• Korean Journal of Poultry Science
• /
• v.40 no.3
• /
• pp.171-178
• /
• 2013

The purpose of this study was to evaluate the sperm viability, normal acrosome and mitochondrial activity in the frozen-thawed fowl semen by different cryoprotectants. The experiment was carried out on 10 sexually adult roosters of Ogye. The semen was collected twice a week and pooled semen was diluted 1:1 EK extender containing no cryoprotectant at $5^{\circ}C$. After equilibration for 30 minutes, diluted chicken semen was diluted 1:1 extender containing either 7% dimethylacetamide (DMA), 7% dimethylformamide (DMF) or 7.5% methylacetamide (MA) at final concentration and was put in 0.5 mL plastic straws and frozen for 30 minutes by exposure to liquid nitrogen vapor 4 cm above the surface of liquid nitrogen, followed by plunging into liquid nitrogen. Frozen semen was thawed in water bath at $5^{\circ}C$ for 2 minutes. For cytometric analysis, the frozen-thawed semen was diluted with EK extender to a final concentration of 90 million spermatozoa per mL. Sperm membrane integrity was evaluated as SYBR-14 and propidium iodide (PI). Acrosome integrity was assessed with fluorescein isothiocyanate-labeled PSA and PI. The percentage of mitochondrial function was estimated by using Rhodamine123 (R123) and PI. In conclusion, freezing rooster semen by using 7% DMF as cryoprotectant was significantly highest in rates of survival and mitochondrial function while its rate of damage of acrosome was significantly lowest. As a result, DMF is the cryoprotectant that has the lowest influences on sperm membranes and acrosome integrity. Therefore it could be used for freezing method of animal genetic conservation method for poultry diversity.

• Korean Journal of Poultry Science
• /
• v.40 no.3
• /
• pp.197-205
• /
• 2013

This study established a method for preserving chicken primordial germ cells (PGCs) that enables long-term storage in liquid nitrogen ($LN_2$) for preservation of the species. The purpose of this study was to compare the effects of Ethylene Glycol (EG) and Propylene Glycol (PG) on viability of cryopreserved PGCs with vitrification in Korean Native Chicken (Ogye), and to fine should be find or to the optimal protocol for PGCs freezing. One of the important components of cryopreservation process is cryopreservation medium that plays a vital role in preventing cellular injury during freeze-thawing. Cryoprotective agents have been known to improve cell viability after freeze-thawing. PGCs obtained from the germinal gonade of 5.5~6 day (stage 28) chick embryos, using the MACS method were suspended in a freezing medium containing a freezing and protecting agents. Gonads were harvested from stage 28 chick embryos and pooled in groups of 10E embryos, contributing gonads to the cell suspension. The gonadal cells, including PGCs, were then frozen in 1 of the following cryoprotectant treatments: 2.5% EG, 5% EG, 10% EG, 2.5% PG, 5% PG, 10% PG, and 0% cryoprotectant as a control. Effects of exposure to vitrification solution and vitrification, with different concentrations of the cryoprotectant solution, were examined. After freezing and thawing, survival rates of the frozen-thawed PGCs from the 0, 2.5, 5, 10 and 15% EG plus FBS treatment were 44.24%, 64.51%, 85.63%, 80.51% and 73.52% (p<0.05), respectively. The viability of PGCs after freeze-thawing was significantly higher for 10% EG plus FBS treatment than for 10% PG + FBS treatment (p<0.05)(85.63% vs 66.81%). Therefore, these systems may contribute in the improvement of cryopreservation for a scarce species in birds preservation. This study established a method for preserving chicken PGC that enables systematic storage and labeling of cryopreserved PGCs in liquid N at a germplasm repository and ease of entry into a database. In the future, the importance for this new technology is that poultry lines can be conserved while work is being conducted on improving the production of germline chimeras.

