Parasites, Hosts and Diseases

"Eun-Kyung Moon"

Original Articles

Exploration of Naegleria-preferentially secreted proteins for identifying diagnostic candidates to detect Naegleria fowleri: Hye-Jeong Jo, Hae-Ahm Lee, Fu-Shi Quan, Hyun-Hee Kong, Eun-Kyung Moon; Received July 31, 2025 Accepted November 12, 2025 Published online January 26, 2026; DOI: https://doi.org/10.3347/PHD.25058 [Epub ahead of print]

Abstract
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Naegleria fowleri is a free-living amoeba that can cause primary amebic meningoencephalitis (PAM), a very serious infection of the central nervous system. Early diagnosis of PAM is challenging, and the condition is almost always fatal. In this study, we conducted 2-dimensional gel electrophoresis (2-DE) analysis using N. fowleri trophozoite lysates and conditioned media to identify preferentially secreted proteins. As a result of the 2-dimensional gel electrophoresis analysis, 1 protein was found to increase, 5 proteins were found to decrease, 3 proteins showed a qualitative increase, and 15 proteins showed a qualitative decrease in the conditioned media compared to the proteins in the trophozoite lysates. Using cDNA from N. fowleri, Acanthamoeba castellanii, and Balamuthia mandrillaris, all of which can cause encephalitis, real-time PCR was performed on 5 genes corresponding to the p23-like domain-containing protein, cystatin-like domain-containing protein, fowlerpain-2, hemerythrin family non-heme iron protein, and an uncharacterized protein. The results showed that all 5 genes were highly expressed in N. fowleri. In animal models infected with N. fowleri resulting in PAM, real-time PCR analysis of brain tissue revealed significant overexpression of the p23-like domain-containing protein and fowlerpain-2. These results suggest that the 2 secreted proteins could provide valuable insights for developing antibody-based or molecular diagnostic methods to detect N. fowleri in patients with PAM.

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Identification of essential genes for Acanthamoeba castellanii excystation during encystation and excystation: Min-Jeong Kim, Hye-Jeong Jo, Fu-Shi Quan, Ki Back Chu, Hyun-Hee Kong, Eun-Kyung Moon; Parasites Hosts Dis 2024;62(4):399-407.
Published online November 22, 2024; DOI: https://doi.org/10.3347/PHD.24062

Acanthamoeba is an opportunistic pathogen that causes Acanthamoeba keratitis, granulomatous amoebic encephalitis, and other cutaneous diseases. The life cycle of Acanthamoeba consists of 2 stages of trophozoites and cysts. Under adverse environmental conditions, Acanthamoeba encysts, while the conditions become favorable for growth, it reverts to the trophozoite form. Acanthamoeba excystation is crucial for its proliferation and can lead to recurrent infections after incomplete treatment. To identify the factors involved in excystation, A. castellanii was subjected to either encystation- or excystation-inducing conditions, and gene expression profiles were compared using mRNA sequencing. A. castellanii samples were collected at 8 h intervals for analysis under both conditions. Differentially expressed gene analysis revealed that 1,214 and 1,163 genes were upregulated and downregulated, respectively, by more than 2-fold during early excystation. Five genes markedly upregulated in early excystation (ACA1_031140, ACA1_032330, ACA1_374400, ACA1_275740, and ACA1_112650) were selected, and their expression levels were confirmed via real-time PCR. Small interfering RNA (siRNA) targeting these 5 genes was transfected into Acanthamoeba and gene knockdown was validated through real-time PCR. The silencing of ACA1_031140, ACA1_032330, ACA1_374400, and ACA1_112650 inhibited excystation and suggested that these genes might be essential for excystation. Our findings provide valuable insights for suppressing Acanthamoeba proliferation and recurrence.

