• Molecules and Cells
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• v.38 no.2
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• pp.138-144
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• 2015

Raloxifene is a selective estrogen receptor modulator (SERM) that binds to the estrogen receptor (ER), and exhibits potent anti-tumor and autophagy-inducing effects in breast cancer cells. However, the mechanism of raloxifene-induced cell death and autophagy is not well-established. So, we analyzed mechanism underlying death and autophagy induced by raloxifene in MCF-7 breast cancer cells. Treatment with raloxifene significantly induced death in MCF-7 cells. Raloxifene accumulated GFP-LC3 puncta and increased the level of autophagic marker proteins, such as LC3-II, BECN1, and ATG12-ATG5 conjugates, indicating activated autophagy. Raloxifene also increased autophagic flux indicators, the cleavage of GFP from GFP-LC3 and only red fluorescence-positive puncta in mRFP-GFP-LC3-expressing cells. An autophagy inhibitor, 3-methyladenine (3-MA), suppressed the level of LC3-II and blocked the formation of GFP-LC3 puncta. Moreover, siRNA targeting BECN1 markedly reversed cell death and the level of LC3-II increased by raloxifene. Besides, raloxifene-induced cell death was not related to cleavage of caspases-7, -9, and PARP. These results indicate that raloxifene activates autophagy-dependent cell death but not apoptosis. Interestingly, raloxifene decreased the level of intracellular adenosine triphosphate (ATP) and activated the AMPK/ULK1 pathway. However it was not suppressed the AKT/mTOR pathway. Addition of ATP decreased the phosphorylation of AMPK as well as the accumulation of LC3-II, finally attenuating raloxifene-induced cell death. Our current study demonstrates that raloxifene induces autophagy via the activation of AMPK by sensing decreases in ATP, and that the overactivation of autophagy promotes cell death and thereby mediates the anti-cancer effects of raloxifene in breast cancer cells.

• Molecules and Cells
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• v.37 no.3
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• pp.257-263
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• 2014

A mammalian cell renovates itself by autophagy, a process through which cellular components are recycled to produce energy and maintain homeostasis. Recently, the abundance of gap junction proteins was shown to be regulated by autophagy during starvation conditions, suggesting that transmembrane proteins are also regulated by autophagy. Transient receptor potential vanilloid type 1 (TRPV1), an ion channel localized to the plasma membrane and endoplasmic reticulum (ER), is a sensory transducer that is activated by a wide variety of exogenous and endogenous physical and chemical stimuli. Intriguingly, the abundance of cellular TRPV1 can change dynamically under pathological conditions. However, the mechanisms by which the protein levels of TRPV1 are regulated have not yet been explored. Therefore, we investigated the mechanisms of TRPV1 recycling using HeLa cells constitutively expressing TRPV1. Endogenous TRPV1 was degraded in starvation conditions; this degradation was blocked by chloroquine (CLQ), 3MA, or downregulation of Atg7. Interestingly, a glucocorticoid (cortisol) was capable of inducing autophagy in HeLa cells. Cortisol increased cellular conversion of LC3-I to LC-3II, leading autophagy and resulting in TRPV1 degradation, which was similarly inhibited by treatment with CLQ, 3MA, or downregulation of Atg7. Furthermore, cortisol treatment induced the colocalization of GFP-LC3 with endogenous TRPV1. Cumulatively, these observations provide evidence that degradation of TRPV1 is mediated by autophagy, and that this pathway can be enhanced by cortisol.

• Journal of Applied Biological Chemistry
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• v.60 no.4
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• pp.313-319
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• 2017

Strawberries are one of the main commodities in Korea and have been exported over 30 million dollar. And the safety has recently become an important issue in the agro-trade, pesticide residue is most important matter of safe agricultural products. As strawberries can be cultivated using elevated production system, so it is possible to use the drenching. When spraying the pesticides, the use of drenching can be lower amount of residual than using foliar treatment. In this study, time sequential residual amount of pesticides were compared between drenching and foliar application in strawberries. Pesticides were treated with the standard and double dosage and samples were collected at 0, 3, 6, 9, 13, 16 and 21 day and analyzed by LC and LC-MS/MS after liquid-liquid extraction and QuEChERS. Two pesticides were detected less than 1 mg/kg in all samples for drenching treatment. Carbendazim residues in drenching treatment were 5 to 25 times lower than foliar application. And azoxystrobin was not detected in strawberries of drenching treatment.

