• The Korean Journal of Pharmacology
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• v.29 no.2
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• pp.275-282
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• 1993

Liver microsomal epoxide hydrolase (mEH) is active in the detoxification of epoxide-containing reactive intermediate. Previous studies in this laboratory have shown that thiazole and pyrazine are efficacious inducers of mEH in rats with large increases in mEH mRNA levels (Carcinogensis, Kim et al, 1993). mEH was purified to electrophoretic homogeneity from thiazole-induced rat hepatic microsomes using DEAE-cellulose column chromatography whereas another protein $({\sim}43\;kDa)$ was co-purified with mEH from pyrazine-induced rat hepatic micrsomes (200 mg/kg body weight/day, ip, 3d). The antibody raised from a rabbit against mEH protein purified from thiazole-induced rat hepatic microsomes appeared to specifically recognize mEH protein in rat hepatic microsomes, as assessed by immunoblotting analysis. Immunoblotting analyses revealed a 10- and 7-fold increase in mEH levels in the hepatic microsomes isolated from thiazole- and pyrazine-treated rats, respectively. Moreover, immunoblotting analysis showed cross-reactivity of the mEH antibody with a 43 kDa protein in pyrazine-induced rat hepatic microsomes and with co-purified 43 kDa protein in purified fractions. The ratio between the 43 kDa protein and mEH in pyrazine-induced rat microsomes or in purified fractions was ${\sim}1$ to 15. N-terminal amino acid sequence analysis of both purified rat mEH and 43 kDa protein revealed that 10 out of 12 amino acids in N-terminus of the 43 kDa protein were identical with the mEH sequence with two amino acid residues of the 43 kDa protein undetermined. Either thiazole or pyrazine treatment, however, failed to increase the levels of mEH protein in rabbits while pyrazine caused elevation of the 43 kDa protein in this species, as determined by irnrnunoblotting analysis. These results demonstrated that treatment of rats with either thiazole or pyrazine causes elevation in hepatic mEH expiession whereas pyrazine treatment results in induction of another mEH-related 43 kDa protein and that a distinct species difference exists between rats and rabbits in the induction of mEH by these xenobiotics.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.30 no.6
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• pp.465-473
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• 2004

Purpose : The essential triad for nerve regeneration is nerve conduit, supporting cell and neurotrophic factor. In order to improve the peripheral nerve regeneration, we used polyglycolic acid(PGA) tube and brain-derived neurotrophic factor(BDNF) gene transfected Schwann cells in sciatic nerve defects of SD rat. Materials and methods : Nerve conduits were made with PGA sheet and outer surface was coated with poly(lactic-co-glycolic acid) for mechanical strength and control the resorption rate. The diameter of conduit was 1.8mm and the length was 17mm Schwann cells were harvested from dorsal root ganglion(DRG) of SD rat aged 1 day. Schwann cells were cultured on the PGA sheet to test the biocompatibility adhesion of Schwann cell. Human BDNF gene was obtained from cDNA library and amplified using PCR. BDNF gene was inserted into E1 deleted region of adenovirus shuttle vector, pAACCMVpARS. BDNF-adenovirus was multiplied in 293 cells and purified. The BDNF-Adenovirus was then infected to the cultured Schwann cells. Left sciatic nerve of SD rat (250g weighing) was exposed and 14mm defects were made. After bridging the defect with PGA conduit, culture medium(MEM), Schwann cells or BDNF-Adenovirus infected Schwann cells were injected into the lumen of conduit, respectively. 12 weeks after operation, gait analysis for sciatic function index, electrophysiology and histomorphometry was performed. Results : Cultured Schwann cells were well adhered to PGA sheet. Sciatic index of BDNF transfected group was $-53.66{\pm}13.43$ which was the best among three groups. The threshold of compound action potential was between 800 to $1000{\mu}A$ in experimental groups which is about 10 times higher than normal sciatic nerve. Conduction velocity and peak voltage of action potential of BDNF group was the highest among experimental groups. The myelin thickness and axonal density of BDNF group was significantly greater than the other groups. Conclusion : BDNF gene transfected Schwann cells could regenerate the sciatic nerve gap(14mm) of rat successfully.

