• Title/Summary/Keyword: LC3

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Is Autophagy a Prerequisite for Steroidogenesis in Leydig Cells?

  • Ji-Eun Park;Yoon-Jae Kim;Jong-Min Kim
    • Development and Reproduction
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    • v.27 no.3
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    • pp.149-157
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    • 2023
  • We investigated the involvement of autophagy with steroidogenesis in testicular Leydig cells. Human chorionic gonadotropin (hCG)-stimulated T production in Leydig cells was not remarkably altered in the presence of an autophagy inhibitor 3-methyladenine (3-MA). Although pretreatment with 3-MA demonstrated a tendency to decrease hCG-induced T production, the differences were significant only at a higher time point of 24 h following hCG. Microtubule associated protein light chain 3 (LC3)-II was detectable in the control cells in all the experiments. The hCG-induced increase in steroidogenic acute regulatory protein (StAR) and cytochrome P450 side chain cleave (P450scc) protein levels were not significantly altered by 3-MA. Leydig cells isolated from immature rat testes 12 h following hCG treatment showed relatively increased levels of LC3-II protein compared to the control group. Furthermore, LC3-II levels shown in these cells reached almost the identical to those from normal adult testes. However, LC3-II protein levels were almost comparable or even slightly lower than the controls at 48 h following hCG. Expression of StAR and P450scc was upregulated at both 12 and 48 h after hCG. We also used MA-10 cells, the mouse Leydig cell line, in this experiment. When dibutyryl cyclic-AMP was treated with MA-10 cells, P4 levels were significantly increased in the cell culture medium. However, P4 levels tended to decrease in the presence of 3-MA, but the difference was not statistically significant. This was consistent with the results of the rat Leydig cell experiments. Together, we believe that although autophagy participates in steroidogenesis and enhances steroidogenic efficacy of Leydig cells, it may not be a decisive cellular process for steroidogenesis, specifically in the mature Leydig cells.

Comparison of Acute Toxicity of Ammonia in Juvenile Rockfish and Red Sea Bream (조피볼락과 참돔 유어에 대한 암모니아 독성 비교)

  • 조성환;허성범
    • Journal of Aquaculture
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    • v.11 no.4
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    • pp.429-435
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    • 1998
  • This study compared acute ammonia toxicity in juvenile rockfish and red sea bream. Oxygen concentration affected ammonia toxicity to aquatic animals. Without aeration, the 96-hr $LC_50$ values of $NH_4^+$and un-ionhized ammonia for juvenil rockfish(3.6g, 6.6cm) were 2.61 and 0.09 mg/l, respectively. The 3-hr $LC_50$ values of $NH_4^+$ for the size of 1.9g (5.2cm) and 3.6g (6.6cm) rockfish, and 1.0g (3.8cm) and 2.0g (5.0cm) red sea bream without aeration were 3.96, 3.94, 4.35 and 3.75 mg/l, respectively. When oxygen level was low, larger fish were more susceptible to ammonia toxicity than smaller one because of stress resulted from low oxygen. However, with aeration, the 96-hr $LC_50$ values of $NH_4^+$ for the size of 1.2g (3.9cm) and 2.3g (5.1cm) red sea bream with aeration were 3.84 and 3.90mg/l, respectively. The 6-hr $LC_50$values of $NH_4^+$ for the size of 3.1g (5.9cm) and 6.2g (7.0cm) rockfish with aeration were 3.83 and 3.94 mg/l, respectively. When oxygen level was high, larger rockfish and red sea bream were less susceptible to ammonia toxicity than smaller ones. The 6 hr-or 96 hr-$LC_50$ values of $NH_4^+$for rockfish with reduced ammonia toxicity. In comparing 96 hr-$LC_50$ values of $NH_4^+$ and un-ionized ammonia for juvenil rockfish with those for juvenil red sea bream, the values for rockfish were lower than for red sea bream. This indicates that juvenile rockfish is more susceptible to ammonia toxicity than juvenile red sea bream.

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Effect of Sublethal Doses of Imidacloprid on the Green Peach Aphid, Myzus persicae (Imidacloprid의 아치사량이 복숭아혹진딧물의 생물적 특성에 미치는 영향)

  • Choi, Byeong-Ryeol;Lee, Si-Woo;Song, Yoo-Han;Yoo, Jai-Ki;Chung, Bu-Keun
    • The Korean Journal of Pesticide Science
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    • v.9 no.4
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    • pp.374-379
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    • 2005
  • Sublethal dose effect of imidacloprid on green peach aphid (Myzus persicae) was investigated. Nymphs of green peach aphid newly produced were counted on 4 days after sub-lethal dose treatment of imidacloprid. Numbers of nymph were not significantly different between $LC_1$ treatment and untreated control. Their numbers of nymph were 8.8 and 12.7 at $LC_1$ and untreated control, respectively. When they were tented at $LC_{10}$, $LC_{30}$ and $LC_{50}$. their numbers of newly produced nymph were 6.0, 5.1 and 3.9 and reduction rates were 52.7%, 59.8% and 69.3% at each treatment compare to untreated control. Reduction rates of newly laid nymph were proportional to the concentrations of imidacloprid treated. Repellent effect of aphid to imidacloprid was bigger in dipping than in watering method. Secretion amount had negative correlation with imidacloprid concentration in dipping method and suppression of secretion went up to 96%. In watering method, amount of secretion did not show any difference between insecticide concentration, but suppression, compared to untreated control, was over 97%.

