Background: Current vascular prostheses are still inadequate for reconstruction of small-diameter vessels. Autologous pericardium can be a good alternative for this purpose as it already possesses good blood compatibility and shows a mechanical behavior similar to that of natural arteries. However, the clinical use of autologous pericardial tissue as a small-diameter vascular graft has limitations due to mixed outcomes from uncertain biological behavior and difficulty to gain reliable patency results in animal experiments. To study this issue, we implanted fresh and glutaraldehyde-treated autologous pericardium as small-diameter arterial grafts in dogs, and compared their time-related changes histologically. Material and Method: As a form of 5mm-diameter arterial graft, one pair of autologous pericardial tissue was used for comparison between the glutaraldehyde-treated and the glutaraldehyde-untreated grafts in the bilateral carotid arteries in the same dog. The patency of the grafts were evaluated at regular intervals with Doppler ultrasonography. After the predetermined periods of 3 days, 2 weeks, 1 month, 3 months and 6 months, the grafts in each animal were explanted. The retrieved grafts were processed for light and electron microscopic analyses following gross observation. Result: Of 7 animals, 2 were excluded from the study because one died postoperatively due to bleeding and the other was documented as one side of the grafts being obstructed. All 10 grafts in the remaining 5 dogs were patent. Grossly, a variable degree of thromboses were observed in the luminal surfaces of the grafts at 3 days and 2 weeks, despite good patency. Pseudointimal smooth blood-contacting surfaces were developed in the grafts at f month and later. By light microscopy, mesothelial cell layers of the pericardial tissue were absent in all explanted grafts. Newly formed endothelial cell layers on the blood-contacting surface were observed in both the glutaraldehyde-treated and fresh grafts at 3 months and later. The collagen fibers became degraded by fragmentation in the fresh graft at 1 month and In the glutaraldehyde-treated graft at 3 months. At 6 months, the collagen layers were no longer visible in either the glutaraldehyde-treated or fresh grafts. By electron microscopy, a greater amount of coarse fibrin fibers were observed in the fresh grafts than in the glutaraldehyde-treated grafts and, more compact and well-arrayed layers were observed in the glutaraldehyde-treated grafts than in the fresh grafts. Conclusion: The glutaraldehyde-treated small-diameter pericardial arterial grafts showed a better endothelialization of the blood-contacting surface and a slower fragmentation of the collagen layers than the fresh grafts, although it has yet to be proven whether these differences are so significant as to affect the patency results between the groups.
Purpose : To evaluate changes in expression of cell cycle related genes during apoptosis induced in HL60 cells by X-irradiation to understand molecular biologic aspects in mechanism of radiation therapy. Material and Methods : HL-60 cell line (promyelocytic leukemia cell line) was grown in culture media and irradiated with 8 Gr by linear accelerator (6 MV X-ray). At various times after irradiation, ranging from 3 to 48 hours were analyzed apoptotic DNA fragmentation assay for apoptosis and by western blot analysis and semi-quantitative RT-PCR for expression of cell cycle related genes (cyclin A, cyclin B, cyclin C, cyclin Dl, cyclin E, cdc2, CDK2, CDK4, $p16^{INK4a}$, $p21^{WAF1}$, $p27^{KIP1}$, E2F, PCNA and Rb). Results : X-irradiation (8 Gy) induced apoptosis in HL-60 cell line. Cycline A protein increased after reaching its peak 48 h after radiation delivery and cyclin E, E2F, CDK2 and RB protein increased then decreased after radiation. Radiation induced up-regulation of the expression of E2F is due to mostly increase of Phosphorylated retinoblastoma proteins (ppRb). Cyclin Dl, PCNA, CDC2, CDK4 and $p16^{INK4a}$ protein underwent no significant change at any times after irradiation. There was not detected $p21^{WAF1}$ and $p27^{KIP1}$ protein. Cyclin A, B, C mRNA decreased immediately after radiation and then increased at 12 h after radiation. Cyclin Dl mRNA increased immediately and then decreased at 48 h after radiation. After radiation, cyclin E mRNA decreased with the lapse of time. CDK2 mRNA decreased at 3h and increased at eh after radiation. CDK4 mRNA rapidly increased at 6 to 12 h after radiation. There was no change of expression of $p16^{INK4a}$ and not detected in expressin of $p21^{WAF1}$ and $p27^{KIP1}$ mRNA. Conclusion : We suggest that entry into S phase may contribute to apoptosis of HL60 cells induced by irradiation. Increase of ppRb and decrease of pRb protein are related with radiation induced auoptosis of HL60 cells and tosis of HL60 cells induced by irradiation. Increase of ppRb and decrease of PRb protein are related with radiation induced apoptosis of HL60 cells and this may be associated with induction of E2F and cyclinE/CDK2. These results support that $p21^{WAF1}$ and $p27^{KIP1}$ are not related with radiation induced-apoptosis.
