoprecipated with anti-JNKl antibodies. The immune complex was reacted with $32^p-ATP$ and c-Jun as a substrate. The phosphotransferase activity of JNKI was measured by using PhosphoImage analyzer (Fuji Co., Japan). Nuclear extracts were isolated and incubated with oligonucleotide probe of $NF-{\kappa}B$. Transcriptional activation of ${\kappa}B$ was measured by using EMSA and visualized by PhosphoImage analyzer (Fuji Co, Japan). Cell lysates were prepared and analyzed by Western blotting with anti-Bc12 antibodies and anti-Bax antibodies. Cells were pretreated with various doses of methanol extract for 2 hr. Then, the extract was removed by centrifugation. Cells were resuspended with RPMI-1640 media containing 0.3% agarose, 10% FBS, overlayred onto bottom layer agarose and incubated at $CO_2$ incubator for 6 days. The number of colony was counted under light microscopy ($\time100$). Results: The death of HL-60 cells was markedly induced by the addition of methanol extract of Hedyotis diffusa in a dose and time-dependent manners. The apoptotic characteristic ladder pattern of DNA strand break was observed in death of HL-60 cells. In addition, it was shown nucleus chromatin condensation and fragmentation under Hoechst staining. Therefore, Hedyotis diffusa extract-induced death of HL-60 cells is mediated by apoptotic signaling processes. The activity of Caspase 3-like proteases remained in a basal level in HL-60 cells treated with aqueous extract of Hedyotis diffusa. However, it was markedly increased in HL-60 cells treated with methanol extract of Hedyotis diffusa. In addition, the phosphotransferase activity of JNKl was increased in HL-60 cells treated with methanol extract of Hedyotis diffusa. Furthermore, the activation of transcriptional activator, $NF-{\kappa}B$ was markedly induced by methanol extract of Hedyotis diffusa. Anti-apoptotic Bc12 was cleaved into 23Kda fragment by treatment of methanol extract of Hedyotis diffusa. However, expression of proapoptotic Bax protein was increased by treatment of methanol extract of Hedyotis diffusa in a time-dependent manner. Furthermore, methanol extract markedly inhibited the colony forming efficiency of HL-60 cells in semisolid agar culture. Conclusions: Above results suggest that methanol extract of Hedyotis diffusa induces the apoptotic death of human leukemic HL-60 cells via activations of Caspase-3 proteases, JNKI, transcriptional activator $NF-{\kappa}B$, In addition, our results also suggest that methanol extract of Hedyotis diffusa reduces the malignant potential of HL-60 cells via down regulation of colony forming effciency through cleavage of Bc12 as well as induction of Bax.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.48 no.4
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• pp.365-372
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• 2022

Cellobiose is a dissacharide constituted by two glucose units joined by a β-('1,4') glycosidic bond that is produced by the decomposition of cellulose. This product exists naturally in plants and has been utilized in different industries as a food sweetener, and as a cosmetic and pharmaceutical material. In this study, the potential of cellobiose as a whitening cosmetic product was evaluated by analyzing autophagy induction and the inhibition of melanin production. A cytotoxicity test conducted in the human melanin-producing cell line MNT-1 with increasing concentrations of cellobiose revealed that this compound did not cause cytotoxicity at 20 mg/mL or less. Based on this, autophagy was firstly evaluated by immunostaining with the autophagy marker microtubule-associated protein 1 light chain 3 (LC3) after treatment with 20 mg/mL of cellobiose. The subsequent confocal microscopy analysis revealed an increase in LC3 puncta, indicating induction of autophagy. In addition, autophagy was further confirmed by western blot analysis, which demonstrated that cellobiose converted LC3-I to LC3- ∏ in a concentration- and time-dependent manners. An analysis of melanin contents after cellobiose treatment at a concentration of 20 mg/mL during 7 days revealed that melanin production was reduced by more than 50%. Additionally, the expression levels of melanogenesis-related proteins TYR and TYRP1 were markedly decreased after cellobiose treatment. Based on these studies, a cosmetic cream formulation containing cellobiose was prepared and the change in formulation was tested for 4 weeks, and it was confirmed that the appearance changed to liquid form at high temperature, but the pH did not change. In conclusion, the present research demonstrated that cellobiose activates autophagy and inhibits melanin production, and showed the potential of this product as a material for whitening cosmetics.

