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Anticancer Activity of Periplanetasin-5, an Antimicrobial Peptide from the Cockroach Periplaneta americana

  • Kim, In-Woo;Choi, Ra-Yeong;Lee, Joon Ha;Seo, Minchul;Lee, Hwa Jeong;Kim, Mi-Ae;Kim, Seong Hyun;Kim, Iksoo;Hwang, Jae Sam
    • Journal of Microbiology and Biotechnology
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    • v.31 no.10
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    • pp.1343-1349
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    • 2021
  • Cockroaches live in places where various pathogens exist and thus are more likely to use antimicrobial compounds to defend against pathogen intrusions. We previously performed an in silico analysis of the Periplaneta americana transcriptome and detected periplanetasin-5 using an in silico antimicrobial peptide prediction method. In this study, we investigated whether periplanetasin-5 has anticancer activity against the human leukemia cell line K562. Cell growth and survival of K562 cells treated with periplanetasin-5 were decreased in a dose-dependent manner. By using flow cytometric analysis, acridine orange/ethidium bromide (AO/EB) staining and DNA fragmentation, we found that periplanetasin-5 induced apoptotic and necrotic cell death in leukemia cells. In addition, these events were associated with increased levels of the pro-apoptotic proteins Fas and cytochrome c and reduced levels of the anti-apoptotic protein Bcl-2. Periplanetasin-5 induces the cleavage of pro-caspase-9, pro-caspase-8, pro-caspase-3, and poly (ADP-ribose) polymerase (PARP). The above data suggest that periplanetasin-5 induces apoptosis via both the intrinsic and extrinsic pathways. Moreover, caspase-related apoptosis was further confirmed by using the caspase inhibitor carbobenzoxy-valyl-alanyl-aspartyl-[O-methyl]-fluoromethylketone (Z-VAD-FMK), which reversed the periplanetasin-5-induced reduction in cell viability. In conclusion, periplanetasin-5 caused apoptosis in leukemia cells, suggesting its potential utility as an anticancer therapeutic agent.

Molecular Identification and Morphological Descriptions of the Eggs, Larvae and Juvenile of the Previously Unrecorded Species Acanthaphritis unoorum (Perciformes, Percophidae) in Korean Waters (한국산 1미기록종, Acanthaphritis unoorum (농어목, 꼬리점눈퉁이과)의 어란 및 자치어의 분자동정 및 형태기재)

  • Heo, Sung-Hyun;Ban, Tae Woo;Kim, Jin-Koo;Ji, Hwan-Sung;Moon, Seong Yong
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.52 no.1
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    • pp.67-73
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    • 2019
  • We presented detailed morphological descriptions of the eggs, larvae and juvenile of Acanthaphritis unoorum based on specimens collected with bongo nets from Korean waters during the period May 2017-July 2018. We collected 18 individuals including eggs (n= 4, 0.77-0.85 mm in egg diameter), preflexion larvae (n= 6, 4.11-6.31 mm in standard length, SL), flexion larvae (n= 4, 6.60-7.82 mm SL), postflexion larvae (n= 3, 8.94-13.46 mm SL), and one juvenile (n= 1, 14.67 mm SL). The mitochondrial (mt) DNA 16S rRNA sequences of the eggs, and the cytochrome c oxidase subunit I (COI) sequences of the larvae were identical to those of A. unoorum adults (genetic distances <0.01). The A. unoorum larvae and the juvenile that we collected were morphologically similar to those of Dactylopsaron dimorphicum, but the A. unoorum specimens were readily distinguishable by the presence of lateral melanophores. This is the first record of A. unoorum in Korean waters. We propose a new Korean name for A. unoorum: "O-ri-bu-ri-nuntung-i".

Identification and Validation of Four Novel Promoters for Gene Engineering with Broad Suitability across Species