Amphibians are a taxonomic group that ecologically connects terrestrial ecosystems and aquatic ecosystems. They play a very important role in the food chain of the ecosystem. It is known that there are about 5,948 species distributed all over the world, but after the Industrial Revolution, due to industrialization and urbanization, there has been a decrease in species and populations. In particular, it is becoming a factor in exacerbating habitat fragmentation or fragmentation due to artificial canals. In orderto improve the survivalrate of wild animals in artificial canals, escape facilities are installed to reduce it. This study analyzed the slope, height of the escape facility, escape rate, and travel distance in the operating facility for Bombina orientalis, which mainly inhabits near forests. The slope of the escape facility showed a relatively similar escape success rate regardless of height at 50° and 60°, while at 70°, it showed a relatively high escape success rate at only 40cm in height. The success rate of escape from the waterway escape facility in operation was 14.71%, showing a very low utilization rate, and the recognition rate of the artificial canal escape facility was found to be very low as it moved along the side wall of the artificial canal. Therefore, in the case of a waterway escape facility for Bombina orientalis, it is possible to construct it at an angle of 60°, and if the side walls of the artificial canals are built within 60°, Bombina orientalis can move freely in both directions, overcoming the low utilization rate of existing waterway escape facilities. It is expected to minimize the impact of movement and death of artificial canals. In addition, if the spacing between escape facilities is narrowed from the installation standard of 30m and ramps are constructed in both directions upstream and downstream, the escape success rate of amphibians,reptiles, and small mammals otherthan lady frogs is expected to improve.
The wild simulated ginseng (WSG) has been effectively used in folk medicine as a remedy against hepatic disease, hypertension and arthritic disease. However, there is still lack of scientific proof about its antioxidant capability. The present study has been conducted to evaluate the protective role of the WSG ethanol extract in the CCl4-induced oxidative stress and resultant hepatic disfunction in ICR mice. The electron donating abilities and IC50 of WSG etnanol extract were 76.86 ${\pm}$ 1.06% and 33.3 ${\mu}g$/mL (that of ascobic acid was 16.5 ${\mu}g$/mL), respectively. Total antioxidant status of WSG extract was 2.13 ${\pm}$ 0.06 mmoL/mg, while the values of ascorbic acid and BHT were 3.63 ${\pm}$ 0.06 and 3.12 ${\pm}$ 0.02, respectively. ICR mice (aged 3weeks) were fed for 4 weeks on AIN-93M diet and had free access to food and water. The animals were divided into three groups: normal group (intraperitoneally (i.p) injected with PBS at 100 ${\mu}L$/mouse), group C; CCl4-induced and without any treatment. (i.p injected only PBS, 100 ${\mu}L$ /mice), group G; CCl4-induced and treated with WSG (i.p injected with 5 mg WSG extract per mouse, suspended in 100 ${\mu}L$ phosphate buffer). After the i.p. injection of WSG or PBS (5 times for 7weeks), all mice were administered CCl4 in olive oil at the last day of the experiment, except for normal group. The normal group was administered only olive oil. Determination of plasma triglyceride, total cholersterol, fasting glucose and GPT activity was performed using automatic blood analyzer. To evaluate the protective effect against the oxidative stress, DNA fragmentation and TBARS were determined in blood leucocytes and RBC and hepatocyte, respectively. Body and organs weights and food intake did not show significant differences among the groups. Blood total cholesterol of group G was similar to that of normal group, which was the lowest in group C. The fasting blood glucose level was the highest in normal group (205.20 ${\pm}$ 135.24), which were decreased in group C (134.2 ${\pm}$ 79.31) and group G (126.48 ${\pm}$ 77.05). TBARS values in a red blood cell and hepatic tisuue homogenate were lower in group G comparing to the group C. DNA% in tail, tail length (TL) and tail moment (TM) of blood leucoocytes showed the highest values in group C (20.11 ${\pm}$ 2.47, 17.36 ${\pm}$ 2.58, 94.11 ${\pm}$ 12.29) and they were significantly diminished in group G (9.63 ${\pm}$ 1.19, 7.04 ${\pm}$ 1.50, 38.64 ${\pm}$ 7.60). In conclusion, wild simulated ginseng might be a protective agent against the oxidative stress.