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Integrating molecular pathogenesis and predictive modelling in granulomatous amoebic encephalitis due to Acanthamoeba : a digital twin framework for personalised therapy
Ruqaiyyah Siddiqui, David Lloyd, Naveed Ahmed Khan
Infectious Diseases.2026; 58(1): 164. CrossRef
Encystment and Excystment Processes in Acanthamoeba castellanii: An Emphasis on Cellulose Involvement
Mathew Choaji, Ascel Samba-Louaka, Zineb Fechtali-Moute, Willy Aucher, Sébastien Pomel
Pathogens.2025; 14(3): 268. CrossRef
Microarray-based characterization of airway inflammation induced by environmental Acanthamoeba exposure versus the ovalbumin-alum model
Shin Ae Kang, Hak Sun Yu
Parasites, Hosts and Diseases.2025; 63(4): 327. CrossRef

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Virus-like particles expressing microneme-associated antigen of Plasmodium berghei confer better protection than those expressing apical membrane antigen 1: Min-Ju Kim, Ki Back Chu, Keon-Woong Yoon, Hae-Ji Kang, Dong-Hun Lee, Eun-Kyung Moon, Fu-Shi Quan; Parasites Hosts Dis 2024;62(2):193-204.
Published online May 27, 2024; DOI: https://doi.org/10.3347/PHD.24017

Malaria is a global disease affecting a large portion of the world’s population. Although vaccines have recently become available, their efficacies are suboptimal. We generated virus-like particles (VLPs) that expressed either apical membrane antigen 1 (AMA1) or microneme-associated antigen (MIC) of Plasmodium berghei and compared their efficacy in BALB/c mice. We found that immune sera acquired from AMA1 VLP- or MIC VLP-immunized mice specifically interacted with the antigen of choice and the whole P. berghei lysate antigen, indicating that the antibodies were highly parasite-specific. Both VLP vaccines significantly enhanced germinal center B cell frequencies in the inguinal lymph nodes of mice compared with the control, but only the mice that received MIC VLPs showed significantly enhanced CD4⁺ T cell responses in the blood following P. berghei challenge infection. AMA1 and MIC VLPs significantly suppressed TNF-α and interleukin-10 production but had a negligible effect on interferon-γ. Both VLPs prevented excessive parasitemia buildup in immunized mice, although parasite burden reduction induced by MIC VLPs was slightly more effective than that induced by AMA1. Both VLPs were equally effective at preventing body weight loss. Our findings demonstrated that the MIC VLP was an effective inducer of protection against murine experimental malaria and should be the focus of further development.

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Orally Dissolving Film-Based Influenza Vaccines Confer Superior Protection Compared to the Oral Administration of Inactivated Influenza Virus
Keon-Woong Yoon, Jie Mao, Gi-Deok Eom, Su In Heo, Ki Back Chu, Mi Suk Lee, Fu-Shi Quan
Vaccines.2025; 13(6): 600. CrossRef
Protective Efficacy Induced by Virus-like Particles Expressing Dense Granule Protein 5 of Toxoplasma gondii
Su In Heo, Hae-Ji Kang, Jie Mao, Zhao-Shou Yang, Md Atique Ahmed, Fu-Shi Quan
Vaccines.2025; 13(8): 787. CrossRef
Efficacy of Heterologous Vaccination Using Virus-Like Particles and Vaccinia Virus Containing MIC8 and AMA1 Proteins of Toxoplasma gondii
Hae-Ji Kang, Fu-Shi Quan
Vaccines.2025; 13(8): 862. CrossRef
Ivermectin Identified Using a High-Throughput Screening System Exhibits Anti-Clonorchis sinensis Activity in Rats
Soon-Ok Lee, Hyeryon Lee, Ki Back Chu, Jianhua Li, Sung-Jong Hong, Sung Soo Kim, Joo Hwan No, Fu-Shi Quan
Antibiotics.2025; 14(8): 837. CrossRef

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Phagocytosis-associated genes in Acanthamoeba castellanii feeding on Escherichia coli: Min-Jeong Kim, Eun-Kyung Moon, Hye-Jeong Jo, Fu-Shi Quan, Hyun-Hee Kong; Parasites Hosts Dis 2023;61(4):397-404.
Published online November 28, 2023; DOI: https://doi.org/10.3347/PHD.23088

