• 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.

• Radiation Oncology Journal
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• v.16 no.2
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• pp.159-165
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• 1998

Purpose : The best prognosis for hepatocellular carcinoma can be achieved with surgical resection. However, the number of resected cases is limited due to the advanced lesion or associated liver disease. A trial of combined transcatheter arterial chemoembolization(TACE) and local radiotherapy(RT) for unresectable hepatocellular carcinoma(HCC) was prospectively conducted and its efficacy and toxicity were investigated. Materials and Methods : From 1992 to 1994, 30 Patients with unresectable HCC due either to advanced lesion or to associated cirrhosis were entered in the study Exclusion criteria included the presence of extrahepatic metastasis, liver cirrhosis of Child's class C, tumors occupying more than two-thirds of the whole liver, and an ECOG scale of more than 3. Patient cHaracteristics were : mean tumor size $8.95\pm3.4cm$, serum AFP+ in all patients, portal vein thrombosis in all patients, liver cirrhosis in 22 patients, and UICC stage III and IVA in 10 and 20 patients, respectively. TACE was performed with the mixture of Lipiodol(5ml) and Adriamycin(50mg) and Gelfoam embolizatin. RT(mean dose $44.0\pm9.3Gy$) 10 days with conventional fractionation. Results : An objective response was observed in 19 patients($63.3\%$). Survival rates at 1 2, and 3 years were $67\%,\;33.3\%$ and $22.2\%$, respectively. Median survival was 17 months. There were 6 patients surviving more than 3 years. Distant metastasis occurred in 10 patients, with 8 in the lung only and 2 in both lung and bone, Toxicity included transient elevation of liver function test in all patients, fever in 20, thrombocytopenia in 4, and nausea and vomiting in 1. There was no treatment-related death. Conclusion : Combined TACE and RT appear to produce a favorable response and survival results with minimal toxicity.

• Journal of Chest Surgery
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• v.41 no.2
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• pp.239-246
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• 2008

Background: Aortic surgery for high risk patients has high mortality and morbidity rates, and the necessity of performing aortic surgery in cancer patients is questionable because of their short life expectancy. Endovascular repair of aneurysm repair can be considered for high risk patients and cancer patients because it has relatively lower invasiveness and shorter recovery times than aortic surgery does. Especially, percutaneous endovascular stent graft treatment is more useful for high risk patients because it does not require general anesthesia. Material and Method: From July 2003 to September 2007, twelve patients who had inoperable malignancy or who had a high risk of complication because of their combined diseases during aortic surgery underwent endovascular aortic aneurysm repair. he indications for endovascular repair were abdominal aortic aneurysm in 5 patients, descending thoracic aortic aneurysm in 6 patients and acute type B aortic dissection in one patient. The underlying combined disease of these patients were malignancy in 3 patients, respiratory disease in 6 patients, old age with neurologic disease in 6 patients, Behcet's iseae in one patient and chronic renal failure in one patient. Result: Stent grafts were inserted percutaneously in all cases. There were 4 hospital deaths and there were 3 delayed deaths during the follow-up periods. There were no deaths from aortic disease, except one hospital death. There were several complications: a mild cerebrovascular accident occurred in one patient, acute renal failure occurred in 2 patients and ischemic bowel necrosis occurred in one patient. Mild type I endoleak was observed in 2 patients and type II endoleak was observed in a patient after stent graft implantation. Newly developed type I endoleak was observed in a patient during the follow-up period. Conclusion: Percutaneous endovascular stent graft insertion is relatively safe procedure for high risk patients and cancer patients. Yet it seems that its indications and its long term results need to be further researched.

• Proceedings of the Microbiological Society of Korea Conference
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• 2008.05a
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• pp.39-41
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• 2008