Park, Hae-Sun;Jun, Do-Youn;Woo, Hyun-Ju;Rue, Seok-Woo;Kim, Sang-Kook;Kim, Kyung-Min;Park, Wan;Moon, Byung-Jo;Kim, Young-Ho
Journal of Life Science
/
v.19
no.11
/
pp.1529-1537
/
2009
To elucidate further the antitumor effects of a natural L-arginine analogue, L-canavanine, the mechanism underlying apoptogenic activity of L-canavanine and its modulation by protein tyrosine kinase $p56^{lck}$ was investigated in human Jurkat T cells. When the cells were treated with 1.25 to 2.5 mM L-canavanine for 36 h, several apoptotic events including mitochondrial membrane potential (${\Delta\Psi}m$) loss, activation of caspase-9, -3, -8, and -7, poly (ADP-ribose) polymerase (PARP) degradation, and DNA fragmentation were induced without alteration in the levels of Fas or FasL. These apoptotic changes were more significant in $p56^{lck}$-deficient Jurkat clone JCaM1.6 than in $p56^{lck}$-positive Jurkat clone E6.1. The L-canavanine-induced apoptosis observed in $p56^{lck}$-deficient JCaM1.6 cells was significantly reduced by introducing $p56^{lck}$ gene into JCaM1.6 cells by stable transfection. Treatment of JCaM1.6/lck cells with L-canavanine caused a transient 1.6-fold increase in the kinase activity of $p56^{lck}$. Both FADD-positive wild-type Jurkat T cell clone A3 and FADD-deficient Jurkat T cell clone I2.1 exhibited a similar susceptibility to the cytotoxicity of L-canavanine, excluding involvement of Fas/FasL system in triggering L-canavanine-induced apoptosis. The L-canavanine-induced apoptotic sub-$G_1$ peak and activation of caspase-3, -8, and -7 were abrogated by pan-caspase inhibitor (z-VAD-fmk), whereas L-canavanine-induced activation of caspase-9 was not affected. These results demonstrated that L-canavanine caused apoptosis of Jurkat T cells via the loss of ${\Delta\Psi}m$, and the activation of caspase-9, -3, -8, and -7, leading to PARP degradation, and that the $p56^{lck}$ kinase attenuated the ${\Delta\Psi}m$ loss and activation of caspases, and thus contributed as a negative regulator to L-canavanine-induced apoptosis.