• The Korean Journal of Malacology
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• v.25 no.2
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• pp.105-112
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• 2009

We investigated the degree of obesity, histological development stages of gonads and sexual maturation induction rates of comb pen shell, Atrina pectinata, per the type of micro-algae supplied. Terms of maturation by singular or mixed supply of microalgae, it was found that maturation of the female was the quickest at 60.0% by the Tetraselmis tetrathele (Tet). experiment group followed by 57.1% by the Chlorella ellipsoidea (Chl). experiment group and 16.7% by the Phaeodactylum tricornutum (Pha). experiment group. However, there were no significant differences between Tet. experiment group and Chl. experiment group. As for the male, maturation was the quickest at 60.0% by the Tet. experiment group followed by 16.7% by the Chl. experiment group and 14.3% by the Pha. experiment group. In light of these results, Tet. is concluded to be a very useful feed organism in breeding the mother comb pen shells. Upon completion of the experiment, the sexual maturation induction rate for the female was found to be the highest at 82.0% in the Tet. experiment group followed by 72.0% by the Chl. experiment group, 64.0% by the Pha. experiment group and 58.0% by the mixed micro-algae experiment group. During the period of experiment, the survival rate was the highest at 94.4% by the mixed micro-algae experiment group followed by 90.0% by the Pha. experiment group, 83.1% by the Tet. experiment group and 78.8% by the Chl. experiment group.

• Korean Journal of Agricultural Science
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• v.7 no.2
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• pp.169-175
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• 1980

These studies were conducted to induce the available mutant strains in acetic acid bacteria by the irradiation of UV-light and the treatment of N-methyl-N'-nitro-N'-nitrosoguanidine. 109 strains which were capable of producing acid in the ethanol containing medium were isolated from vinegar and Kimchi collected from the region of Daejeon city. From the collection T-50 strain was identified to have a strong fermentation power and selected as a mother strain for the study. Two mutants were obtained by treating T-50 strain with UV and NTG, and these mutants had a rapid acid production in the initial stage. The study was then made to compare the basic condition for acetic acid production of the mother strain and two mutant strains. The summarized results were as follows; 1. The isolated strain (T-50) was identified as Acetobacter aceti by Bergey's manual and Acetic acid bacteria (Tokyo Univ. press). 2. The selected strain was died completely when the strain was irradiated with 15 W UV-light at a distance of 45 cm for 160 seconds. 3. The mutants such as U-48 and N-67 were rapid in the acetic acid production at the initial stage compare to the mother strain. 4. Acetic acid formation by the shaking culture method was maximized in 2 days culture, and the optimal temperature for acetic acid production was $30^{\circ}C$. 5. Acetic acid was effectively produced by the addition of 0.1% ammonium sulfate as a nitrogen source and was also produced rapidly by the addition of 0.1% glucose.

• Horticultural Science & Technology
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• v.19 no.4
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• pp.459-464
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• 2001

Leaf disks from cultivar 'Kennebec' and one selection line (ND 860-2) were cultured on Murashige-Skoog medium with various combinations of indole acetic acid (IAA) and zeatin riboside. Shoots, roots and callus were induced at various combinations of plant growth regulator levels. The medium containing $3.5mg{\cdot}L^{-1}$ IAA and $4.0mg{\cdot}L^{-1}$ zeatin riboside produced the most plantlets. Rooted regenerants were grown in the greenhouse. The growth of regenerated plants obtained from the MS medium supplemented with $7.0mg{\cdot}L^{-1}$ IAA and $3.0mg{\cdot}L^{-1}$ zeatin riboside was significantly greater than those grown from nodal expalnts. In ND 860-2, a leaf chimera with chlorophyll deficient (light yellow) sectors was found in plants regenerated fiom leaf disks (grown on MS medium supplemented with $3.5mg{\cdot}L^{-1}$ IAA and $3.0mg{\cdot}L^{-1}$ zeatin riboside) but not in plants grown from nodal explants. The phenotypic variability was also observed in the tuber number, size and weight.

