• Journal of Acupuncture Research
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• v.21 no.6
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• pp.1-17
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• 2004

Objective : To compare and examine how adjustment of pH and electrolyte can affect the efficacy of cultivated wild ginseng distilled herbal acupuncture, we've administered pure cultivated wild ginseng distilled herbal acupuncture and pH and electrolyte adjusted cultivated wild ginseng distilled herbal acupuncture on A549 human lung cancer lines. Then mRNA and proteins which take parts in apoptosis were examined. Methods : Pure cultivated wild ginseng distilled herbal acupuncture treated group was set as the control group and pH and electrolyte adjusted cultivated wild ginseng distilled herbal acupuncture groups were administered on A549 human lung cancer lines. Cell toxicity was carefully examined and from the analysis of DNA fragmentation, RT-PCR, and Western blot, manifestation of mRNA and proteins which are associated with apoptosis were inspected. Results : The following results were obtained on apoptosis of A549 human lung cancer lines after administering pH and electrolyte adjusted cultivated wild ginseng distilled herbal acupuncture. 1. Measuring cell toxicity of lung cancer cells, higher cell toxicity was detected at pH and electrolyte adjusted groups and the results were concentration dependent. 2. Through DNA fragmentation, we were able to confirm cell destruction in all groups. 3. Experiment groups treated with cultivated wild ginseng distilled herbal acupuncture showed inhibition of Bcl-2 and COX-2 at mRNA and Protein level, whileas increase of Bax was shown. 4. Manifestation of p21, p53, Cyclin E, and Cyclin D1 were confirmed in all groups. 5. Extrication of Cytochrome C was detected at all groups, as well as increased activity of the enzyme caspase-3 and caspase-9, and PARP fragmentation were confirmed. Conclusions : From the above results, we can carefully deduce cell destruction of A549 human lung cancer lines were induced by Apoptosis. At the same concentration level, cell destruction efficacy was better with adjusted pH and electrolyte. Cultivated wild ginseng distilled herbal acupuncture also showed decrease of Bcl-2 and COX-2, as well as increase of Bax. Since cultivated wild ginseng distilled herbal acupuncture increases manifestation of p21, p53, Cyclin E, and Cyclin D1, it affects cellular cycle and through these phenomena, we can consider extrication of Cytochrome C, increase of caspase, and PARP fragmentation are the results.

• Korean Journal of Ecology and Environment
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• v.54 no.3
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• pp.229-239
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• 2021

Thamnaconus modestus, distributed in the Northwest Pacific, has high economic value and is used in various seafood. In this study, the morphological and genetic characteristics of T. modestus and T. septentrionalis were compared and analyzed. We observed the external and internal morphology of T. modestus, sketched skeletal elements, and analyzed phylogenetic evolutionary relationships using the cytochrome c oxidase subunit I (COI) gene on mitochondrial DNA compared to T. septentrionalis. The T. modestus observed in this study had blackish-brown patterns irregularly scattered on the gray-brown body, and the fins were blue-green. Genetic analysis results based on the COI sequences of T. modestus showed seven types of base sequence variation; however, the homology was more than 98.8%. In addition, as a result of comparison of the COI nucleotide sequences and phylogenetic analysis in Tetraodontiformes, two T. septentrionalis sequences (JN813099, MW485059) were similar to T. modestus with 99% homology, and the other two T. septentrionalis sequences (EF607583, KP267619) were similar to those of species belonging to another genus Thamnaconus with 95% homology with T. modestus. It was not easy to classify the species based on morphological characteristics, and phylogenetic analysis between T. modestus and T. septentrionalis confirmed the difference in classification. These results provide the external and internal morphology of T. modestus and will be used as important information for the taxonomic study of T. modestus and T. septentrionalis.

• Molecules and Cells
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• v.46 no.9
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• pp.545-557
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• 2023