  • Wang, Cai-Yun;Liu, Li-Cheng;Wu, Ying-Cai;Zhang, Yi-Xuan
    • Journal of Microbiology and Biotechnology
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    • v.31 no.8
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    • pp.1154-1162
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    • 2021
  • The transcriptional capacities of target genes are strongly influenced by promoters, whereas few studies have focused on the development of robust, high-performance and cross-species promoters for wide application in different bacteria. In this work, four novel promoters (Pk.rtufB, Pk.r1, Pk.r2, and Pk.r3) were predicted from Ketogulonicigenium robustum and their inconsistency in the -10 and -35 region nucleotide sequences indicated they were different promoters. Their activities were evaluated by using green fluorescent protein (gfp) as a reporter in different species of bacteria, including K. vulgare SPU B805, Pseudomonas putida KT2440, Paracoccus denitrificans PD1222, Bacillus licheniformis and Raoultella ornithinolytica, due to their importance in metabolic engineering. Our results showed that the four promoters had different activities, with Pk.r1 showing the strongest activity in almost all of the experimental bacteria. By comparison with the commonly used promoters of E. coli (tufB, lac, lacUV5), K. vulgare (Psdh, Psndh) and P. putida KT2440 (JE111411), the four promoters showed significant differences due to only 12.62% nucleotide similarities, and relatively higher ability in regulating target gene expression. Further validation experiments confirmed their ability in initiating the target minCD cassette because of the shape changes under the promoter regulation. The overexpression of sorbose dehydrogenase and cytochrome c551 by Pk.r1 and Pk.r2 resulted in a 22.75% enhancement of 2-KGA yield, indicating their potential for practical application in metabolic engineering. This study demonstrates an example of applying bioinformatics to find new biological components for gene operation and provides four novel promoters with broad suitability, which enriches the usable range of promoters to realize accurate regulation in different genetic backgrounds.

Morphological and Molecular Diagnosis of Necator americanus and Ancylostoma ceylanicum Recovered from Villagers in Northern Cambodia

  • Chang, Taehee;Jung, Bong-Kwang;Sohn, Woon-Mok;Hong, Sooji;Shin, Hyejoo;Ryoo, Seungwan;Lee, Jeonggyu;Lee, Keon Hoon;Khieu, Virak;Huy, Rekol;Chai, Jong-Yil
    • Parasites, Hosts and Diseases
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    • v.58 no.6
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    • pp.619-625
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    • 2020
  • Human hookworm infections caused by adult Ancylostoma spp. and Necator americanus are one of the most important tropical diseases. We performed a survey of intestinal helminths using the Kato-Katz fecal examination technique targeting 1,156 villagers residing in 2 northern provinces (Preah Vihear and Stung Treng) of Cambodia in 2018. The results revealed a high overall egg positive rate of intestinal helminths (61.9%), and the egg positive rate of hookworms was 11.6%. Nine of the hookworm egg positive cases in Preah Vihear Province were treated with 5-10 mg/kg pyrantel pamoate followed by purging with magnesium salts, and a total of 65 adult hookworms were expelled in diarrheic stools. The adult hookworms were analyzed morphologically and molecularly to confirm the species. The morphologies of the buccal cavity and dorsal rays on the costa were observed with a light microscope, and the nucleotide sequences of mitochondrial cytochrome c oxidase subunit 1 (cox1) gene were analyzed. The majority of the hookworm adults (90.7%) were N. americanus, whereas the remaining 9.3% were Ancylostoma ceylanicum, a rare hookworm species infecting humans. The results revealed a high prevalence of hookworm infections among people in a northern part of Cambodia, suggesting the necessity of a sustained survey combined with control measures against hookworm infections.

Apigenin Ameliorates Oxidative Stress-induced Neuronal Apoptosis in SH-SY5Y Cells

  • Kim, Yeo Jin;Cho, Eun Ju;Lee, Ah Young;Seo, Weon Taek
    • Microbiology and Biotechnology Letters
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    • v.49 no.2
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    • pp.138-147
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    • 2021
  • The overproduction of reactive nitrogen species (RNS) and reactive oxygen species (ROS) causes oxidative damage to neuronal cells, leading to the progression of neurodegenerative diseases. In this study, we determined the nitric oxide radical (NO), hydroxyl radical (·OH), and superoxide anion radical (O2-) scavenging activities of apigenin. Our results showed that apigenin exhibited remarkable, concentration-dependent ·OH, O2-, and NO radical scavenging activities. Particularly, apigenin indicated the strongest ·OH radical scavenging activity with 93.38% in the concentration of 100 µM. Furthermore, we also investigated the protective effects of apigenin against hydrogen peroxide (H2O2)-induced oxidative stress in SH-SY5Y cells. The H2O2 treatment resulted in a significant decrease in cell viability, as well as an increase in lactate dehydrogenase (LDH) release and ROS production compared with the H2O2-nontreated SH-SY5Y cells. However, the cell viability significantly increased in the apigenin-treated group, as well as inhibited ROS generation and LDH release compared with the H2O2-induced control group. To elucidate the protective mechanisms of apigenin against oxidative stress in SH-SY5Y, we analyzed the apoptosis-related protein expression. The apigenin treatment resulted in the downregulated expression of apoptosis-related protein markers, such as cytochrome C, cleaved caspase-3, poly (ADP)-ribose polymerase (PARP), and B-cell lymphoma 2-associated X (Bax), as well as the upregulated expression of anti-apoptosis markers such as B-cell lymphoma 2 (Bcl-2). In this study, we report that apigenin exhibits a neuroprotective effect against oxidative stress in SH-SY5Y cells. These results suggest that apigenin may be considered as a potential agent for neurodegenerative disease prevention.

