Lee Bom Bi;Chae Yun Byung;Kwon Young Kyu;Yang Cae Ha;Kim Mi Ryo;Kim Kwang Jung;Hahm Dae Hyun;Lee Hye Jeong;Shim In Sub
Journal of Physiology & Pathology in Korean Medicine
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v.18
no.3
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pp.767-773
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2004
Substantial evidence suggests that repeated injections of nicotine produce increase in locomotor activity and expression of the immediate-early gene, c-fos in the dopaminergic target areas. Herbal medicine as a therapeutic intervention has been widely used for the treatment of mental dysfunction. Many studies have shown that Cortex Phellodendris (CP) can affect the biochemical balance in the central nervous system. In order to investigate whether CP have an influence on their nicotine-induced behavioral sensitization, we examined the effect of CP on nicotine-induced locomotor activity and c-Fos expression in the striatum and nucleus accumbens utilizing the Fos-like immunohistochemistry (FLI). Male SD rats received CP (200㎎/㎏, i.p.) 30 min before repeated daily injections of nicotine (0.4㎎/㎏, s.c.) for 7 days. Rats were followed withdrawal for 3 days and one challenge for 1 day. System challenge with nicotine produced a much larger increase in locomotor activity and accumbal FLI. Pretreatment with CP significanly inhibited nicotine-induced locomotor activity and FLI in the striuatum and nucleus accumbens. These results demonstrated that reduction in locomotor activity by CP may be reflected by reduction of dopamine release and postsynaptic neuronal activity in the striatum and nucleus accumbens. Our results suggest that CP may have therapeutic effect on nicotine addiction. Supported by a fund (99-PJ9-PG1-002-0004).
Capsaicin is known to be an analgesic agent, affecting the synthesis, storage, , transport and release of substance p, the principal neurotransmitter of pain from periphery to the central nervous system(CNS). DA-5018, a newly synthesized capsaicin derivative has shown potent analgesic effect comparable to that of morphine in various rat models of experimentally inducted acute pairs. In this study the mechanism of analgesic actlvity of DA-5018 was examined. First, the electrically-evoked contraction of guinea pig trachea was inhibited by DA-5018 and these inhibition was recovered by incubation with capsafepine(3$\muM$), capsaicin receptor antagonist and this result suggested that DA-5018 has affinity on capsaicin receptor. The correlation between the norciceptive threshold and the release of substance P was evaluated. In vivo perfusion of slices of the rat spinal cord with DA-5018(10, 100$\muM$) produced a significant increase of the release of substance P and this increase was less than that of capsaicin(10$\muM$). The norciceptive threshold of rat treated with DA-5018(1 mg/kg, p.o) in tall pinch test increased from 2.9$\pm$0.3 to 23.5 $\pm$6.61. Tail pinch latency increased to a maximun at 15 min after DA-5018 treatment and then declined to control values by 120 min. The capsaicin-evoked release ot substance P from the spinal cord slices of rat treated with DA-5018 reduced from 2.38$\pm$ 0.79 to 0.69$\pm$ 0.26 pg/mg wet weight. This reduction reached to a minium at 15 min after DA-5018 treatment and then recovered to control value by 120 min. These results mean that analgesic activity of DA-5018 is due to release of substance P The effect of DA-5018 cream on electrically-evoked neurogenic inflammation of rat saphenous nerve was compared with capsaicin (zostrix-HP). DA-5018 showed 34% inhibition of the neurogenic extravasation while capsaicin showed significant 67% inhibition. This result indicates that the potency of DA-5018 in the release of substance P is less than that of capsaicin. These results suggest that the release of substance P is partially involved in the mechanism of analgesic action of DA-50l8.