• The Korean Journal of Zoology
• /
• v.16 no.2
• /
• pp.79-96
• /
• 1973

In order to elucidate the effects of copper on Corassius carassius, the following were studied: 1) lactate dehydrogenase isozyme patterns by cellulose acetate electrophoresis, 2) LDH activity and copper effect on LDH enzyme system y spectrophotometry, 3) esterase isozyme patterns by agar thin layer electrophoresis, 4) hemoglobin patterns by starch gel electrophoresis, and 5) histological study. 1. There were two bands of LDH isozymes (LDH-3 and LDH-5) in the gill, three bands (LDH-2, LDH-4, and LDH-5) in the liver, and two bands (LDH-3 and LDH-4) in the muscle of the normal fish. The LDH-1 bond was not found in the above three tissues. When the fish were exposed to copper, LDH-3 appeared in the liver, LDH-5 in the muscle, but no new LDH band appeared in the gill. 2. The sepcific activities of the LDH were lowest in the gill and highest in the muscle of the normal fish, and they were gradually decreassed in the gill and highest in the muscle of the normal fish, and they were gradually decreased in the liver and mucle except in the gill from 1-day to 10-day exposure to copper. It indicates that LDH activities in the liver and muscle of the fish were inhibited by copper. 3. Through in vitro experiment, it is clear that the decrease of the LDH activities of the liver and muscle of the fish exposed to copper is mainly caused by the inhibition on the M-LDH in the fish. 4. The numbers of the esterase isozyme bands of the gill, liver, muscle, blood, brain, and kidney of the normal fish were 3, 6, 2, 2, 2, and 2 respectively, and these numbers were the same as those exposed to copper. The relative mobilities of the esterase bands in the gill, liver, blood, and kidney of the exposed group were different from those of the control. 5. There was one hemoglobin band on the anode in the normal fish. It seems that the nobility of hemoglobin band of the fish exposed to copper was slightly faster than that of the normal fish. 6. The normal gill lamellae of the fish consisted of centrally located pillar cells and a number of mucus cells. When the fish were exposed to copper, the epithelial layer was divorced first, disintegrated, and then destroyed completely. 7. The liver of the normal fish had prominent central veins, cords of hepatic cells, and sinusoids. When the fish were exposed to copper, numerous droplets of fat appeared in the cells around the central vein of the liver. It is assumed that the fatty droplets were accumulated by the lesion due to fatty metamorphosis of the liver caused by copper. 8. There was no histological difference between the muscle of the normal fish and that of the fish exposed to copper. 9. In the normal fish, the tubules of the kidney were surrounded by hemopoetic tissues. However, the kidney tissue of the fish exposed to copper received some damage on the proximal tubules. Since the tubule cells were reduced in height, the lumens of the tubules were enlarged. Consequently many proximal tubules exhibited some pink-stained granular casts and various stages of degeneration.

• The Journal of Dong Guk Oriental Medicine
• /
• v.7 no.1
• /
• pp.33-41
• /
• 1998

Lymph node tissues of BALB/C mouse treated with DNCB were immunohistochemically observed to investigate the activation of cell mediated immunity in lymph node of murine with allergic contact dermatitis. The inguinal region of BALB/C mice were sensitized by one application of $25{\mu}l$ of 5% 2,4-dinitrochlorobenzene(DNCB) onto an abdominal skin and 2 weeks later, the mice were challenged with $4{\mu}l$ of 2.5% DNCB. The inguinal lymph node were obtained at hour 24, 48, and 72 after 2nd DNCB treatment and embedded with paraffin, and then stained by following ABC method that used monoclonal antibody including L3T4(CD4), Ly2(CD8), IL-2R(CD25). The distribution of helper T lymphocytes, cytotoxic T lymphocytes and IL-2 receptors began to increase at hour 24 after after 2nd DNCB treatment and these increase appeared in paracortical area and medullary sinius. These increase were greatest at hour 48. These results indicated that the IL-2 secretion began to increase by activation of helper T lymphocytes in lymph node of DNCB re-exposure area and subsequently to activate suppress T lymphocytes.