Silencing of Rac3 Inhibits Proliferation and Induces Apoptosis of Human Lung Cancer Cells

  • Liu, Tie-Qin;Wang, Ge-Bang;Li, Zheng-Jun;Tong, Xiang-Dong;Liu, Hong-Xu
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.7
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    • pp.3061-3065
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    • 2015
  • Background: Rac3, a member of the Rac family of small guanosine triphosphatases (GTPases), regulates a variety of cell functions, including the organization of the cytoskeleton, cell migration, and invasion. Overexpression of Rac3 has been reported in several human cancers. However, the role of Rac3 in lung cancer (LC) has not been determined in detail. The purpose of this study was to investigate the effect of silencing of Rac3 expression in human LC cells and the consequences for cell survival. Materials and Methods: Lentivirus small hairpin RNA (shRNA) interference techniques were utilized to knock down the Rac3 gene. Gene and protein expression was quantified by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blotting. LC cell apoptosis was examined by annexin V-APC /propidium iodide staining. Results: Efficient silencing of Rac3 strongly inhibited A549 cell proliferation and colony formation ability, and significantly decreased tumor growth. Moreover, flow cytometry analysis showed that knockdown of Rac3 led to G2/M phase cell cycle arrest as well as an excess accumulation of cells in the G1 and S phase. Conclusions: Thus, functional analysis using shRNAs revealed a critical role for Rac3 in the tumor growth of LC cells. shRNA silencing of Rac3 could provide an effective strategy to treat LC.

Median Lethal Concentration ($LC_{50}$) for 24 Hours of Formalin to Eel, Anguilla japonica (뱀장어에 대한 포르말린의 24시간 반수치사농도 ($LC_{50}$))

  • Jung, Sung-Hee;Lee, Joo-Seok;Jee, Bo-Young;Seo, Jung-Soo;Kim, Jin-Woo;Kim, Eung-Oh
    • Journal of fish pathology
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    • v.20 no.2
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    • pp.161-167
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    • 2007
  • Acute toxicity of formalin (37% formaldehyde) was conducted to determine the median lethal concentration (LC50) on eel (avarage weight 96 ± 3.6 g, average total length 43 cm), Anguilla japonica at concentrations ranging from 0 to 500 ppm. In particular, this study was designed to estimate the safety concentrations of formalin in testing eels to eradicate Pseudodactylogyrus. All fish died after 10 hours and 24 hours at 500 ppm and 400 ppm, respectively. After 24 hours, cumulative mortality was 96.6% and 13.3% at 300 ppm and 200 ppm formalin, respectively. However, all experimental fish were alive after 24 hours at 100 ppm. The lethal concentration values were computed by using non-linear least square method. At the start of the test, water temperature, pH and dissolved oxygen level were 27~28℃, 7.4 and 5.55 ppm, respectively. The 24 hr-LC50 were 269 ppm.

Do-It-Yourself (DIY) manufacture of a Nano-LC MALDI spotter robot using 3D printing technology

  • Lee, Jae-ung;Oh, Han Bin
    • Analytical Science and Technology
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    • v.30 no.4
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    • pp.167-173
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    • 2017
  • In the era of the forth Industrial Revolution, open source code and open source hardware have gained much attention. In particular, 3D printing technology is expanding into the realms of classical science, technology and our daily lives. Relatedly, in the present study, we demonstrate the manufacture of a nano-LC MALDI spotter robot using 3D printing technology. The parts of the spotter robot were either made using a 3D printer or purchased as 3D printer parts from the 3D printer online market, so that anyone can make the robot without a deep knowledge of engineering or electronics, i.e., DIY (do-it-yourself) product. In the nano-LC MALDI spotter, the nano-LC eluent and MALDI matrix were mixed in a T-union and discharged from the capillary outlet. The eluent and matrix mixture could be spotted onto the movable MALDI plate. The MALDI plate was designed to translate in a two-dimensional space (xy plane), which was enabled by the movements of two stepper motors. In the paper, all computer-aided design (CAD) files for the parts and operation software are provided to help the reader manufacture their own spotter robot.