Acanthamoeba species are free-living amoebae those are widely distributed in the environment. They feed on various microorganisms, including bacteria, fungi, and algae. Although majority of the microbes phagocytosed by Acanthamoeba spp. are digested, some pathogenic bacteria thrive within them. Here, we identified the roles of 3 phagocytosis-associated genes (ACA1_077100, ACA1_175060, and AFD36229.1) in A. castellanii. These 3 genes were upregulated after the ingestion of Escherichia coli. However, after the ingestion of Legionella pneumophila, the expression of these 3 genes was not altered after the consumption of L. pneumophila. Furthermore, A. castellanii transfected with small interfering RNS (siRNA) targeting the 3 phagocytosis-associated genes failed to digest phagocytized E. coli. Silencing of ACA1_077100 disabled phagosome formation in the E. coli-ingesting A. castellanii. Alternatively, silencing of ACA1_175060 enabled phagosome formation; however, phagolysosome formation was inhibited. Moreover, suppression of AFD36229.1 expression prevented E. coli digestion and consequently led to the rupturing of A. castellanii. Our results demonstrated that the ACA1_077100, ACA1_175060, and AFD36229.1 genes of Acanthamoeba played crucial roles not only in the formation of phagosome and phagolysosome but also in the digestion of E. coli.

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Microarray-based characterization of airway inflammation induced by environmental Acanthamoeba exposure versus the ovalbumin-alum model
Shin Ae Kang, Hak Sun Yu
Parasites, Hosts and Diseases.2025; 63(4): 327. CrossRef
Phylogenomic, structural, and cell biological analyses reveal that Stenotrophomonas maltophilia replicates in acidified Rab7A-positive vacuoles of Acanthamoeba castellanii
Javier Rivera, Julio C. Valerdi-Negreros, Diana M. Vázquez-Enciso, Fulvia-Stefany Argueta-Zepeda, Pablo Vinuesa, Michael L. Ginger, Monica Crary, Sutherland K. Maciver
Microbiology Spectrum.2024;[Epub] CrossRef

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The detection of Toxoplasma gondii ME49 infections in BALB/c mice using various techniques: Hae-Ji Kang, Jie Mao, Min-Ju Kim, Keon-Woong Yoon, Gi-Deok Eom, Ki-Back Chu, Eun-Kyung Moon, Fu-Shi Quan; Parasites Hosts Dis 2023;61(4):418-427.
Published online November 28, 2023; DOI: https://doi.org/10.3347/PHD.23048

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Toxoplasma gondii infections are primarily diagnosed by serological assays, whereas molecular and fluorescence-based techniques are garnering attention for their high sensitivity in detecting these infections. Nevertheless, each detection method has its limitations. The toxoplasmosis detection capabilities of most of the currently available methods have not been evaluated under identical experimental conditions. This study aimed to assess the diagnostic potential of enzyme-linked immunosorbent assay (ELISA), real-time polymerase chain reaction (RT-PCR), immunohistochemistry (IHC), and immunofluorescence (IF) in BALB/c mice experimentally infected with various doses of T. gondii ME49. The detection of toxoplasmosis from sera and brain tissues was markedly enhanced in mice subjected to high infection doses (200 and 300 cysts) compared to those subjected to lower doses (10 and 50 cysts) for all the detection methods. Additionally, increased B1 gene expression levels and cyst sizes were observed in the brain tissues of the mice. Importantly, IHC, IF, and ELISA, but not RT-PCR, successfully detected T. gondii infections at the lowest infection dose (10 cysts) in the brain. These findings may prove beneficial while designing experimental methodologies for detecting T. gondii infections in mice.