Giardia intestinalis infections arise primarily from contaminated food or water Zoonotic transmission is possible, and at least 7 major assemblages including 2 assemblages recovered from humans have been identified. The determination of the genotype of G. intestinalis is useful not only for assessing the correlation of clinical symptoms and genotypes, but also for finding the infection route and its causative agent in epidemiological studies. In this study, methods to identify the genotypes more specifically than the known 2 genotypes recovered from humans have been developed using the intergenic spacer (IGS) region of rDNA. The IGS region contains varying sequences and is thus suitable for comparing isolates once they are classified as the same strain. Genomic DNA was extracted from cysts isolated from the feces of 5 Chinese, 2 Laotians and 2 Koreans infected with G. intestinalis and the trophozoites of WB, K1, and GS strains cultured in the laboratory, respectively. The rDNA containing the IGS region was amplified by PCR and cloned. The nucleotide sequence of the 3' end of IGS region was determined and examined by multiple alignment and phylogenetic analysis. Based on the nucleotide sequence of the IGS region, 13 G. intestinalis isolates were classified to assemblages A and B, and assemblage A was subdivided into A1 and A2. Then, the primers specific to each assemblage were designed, and PCR was peformed using those primers. It detected as little as 10 pg of DNA, and the PCR amplified products with the specific length to each assemblage (A1, 176bp; A2, 261 bp; B, 319 bp) were found. The PCR specific to 3 assemblages of G. intestinalis did not react with other bacteria or protozoans, and it did not react with G. intestinalis isolates obtained from dogs and rats. It was thus confirmed that by applying this PCR method amplifying the IGS region, the detection of G. intestinalis and its genotyping can be determined simultaneously.
Kim, Yeongsam;Yun, Jiyeon;Jeong, Seungho;Kim, Kwangwoo
International Journal of Highway Engineering
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v.18
no.3
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pp.1-9
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2016
PURPOSES : This study aims to show the difference of the binder aging level in the hot-mix asphalt (HMA) mixture after short-term aging (SA) under different aging conditions, such as mixture temperature and duration in hour. METHODS : Three SA times (i.e., 1 h, 2 h, and 4 h) at two temperatures (i.e., $160^{\circ}C$ and $180^{\circ}C$) were used for the normal mixtures prepared using a PG64-22 asphalt. The field long-term aging (LA) was simulated by applying the same LA procedure (65 h at $110^{\circ}C$) to all compacted specimens, prepared at the air void of 7% using each SA-treated mixture, in a convection oven. The binder aging level was measured in terms of large molecular size by gel-permeation chromatography (GPC) from the mixture and the absolute viscosity (AV) from the recovered binder. The aging levels were evaluated using those two properties after SA and LA, and then compared based on the normal SA (NSA) mixture (1 h at $160^{\circ}C$). The service life reduction caused by SA in various conditions was estimated based on the aging level of the field cores from different locations in various service lives. RESULTS : The results of the laboratory evaluation indicated that the binder of the mixture, which was treated at longer SA time and higher temperature, showed a significantly higher aging level than the NSA mixture. The binder aging level from a longer time, such as 2 h and 4 h SA, or at a higher temperature ($180^{\circ}C$), were estimated to be similar to that of the mixtures, which had already been in field service for several years. CONCLUSIONS : The HMA mixture should be produced at a moderate temperature, such as $160^{\circ}C$, and placed within a limited hauling and queuing time to avoid a significant short-term aging of the binder before placement in the field pavement. The SA for a longer time at a higher temperature than the NSA condition was found to be detrimental to the service life of the asphalt pavement.
Valizadeh, Armita;Ahmadzadeh, Ahmad;Saki, Ghasem;Khodadadi, Ali;Teimoori, Ali
Asian Pacific Journal of Cancer Prevention
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v.16
no.18
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pp.8533-8539
/
2016
Background: B-cell chronic lymphocytic leukemia B (B-CLL), the most common type of leukemia, may be caused by apoptosis deficiency in the body. Adipose tissue-derived mesenchymal stem cells (AD-MSCs) as providers of pro-apoptotic molecules such as tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), can be considered as an effective anti-cancer therapy candidate. Therefore, in this study we assessed the role of tumor necrosis factor-producing mesenchymal stem cells oin apoptosis of B-CLL cells resistant to fludarabine-based chemotherapy. Materials and Methods: In this study, after isolation and culture of AD-MSCs, a lentiviral LeGO-iG2-TRAIL-GFP vector containing a gene producing the ligand pro-apoptotic with plasmid PsPAX2 and PMDG2 virus were transfected into cell-lines to generate T293HEK. Then, T293HEK cell supernatant containing the virus produced after 48 and 72 hours was collected, and these viruses were transduced to reprogram AD-MSCs. Apoptosis rates were separately studied in four groups: group 1, AD-MSCs-TRAIL; group 2, AD-MSCs-GFP; group 3, AD-MSCs; and group 4, CLL. Results: Observed apoptosis rates were: group 1, $42{\pm}1.04%$; group 2, $21{\pm}0.57%$; group 3, $19{\pm}2.6%$; and group 4, % $0.01{\pm}0.01$. The highest rate of apoptosis thus occurred ingroup 1 (transduced TRAIL encoding vector). In this group, the average medium-soluble TRAIL was 72.7pg/m and flow cytometry analysis showed a pro-apoptosis rate of $63{\pm}1.6%$, which was again higher than in other groups. Conclusions: In this study we have shown that tumor necrosis factor (TNF) secreted by AD-MSCs may play an effective role in inducing B-CLL cell apoptosis.