The yeast Saccharomyces cerevisiae has been shown to contain three isoforms of citrate synthase (CS). The mitochondrial CS, Cit1, catalyzes the first reaction of the TCA cycle, i.e., condensation of acetyl-CoA and oxaloacetate to form citrate [1]. The peroxisomal CS, Cit2, participates in the glyoxylate cycle [2]. The third CS is a minor mitochondrial isofunctional enzyme, Cit3, and related to glycerol metabolism. However, the level of its intracellular activity is low and insufficient for metabolic needs of cells [3]. It has been reported that ${\Delta}cit1$ strain is not able to grow with acetate as a sole carbon source on either rich or minimal medium and that it shows a lag in attaining parental growth rates on nonfermentable carbon sources [2, 4, 5]. Cells of ${\Delta}cit2$, on the other hand, have similar growth phenotype as wild-type on various carbon sources. Thus, the biochemical basis of carbon metabolism in the yeast cells with deletion of CIT1 or CIT2 gene has not been clearly addressed yet. In the present study, we focused our efforts on understanding the function of Cit2 in utilizing $C_2$ carbon sources and then found that ${\Delta}cit1$ cells can grow on minimal medium containing $C_2$ carbon sources, such as acetate. We also analyzed that the characteristics of mutant strains defective in each of the genes encoding the enzymes involved in TCA and glyoxylate cycles and membrane carriers for metabolite transport. Our results suggest that citrate produced by peroxisomal CS can be utilized via glyoxylate cycle, and moreover that the glyoxylate cycle by itself functions as a fully competent metabolic pathway for acetate utilization in S. cerevisiae. We also studied the relationship between Cit1 and apoptosis in S. cerevisiae [6]. In multicellular organisms, apoptosis is a highly regulated process of cell death that allows a cell to self-degrade in order for the body to eliminate potentially threatening or undesired cells, and thus is a crucial event for common defense mechanisms and in development [7]. The process of cellular suicide is also present in unicellular organisms such as yeast Saccharomyces cerevisiae [8]. When unicellular organisms are exposed to harsh conditions, apoptosis may serve as a defense mechanism for the preservation of cell populations through the sacrifice of some members of a population to promote the survival of others [9]. Apoptosis in S. cerevisiae shows some typical features of mammalian apoptosis such as flipping of phosphatidylserine, membrane blebbing, chromatin condensation and margination, and DNA cleavage [10]. Yeast cells with ${\Delta}cit1$ deletion showed a temperature-sensitive growth phenotype, and displayed a rapid loss in viability associated with typical apoptotic hallmarks, i.e., ROS accumulation, nuclear fragmentation, DNA breakage, and phosphatidylserine translocation, when exposed to heat stress. Upon long-term cultivation, ${\Delta}cit1$ cells showed increased potentials for both aging-induced apoptosis and adaptive regrowth. Activation of the metacaspase Yca1 was detected during heat- or aging-induced apoptosis in ${\Delta}cit1$ cells, and accordingly, deletion of YCA1 suppressed the apoptotic phenotype caused by ${\Delta}cit1$ mutation. Cells with ${\Delta}cit1$ deletion showed higher tendency toward glutathione (GSH) depletion and subsequent ROS accumulation than the wild-type, which was rescued by exogenous GSH, glutamate, or glutathione disulfide (GSSG). Beside Cit1, other enzymes of TCA cycle and glutamate dehydrogenases (GDHs) were found to be involved in stress-induced apoptosis. Deletion of the genes encoding the TCA cycle enzymes and one of the three GDHs, Gdh3, caused increased sensitivity to heat stress. These results lead us to conclude that GSH deficiency in ${\Delta}cit1$ cells is caused by an insufficient supply of glutamate necessary for biosynthesis of GSH rather than the depletion of reducing power required for reduction of GSSG to GSH.

• Journal of The Korean Society of Grassland and Forage Science
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• v.3 no.2
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• pp.67-76
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• 1983

This experiment was carried out to investigate the effects of the different nitrogen rates and cutting heights on the dead stubble after cutting, and the carbohydrate reserved in stubble of sorghum-sudangrass hybrid (Sorghum bicolor (L.) Moench) Pioneer 988. The experiment was undertaken in the Experimental Livestock Farm of Agriculture Coll., Seoul Nat'l Univ. In Suweon, 1981 and 1982. The results obtained are summarized as follows: 1. The dead stubble after cutting was found to be great with high rate of nitrogen fertilizer and low cutting height. It was also learned that the rainfall during forage cutting period made the dead stubble increased. 2. A significant higher (p<0.05) carbohydrate reserves in the stubble was observed in the high stubble height at the first cutting time and 6th through 7th day after the first cutting. The results indicate that the high stubble height reserves more carbohydrate for the early regrwoth stage after the first cutting when comparing with the low stubble. 3. The content of carbohydrate reserves was influenced by climete. Drought is caused to high content of carbohydrate, whereas, rain is caused to low content of that. The critical soluble carbohydrate content causing the death of stubble supposed to be 3 to 6% at least. 4. It is suggest that carbohydrate reserves in plant do not play a distinctive role for the regrowth in a summer annual forage like sorghum-sudangrass hybrid, but it might be rather influenced by the other factors, for example, environmental conditions at harvest and new bud.