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Protection Against Toxoplasma gondii Lethal ME49 Challenge Induced by Influenza Virus-like Particles Containing Dense Granule Protein 14
Jie Mao, Hae-Ji Kang, Gi-Deok Eom, Su In Heo, Hynnu Nam, Ji-Hyun Lee, Ki-Ho Park, Mi Suk Lee, Sung Soo Kim, Fu-Shi Quan
Pharmaceutics.2026; 18(1): 93. CrossRef
Spatiotemporal Diffusion, Colonization, and Antibody Responses in Susceptible C57BL/6J Mice Orally Infected with Toxoplasma gondii Cysts
Zhao Li, Qi-Shuai Liu, Jun-Jie Hu, Cai-Qin Deng, Tao Li, Wen-Bin Zheng, Xing-Quan Zhu, Feng-Cai Zou
Veterinary Sciences.2025; 12(3): 212. CrossRef
Ivermectin Identified Using a High-Throughput Screening System Exhibits Anti-Clonorchis sinensis Activity in Rats
Soon-Ok Lee, Hyeryon Lee, Ki Back Chu, Jianhua Li, Sung-Jong Hong, Sung Soo Kim, Joo Hwan No, Fu-Shi Quan
Antibiotics.2025; 14(8): 837. CrossRef
Recombinant vaccinia virus expressing MIC8, AMA1, or RON4 induce protection against Toxoplasma gondii ME49 strain infection
Hae-Ji Kang, Fu-Shi Quan
Acta Tropica.2025; 270: 107812. CrossRef
Protective Efficacy Induced by Virus-like Particles Expressing Dense Granule Protein 5 of Toxoplasma gondii
Su In Heo, Hae-Ji Kang, Jie Mao, Zhao-Shou Yang, Md Atique Ahmed, Fu-Shi Quan
Vaccines.2025; 13(8): 787. CrossRef
Vaccinia virus expressing MIC8 and AMA1 provides protection against Toxoplasma gondii ME49 infection
Hae-Ji Kang, Yan Jin, Zhao-Shou Yang, Md Atique Ahmed, Fu-Shi Quan
Parasites, Hosts and Diseases.2025; 63(4): 340. CrossRef

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Brief Communication

Specific Detection of Acanthamoeba species using Polyclonal Peptide Antibody Targeting the Periplasmic Binding Protein of A. castellanii: Min-Jeong Kim, Fu-Shi Quan, Hyun-Hee Kong, Jong-Hyun Kim, Eun-Kyung Moon; Korean J Parasitol 2022;60(2):143-147.
Published online April 20, 2022; DOI: https://doi.org/10.3347/kjp.2022.60.2.143

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Acanthamoeba keratitis (AK) is a rare ocular disease, but it is a painful and sight-threatening infectious disease. Early diagnosis and adequate treatment are necessary to prevent serious complications. While AK is frequently diagnosis via several PCR assays or Acanthamoeba-specific antibodies, a more specific and effective diagnostic method is required. This study described the production of a polyclonal peptide antibody against the periplasmic binding protein (PBP) of A. castellanii and investigated its diagnostic potential. Western blot analysis showed that the PBP antibody specifically reacted with the cell lysates of A. castellanii. However, the PBP antibody did not interact with human corneal epithelial (HCE) cells and the other 3 major causative agents of keratitis. Immunocytochemistry (ICC) results revealed the specific detection of A. castellanii trophozoites and cysts by PBP antibodies when A. castellanii were co-cultured with HCE cells. PBP antibody specificity was further confirmed by co-culture of A. castellanii trophozoites with F. solani, S. aureus, and P. aeruginosa via ICC. The PBP antibody specifically reacted with the trophozoites and cysts of A. polyphaga, A. hatchetti, A. culbertsoni, A. royreba, and A. healyi, thus demonstrated its genus-specific nature. These results showed that the PBP polyclonal peptide antibody of A. castellanii could specifically detect several species of Acanthamoeba, contributing to the development of an effective antibody-based AK diagnostics.

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Evaluation of the potential for diagnosis of fungal keratitis using a Fusarium-specific antibody
Hye-Jeong Jo, Min-Jeong Kim, Hae-Ahm Lee, Fu-Shi Quan, Hyun-Hee Kong, Eun-Kyung Moon
Scientific Reports.2025;[Epub] CrossRef
Detection of Fusarium solani using cutinase antibody and its application in diagnosing fungal keratitis in an animal model
Hye-Jeong Jo, Min-Jeong Kim, Hae-Ahm Lee, Fu-Shi Quan, Hyun-Hee Kong, Eun-Kyung Moon, Olaf Kniemeyer
PLOS One.2025; 20(8): e0330455. CrossRef
A chorismate mutase-targeted, core-shell nanoassembly-activated SERS immunoassay platform for rapid non-invasive detection of Acanthamoeba infection
Hyerin Lee, Min-Jeong Kim, Junkyu Chung, Wansun Kim, Hye-Jeong Jo, Tae Gi Kim, Jae-Ho Shin, Gi-Ja Lee, Fu-Shi Quan, Hyun-Hee Kong, Sang Woong Moon, Eun-Kyung Moon, Samjin Choi
Nano Today.2024; 59: 102506. CrossRef
Evaluating the Diagnostic Potential of Chorismate Mutase Poly-Clonal Peptide Antibody for the Acanthamoeba Keratitis in an Animal Model
Min-Jeong Kim, Hye-Jeong Jo, Hae-Jin Sohn, Ho-Joon Shin, Fu-Shi Quan, Hyun-Hee Kong, Eun-Kyung Moon
Pathogens.2023; 12(4): 526. CrossRef
Detection of Acanthamoeba from Acanthamoeba Keratitis Mouse Model Using Acanthamoeba-Specific Antibodies
Min-Jeong Kim, A-Jeong Ham, A-Young Park, Hae-Jin Sohn, Ho-Joon Shin, Fu-Shi Quan, Hyun-Hee Kong, Eun-Kyung Moon
Microorganisms.2022; 10(9): 1711. CrossRef

