• Journal of Life Science
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• v.25 no.1
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• pp.53-61
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• 2015

In this study, we tried to separate the photosensitizer that induces apoptosis of leukemia cells (U937) from perilla leaves. Perilla leaves (Perilla frutescens Britt var. japonica Hara) are a popular vegetable in Korea, being rich in vitamins (A and E), GABA, and minerals. Dried perilla leaves were extracted with methanol to separate the photosensitizer by various chromatographic techniques. The structure of the isolated compound (PL9443) was identified by 1D-NMR, 2D-NMR, and FAB-mass spectroscopy. Absorbance of the UV-Vis spectrum was highest at 410 nm and was confirmed by the 330, 410, and 668 nm. PL9443 compound was determined to be pheophorbide, an ethyl ester having a molecular weight of 620. It was identified as a derivative compound of pheophorbide structure when magnesium comes away from a porphyrin ring. Observation of morphological changes in U937 cells following cell death induced by treated PL9443 compound revealed representative phenomena of apoptosis only in light irradiation conditions (apoptotic body, vesicle formation). Results from examining the cytotoxicity of PL9443 substance against U937 cells showed that inhibition rates of the cell growth were 99.9% with the concentration of 0.32 nM PL9443. Also, the caspase-3/7 activity was 99% against U937 cells with the concentration of 0.08 nM of PL9443 substance. The result of the electrophoresis was that a DNA ladder was formed by the PL9443. The PL9443 compound is a promising lead compound as a photosensitizer for photodynamic therapy of cancer.

• Journal of Life Science
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• v.19 no.6
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• pp.770-780
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• 2009

A new photosensitizer, 9-Hydroxypheophorbide-a (9-HpbD-a), was derived from Spirulina platensis. We conducted a series of experiments, in vitro and in vivo, to evaluate the anticancer effect and mechanism of photodynamic therapy using 9-HpbD-a and 660 nm diode lasers on a squamous carcinoma cell line. We studied the cytotoxic effects of pheophytin-a, 9-HpbD-a, 9-HpbD-a red and 660 nm diode lasers in a human head and neck cancer cell line (SNU-1041). Cell growth inhibition was determined by using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction assay. The effects of 9-HpbD was higher than those of 9-HpbD-a red or pheophytin-a in PDT. We then tested the cytotoxic effects of 9-hydroxypheophorbide-a (9-HpbD-a) in vitro. The cultured SNU-I041 cells were treated with serial concentrations of 9-HpbD-a followed by various energy doses (0, 0.1, 0.5, 3.2 J/$cm^{2}$) and by various interval times (0, 3, 6, 9, 12 hr) until laser irradiation, then MTT assay was applied to measure the relative inhibitory effects of photodynamic therapy (PDT). Optimal laser irradiation time was 30 minutes and the cytotoxic effects according to incubation time after 9-HpbD-a treatment increased until 6 hours, after which it then showed no increase. To observe the cell death mechanism after PDT, SUN-I041 cells were stained by Hoechst 33342 and propidium iodide after PDT, and observed under transmission electron microscopy (TEM). The principal mechanism of PDT at a low dose of 9-HpbD-a was apoptosis, and at a high dose of 9-HpbD-a it was necrosis. PDT effects were also observed in a xenografted nude mouse model. Group I (no 9-HpbD-a, no laser irradiation) and Group II (9-HpbD-a injection only) showed no response (4/4, 100%), and Group III (laser irradiation only) showed recurrence (1/4,25%) or no response (3/4, 75 %). Group IV (9-HpbD-a + laser irradiation) showed complete response (10/16, 62.5%), recurrence (4/16, 25%) or no response (2/16, 12.5%). Group IV showed a significant remission rate compared to other groups (p<0.05). These results suggest that 9-HpbD-a is a promising photosensitizer for the future and that further studies on biodistribution, toxicity and mechanism of action would be needed to use 9-HpbD-a as a photosensitizer in the clinical setting.