Localization of Autophagosome in Porcine Follicular Cumulus-oocyte Complex

  • Lee, Seunghoon;Kim, Dong-Hoon;Im, Gi-Sun;Ock, Sun-A;Ullah, Imran;Hur, Tai-Young
    • Journal of Embryo Transfer
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    • v.32 no.3
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    • pp.105-109
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    • 2017
  • Autophagy is an intracellular degradation and recycling system. Oocyte maturation is dynamic process, in which various proteins should be synthesized and degraded. In our previous study, we reported the loci of autophagosome and dynamics of autophagic activity in porcine oocytes during in vitro maturation. In this study, we verified loci of autophagosome in porcine follicular cumulus-oocyte complex by detection of microtubule-associated protein 1A/1B-light chain 3 (LC3) which is the reliable marker of autophagosome. Porcine ovary including various sizes of follicles was fixed within 1 hour after collection from slaughterhouse. After fixation, immunohistochemistry was conducted on sliced ovary tissue containing various sizes of follicles by using LC3 antibody. As a result, LC3 signal was clearly detected in both cumulus and oocytes of various sizes of follicles. We also found ring shaped signal which represent autophagosome near oocyte membrane. Most of the signals in oocytes were localized nearby cellular membrane while evenly dispersed in cumulus cells. Therefore, this result suggests that autophagy occurs in porcine COCs (cumulus-oocyte complexes) at follicular stage.

Diamondback moth (Plutella xylostella L.) resistance to organophosphorus and carbamate insecticides in Kangwon alpine vegetable croplands (강원도 고랭지대 배추경작지 배추좀나방(Plutella xylostella L.)의 유기인계 및 카바메이트계 살충제에 대한 저항성 발달)

  • Cho, Jun-Mo;Kim, Kyoung-Ju;Kim, Song-Mun;Han, Dae-Sung;Hur, Jang-Hyun
    • The Korean Journal of Pesticide Science
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    • v.5 no.1
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    • pp.30-35
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    • 2001
  • Diamondback moth (Plutella xylostella L.) recently became a single worst insect which is not controlled effectively by organophosphorus and carbamate insecticides in Kangwon alpine croplands. The objective of this study was to determine if diamondback moth has developed a resistance to organophosphorus and carbamate insecticides. Resistance of diamondback moth, collected at Keichon, Jangpyong Taebaek, Chunchon, and Hongchon, was determined by the concentration required to kill fifty percent of population, $LC_{50}$. Their response of resistance varied to insecticides and locations: Taebaek populations were 35 and 70 times more resistant to chlorpyrifos and fenitrothion, respectively, than susceptible(S) population. Hongchon populations were 94 and 254 times more resistant to chlorpyrifos and fenitrothion, respectively, than S population. In addition, Chunchon populations were 37 and 19 times more resistant to profenofos and benfuracarb, respectively, than S population. However, the field populations did not differ in resistance to diazinon, phenthoate, flupyrazofos, carbofuran, and furathiocarb. This study show that field populations of diamondback moth found in Kangwon alpine vegetable croplands have developed a resistance and/or multiple resistance to some insecticides, implying that farmers are losing organophosphorus and carbamate insecticide options for selective control in vegetable crops.

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Interconnections between the Rat Dorsal Raphe and the Locus Coeruleus Nuclei Demonstrated by Anterograde Tracing with Phaseolus Vulgaris Leucoagglutinin

  • Lee, Hyun S.
    • Animal cells and systems
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    • v.8 no.3
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    • pp.221-229
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    • 2004
  • The projections from the dorsal raphe (DR) to the locus coeruleus (LC) or vice versa were analyzed in the rat using an anterograde tracer, Phaseolus vulgaris leucoagglutinin (PHA-L) combined with serotonin (5-hydroxytryptamine, 5-HT) or dopamine-beta-hydroxylase (DBH) immunostaining. Following the injection of PHA-L into the middle DR, DR-originating fibers with varicosities have contacted DBH-immunolabeled cells in the rostral, middle, and caudal LC. Axon terminals were also observed in the subcoeruleus nucleus. When the PHA-L injection was confined within the caudal DR, axonal fibers with varicosities were observed mainly at the rostral pole of the LC. Following the injection of PHA-L into the caudal, principal LC, labeled fibers with varicosities have contacted 5-HT-immunolabeled neurons at dorsomedial, ventromedial, lateral wing, and caudal sub-divisions of the DR. The present anterograde study suggests that the DR or the LC nuclei communicate with each other in order to perform a variety of functions including vigilance, analgesia, and stress responses.

A Study on Effect of Photo-alignment in Nematic Liquid Crystal on Polyimide Film (폴리이미드막을 이용한 광배향 효과에 관한 연구)

  • 서대식;황율연
    • Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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    • 1997.04a
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    • pp.116-119
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    • 1997
  • We investigated new type non-robbing liquid crystal (LC) alignment techniques in the cell with slanted non-polarized ultraviolet (UV) irradiation on polyimide (Pl) film. It is shown that the uniform alignment for nematic (N) LC is obtained by using slanted non-polarized UV irradiation on Pl surface. We successfully obtained that the pretilt ang1e of NLC is generated about 3 degree in the LC cells by using slanted non-polarized UV irradiation with 70 degree on Pl surface. We consider that the pretilt angle generation for NLC is attributed to interaction between the LC molecular and the PI surface. We conclude that the uniform LC alignment is attributed to anisotropic dispersion force effect due to photo depolymerization of polymer on Pl surface.

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