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Original Articles

Characterization of a Peptide Antibody Specific to the Adenylyl Cyclase-Associated Protein of Acanthamoeba castellanii: Min-Jeong Kim, Hae-Ahm Lee, Fu-Shi Quan, Hyun-Hee Kong, Eun-Kyung Moon; Korean J Parasitol 2022;60(1):7-14.
Published online February 23, 2022; DOI: https://doi.org/10.3347/kjp.2022.60.1.7

Abstract
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Acanthamoeba keratitis (AK) is a rare infectious disease and accurate diagnosis has remained arduous as clinical manifestations of AK were similar to keratitis of viral, bacterial, or fungal origins. In this study, we described the production of a polyclonal peptide antibody against the adenylyl cyclase-associated protein (ACAP) of A. castellanii, and evaluated its differential diagnostic potential. Enzyme-linked immunosorbent assay revealed high titers of A. castellanii-specific IgG and IgA antibodies being present in low dilutions of immunized rabbit serum. Western blot analysis revealed that the ACAP antibody specifically interacted with A. castellanii, while not interacting with human corneal epithelial (HCE) cells and other causes of keratitis such as Fusarium solani, Pseudomonas aeruginosa, and Staphylococcus aureus. Immunocytochemistry (ICC) results confirmed the specific detection of trophozoites and cysts of A. castellanii co-cultured with HCE cells. The ACAP antibody also specifically interacted with the trophozoites and cysts of 5 other Acanthamoeba species. These results indicate that the ACAP antibody of A. castellanii can specifically detect multiple AK-causing members belonging to the genus Acanthamoeba and may be useful for differentially diagnosing Acanthamoeba infections.

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The pathogenesis, risk factors, diagnosis and treatment of Acanthamoeba keratitis
Mingliang Bao, Hai Bao, Shuqing Wang, Hongyan Zhou
Frontiers in Medicine.2025;[Epub] CrossRef
A chorismate mutase-targeted, core-shell nanoassembly-activated SERS immunoassay platform for rapid non-invasive detection of Acanthamoeba infection
Hyerin Lee, Min-Jeong Kim, Junkyu Chung, Wansun Kim, Hye-Jeong Jo, Tae Gi Kim, Jae-Ho Shin, Gi-Ja Lee, Fu-Shi Quan, Hyun-Hee Kong, Sang Woong Moon, Eun-Kyung Moon, Samjin Choi
Nano Today.2024; 59: 102506. CrossRef
Evaluating the Diagnostic Potential of Chorismate Mutase Poly-Clonal Peptide Antibody for the Acanthamoeba Keratitis in an Animal Model
Min-Jeong Kim, Hye-Jeong Jo, Hae-Jin Sohn, Ho-Joon Shin, Fu-Shi Quan, Hyun-Hee Kong, Eun-Kyung Moon
Pathogens.2023; 12(4): 526. CrossRef
New Frontiers in Acanthamoeba Keratitis Diagnosis and Management
Omar Shareef, Sana Shareef, Hajirah N. Saeed
Biology.2023; 12(12): 1489. CrossRef
Detection of Acanthamoeba from Acanthamoeba Keratitis Mouse Model Using Acanthamoeba-Specific Antibodies
Min-Jeong Kim, A-Jeong Ham, A-Young Park, Hae-Jin Sohn, Ho-Joon Shin, Fu-Shi Quan, Hyun-Hee Kong, Eun-Kyung Moon
Microorganisms.2022; 10(9): 1711. CrossRef