• Journal of Life Science
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• v.24 no.4
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• pp.343-351
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• 2014

Phosphoinositide 3-kinase (PI3K) plays a central role in cell signaling and leads to cell proliferation, survival, motility, exocytosis, and cytoskeletal rearrangements, as well as specialized cell responses, superoxide production, and cardiac myocyte growth. PI3K is divided into three classes; type I PI3K is preferentially expressed in leukocytes and activated by ${\beta}{\gamma}$ subunits of heterotrimeric G-proteins. In this study, the cDNAs encoding the $PI3K{\gamma}$ gene were isolated from a brain cDNA library constructed using the flounder (Paralichthys olivaceus). The sequence of the isolated $PI3K{\gamma}$ was 1341 bp, encoding 447 amino acids. The nucleotide sequence of the $PI3K{\gamma}$ gene was analyzed with that of other species, including Oreochromis niloticus and Danio rerio, and it turned out to be well conserved during evolution. The $PI3K{\gamma}$ gene was subcloned into the expression vector pET-44a(+), and expressed in the E. coli BL21 (DE3) codon plus cell. The resulting protein was expressed as a fusion protein of approximately 49 kDa containing a C-terminal six-histidine extension that was derived from the expression vector. The expressed protein was purified to homogeneity by His-tag affinity chromatography and showed enzymatic activity corresponding to $PI3K{\gamma}$. The binding of wortmannin to $PI3K{\gamma}$, as detected by anti-wortmannin antisera, closely followed the inhibition of the kinase activities. The results obtained from this study will provide a wider base of knowledge on the primary structure and characterization of the $PI3K{\gamma}$ at the molecular level.

• Korean Journal of Weed Science
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• v.18 no.3
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• pp.225-236
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• 1998

KSC-13906 [Erythro N-{(4,6-dimethoxypyrimidin-2-yl)aminocarbonyl} -2-(2-fluoro-1-hydroxy-n-propyl) benzenesulfonamide, US Patent 5,461,025] was investigated how can control phytotoxicity fluctuation and what a good method apply to new rice herbicide. The growth inhibition was observed when the rice plants was transplanted at a shallow depth(0 - 1cm) and leaching was low(0 - 1cm/ day) from the paddy soil. KSC-13906 appeared to move readily down into the paddy soil with water by 3cm depth in the soil column(${\phi}$ 10cm) filled with loamy sand soil under 3cm/day of leaching condition. Artificial control releasing pattern, designed as treated with KSC-13906 of 9 or 18g ai/ha either at a once or daily treated dividing volume of 1/20, 1/25 and 1/30 of the total volume, increased the safety of KSC-13906 to direct seeded and transplanted rice. The safety of KSC-13906 was also enhanced when KSC-13906 was mixed with dymron. For example, the mixture of KSC-13906 and dymron effectively reduced injury of direct seeded rice plants at 18 and 500g ai/ha, respectively, treated 7 days after transplanting. However, combination of KSC-13906 and several herbicides didn't show any synergistic effetct on herbicidal activity and safening effect on rice. However, the combination of KSC-13906+dymron (9~12+250~500g ai/ha) or KSC-13906+mefenacet+dymron(9+250+250g ai/ha) controlled almost all weeds in paddy field without causing any injury to rice and thus the combination would successfully be used as an oneshot herbicide in rice culture.

• Korean Journal of Medicinal Crop Science
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• v.9 no.2
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• pp.156-165
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• 2001

Exogeneous application of pokeweed antiviral protein (PAP), a ribosomal-inacivating protein in the cell wall of Phytolacca americana (pokeweed) protects heterologous plants from viral and fungal infection. A cDNA clone of PAP introduced into Scrophularia buergeriana Miquel by thransformation with Agrobacterium tumefaciences. For plant transformation, explants were precultured on shoot induction medium without kanamycin for 2-5 day, and then they were cocultured with Agrobacterium for 10 minutes. The explants were placed on co culture medium in dark condition, $28^{\circ}C$ for 2days. After explants were washed in MS liquid medium, they were transferred into selection medium including kanamycin 50mg/L (MS salts+1mg/ l BAP+2mg/ l TDZ+0,2mg/ l NAA+MS vitamin+3% sucrose+0.8% agar, pH5.8). From PCR analysis, NPT II band was confirmed in transgenic plant genome and showed resistance against fungi in antifungal activity test. Micro assay to which protein extracted from transgenic line were added, revealed hyphae growth inhibition and no spore germination at high concentration. The characteristics of inhibited hyphae was represented transparent and thin. Expression of PAP in transgenic plants offers the possibility of developing resistance to viral and fungal infection.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.53 no.1
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• pp.75-84
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• 2008