• Journal of the Korean Applied Science and Technology
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• v.35 no.2
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• pp.423-432
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• 2018

Arthrospira platensis is one of the oldest algae in the world and has been reported to have anti-aging properties, including phycocyanin, tocopherol and beta-carotene. In this study, we tried to search protective activities against UVB-induced reactive oxygen species(ROS) of Arthrospira platensis under indoor cultivation ethanol extracts(ICAE) and outdoor cultivation ethanol extracts(OCAE). The anti-oxidative capacities were evaluated by DPPH radical scavenging activity and SOD-like activities at various concentrations(0.1, 0.5, $1mg/m{\ell}$) of ICAE and OCAE. Zebrafish embryos and HaCaT cells were exposed to UVB radiation and treated with various concentrations(0, 0.01, 0.05, 0.1, 0.5, $1mg/m{\ell}$) of ICAE and OCAE. ROS levels of zebrafish and HaCaT cells were generated by UVB radiation. ROS levels were detected using a fluorescent microscope after DCFH-DA staining. The DPPH radical scavenging activity of ascorbic acid was 73% and SOD-like activity was 86% in the positive control group. ICAE and OCAE at $1mg/m{\ell}$ concentration showed 43, 57% DPPH radical scavenging activity and 20, 19% SOD-like activity. Anti-oxidative of ICAE and OCAE had lower effects than the positive control ascorbic acid but significant results. ROS of UVB-induced zebrafish embryos and HaCaT cells were higher than negative control. ICAE and OCAE treated group decreased ROS concentration dependently than UVB-induced positive control group. These results suggest that Arthrospira platensis ethanol extract may have usability value as a cosmetic material for skin protection.

• Applied Microscopy
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• v.35 no.3
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• pp.135-152
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• 2005

Prefrontal cortex is a psychological and metaphysical cortex, which deals with feeling, memory, planning, attention, personality, etc. And it also integrates above-mentioned events with motor control and locomotor activities. Prefrontal cortex works as a highest CNS center, since the above mentioned functions are very important for one's successful life, and further more they are upgraded every moments through memory and learning. Many of these highest functions are supposed to be generated via forebrain basal nuclei (caudate nucleus, fundus striati nucleus, accumbens septi nucleus, septal nucleus, etc.). In this experiment, prefrontal efferent terminals within basal forebrain nuclei were ultrastructurally studied. Spraque Dawley rats, weighing $250{\sim}300g$ each, were anesthetized and their heads were fixed on the stereotaxic apparatus (experimental model, David Kopf Co.). Rats were incised their scalp, perforated a 3mm-wide hole on the right side of skull at the 11mm anterior point from the frontal O point (Ref. 13, Fig. 1), suctioned out the prefrontal cortex including cortex of the frontal pole, with suction instrument. Two days following the operations, small tissue blocks of basal forebrain nuclei were punched out, fixed in 1% glutaraldehyde-1% paraformaldehyde solution followed by 2% osmium tetroxide solutions. Ultrathin sections were stained with 1% borax-toluidin blue solution, and the stained sections were obserbed with an electron microscope. Degenerating axon terminals were found within all the basal forbrain nuclei. Numbers of degenerated terminals were largest in the caudate nucleus, next in order, in the fundus striati nucleus, in the accumbens septi nucleus, and the least in the septal nucleus. Only axospinous terminals were degenerated within the caudate nucleus and the fundus striati nucleus, and they showed the characters of striatal motor control system. Axodendritic and axospinous terminals were degenerated within the accumbens septi nucleus and the lateral septal nucleus, and they showed the characters of visceral limbic system. Prefrontal role in integrating the limbic system with the striatal system, en route basal forebrain nuclei, was discussed.