Sphingomyelinase (SMase) catalyzes ceramide production from sphingomyelin. Ceramides are critical in cellular responses such as apoptosis. They enhance mitochondrial outer membrane permeabilization (MOMP) through self-assembly in the mitochondrial outer membrane to form channels that release cytochrome c from intermembrane space (IMS) into the cytosol, triggering caspase-9 activation. However, the SMase involved in MOMP is yet to be identified. Here, we identified a mitochondrial Mg²⁺-independent SMase (mt-iSMase) from rat brain, which was purified 6,130-fold using a Percoll gradient, pulled down with biotinylated sphingomyelin, and subjected to Mono Q anion exchange. A single peak of mt-iSMase activity was eluted at a molecular mass of approximately 65 kDa using Superose 6 gel filtration. The purified enzyme showed optimal activity at pH of 6.5 and was inhibited by dithiothreitol and Mg²⁺, Mn²⁺, Ni²⁺, Cu²⁺, Zn²⁺, Fe²⁺, and Fe³⁺ ions. It was also inhibited by GW4869, which is a non-competitive inhibitor of Mg²⁺-dependent neutral SMase 2 (encoded by SMPD3), that protects against cytochrome c release-mediated cell death. Subfractionation experiments showed that mt-iSMase localizes in the IMS of the mitochondria, implying that mt-iSMase may play a critical role in generating ceramides for MOMP, cytochrome c release, and apoptosis. These data suggest that the purified enzyme in this study is a novel SMase.

• Applied Biological Chemistry
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• v.25 no.4
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• pp.248-251
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• 1982

Commassie blue G250 produced metachromatic effect with some solvents. The absorptivity and molar absorptivity of the dye in ethanol were 82.4 and $70.4{\times}10^3$ at maximum absorption wavelength 610nm, respectively. The dye had a red from$({\lambda}_m=465nm)$ in ethanol-phosphoric acid-water solution and converted to a blue form$({\lambda}_m=590nm)$ after binding to protein. Absorbance at 590nm gave linear responses with respect to protein contents. The dye-binding capacities of proteins varied considerably with the content and source of proteins. Under the experimental condition the dye-binding capacities of bovine serum albumin, cytochrome C and ${\gamma}-globulin$ were 110, 103, and $88{\mu}g$ commassie blue G250 bound per $100{\mu}g$ protein, respectively.

• Korean Journal of Ichthyology
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• v.24 no.2
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• pp.101-109
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• 2012

The egg development and early life history of Acheilognathus macropterus from Japan which is an exotic bitterling from China were observed under the controlled water temperature, $20.0{\pm}1.0^{\circ}C$. Fertilized eggs are opaque yellow in color and long elliptic globe shaped measuring $2.78{\pm}0.12mm$ in length and $1.44{\pm}0.04mm$ in breadth. The number of egg averaged 151 per an oviposition. The eggs of this species began to hatch about seventy eight hours after insemination and the mean of total length of larvae were 3.8 mm. S form moving of larvae were observed from three days after hatching. The larvae reached at the heterotrophic stage about twenty-five days after hatching. Morphological character and analysis of cytochrome DNA of this species from Japan were relatively similar to Korean but spawned egg shape was different remarkably. Taxonomical research is necessary in the future.

• Advances in Traditional Medicine
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• v.9 no.2
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• pp.186-191
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• 2009

Eriocaulon sieboldianum (ES) is used in traditional oriental medicine for various medicinal purposes including headache, toothache, and inflammation. However, the anti-cancer effect of the ES is still not fully understood. In the present study, the human leukemia cell line HL-60 was used to characterize the apoptotic effects of ES. ES induced cytotoxicity of HL-60 cells in a dose- and time-dependent manner. ES induced the generation of reactive oxygen species, and the release of cytochrome c in a dose-dependent manner. In addition, we showed that ES-induced apoptosis was accompanied by activation of caspase-3. Taken together, our results demonstrate that ES possesses anti-cancer activity in HL-60 cells.

• Journal of Microbiology and Biotechnology
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• v.31 no.3
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• pp.464-474
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• 2021

Bacterial cytochrome P450 (CYP) enzymes are responsible for the hydroxylation of diverse endogenous substances with a heme molecule used as a cofactor. This study characterized two CYP154C3 proteins from Streptomyces sp. W2061 (CYP154C3-1) and Streptomyces sp. KCCM40643 (CYP154C3-2). The enzymatic activity assays of both CYPs conducted using heterologous redox partners' putidaredoxin and putidaredoxin reductase showed substrate flexibility with different steroids and exhibited interesting product formation patterns. The enzymatic characterization revealed good activity over a pH range of 7.0 to 7.8 and the optimal temperature range for activity was 30 to 37℃. The major product was the C16-hydroxylated product and the kinetic profiles and patterns of the generated hydroxylated products differed between the two enzymes. Both enzymes showed a higher affinity toward progesterone, with CYP154C3-1 demonstrating slightly higher activity than CYP154C3-2 for most of the substrates. Oxidizing agents (diacetoxyiodo) benzene (PIDA) and hydrogen peroxide (H2O2) were also utilized to actively support the redox reactions, with optimum conversion achieved at concentrations of 3 mM and 65 mM, respectively. The oxidizing agents affected the product distribution, influencing the type and selectivity of the CYP-catalyzed reaction. Additionally, CYP154C3s also catalyzed the C-C bond cleavage of steroids. Therefore, CYP154C3s may be a good candidate for the production of modified steroids for various biological uses.