Kim, Han-Kyun;Zhao, Rong-Jie;Jo, Mi-Jeong;Choi, Sun-Mi;Park, Sook-Jahr;Kim, Mi-Ryeo;Kwon, Young-Kyu;Kim, Sang-Chan
Journal of Physiology & Pathology in Korean Medicine
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v.21
no.1
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pp.158-164
/
2007
Uterine leiomyoma is the most common tumor in the female genital tract. Although the tumor is benign, it is a matter of paramount importance since it often causes profuse menstrual bleeding, pressure symptoms and infertility. Nevertheless, the etiology and pathophysiology of this abnormality remain poorly understood. The traditional definitive treatment for uterine leiomyomas is hysterectomy and, even today, symptomatic leiomyomas are the leading cause of hysterectomy in Korea. Clearly, the development of a safe, effective, and nonsurgical method of treatment for leiomyoma would be of great benefit to many women. This study demonstrated growth inhibition of uterine leiomyoma cells using Haeohyultang (HT). When human leiomyoma cells were treated with Haeohyultang, cells showed dose-dependent growth inhibitory effect. Cell growth was inhibited by over 40% as determined by both cell counts and MTS assay. Reduction of cellular viability as a consequence of exposure to Haeohyultang resulted from induction of apoptosis, as assessed by DNA fragmentation, PARP cleavage, caspase 9 and caspase 3 assay. Flow cytometry analysis with uterine leiomyoma cells demonstrated sub G1 cell cycle arrest after treatment with drug Haeohyultang. But, the expression levels of p27 and p21 were not changed in Haeohyultang treated cells compared with control. However, the expression levels of clAP1 were reduced by Haeohyultang compared with control. This reduction of clAP1 data means activation of the caspase family, and then induction of PARP cleavage and apoptosis. These results suggest that Haeohyultang may be potential therapeutic approach in the clinical management of uterine leiomyoma.
Park, Cheol-Ho;Kim, Sung-Won;Hwang, You-Jin;Kim, Dae-Young
Journal of Embryo Transfer
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v.27
no.2
/
pp.107-111
/
2012
Miniature pig sperm cryopreservation is continually researched in biotechnology for breed conservation and reproduction. It is important to control the temperature at each stage of cryopreservation and cryoprotectant. It is also necessary to find the optimal cryoprotectant concentration and chemical elements of the extender. Recently, many studies have used various cryoprotectant materials, such as dimethyl sulphoxide (DMSO), ethylene glycol (EG), antifreeze protein (AFP), amides, and glycerol. Glycerol is a commonly used cryoprotectant. However, glycerol has critical cytotoxic properties, including osmotic pressure and it can cause irreversible damage to live cells. Therefore, We focused on membrane fluidity modifications can reduce cell damage from freezing and thawing procedures and evaluated on the positive effects of trehalose to the viability, chromatin integrity, and motility of boar sperm. Miniature pig sperm was separated from semen by washing with modified- Modena B (mMB) extender. After centrifugation, the pellet was diluted with the prepared first extender. This experiment was designed to compare the effects that sperm cryopreservation using two different extenders has on sperm chromatin. The control group used the glycerol only and it was compared with the glycerol and glycerol plus trehalose extender. Sperm viability and motility were evaluated using WST1 assays and computer-assisted semen assays (CASA). Chromatin structure was examined using acridine orange staining. For the motility descriptors, trehalose caused a significant (p<0.01) increase in total motility ($57.80{\pm}4.60%$ in glycerol vs. $75.50{\pm}6.14%$ in glycerol + trehalose) and progressive ($51.20{\pm}5.45%$ in glycerol vs. $70.74{\pm}8.06%$ in glycerol + trehalose). A significant (p<0.05) increase in VAP ($42.70{\pm}5.73{\mu}m/s$ vs. $59.65{\pm}9.47{\mu}m/s$), VSL ($23.06{\pm}3.27{\mu}m/s$ vs. $34.60{\pm}6.58{\mu}m/s$), VCL ($75.36{\pm}11.36{\mu}m/s$ vs. $99.55{\pm}12.91{\mu}m/s$), STR ($54.4{\pm}2.19%$ vs. $58.0{\pm}1.63%$), and LIN ($32.2{\pm}2.05%$ vs. $36.0{\pm}2.45%$) were also detected, respectively. The sperm DNA fragmentation index was 48.8% to glycerol only and 30.6% to glycerol plus trehalose. Trehalose added group showed higher percentages of sperm motility, stability of chromatin structure than glycerol only. In this study, we suggest that trehalose is effective in reducing freezing damage to miniature pig sperm and can reduce chromatin damage during cryopreservation.