• 보존과학연구
• /
• s.17
• /
• pp.161-182
• /
• 1996

In natural atmosphere, copper and copper alloy have been used to make buddha statues and ornaments of historic buildings since the abovementioned metals have corrosion resistance in some extent, and the patinaformed on the surface of the metals has provided the people aesthetic satisfaction with its beauty. But in atmosphere polluted by $SO_x$and $NO_x$, the patina layer does not work as a protective film, and it allows damages of the metal. Since 1992, Tokyo National Research Institute of Cultural Properties(TNRICP)has conducted studies on the influence of atmospheric pollution on metal cultural property held under open air. The Great Buddha Image which is located in Kamakura about 50km west from Tokyo, has been selected as one of the objects to study because it is made by copper alloy and it has stood exposed in the air for about a few hundreds years. Furthermore it is also the reason to study on it that there are many cultural properties in the surroundings of it. We have analysed the components and the structure of the corrosion products formed on the surface of the Buddha, have carried out exposure tests using the alloy samples which have simulated the components of the Great Image, and have observed climated and polluted air in order to discuss the relation between corrosion of metals in open air and conditions of the atmosphere. In this paper, the authors have described the components and the structure of the corrosion product formed on the surface of the Great Image by means of X-ray fluorescence spectroscopy and X-ray diffraction. The conclusions are as follows. (1) Sulfate patina composed mainly with brochantite were detected on the all sides of the Image and the amount of the patina is found more on the back of the Image facing to north. (2) Antlerite were detected on the back and a park of the left side facing to west, and formation of it was considered to have close relation with malignant atmosphere. (3) A big amount of chloride patina which mainly composed of atacamite were observed on the front facing to south. (4) Carbonate patina mainly composed of malachite were detected on the area where brochantite was often detected as well. It suggested that malachite had been transformed into brochantite by deteriorated atmosphere. (5) On the all sides of the Image, patina were observed together with copper oxides mainly composed of cuprous oxide. It showed that the surface layer of the Image consists of two layers : inner layer of oxide and outer layer of patina. (6) Corrosion products of lead which was a component of copperalloy were detected on the all sides : the main lead product found on the front was chlorophosphate whereas the one on the back was sulfate.

• Radiation Oncology Journal
• /
• v.25 no.3
• /
• pp.185-191
• /
• 2007

Purpose: This study was performed to investigate the effects of immune cell activation and the antitumor effect for the combination of treatment with X-irradiation and E/eutherococcus senticosus Maxim Root (ESMR) on mouse tumor cells. Materials and Methods: ESMR (250g) was extracted with 80% methanol, concentrated under decompression and lyophilized. To determine whether ESMR is able to activate the immune cells or not, the proliferation of splenocytes in vitro and the number of B cells and T cells in splenic lymphocytes in ESMR-pretreated mice were evaluated. X-irradiation was given to the mouse fibrosarcoma tumor cells (FSa II) by 250 kv X-irradiation machine. The cytotoxicity of ESMR was evaluated from its ability to reduce the clonogenecity of FSa II cells. In X-irradiation alone group, each 2, 4, 6 and 8 Gy was given to FSa II cells. In X-irradiation with ESMR group, 0.2 mg/ml of ESMR was exposed to FSa II cells for 1 hour before X-irradiation. Results: The proliferation of cultured mouse splenocytes and thymocytes were enhanced by the addition of ESMR in vitro. The number of B cells and T cells in mouse splenic lymphocytes was significantly increased in ESMR pretreated mice in vivo. In FSa II cells that received a combination of 0.2 mg/ml of ESMR with X-irradiation exposure, the survival fraction with a dose of 2, 4 and 6 Gy was $0.39{\pm}0.005$, $0.22{\pm}0.005$ and $0.06{\pm}0.007$, respectively. For FSa II cells treated with X-irradiation alone, the survival fraction with a dose of 2, 4 and 6 Gy was $0.76{\pm}0.02$, $0.47{\pm}0.008$ and $0.37{\pm}0.01$. The difference in the survival fraction of the mouse FSa II cells treated with and without ESMR was statistically significant (p<0.05). Conclusion: Treatment with ESMR increased cell viability of mouse splenocytes in vitro and especially the subpopulation of B cells and T cells in splenocytes in ESMR-pretreated mice. However, treatment with ESMR did not increase the level of Th and Tc subpopulations in the thymocytes. Treatment with the combination of ESMR and X-irradiation was more cytotoxic to mouse tumor cells than treatment with X-irradiation alone; this finding was statistically significant.