• Journal of the korean academy of Pediatric Dentistry
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• v.29 no.2
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• pp.243-254
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• 2002

For the purpose of evaluating the biocompatability of 3 kinds of metallic materials frequently used in pediatric dentistry (stainless steel crown, orthodontic band, orthodontic wire), cellular and molecular studies, including cell growth and proliferation, screening of cell death with determination of types whether necrosis or apoptosis and changes in expressions of related signaling molecules were examined, using cultured human gingival fibroblasts (HGF-1), HGF-1 was cultured in Dulbecco's modified Eagle's medium. among which the 3rd to 6th generations of HGF-1 were used. The specimen were divided into stainless steel crown (R), band (B) and wire (W). The immunocytochemical study was done for the detection of anti-PCNA (proliferating cell nuclear antigen) labeling. With extracted protein, western blot was done for the detection of ERK1/2, JNK, and p38, using individual antibodies. Cultured cells proliferated, remarkably till 7 day and slightly at 11 day. There was no statistical significance in the counts of proliferating HGF-1 between control and experimental groups (p>0.05). Relative growth rates were no statistically significant difference between control and experimental groups (p>0.05). PCNA labeling indexes showing similar patterns in control and experimental groups. The expressions of ERK1 and ERK2, p38 were similar in control and experimental groups. The expression of JNK increased at 1st day, slightly decreased at 4th day and markedly increased at 7th and 11 day. Although the patterns of control and experimental groups were similar, the increased expressions of JNK at late period suggest a possible stress due to inhibited cell growth and proliferation, and worse culture condition. Conclusively, the 3 kinds of metal specimens used in this study did not induce cellular and molecular hazards during short term culture of HGF-1. But, for the better clinical stability, the establishment of long period culture and animal experiment was thought necessary.

• Journal of Life Science
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• v.24 no.3
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• pp.242-251
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• 2014

The objective of this study was to isolate a photosensitizer from Pueraria thunbergiana leaves that induces apoptosis in SK-HEP-1 cells. Column chromatography and thin layer chromatography were used to isolate active compounds from extracts of P. thunbergiana leaves. The structures of the isolated compounds were determined by 1D-NMR, 2D-NMR, and FAB-mass spectroscopy. A substance, named M4-3, was purified from the leaves of P. thunbergiana using various chromatography methods, and the absorbance of the substance was measured. The absorbance was highest at 410 nm, suggesting that the M4-3 substance was a different compound from chlorophyll a and b, which absorb at 410, 502, 533, and 607 nm. Further analyses revealed that the M4-3 compound was a $13^2$-hydoxy pheophorbide, a methyl ester with a molecular weight of 662. M4-3 was identified as a derivative compound of pheophorbide, with a structure that magnesium comes away from the porphyrin ring. The results of the analysis of the cytotoxicity of the M4-3 substance against the SK-HEP-1 cells revealed that it inhibited rates of cell growth by 40% and 80% at a concentration of 0.04 ${\mu}M$ and 0.08 ${\mu}M$, respectively. The M4-3 compound was found to be a photosensitizer for cytotoxicity because it was appeared only in light condition as examining activity in different irradiation conditions (light condition and nonlight condition) under the same concentration. Analysis of morphological changes in the cells following cell death induced by exposure to the M4-3 substance reveled representative phenomena of apoptosis (nuclear condensation, vesicle formation, and fragmentation of DNA). The induction of apoptosis was attributed to the compound's photodynamic activity.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.18 no.1
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• pp.52-56
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• 1985

For the fundamental studies of radiation breeding in edible marine algae, the biological effects on conchospores of Porphyra species by gamma-irradiation were examined. Two varieties, Keun-cham-gim (Porphyra tenera Kjell. form tamatsuensis Miura) and Saga No.5, were chosen for this study, and their conchospores after r-irradiation($5.0{\sim}20.0$ KR) were cultured for 50 days. The results obtained were summarized as follows. 1. Gamma-irradiation in less than the dose of 20KR did not affect germination of conchospores, and almost all spores grew into two cells germ in 24 hours after irradiation, but withering germs were gradually increased in number according to higher exposure within 5 days old culture. 2. High irradiation caused the induction of giant cells, abnormal useless growth of hold-fast, lumpish thalli and callus-like lumpy tissues. 3. The liberation of neutral spores from young germs and carpospores from mature thalli were observed on the frond exposed at $10{\sim}20$ KR irradiation. All spores were normal in division and its size. 4. The best irradiation effect on growth of Keun-cham-gim was observed at 10 KR dose, whose growth-rates were $140\%$ in wet weight and $108\%$ in mean frond area, but only $48\%$ was recorded in wet weight at 20 KR exposure. Saga No.5 were in contrast with Keun-cham-gim, and their most growth-rate was $400\%$ in wet weight ($258\%$ in frond area) at 10 KR irradiation and the worst was $20\%$ at the dose of 20 KR. 5. The withering phenomenon to death by treatment of gamma-ray presented substantial difference between two varieties. Survival rate compared with control in Keun-cham-gim was $70.7\%$ at 20 KR, but that in Saga No.5 recorded $47.0\%$ at same dose. 6. Synthesizing the results of high and low r-irradiation, it was suggested tat high r-irradiation in more than 5.0 KR inhibited conspicuously the growth of germs derived from conchospores, and about half of them withered at 15.0 KR dose or more.