Changes in microbial count, enzyme, texture and color during storage of dongchimi (pickled radish roots) were investigated. Dongchimi was pressurized at 400 MPa for 5 min. Pressurized dongchimi stored at $4^{\circ}C\;(PS4)\;and\;37^{\circ}C\;(PS37)$ were compared with control $(stored\;at\;4^{\circ}C)$. PS4 and PS37 maintained their initial pH values during storage. Lactic acid bacteria, yeast and mold were not detected during storage in pressurized dongchimi. Pectinesterase, polygalacturonase activities and hardness of pressurized dongchimi decreased during storage. PS4 maintained its hardness longer than both control and PS37 during storage. The yellowness of PS37 was too high compared to control and PS4, indicating that storage of pressurized dongchimi at $37^{\circ}C$ was undesirable.
The effects of reaction temperature and the level of hydration(water activity) were studied for gas phase reactions of alcohol oxidase and alcohol dehydrogenase immobilized on DEAE-cellulose and controlled pore glass(CPG). Optimum reaction temperature zone of gas phase reaction was similar to that of aqueous phase reaction. The activity of alcohol oxidase increased dramatically and the stability decreased when the water activity was increased from 0.3 to 0.8. The apparent activation energies of the gas phase reaction decreased approaching the values obtained in the aqueous phase reaction as the water activity increased. In the both cases of alcohol oxidase and alcohol dehydrogenase, the rate constants of the gas phase reaction were lower than those of aqueous phase reaction by two orders of magnitude and these results could be correlated to the vapor-liquid equilibrium data of the substrate, ethanol.
Kim, Boing-Soon;Naidansuren, Purevjargal;Min, Kwan-Sik
Journal of Life Science
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v.17
no.11
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pp.1497-1504
/
2007
To investigate the function and secretion of human thrombopoietin (TPO) in mammalian cells, hTPO cDNA was cloned using human liver cDNA, and recombinant hTPO (rec-hTPO) was produced in CHO cell lines. In addition, six N-linked glycosylation sites were substituted for Ala to elucidate the role of each carbohydrate chain. To analyze the biological activity, rec-hTPO protein was injected subcutaneously. Blood was withdrawn for platelet determination. The metabolic clearance rate (MCR) was also analyzed at the 1, 4, 10 and 24 hr after tail vein injection. Wild-type TPO (WT) was efficiently secreted into the medium. However, a hTPO mutant with 116 deleted nucleotides detected by PCR cloning was not secreted. The N-linked glycosylation sites had nearly the same expression quantity as rec-hTPO WT apart from mutants 3 and 4. The glycosylation site of mutant 4 appeared to be an indispensable site for hTPO secretion. Also characterized was the biological activity through an injection with rec-hTPO (10 ng) to ICR mice (7 weeks). The result of the blood analysis showed a considerable increase in the platelet number six days after He injection. To analyze the pharmacokinetics, rec-hTPO was injected into the tail vein (5 ng). The result was 200 pg/ml 1hr after this injection. Following this, it dramatically decreased and virtually disappeared 10 hours after the injection. Thus, rec-hTPO may be a treatment for thrombopenia by the production of the high active rec-hTPO. In addition, hTPO can permit the development of potent new analogues that stimulate the platelet value.