• Journal of Marine Life Science
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• v.9 no.1
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• pp.9-21
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• 2024

Paralytic shellfish poisoning (PSP) including Saxitoxin (STX) is caused by harmful algae, and poisoning occurs when the contaminated seafood is consumed. The mouse bioassay (MBA), a standard test method for detecting PSP, is being sanctioned in many countries due to its low detection limit and the animal concerns. An alternative to the MBA is the Neuro-2a cell-based assay. This study aimed to establish various test conditions for Neuro-2a assay, including cell density, culture conditions, and STX treatment conditions, to suit the domestic laboratory environment. As a result, the initial cell density was set to 40,000 cells/well and the incubation time to 24 hours. Additionally, the concentration of Ouabain and Veratridine (O/V) was set to 500/50 μM, at which most cells died. In this study, we identified eight concentrations of STX, ranging from 368 to 47,056 fg/μl, which produced an S-shaped dose-response curve when treated with O/V. Through inter-laboratory variability comparison of the Neuro-2a assay, we established five Quality Control Criteria to verify the appropriateness of the experiments and six Data Criteria (Top and Bottom OD, EC₅₀, EC₂₀, Hill slop, and R² of graph) to determine the reliability of the experimental data. The Neuro-2a assay conducted under the established conditions showed an EC₅₀ value of approximately 1,800~3,500 fg/μl. The intra- & inter-lab variability comparison results showed that the coefficients of variation (CVs) for the Quality Control and Data values ranged from 1.98% to 29.15%, confirming the reproducibility of the experiments. This study presented Quality Control Criteria and Data Criteria to assess the appropriateness of the experiments and confirmed the excellent repeatability and reproducibility of the Neuro-2a assay. To apply the Neuro-2a assay as an alternative method for detecting PSP in domestic seafood, it is essential to establish a toxin extraction method from seafood and toxin quantification methods, and perform correlation analysis with MBA and instrumental analysis methods.

• The Korean Journal of Malacology
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• v.22 no.2
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• pp.125-134
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• 2006

We investigated the reproductive cycle of the hard clam, Meretrix petechialis with its gonadal development by histological observations. The seasonal changes in biochemical component of the adductor muscle, visceral mass, foot muscle and mantle of the clam were studied by biochemical analysis, from January to December, 2002. The reproductive cycle of this species can be divided into five successive stages: early stage (January to March), late active stage (February to May), ripe stage (April to August), partially spawned stage (July to August) and spent/inactive stage (September to January). Total protein content in the visceral mass was over two times higher than that in the adductor muscle. Monthly changes of total protein content in the adductor muscle were not statistically significant (ANOVA, p = 0.071), while the changes in the visceral mass were significant (p < 0.001). Total protein content in visceral mass was higher during the early active, late active, and ripe stages (from January to May), while the lowest in July. Glycogen content in the adductor muscle was higher than that in the visceral mass. Monthly changes in glycogen contents were statistically significant in both adductor muscle (F = 237.2, p < 0.001) and the visceral mass (F = 64.04, p < 0.001). Glycogen content in the adductor muscle was the highest in the ripe stage (April). Its content was lower in the partially spawned and the spent/inactive stages (June-September). Glycogen contents in the visceral mass were relatively lower until the early active stage, while the highest in the late active stage. RNA content was higher in visceral mass than that in the adductor muscle. Monthly changes in RNA contents were significant in both adductor muscle (F = 195.2, p < 0.001) and visceral mass (F = 78.85, p < 0.001). RNA content in the adductor muscle was high in the early active stage (January-February), and then it decreased rapidly in the late active stage (March-April), thereafter, slightly increased during the partially spawned stage (June-July). RNA content in the visceral mass reached a maximum during the ripe stage (May), and then it decreased rapidly during the partially-spawned stage (June-July). There was significant positive correlation in total protein contents between adductor muscle and visceral mass (r = 0.715, p = 0.020). However, there was no correlation between adductor muscle and visceral mass in glycogen (p = 0.550), while a negative correlation was found between the adductor muscle and visceral mass in RNA (p = 0.518) contents. Especially, changes in RNA content showed a negative correlation between the adductor muscle tissue and visceral mass. Therefore, these results suggest that the nutrient content of the adductor muscle, visceral muscle and foot muscle changed in response to gonadal energy needs.