The purpose of this study is to predict factors influencing participant demand for the temple stays and to help find alternatives for temple stay marketing strategies. Specifically, the study sought to examine input variables on the visit frequency of temple visitors who partook in temple food. Research subjects were temple stay participants with experience with temple food. Through convenience sampling method, 300 self-administered questionnaires were distributed to participants at 4 temple stays in Seoul. Of the 278 questionnaires collected, 232 (83%) were used for research analysis. Given that the requirement that proper model for analysing the collected data be applied, the Truncated Negative Binomial(TNB) Poisson model, which is useful for analysing count data that are truncated at '0' and overcrowded with a certain value, was selected fort his study. Study results found that, for temple stay food revitalization, the most crucial item for temple food proponents to recognize is natural food ingredients. The degree of affection was higher among respondents over 40 years of groups and with incomes over 40 million won or more than others. In addition, unmarried and male were higher than married and female, and the Christian population in the temple food demand higher impact than Shamanism community. This match should be a priority to establish an in-depth public relations policy of targeted marketing of consumers according to various demographic characteristics. Active and aggressive efforts to expand food inspection are required to promote the healthy image of the temple food to the fragmentation of consumer marketing hierarchy.
The ecosystem fragmentation due to transportation infrastructure causes road-kill phenomenon. When making policy for reducing road-kill it is important to select target-species in order to enhance its efficiency. However, many corridors installed in Korea have been raised issues about their effectiveness due to lack of considerations such as target-species selection, site selection, and management, etc. In this study, we are to grasp relationship between road-kill area and habitat patches and suggest that spatial location of habitat patches should be considered as one of the important factors when making policy for reducing road-kill. Habitat patches were presumed from overlying suitability index that chosen by literature review and road-kill hotspot was determined by Getis-Ord $G_i^*$ analysis. Afterwards, we performed a correlation analysis between $G_i$ Z-score and the distance from habitat patches to the roads. As a result, there is a negative correlation between two variables, It affects the $G_i^*$ Z-score going up if the habitat patches and the roads become closer.
BACKGROUND: This study was performed for the identification and quantification of glucosinolate (GSL) contents in seven varieties of rapeseed (Brassica napus L.) sprouts cultivated under dark and light conditions. METHODS AND RESULTS: Crude glucosinolates (GSLs) were desulfated by treating with aryl sulfatase and purified using diethylaminoethyl sepharose (DEAE) anion exchange column. Individual GSLs were quantified using high-performance liquid chromatography (HPLC) with electrospray ionization-tandem mass spectrometry (ESI-MS/MS). Eleven GSLs including six aliphatic (progoitrin, sinigrin, glucoalyssin, gluconapoleiferin, gluconapin, and glucobrassicanapin), four indolyl (4-hydroxyglucobrassicin, glucobrassicin, 4-methoxyglucobrassicin, and neoglucobrassicin) and one aromatic (gluconasturtiin) were identified based on the fragmentation patterns of MS spectrum. Aliphatic GSLs were noted as the predominant group with average 85.2% of the total contents. The most abundant GSLs were progoitrin which was ranged at $8.14-118.68{\mu}mol/g$ dry weight (DW). The highest total GSL amounts were documented in 'Hanra' ($146.02{\mu}mol/g$ DW) under light condition and 'Mokpo No. 68' ($86.67{\mu}mol/g$ DW) in dark condition, whereas the lowest was in 'Tamra' (30.13 and $14.50{\mu}mol/g$ DW) in both conditions. The sum of aliphatic GSLs attributed > 80% in all varieties, except 'Tamra' (67.7% and 64.9% in dark and light conditions, respectively) in the total GSL accumulation. Indolyl GSLs were ranged $2.41-15.73{\mu}mol/g$ DW, accounted 2.78-33.6% of the total GSLs in rapeseed varieties. CONCLUSION(S): These results provide valuable information regarding potential beneficial GSL contents individually. This study attempts to contribute to knowledge of the nutritional properties of the different varieties of rapeseed plants. These results may be useful for the evaluation of dietary information.
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