Colored potatoes are an excellent source of dietary polyphenols including anthocyanins. Generally, anthocyanins from fruits and vegetables exhibit anti-carcinogenesis and anti-cancer properties in vitro test. This experiment was conducted to know the effects of colored potato extracts contained anthocyanins on antimutagenic activity and anticancer activity to six human cancer cell lines containing LNCaP (androgen-dependent) prostate cancer cells. Extracts of three colored potatoes ('Hongyoung', 'Jayoung' and 'Jasim') and the white potato ('Superior') cultivars were used in this study. The extracts of three colored potatoes inhibited the mutagenicities induced by direct mutagen such as 4-nitro-quinoline-1-oxide (4-NQO) and another indirect mutagens of bezo(a)pyrene (BaP). Also, the extracts of 'Hoyoung' and 'Jayoung' showed higher antimutagenic activity than 'Jasim' and 'Superior' against to direct or indirect mutagen on both strains of TA98 and TA100. The activity of growth-inhibitory of extract of four potato cultivars were screened by SRB (sulphorhodamine B) method on diverse human cancer cells representing different types of cancers. Among the extract of four potato cultivars, the extract of 'Jasim' showed moderate inhibition on proliferation of LNCaP, ACHN and MOLT-4F cells and did not inhibit the proliferation of other cancer cells. On the other hand, extract of 'Superior' did not inhibit the proliferation of any tested cancer cell lines. However, the extracts of 'Hongyoung and Jayoung' inhibited the proliferation of cancer cells with $GI_{50}$ values ranging from 2.5 to $30\;{\mu}g/mL$. On the basis of the $GI_{50}$ values, it is clear that LNCaP cells were more sensitive to extracts of colored potato cultivars than other cancer cells. The extract of 'Jayoung' at $30\;{\mu}g/mL$ were more active and inhibited cell proliferation, and induced apoptosis in LNCaP cells. This result revealed that the extracts of colored potatoes are expected to be good candidate for development into source of antimutagenic and anticancer agent.

• Horticultural Science & Technology
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• v.28 no.5
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• pp.871-876
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• 2010

Seven native and four introduced herbs namely $Thymus$ $quinquecostatus$, $Chrysanthemum$ $zawadskii$ var. $latilobum$, $Rosmarinus$ $officinalis$, etc. were selected for analysis of the DPPH radical scavenging and anti-microbial activity of their extracts. These perennial herbs are classified as $Labiatae$ and $Compositae$ except for $Saururus$ $chinensis$ and can be propagated through seedling and cuttage. These edible herbs are used as medicinal as well as ornamental plants. Their extract has electron donating ability which ranges from 69.7 to 78.7% for native herbs and 67.4 to 75.4% for introduced herbs. Native herbs have higher (3.54%) average DPPH radical scavenging than introduced herbs. In native herbs, maximum DPPH radical scavenging activity was observed in $Agastache$ $rugosa$ (78.7%) followed by $Saururus$ $chinensis$ while $Chamaemelum$ $nobile$ showed highest activity among the introduced herbs. Many herbs viz. $Saururus$ $chinensis$, $Chrysanthemum$ $zawadskii$ var. $latilobum$ and $Solidago$ $virga-aurea$ var. $gigantean$ showed excellent anti-microbial activity against gram positive $Enterococcus$ $faecalis$, maximum (80.0%) by $Saururus$ $chinensis$. Other herbs viz. $Solidago$ $virga-aurea$ var. $gigantea$, $Chrysanthemum$ $zawadskii$ var. $latilobum$, $Salvia$Salvia $plebeia$, $Chrysanthemum$ $indicum$, $Rosmarinus$ $officinalis$, $Chamaemelum$ $nobile$ and $Lavandula$ $stoechas$ showed anti-microbial activity against gram negative $Citrobacter$ $freundii$. Especially, the inhibition of colony growth of $Citrobacter$ $freundii$ was highest in the extract of $Chrysanthemum$ $zawadskii$ var. $latilobum$, and $Chamaemelum$ $nobile$.

• Journal of Animal Environmental Science
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• v.13 no.1
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• pp.45-52
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• 2007