• Reproductive and Developmental Biology
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• v.35 no.3
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• pp.287-294
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• 2011

Autophagy is a process of intracellular bulk protein degradation, in which the accumulated proteins and cytoplasmic organelles are degraded. It plays important roles in cellular homeostasis, apoptosis, and development, but its role during early embryo development remains contentious. Therefore, in the present study, we investigated the effects of 3-methyladenine (3-MA) on early embryonic development in pigs, we also investigated several indicators of developmental potential, including mitochondrial distribution, genes expressions (autophagy-, apoptosis- related genes), apoptosis and ER-stress, which are affected by 3-MA. After in vitro maturation and fertilization, presumptive pig embryos were cultured in PZM-3 medium supplemented with 3-MA for 2 days at $39^{\circ}C$ 5% $CO_2$ in air. Developmental competence to the blastocyst stage in the presence of 3-MA was gradually decreased according to increasing concentration. Thus, all further experiments were performed using 2 mM 3-MA. Blastocysts that developed in the 3-MA treated group decreased LC3-II intensity and expressions of autophagy related genes than those of the untreated control, resulting in down-regulates the autophagy. Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) showed that the number of containing fragmented DNA at the blastocyst stage increased in the 3-MA treated group compared with control ($6.0{\pm}1.0$ vs $3.3{\pm}0.6$, p<0.05). Also, the expression of the pro-apoptotic gene Bax increased in 3-MA treated group, whereas expression of the anti-apoptotic gene Bcl-XL decreased. Mito Tracker Green FM staining showed that blastocysts derived from the 3-MA treated group had lower mitochondrial integrity than that of the untreated control, resulting in decrease the embryonic qualities of preimplantation porcine blastocysts. Then, the expression of the spliced form of pXBP-1 product (pXBP-1s) increased in 3-MA treated group, resulting increase of ER-stress. Taken together, these results indicate that inhibition of autophagy by 3-MA is closely associated with apoptosis and ER-stress during preimplantation periods of porcine embryos.

• BMB Reports
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• v.48 no.9
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• pp.487-488
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• 2015

The ubiquitin-proteasome system and the autophagy lysosome system are the two major protein degradation machineries in eukaryotic cells. These two systems coordinate the removal of unwanted intracellular materials, but the mechanism by which they achieve this synchronization is largely unknown. The ubiquitination of substrates serves as a universal degradation signal for both systems. Our study revealed that the amino-terminal Arg, a canonical N-degron in the ubiquitin-proteasome system, also acts as a degradation signal in autophagy. We showed that many ER residents, such as BiP, contain evolutionally conserved arginylation permissive pro-N-degrons, and that certain inducers like dsDNA or proteasome inhibitors cause their translocation into the cytoplasm where they bind misfolded proteins and undergo amino-terminal arginylation by arginyl transferase 1 (ATE1). The amino-terminal Arg of BiP binds p62, which triggers p62 oligomerization and enhances p62-LC3 interaction, thereby stimulating autophagic delivery and degradation of misfolded proteins, promoting cell survival. This study reveals a novel ubiquitin-independent mechanism for the selective autophagy pathway, and provides an insight into how these two major protein degradation pathways communicate in cells to dispose the unwanted proteins. [BMB Reports 2015; 48(9): 487-488]

• BMB Reports
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• v.53 no.4
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• pp.229-233
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• 2020

The anti-colorectal cancer monoclonal antibody CO17-1A (mAb CO), which recognizes the tumor-associated antigen EpCAM, was expressed in transgenic Arabidopsis plants. PCR and western blot analyses showed the insertion and expression of heavy chain (HC)/HC fused to the KDEL ER retention modif (HCK) and light chain (LC) of mAb CO and mAb CO with HCK (mAb COK) in Arabidopsis transformants. Both plant-derived mAb^P CO and mAb^P COK were purified from a biomass of approximately 1,000 seedlings grown in a greenhouse. In sandwich ELISA, both mAb^P CO showed a slightly higher binding affinity for the target, EpCAM, compared to mAb^M CO. In cell ELISA, both mAbs^P COs showed binding affinity to the human colorectal cancer cell line SW480. Furthermore, mAb^M CO, mAb^P CO, and mAb^P COK exhibited dose and timedependent regression effects on SW480 cells in vitro. In summation, both mAb^P CO and mAb^P COK, expressed in Arabidopsis, recognized the target antigen EpCAM and showed anti-proliferative activity against human colorectal cancer cells.