• Journal of Applied Biological Chemistry
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• v.60 no.3
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• pp.271-277
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• 2017

Ethyl formate has been used for the control of insect pests by fumigation. However, there were not many reports to show its target site of fumigant toxicity on insect pests since its first use in the agricultural industry. In the present study, we showed the presumable target sites of ethyl formate fumigation in insect pests using Myzus persicae nymphs. After ethyl formate fumigation, the nymphs of this species were collected and the changes at the biochemical and molecular level were determined. The activity of cytochrome c oxidase (COX) was approximately two-fold higher after ethyl formate fumigation. In addition, the expression levels of acetylcholinesterase (AChE) decreased gradually with increasing ethyl formate concentration. These two findings suggested that COX and AChE might be the major target sites of ethyl formate fumigation. In addition to these results, the analysis of lipid content using MALDI-TOF MS/MS identified 9 phospholipids differently generated 2-fold higher in the ethyl formate-treated nymphs than that in the control nymphs, thereby leading to changes in cell membrane composition in M. persicae nymphs. Therefore, the ethyl formate fumigation caused lethal effects on M. persicae nymphs by changing COX activity, AChE gene expression, and phospholipid production.

• Radiation Oncology Journal
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• v.22 no.4
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• pp.298-306
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• 2004

Purpose: The purpose of this study was to identify Radiosensitivity of proteins in tissues with different radiosensitivity. Materials and Methods: C3H/HeJ mice were exposed to 10 Gy. The mice were sacrifiud 8 hrs after radiation. Their spleen and liver tissues were collected and analyzed histologicaly for apoptosis. The expressions of radiosusceptibillty protein were analyzed by 2-dimensional electrophoresis and matrix-assisted laser desorption ionization time-of-flight mass spectrometry. Resilts: The Peak of apoptosis levels were $35.3{\pm}1.7{\%}$ in spleen and $0.6{\pm}0.2{\%}$ in liver at 8 hrs after radiation. Liver, radioresistant tissues, showed that the levels of ROS metabolism related to proteins such as cytochromm c, glutathione S transferase, NADH dehydrogenase, riken cDNA and peroxiredoxin Vl increased after radiation. The expression of cytochrome c increased significantly in spleen and liver tissues after radiation. In spleen, radiosensitivity tissue, the identified proteins showed a significantly quantitative alteration following radiation. It was categorized as signal transduction, apoptosis, cytokine, Ca signal related protein, stress-related protein, cytoskeletal regulation, ROS metabolism, and others. Conclusion: Differences of radiation-induced apoptosis by tissues specifted were coupled with the induction of related radiosensitivity and radioresistant proteins. The result suggests that apoptosis relate protein and redox proteins play important roles in this radiosusceptibility.

• Biomolecules & Therapeutics
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• v.17 no.3
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• pp.299-304
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• 2009

Rosiglitazone maleate (RGM) is widely used for improving insulin resistance. RGM is a moderate inhibitor of cytochrome P450 2C8 (CYP2C8) and is also mainly metabolized by CYP2C8. The aim of this study was to determine whether the effect of RGM on CYP2C8 is altered by co-treatment with other drugs, and whether amlodipine camsylate (AC) changes the pharmacokinetics (PK) of RGM. Of the 11 drugs that are likely to be co-administered with RGM in diabetic patients, seven drugs lowered the $IC_{50}$ value of RGM on CYP2C8 by more than 80%. In vitro CYP2C8 inhibitory assays of RGM in combination with drugs of interest showed that the $IC_{50}$ of RGM was decreased by 98.9% by AC. In a pharmacokinetic study, Sprague-Dawley (SD) rats were orally administered 1 mg/kg of RGM following by single or 10-consecutive daily administrations of 1.5 mg/kg/day of AC. No significant changes in the pharmacokinetic parameters of RGM were observed after a single administration of AC, but the AUC and $C_{max}$ values of RGM were significantly reduced by 36% and 31%, respectively, by multiple administrations of AC. In conclusion, RGM was found to be affected by AC by in vitro CYP2C8 inhibition testing, and multiple dosing of AC appreciably changed the pharmacokinetics of RGM. These findings suggest that a drug interaction exists between AC and RGM.