• Clinical and Experimental Reproductive Medicine
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• v.43 no.1
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• pp.9-14
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• 2016

Objective: Autophagy contributes to the clearance and recycling of macromolecules and organelles in response to stress. We previously reported that vitrified mouse oocytes show acute increases in autophagy during warming. Herein, we investigate the potential role of Atg7 in oocyte vitrification by using an oocyte-specific deletion model of the Atg7 gene, a crucial upstream gene in the autophagic pathway. Methods: Oocyte-specific Atg7 deficient mice were generated by crossing Atg7 floxed mice and Zp3-Cre transgenic mice. The oocytes were vitrified-warmed and then subjected to in vitro fertilization and development. The rates of survival, fertilization, and development were assessed in the Atg7 deficient oocytes in comparison with the wildtype oocytes. Light chain 3 (LC3) immunofluorescence staining was performed to determine whether this method effectively evaluates the autophagy status of oocytes. Results: The survival rate of vitrified-warmed $Atg7^{f/f}$;Zp3-Cre ($Atg7^{d/d}$) metaphase II (MII) oocytes was not significantly different from that of the wildtype ($Atg7^{f/f}$) oocytes. Fertilization and development in the $Atg7^{d/d}$ oocytes were significantly lower than the $Atg7^{f/f}$ oocytes, comparable to the $Atg5^{d/d}$ oocytes previously described. Notably, the developmental rate improved slightly in vitrified-warmed $Atg7^{d/d}$ MII oocytes when compared to fresh $Atg7^{d/d}$ oocytes. LC3 immunofluorescence staining showed that this method can be reliably used to assess autophagic activation in oocytes. Conclusion: We confirmed that the LC3-positive signal is nearly absent in $Atg7^{d/d}$ oocytes. While autophagy is induced during the warming process after vitrification of MII oocytes, the Atg7 gene is not essential for survival of vitrified-warmed oocytes. Thus, induction of autophagy during warming of vitrified MII oocytes seems to be a natural response to manage cold or other cellular stresses.

• Journal of The Korean Society of Grassland and Forage Science
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• v.18 no.4
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• pp.351-354
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• 1998

Centipedegrass (Eremochloa ophiuroides) is a popular lawn grass in the southeastern USA. It has a naturally light green color and grows well on a wide range of soil types. Studies show limited morphological variation present in centipedegrass germplasm. To obtain the high morphological variation, plants were established from the irradiated seed at 10 Kr, allowed to interpollinate and harvested bulk seed, and then irradiated again for the next cycles. Morphological characteristics were measured in the 5 genetic varition lines (TC201 : cv. Common and non irradiated, TC202 : 4th cycles, TC241 : 6th cycles, TC306 : 8th cycles, and TC318 : 5th cycles) induced by recurrent gamma radiation. The ranges of variation of recurrently radiated centipedegrass lines < TC202, TC241 and TC306 except TC318(TifBlair) > for the stolons per plant, total stolon length per plant, longest stolon length, leaf length and width at top-most exposed internode were wider than those of non-irradiated line (TC201). Recurrent gamma radiation was very effective to enlarge the ranges of variation of morphological characteristics in reproductive organ like stolons of centipedegrass. The effect of quantity of gamma ray irradiation cycles on the means and ranges of variation in the morphological characteristics of centipedegrass was not regularly tended.