To evaluate the feed value of starfish, antimicrobial effects of its extract, nutrients contents, concentration of amino acids and its bioavailability were tested. Steaming and ether processes were applied to obtain the extract from starfish for antimicrobial effects examination. The starfish was dried at $60^{\circ}C$ for 3 days before grinding for processing and fermentation. Ground starfish(GS), extruded starfish(ES), fermented starfish(EFS) were added with enzyme and without enzyme(Non enzyme fermented starfish : NEFS). Then the nutrient composition and bioavailability of those were analyzed. The extract from starfish showed no inhibition of the growth of lactobacillus and pathogenic bacteria. Protein content showed significantly higher 62.86% and 52.82%, respectively in EFS and NEFS than GS and EGS(p<0.05). The Ca content of GS, EGS, EFS and NEFS was 17.26%, 18.26%, 5.37% and 8.55%, respectively. It was low in EFS and NEFS due to measure the Ca content after fermentation. Total amino acid was 17.17 mg/g, 20.28 mg/g, 36.30 mg/g and 29.96 mg/g in GS, EGS, EFS and NEFS, respectively. The ratio of total amino acid to protein tended to show the similar tendency as total amino acid. Both total amino acid and its ratio to protein were increased by the fermentation. Bioavailability of the protein and Ca showed more 80% in EFS and NEFS. The nutrients availability of EFS were significantly higher in laying hens than other treatments. The results of these experiments indicate that starfish would be applied as a feed ingredients if it was properly treated.

• Asian Journal of Turfgrass Science
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• v.22 no.2
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• pp.161-170
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• 2008

Recently irregular dark-colored patches were found on the Kentucky teeing ground in a golf course in Gyunggi providence. Interestingly, blue-green algae from the leaf tissue sample containing black spot-stained symptoms were largely observed through microscopic study. In general, algae present on the upper soil surface or in the upper layer of root zone form dark brown layers of scum or crust, which invoked harmful effects to turf growth such as poor drainage, inhibition of new root development. In this observation, unlike the algae were sometime found in senescing leaves on contacted soil in July and August, the blue-green algae were detected within black spot-stained Kentucky bluegrass leaf tissues including leaf blade, ligule, auriclea as well as leaf sheath. The blue-green algae were also detected on the leaf and stem tissue adjacent to the symptomatic leaf tissues. Two species of blue-green algae, Phomidium and Oscillatoria, were greatly observed. Oscillatoria species was more commonly notified in all samples. In addition, the two species were found on a putting green showing yellow spot disease at another golf course in Gyunggi providence. The data from chemical control assay revealed that chemicals such as propiconazole, iprodione, and azoxystrobin decreased blue-green algae population and leaf spots, which finally resulted in enhanced leaf quality. All taken together, we strongly suggested that the disease-like phenomenon by blue-green algae might be very closely mediated with infection/translocation process in relation with turfgrass. It indicates that blue-green algae in turf management may play an adverse role as a secondary barrier as well as a pathogenic agent. This report may be helpful for superintendents to recognize and understand the fact that algae control should be provided more cautiously and seriously than we did previously in upcoming golf course management.

• Food Science and Preservation
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• v.21 no.5
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• pp.718-726
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• 2014

The objective of this study was to investigate the physiological activities of Aronia melanocarpa extracts on extraction solvents (through hot water extraction, 50% ethanol extraction, and 50% methanol extraction). The yield of 50% ethanol extract, 84.50%, was higher than that of the hot water extract (84.05%) and of the 50% methanol extract (76.20%). The total sugar content of the extraction solvent, 35.56~37.68 g/100 g, did not significantly differ. The total anthocyanin content of the 50% methanol extract, 395.10 mg/100 g, was higher than of 50% ethanol extract (318.61 mg/100 g) and of the hot water extract (252.82 mg/100 g). The anthocyanin composition of the cyanidin-3-galactoside, 364.65 mg/100 g, was higher than that of the cyanidin-3-arabinoside (163.06 mg/100 g) and of the cyanidin-3-glucoside (35.69 mg/100 g) in the 50% methanol extract. The DPPH radical scavenging activities of the 50% ethanol and the 50% methanol extracts at $100-1,000{\mu}g/mL$ were 7.96-70.01%, and 8.90-69.21%, respectively. The superoxide radical scavenging activities of all the extracts improved with an increase in the treatment concentration. The FRAP of the 50% ethanol extract and the 50% methanol extract at $100-1,000{\mu}g/mL$ were $57.14-817.87{\mu}M$ and $67.32-812.78{\mu}M$, respectively. The tyrosinase inhibitory activity of the 50% ethanol extract, 23.03-33.82% ($100-1,000{\mu}g/mL$), was higher than that of the other extracts. The cancer cell growth inhibition activity of the 50% ethanol extract (76.86% at $1,000{\mu}g/mL$) on HeLa cell line was significantly higher than of the hot water and of the 50% methanol extracts. There results suggest that the 50% ethanol extract from Aronia melanocarpa may be a useful for functional food material in the food industry.