• Journal of Home Health Care Nursing
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• v.6
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• pp.33-45
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• 1999

The purpose of this study was to develop a basic framework and criteria for evaluation of quality care provided to patients with the attributes of disease in the home care nursing field, and to provide measurement tools for home health care in the future. The study design was a developmental study for evaluation of hospital-based HCN(home care nursing) in Korea. The study process was as follows: a home care nursing study team of College of Nursing. Yonsei University reviewed the nursing records of 47 patients who were enrolled at Yonsei University Medical Center Home Care Center in March, 1995. Twenty-five patients were insured at that time, were selected from 47 patients receiving home care service for study feasibility with six disease groups; Caesarean Section (C/S), simple nephrectomy, Liver cirrhosis(LC), chronic obstructive pulmonary disease(COPD), Lung cancer or cerebrovascular accident(CVA). In this study, the following items were selected : First step : Preliminary study 1. Criteria and items were selected on the basis of related literature on each disease area. 2. Items were identified by home care nurses. 3. A physician in charge reviewed the criteria and content of selected items. 4. Items were revised through preliminary study offered to both HCN patients and discharged patients from the home care center. Second step : Pretest 1. To verify the content of the items, a pretest was conducted with 18 patients of which there were three patients in each of the six selected disease groups. Third step : Test of reliability and validity of tools 1. Using the collected data from 25 patients with either cis, Simple nephrectomy, LC, COPD, Lung cancer, or CVA. the final items were revised through a panel discussion among experts in medical care who were researchers, doctors, or nurses. 2. Reliability and validity of the completed tool were verified with both inpatients and HCN patients in each of field for researches. The study results are as follows: 1. Standard for discharge with HCN referral The referral standard for home care, which included criteria for discharge with HCN referral and criteria leaving the hospital were established. These were developed through content analysis from the results of an open-ended questionnaire to related doctors concerning characteristic for discharge with HCN referral for each of the disease groups. The final criteria was decided by discussion among the researchers. 2. Instrument for measurement of health statusPatient health status was measured pre and post home care by direct observation and interview with an open-ended questionnaire which consisted of 61 items based on Gorden's nursing diagnosis classification. These included seven items on health knowledge and health management, eight items on nutrition and metabolism, three items on elimination, five items on activity and exercise, seven items on perception and cognition, three items on sleep and rest, three items on self-perception, three items on role and interpersonal relations, five items on sexuality and reproduction, five items on coping and stress, four items on value and religion, three items on family. and three items on facilities and environment. 3. Instrument for measurement of self-care The instrument for self-care measurement was classified with scales according to the attributes of the disease. Each scale measured understanding level and practice level by a Yes or No scale. Understanding level was measured by interview but practice level was measured by both observation and interview. Items for self-care measurement included 14 for patients with a CVA, five for women who had a cis, ten for patients with lung cancer, 12 for patients with COPD, five for patients with a simple nephrectomy, and 11 for patients with LC. 4. Record for follow-up management This included (1) OPD visit sheet, (2) ER visit form, (3) complications problem form, (4) readmission sheet. and (5) visit note for others medical centers which included visit date, reason for visit, patient name, caregivers, sex, age, time and cost required for visit, and traffic expenses, that is, there were open-end items that investigated OPD visits, emergency room visits, the problem and solution of complications, readmissions and visits to other medical institution to measure health problems and expenditures during the follow up period. 5. Instrument to measure patients satisfaction The satisfaction measurement instrument by Reisseer(1975) was referred to for the development of a tool to measure patient home care satisfaction. The instrument was an open-ended questionnaire which consisted of 11 domains; treatment, nursing care, information, time consumption, accessibility, rapidity, treatment skill, service relevance, attitude, satisfaction factors, dissatisfaction factors, overall satisfaction about nursing care, and others. In conclusion, Five evaluation instruments were developed for home care nursing. These were (1)standard for discharge with HCN referral. (2)instrument for measurement of health status, (3)instrument for measurement of self-care. (4)record for follow-up management, and (5)instrument to measure patient satisfaction. Also, the five instruments can be used to evaluate the effectiveness of the service to assure quality. Further research is needed to increase the reliability and validity of instrument through a community-based HCN evaluation.