The effects of water extract from Dictamnus dasycarpus Turcz on Hepatocellular Damage in vitro (백선 추출물의 간세포 손상에 대한 연구)

  • Ha, Hun-Yong
    • The Korea Journal of Herbology
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    • v.29 no.5
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    • pp.91-95
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    • 2014
  • Objectives : This study was carried out to evaluate whether the water extract from cause the cellular damage in HepG2 cell line. It was reported that Dictamnus dasycarpus Turcz(DDT) intake induce poisoning symptoms in human population. These symptoms was closely related to liver toxicity, however, mechanisms for liver toxicity caused by DDT have not been elucidated exactly. Here, hepatotoxicity caused by DDT was evaluated using HepG2 cell line. Methods : Water extract of DDT was treated into HepG2 cell with various doses such as 0, 0.1, 0.5, 1.0 and $5.0mg/m{\ell}$. In order to cell viability, both MTT and LDH assay were carried out. Also, apoptosis array kit was used to identify whether cell death caused by DDT is due to apoptosis or not. In addition, reactive oxygen species (ROS) was measured after treatment of water extract. Results : We found out significant changes in the apoptosis related factors of hepatocyte. The cell viability of HepG2 treated with DDT water extract was decreased in dose-dependent. Also most of the apoptosis related factors were significantly increased. We found out that Caspase 3, Cytochrome C and ROS had increased in dose-dependent. In addition, other apoptosis related factors Bcl 2 and Bax, which were also constant changes. However, there was no significance. Conclusions : These results suggest that water soluble extract of DDT is expected to have oral toxicity, including hepatocellular damage Therefore, it is suggested that DDT could cause various side effects and toxicity of clinical conditions.

Taxonomic Review of a Rare Butterfly Ray Gymnura japonica (Gymnuridae, Chondrichthyes), in Korea (한국의 희귀 나비가오리[Gymnura japonica (나비가오리과, 연골어강)]의 분류학적 재검토)

  • Kim, Jin-Koo;Ryu, Jung-Hwa;Jang, Seo-Ha;Han, Kyeong-Ho;Kim, Byeong-Yeob
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.55 no.1
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    • pp.30-36
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    • 2022
  • We collected a total of four butterfly ray specimens (Gymnura japonica, 213.4-695.0 mm in total length) in Korea from 2016 to 2021 and investigated their morphological and molecular characteristics in order to clarify their taxonomic status. These features are summarized as follows. Disc lozenge-shaped, 1.8-2.0 times broader than long. Tail very short, post-cloaca length 23.9-28.2% in disc width. Snout short, no rostral cartilage. Clasper short, no hook. Dorsal surface uniform yellow or brownish grey, with or without rounded light yellow spots. An analysis of 434 base-pair sequences of mitochondrial DNA cytochrome c oxidase subunit I showed that all four specimens corresponded to G. japonica from Japan (Kimura-2-parameter distance = 0-0.2%), suggesting that the color patterns found may be due to intraspecific color variation. G. japonica resembles Gymnura poecilura but differs in that it has a shorter tail length to disc width (23.9-28.2% in G. japonica vs. 40.1-48.3% in G. poecilura). This study revealed that G. japonica occurred in areas affected by the Tsushima Warm Current, tentatively suggesting that G. japonica may be an indicator species for monitoring marine ecosystem changes due to climate change.

Nelumbo nucifera Leaves Inhibit HASMC Proliferation and Migration Activated by TNF-$\alpha$ (Human Aortic Smooth Muscle Cell에서 하엽(荷葉)의 항동맥경화 활성 연구)