Purpose: This study was performed to investigate the effects of Dangguijakyaksan on the development of experimentally-induced endometriosis in rats. Methods: Endometriosis was induced in rats by autotransplanting uterine tissue to the peritoneum and divided them into three groups: (1) sham-operated group(n=8). (2) surgically induced endometriosis and untreated control group(n=8). (3) surgically induced endometriosis and Dangguijakyaksan treated group. Dangguijakyaksan was orally administrated for 15 days after operation. Then we measured the body weight. the volume of endometriotic implants. the weight of uterus and ovary. and analysed the concentration of cytokines (MCP-1, TNF-$\alpha$, IL-1$\beta$, IL-6) in peritoneal fluids. Histopathology, immunohistochemistry for COX-2 and VEGF. and histochemistry for mast cell in transplanted uterine tissue were also performed. Results: - The volume($mm^3$) of endometriotic implants in Dangguijakyaksan treated group was significantly decreased compared with control group. - The concentration(pg/ml) of MCP-1, TNF-$\alpha$ and IL-l$\alpha$ in peritoneal fluids in Dangguijakyaksan treated group was significantly decreased compared with control group. - Histopathologically, proliferation of endometriotic epithelia. infiltration of inflammatory cells and angiogenesis in transplanted uterine tissue of Dangguijakyaksan treated group were weakly observed than those of control group. - The percentage of positive epithelial layers for COX-2 in Dangguijakyaksan treated group was significantly decreased compared with control group. - The VEGF expression of endometriotic epithelia, neovascular endothelia and stromal cells in transplanted uterine tissue of Dangguijakyaksan treated group were weakly observed than those of control group. - The number of mast cells in transplanted uterine tissue in Dangguijakyaksan treated group was significantly decreased compared with control group. Conclusion: On the basis of these results. we concluded that Dangguijakyaksan has inhibiting effects on the development of transplanted uterine tissue. And these effects may be related with decreased production of MCP-1, TNF-$\alpha$ and IL-1$\beta$, and decreased expression of COX-2 and VEGF, and inhibition of mast cell infiltration by administration of Dangguijakyaksan.
Proceedings of the Korean Society of Developmental Biology Conference
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2003.10a
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pp.72-72
/
2003
The present study examined the possibility of cryopreservation of the D-shaped and umbo larvae of arkshell (Scapharca broughtonii), in terms of the survival rates after freezing and thawing. D-shaped and umbo larvae of arkshells were obtained from a shellfish farming on Yosu city. The average shell lengths were $93.3 \pm 10.1 \mu$m and $201.7 \pm 13.5 \mu$, respectively. Five cryoprotectants (CPAs), dimethyl sulfoxide (DMSO), glycerol, ethylene glycol (EG), propylene glycol (PG), and methanol, were tested at the concentrations of 1.5, 2.0 and 2.5 M. After larvae suspended in CPAs, cryoprotectants were loaded in 0.5 ml straws at a larval density of 50-100 larvae per straw, and epuilibrated for 10 and 20 minute at room temperature ($23^{\circ}C$), repectively. Straws were cooled at a rate of $1^{\circ}C$/min from $0^{\circ}C$ to $-12^{\circ}C$, held for 5 min at $-12^{\circ}C$, and then cooled at $2^{\circ}C$/min to $-35^{\circ}C$ and equilibrated for 5 min followed by plunging in liquid nitrogen. After storage in liquid nitrogen for 1 day, straws were thawed in a $30^{\circ}C$ water. As soon as straws were observed to melt, larvae were diluted with an equal volume of ASW and then washed twice with a large volume of ASW at an interval of 2 min to unload the CPAs. The results showed that after equilibration for 10 and 20 minute at room temperature, no larvae survived using methanol as CPAs, and it was observed that larval shells all open slightly, and larval flesh broke down and slopped over the shells. The highest survival rates (D-shaped larvae: 77.6%, umbo larvae: 59.3%) were obtained with 2M DMSO, and 1.5M glycerol yielded survival rates of 53.8% for D-shaped larvae and 37.5% for umbo larvae. The surviving D-shaped larvae showed active rotary motion and perfect membrane integrity and cytoplasmic normality, and the vigorous movement of veliger cilia was observed inside the closed shells. The breakdown of tissue occurred in the abnormal larvae, and the isolated cell often run out of shells.
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