• Journal of Korean Society of Forest Science
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• v.85 no.2
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• pp.271-279
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• 1996

This study was conducted to find out the appropriate rates of mixed-seeding of native woody and herb species. It was carried out in greenhouse from 1994 to 1995. The early days germination individuals were $60no./m^2$ in case of woody species for mixed seeding treatment and the treatments were fertilization, soil surface treatment, seeding amount. The investigation factors are the number of individuals of woody and herb species, biomass, height, number of tillers, etc. The mean individuals of Lespedeza bicolor and Amorpha frutzcosa were investigated $1.14no./m^2$, $0.496no./m^2$ in 1995, and Spiraea prunifolia var. simpliciflora and Alnus hirsuta were withering to death. In 1995, the Individuals of Oenothera odorata, Themeda triandra var. japonica, Cymbopogon tortilis var. goeringii, Miscanthus sinensis, Artemisia princeps var. orientalis, Lespedeza cuneata, and Arundinella hirta were investigated $5.06no./m^2$, $1,072no./m^2$, $0no./m^2$, $412.53no./m^2$, $88.6no./m^2$, $8.9no./m^2$, $57.46no./m^2$, respectively, in case of herb species. The height and biomass of Lespedeza bicolor and Amorpha fruticosa showed very fast growth, and those of Themeda triandra var. japonica, Miscanthus sinensis had a similar tendency. For the changes in woody species, according to the seeding amount, there was no significant relationship between species, but It was highly significant in 1995. In case of the herb species, the relationship between the number of individuals and seeding amount was significant for both 1994 and 1995. The most number of tiller was Miscanthus sinensis(21), and showed Themeda triandra var. japonica, Arundinella hirta in descending order. For the mixed seeding of wood and herb species, the number of individuals, height, biomass, tiller of herb species diminished as the seeding amount of herb species increased. It can be concluded that using Lespedeza bicolor and Amorpha fruticosa for woody species and like Arundinella hirta in herb species will be efficient for revegetation measures. It seems, therefore, that the plants of revegetation methods will be used to Lespedeza bicolor, Amorpha fruticosa and Arundinella hirta, and further study is needed on the seeding amount.

• Korean journal of applied entomology
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• v.24 no.3 s.64
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• pp.117-121
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• 1985

Through the laboratory and vinyl house experiments, the effects of inoculum density, plant age and temperature on the incidence of Phytophthora crown rot of pepper (Capsicum annum L.) were investigated. The propagule survival was greater in the natural soil than in autoclaved soil within first 2 weeks when the sporangial suspension of the pathogenic fungus was incorporated into soil, thereafter the survivability reduced rapidly. The propagule was not detectable in 35 days by means of Papavizas selective medium neither in natural nor in autoclaved soil. At least 5 sporangia per gram soil were required to induce crown rot for 30 days old pepper seedlings. Further increase in inoculum concentration above this threshold level resulted in higher disease incidence and shorter incubation period. When the same amount of inoculum was infested, higher disease incidence was observed for younger plants until 3 weeks after inoculation. On the other hand after 4 weeks this tendency was not extended any more. Younger plants were recognized as having shorter incubation period upon infection, however, the days from first symptom appearance to complete death were not significantly different among differently aged seedlings. Exposure of inoculated pepper seedlings to $25^{\circ}C$ resulted in highest infection rates and followed by those to $30^{\circ}C\;and\;20^{\circ}C$ but no disease was found at $15^{\circ}C\;and\;35^{\circ}C$ for 10 days. When the plants previously incubated at different temperature for 10 days were moved to $25^{\circ}C$ room temperature, prior exposure to $20^{\circ}C\;and\;30^{\circ}C$ brought continuous disease development. Even those plants preincubated at $15^{\circ}C$ were diseased up to 50%. But the prior exposure to $35^{\circ}C$ induced no symptom developed, indicating no seedlings infected at all.