  • Kim, Sun-Mo;Yun, Hyun-Jeong;Yi, Hyo-Seung;Won, Chan-Wook;Kim, Jai-Eun;Park, Sun-Dong
    • The Korea Journal of Herbology
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    • v.24 no.4
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    • pp.77-86
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    • 2009
  • Objectives : The proliferation and migration of human aortic smooth muscle cells (HASMC) in response to activation by various stimuli plays a critical role in the initiation and development of atherosclerosis. This study was conducted to examine the effects of Nelumbo nucifera leaves (NNL) on the proliferation and migration of HASMC. Additionally, the mechanisms involved in any observed effects were also evaluated. Methods : Apoptotic cells were measured by staining with FITC-labeled annexin V, followed by flow cytometric analysis. The expression level of apoptosis related proteins was confirmed by western blot. And MMP-9 activity was measured by gelatin zymography and MMP-9 expression was measured by ELISA Results : NNL completely inhibited the proliferation of HASMC via induction of the expression of apoptotic proteins including annexin V, cleaved poly ADP-ribose polymerase (PARP), and caspase-3 and -8. NNL treatment resulted in the release of cytochrome c into cytosol, a loss of mitochondrial membrane potential, a decrease in Bcl-2 and Bcl-xL and an increase in Bax expression. NNL also blocked HASMC migration via suppression of MMP-9. Conclusions : Taken together, these results indicate that NNL has the potential for use as an anti-artherosclerosis agent.

First Record of the Fivestripe Wrasse, Thalassoma quinquevittatum (Perciformes: Labridae) Based on Postlarval Specimen from the Southern Coastal Waters of Jeju-do Island, Korea (제주도 남부해역에서 채집한 놀래기과(Labridae) 후기자어 Thalassoma quinquevittatum 한국 첫기록)

  • Jae-Kyung Bae;Hwan-Sung Ji;Jin-Koo Kim
    • Korean Journal of Ichthyology
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    • v.34 no.4
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    • pp.225-230
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    • 2022
  • The single postlarval specimen (7.53 mm in standard length) of Thalassoma quinquevittatum (Lay & Bennett, 1839), belongs to the family Labridae, was collected by a bongo net from the southern coastal waters of Jeju-do Island, Korea in November 2020. T. quinquevittatum has a deeply curved dorsal contour before the dorsal fin, the oval eyes, and no melanophores throughout the body. While T. amblycephalum has a slightly curved dorsal contour before the center of the dorsal fin, the circular eyes, and few melanophores on the body. A molecular analysis based on 548 base pairs sequences in the mitochondrial DNA cytochrome c oxidase subunit I region shows that the specimen was closely matched to adult T. quinquevittatum (K2P distance=0.002-0.005). We report the first record of T. quinquevittatum in Korean waters, and suggest its new Korean name "Da-seot-jul-saek-dong-nol-rae-gi".

Transcriptome Profiling Identifies Genes of Waterlogging-Tolerant and -Sensitive Rapeseeds Differentially Respond to Waterlogging Stress at the Flowering Stage

  • Ji-Eun Lee;Da-Hee An;Kwang-Soo Kim;Young-Lok Cha;Dong-Chil Chang
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2022.10a
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    • pp.229-229
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    • 2022
  • Rapeseed is a crop that is waterlogging sensitive, and it is necessary to breed waterlogging tolerance varieties. Our study presents the comparative transcriptome changes in two rapeseed lines, i.e., waterlogging-tolerant (tJ8634-B-30,) and - sensitive ('EMS26') lines under control and waterlogging stress treatments at the flowering stage. RNA-sequencing analysis revealed 13,279 differentially expressed genes (DEGs) for 'J8634-B-30' and 8,682 DEGs for 'EMS26' under waterlogging stress condition compared to control. Among DEGs of 'J8634-B-30', 6,818 were up-regulated and 6,461 were down-regulated. On the other hand, among the DEGs of 'EMS26', the number of down-regulated genes (5,240) were higher than that of up-regulated genes (3,442). Gene ontology enrichment analysis showed that DEGs related to glucan metabolic, cell wall, and oxidoreductase activity were significantly changed in 'J8634-B-30'. Kyoto Encyclopedia of Genes and Genomes (KEGG)-based analysis in 'J8634-B-30' identified up-regulated DEGs being involved in MAPK signaling pathways. In addition, the DEGs belonging to mechanisms responding to waterlogging stress, i.e., plant hormones, carbon metabolism, Reactive oxygen species (ROS), Nitric oxide (NO) etc. were compared in rapeseed lines. Several DEGs including ethylene-responsive transcription factor (ERF), constitutive triple response (CTR) (in ethylene signaling pathway), monodehydroascorbate Reductase (MDAR), NADPH oxidase (in ROS pathway), cytochrome c oxidase assembly protein (COX) (in NO pathway) up-regulated in 'J8634-B-30'. These outcomes provided the valuable information for further exploring the genetic mechanism of waterlogging tolerance in rapeseed.

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