• Korean Journal of Weed Science
• /
• v.6 no.2
• /
• pp.168-173
• /
• 1986

The effects of varying concentrations and duration of fluazifop-butyl [(${\pm}$)-butyl [2- [4- [(5-(trifluoro methyl)-2-pyridinyl] oxy] phenoxy] propanate] treatment on cell division, cell enlargement, and protein synthesis were studied. Oat (Avena staiva L.) were treated from 0 to 48 hr with concentration ranging from $1{\times}10^{-6}M$ to $1{\times}10^{-3}M$ of fluazifop-butyl in the cell division study. There was a significant reduction in the mitotic indices of oat roots treated with $1{\times}10^{-4}M$ after 6 hr. After 18 hr treatment, All herbicide treatment inhibited cell division significantly. After 24 hr treatment almost 100% inhibition of cell division occurred at $1{\times}10^{-4}M$ to $1{\times}10^{-3}M$ while 20% inhibition of cell division occurred at $1{\times}10^{-6}M$ concentration at same exposure period. The greatest inhibition of cell division occurred between 0 to 18 hr. The avena coleoptile straight- growth test were used to determine the influence of fluazifop-butyl on eoleoptile growth. Significant inhibition of elongation of oat coleoptiles were observed at $1{\times}10^{-7}M$ to $1{\times}10^{-3}M$ after 24 hr incubation. Protein incorporation study showed that the $1{\times}10^{-4}M$ of fluazifop-butyl caused 60% inhibition of protein synthesis. It was concluded that the growth of inhibition of plants caused by fluazifop-butyl results from inhibition of cell division, cell enlargement, and protein synthesis.

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

• Journal of Life Science
• /
• v.23 no.5
• /
• pp.689-697
• /
• 2013

In the United States, about 40,000 new cases of oral cancer are diagnosed each year and nearly 7,800 patients died from it in 2012. Omega-3 polyunsaturated fatty acids have been found to have anticancer effects in a variety of cancer cell lines and animal models, but their effect in oral cancer remains unclear. This study was designed to examine the effect of docosahexaenoic acid (DHA, a kind of omega-3 fatty acid) on oral cancer cells and the molecular mechanism of its action. We found that exposure of squamous cell carcinoma-4 (SCC-4) and squamous cell carcinoma-9 (SCC-9) human oral cancer cells to DHA induced growth inhibition in a dose- and time-dependent manner. Meanwhile, in addition to the elevated levels of apoptotic markers, such as cleaved PARP, subG1 portion and TUNEL-positive nuclei, DHA led to autophagic vesicle formation and an increase in autophagic flux, indicating the involvement of both apoptosis and autophagy in the inhibitory effects of DHA on oral cancer cells. Further experiments revealed that the apoptosis and autophagy induced by DHA were linked to inhibition of mammalian target of rapamycin (mTOR) signaling by AKT inhibition and AMP-activated protein kinase (AMPK) activation in SCC-9 cells. Together, our results suggest that DHA induces apoptosis- and autophagy-associated cell death through the AMPK/AKT/mTOR signaling pathway in oral cancer cells. Thus, utilization of omega-3 fatty acids may represent a promising therapeutic approach for chemoprevention and treatment of human oral cancer.

• The Journal of Korean Society for Radiation Therapy
• /
• v.26 no.1
• /
• pp.91-98
• /
• 2014

Purpose : To compare the dosimetry for the left breast cancer treatment between three dimensional conformal radiation radiotherapy (3D-CRT) and Hybrid planning and to estimate usefulness of Hybrid planning Materials and Methods : Five patients with left breast cancer were included in the study. They were planned using several different radiotherapy techniques including: 1)open rectangular field, 2)tangential wedge-based field 3)field in field, 4)hybrid planning(energy, wedge combine). For each patient planning was using Light Speed RT-16 CT and PINNACLE planning system-ver.9.2. Hybrid plan was made using same system and using the same targets and optimization goals. We comparing the Homogeneity Index(HI), normal organs at the does-volume histogram(DVH) Results : In all plans, the Homogeneity Index(HI) of Hybrid planning was significantly better than other. Dose comparison of HI= 2D-RT:38.32, TW:38.32, FIF:29.22, HYBRID:30.57. 2D-RT, TW, FIF Hybrid$V_{75_-lung}$=112.33, 125.14, 121.3, 123.78. $V_{50_-lung}$=155.43, 159.62, 157.96, 159.06. $V_{25_-lung}$=199.86, 200.22, 198.65, 200.31. $V_{50_-heart}$=26.07, 27.1, 26.85, 27.17 $V_{30_-heart}$=33.71, 34.37, 34.15, 34.65 Conclusion : In summary, 3D-CRT, Hybrid planning techniques were found to have acceptableCTV coverage in our study. However the Hybrid planning increased radiation dose exposure to normal tissue. If you apply for treatment of inhomogeneity areas like lung, For best results will be achieved.