• The Korean Journal of Pesticide Science
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• v.13 no.1
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• pp.1-12
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• 2009

To assess the effect of butachlor on freshwater aquatic organisms, acute toxicity studies for algae, invertebrate and fishes were conducted. The algae grow inhibition studies were carried out to determine the growth inhibition effects of butachlor (Tech. 93.4%) in Pseudokirchneriella subcapitata (formerly knows as Selenastrum capriconutum), Desmodesmus subspicatus (formerly known as Scendusmus subspicatus), and Chlorella vulgaris during the exposure period of 72 hours. The toxicological responses of P. subcapitata, D. subspicatus, and C. vulgaris to butachlor, expressed in individual $ErC_{50}$ values were 0.002, 0.019, and $10.4mgL^{-1}$, respectively and NOEC values were 0.0008, 0.0016, and $5.34mg\;L^{-1}$, respectively. P. subcapitata was more sensitive than any other algae species. Butachlor has very high toxicity to the algae, such as P. subcapitata and D. subspicatu. In the acute immobilisation test for Daphnia magna, the 24 and $48h-EC_{50}$ values were 2.55 and $1.50mg\;L^{-1}$, respectively. As the results of the acute toxicity test on Cyprinus carpio, Oryzias latipes and Misgurnus anguillicaudatus, the $96h-LC_{50}s$ were 0.62, 0.41 and $0.24mg\;L^{-1}$, respectively. The following ecological risk assessment of butachlor was performed on the basis of the toxicological data of algae, invertebrate and fish and exposure concentrations in rice paddy, drain and river. When a butachlor formulation is applied in rice paddy field according to label recommendation, the measured concentration of butachlor in paddy water was $0.41mg\;L^{-1}$ and the predicted environmental concentration (PEC) of butachlor in drain water was $0.03 mg\;L^{-1}$. Residues of butachlor detected in major rivers between 1997 and 1998 were ranged from $0.0004mg\;L^{-1}$ to $0.0029mg\;L^{-1}$. Toxicity exposure ratios (TERs) of algae in rice paddy, drain and river were 0.004, 0.05 and 0.36, respectively and indicated that butachlor has a risk to algae in rice paddy, drain and river. On the other hand, TERs of invertebrate in rice paddy, drain and river were 3.6, 50 and 357, respectively, well above 2, indicating no risk to invertebrate. TERs of fish in rice paddy, drain and river were 0.58, 8 and 57, respectively. The TERs for fish indicated that butachlor poses a risk to fish in rice paddy but has no risk to fish in agricultural drain and river. In conclusion, butachlor has a minimal risk to algae in agricultural drain and river exposed from rice drainage but has no risk to invertebrate and fish.

• Korean Journal of Ecology and Environment
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• v.44 no.3
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• pp.272-279
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• 2011

Copper cyanide is a chemical produced in large quantities with 2,500 tonnes being produced in 2006. It is mainly used for electroplating copper, particularly alkali-Cu plate and brass plating. The purpose of this study is to reassess the physicochemical properties and environmental fate of copper cyanide based on reliable data and and to conduct an ecotoxicity test according to the OECD test guidelines as an initial environmental risk assessment (need to state where this was done). Metal containing inorganic substances are not subject to degradation, biodegradation or hydrolysis. Aquatic toxicity tests of copper cyanide were conducted according to OECD test guideline 201, 202 and 203 for green algae, daphnia, and fish, respectively. The following acute toxicity test results were obtained for aquatic species: 0.089 mg $L^{-1}$ (Algae, 72 Hr-$EC_{50}$); 0.21 mg $L^{-1}$ (flea, 48 Hr-$LC_{50}$); 0.62 mg $L^{-1}$ (Fish, 96 Hr-$ErC_{50}$). The chemical possesses properties indicating a hazard for the aquatic environment (acute toxicity in fish, daphnia and algae below 1.0 mg $L^{-1}$). As a result of this study, copper cyanide has become a candidate for detailed risk assessment. Countries that produce this chemical in significant quantities are recommended to perform specific assessments.