• Korean Journal of Medicinal Crop Science
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• v.7 no.3
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• pp.155-161
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• 1999

This study was carried out to develop the propagation system using in vitro induced- microtubers of yams (Dioscorea alata L.). Effects of kinds of media, mineral composition, sucrose concentration (0, 2, 4, 6, 8, 10%), photoperiod (0, 8, 12, 16, 24h), and growth regulators (NAA, IAA, ZR, JA-Me, ABA) on the development of microtubers, roots, and shoots in nodal stem segment cultures of D. alata L. were evaluated. Microtuberization in nodal stem segment occurred on all the media supplemented with growth regulator and sucrose. Among basic media, 1/2MS medium was the best in microtuber induction. NAA was shown to be the most effective among the growth regulators. Optimal NAA concentration was 1mg/l. The microtuberization was the highest at the concentration of 6% sucrose. When the nodal stem segment were cultured under darkness, the tuberization was increased markedly compared to those cultured under light condition. It was also noticeable that the culture in medium with NAA produced only microtubers and roots, but no shoots, in nodal segments. In this study, the optimal medium composition for microtuberization in nodal stem segment was found to be 1/2MS medium supplemented with 1mg/l NAA and 6% sucrose under dark condition at $25^{\circ}C$.

• The Korea Journal of Herbology
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• v.25 no.4
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• pp.39-45
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• 2010

Objectives : This study was undertaken to compare the effect of Foeniculi Fructus and Lonicerae Flos extract on the cerulein-induced acute pancreatitis in rats. Methods : Acute pancreatitis was induced by intraperitoneal injection of cerulein. Foeniculi Fructus extract (FE; 300 mg/kg) and Lonicerae Flos extract (LE; 300 mg/kg) were injected 2hr before induction of acute pancreatitis. Rats were sacrificed 6hr after first injection of cerulein. The severity of pancreatitis was assessed by measuring pancreatic weight/body weight ratio, neutrophil, lymphocyte, serum amylase activity, platelet activating factor (PAF) activity, tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) activity, interleukin 6 (IL-6) activity and by histological assessments of inflammatory cell infiltration. Results : 1. The pancreatic weight/body weight ratios of FE and LE group compared with the control group were decreased significantly. 2. The neutrophil content ratio of FE and LE group compared with the control group were decreased. 3. The lymphocyte content ratio of FE and LE group compared with the control group were increased significantly. 4. The activities of serum amylase of FE and LE group compared with the control group were decreased significantly. 5. The activities of serum PAF of FE and LE group compared with the control group were decreased significantly. 6. The activities of TNF-${\alpha}$ of FE and LE group compared with the control group were decreased significantly. 7. The activities of IL-6 of FE and LE group compared with the control group were decreased significantly. 8. The pancreas injected with FE and LE showed reduction of swelling of acinar cells, inflammation and vacuolization than the control group on light microscopic observation. Conclusions : These results suggest that Foeniculi Fructus and Lonicerae Flos extract have an effect to suppress inflammation on cerulein-induced acute pancreatitis in rats. But there are no significant differences between Foeniculi Fructus extract and Lonicerae Flos extract.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.34 no.4
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• pp.370-377
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• 1989

Jerusalem artichoke (Helianthus tuberosus L.) as a biomass potential crop has several distinct advantages such as vigorous growth on poor land and high yield of carbohydrate. In this crop, seed dormancy has hampered the efforts of seed-propagration and to use them in breeding programs for improving jerusalem artichoke. Several seed treatments were tested to determine their effectiveness in overcoming the seed dormancy found in five collected varietes of jerusalem artichoke. The first results showed that the seed fertilities of five collected varieties ranged from 2.4％ to 14.7％ and the number of seed produced by one plant ranged from 88 to 1058. Germinability of seeds stored for 3 months at room temperature after harvest was almost 0％ and it was not improved by addition to the treatments of temperature, light and GA3, while germinability of seeds stored for 27 months at room temperature after harvest increased to 47.5％ in germination rate. But the removal and pin-pricking of seedcoat were very effective in breaking the seed dormancy, giving germination of 96.8％ and 82.3％, respectively. These results showed that the seed dormancy of jerusalem artichoke was induced by the seedcoat. Besides the treatment of seedcoat removal and seedcoat-pinpricking, the treatment of low and wet stratification was also effective in breaking the seed dormancy of jerusalem artichoke. Whole dormant seeds incubated for 70 days in low and wet condition germinated over 85％.