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Sirtinol Supresses Trophozoites Proliferation and Encystation of Acanthamoeba via Inhibition of Sirtuin Family Protein: So-Young Joo, Ja Moon Aung, Minsang Shin, Eun-Kyung Moon, Hyun-Hee Kong, Youn-Kyoung Goo, Dong-Il Chung, Yeonchul Hong; Korean J Parasitol 2022;60(1):1-6.
Published online February 23, 2022; DOI: https://doi.org/10.3347/kjp.2022.60.1.1

Abstract
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The encystation of Acanthamoeba leads to the development of metabolically inactive and dormant cysts from vegetative trophozoites under unfavorable conditions. These cysts are highly resistant to anti-Acanthamoeba drugs and biocides. Therefore, the inhibition of encystation would be more effective in treating Acanthamoeba infection. In our previous study, a sirtuin family protein—Acanthamoeba silent-information regulator 2-like protein (AcSir2)—was identified, and its expression was discovered to be critical for Acanthamoeba castellanii proliferation and encystation. In this study, to develop Acanthamoeba sirtuin inhibitors, we examine the effects of sirtinol, a sirtuin inhibitor, on trophozoite growth and encystation. Sirtinol inhibited A. castellanii trophozoites proliferation (IC50=61.24 μM). The encystation rate of cells treated with sirtinol significantly decreased to 39.8% (200 μM sirtinol) after 24 hr of incubation compared to controls. In AcSir2-overexpressing cells, the transcriptional level of cyst-specific cysteine protease (CSCP), an Acanthamoeba cysteine protease involved in the encysting process, was 11.6- and 88.6-fold higher at 48 and 72 hr after induction of encystation compared to control. However, sirtinol suppresses CSCP transcription, resulting that the undegraded organelles and large molecules remained in sirtinol-treated cells during encystation. These results indicated that sirtinol sufficiently inhibited trophozoite proliferation and encystation, and can be used to treat Acanthamoeba infections.

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Pterostilbene: A natural neuroprotective stilbene with anti-Alzheimer's disease properties
Songlan Gao, Honglei Zhang, Na Li, Lijuan Zhang, Zhe Zhu, Changlu Xu
Journal of Pharmaceutical Analysis.2025; 15(4): 101043. CrossRef
Alzheimer’s Disease: A Review of Molecular Mechanisms and Therapeutic Implications by Targeting Sirtuins, Caspases, and GSK-3
Kalpana Pandya, Krishnashish Roul, Avanish Tripathi, Sateesh Belemkar, Anshuman Sinha, Meryem Erol, Devendra Kumar
ACS Chemical Neuroscience.2025; 16(12): 2178. CrossRef
Human Conjunctival Transcriptome in Acanthamoeba Keratitis: An Exploratory Study
Gerami D. Seitzman, Jeremy D. Keenan, Thomas M. Lietman, Kevin Ruder, Lina Zhong, Cindi Chen, YuHeng Liu, Danny Yu, Thomas Abraham, Armin Hinterwirth, Thuy Doan
Cornea.2024; 43(10): 1272. CrossRef
Comparative cytotoxicity of Acanthamoeba castellanii-derived conditioned medium on human corneal epithelial and stromal cells
Abdullah Alhazmi, Laura E. Sidney, Andy Hopkinson, Hany M. Elsheikha
Acta Tropica.2024; 257: 107288. CrossRef
Biological characteristics and pathogenicity of Acanthamoeba
Yuehua Wang, Linzhe Jiang, Yitong Zhao, Xiaohong Ju, Le Wang, Liang Jin, Ryan D. Fine, Mingguang Li
Frontiers in Microbiology.2023;[Epub] CrossRef

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Brief Communication

Differentially Expressed Gene Profile of Acanthamoeba castellanii Induced by an Endosymbiont Legionella pneumophila: Eun-Kyung Moon, So-Min Park, Ki-Back Chu, Fu-Shi Quan, Hyun-Hee Kong; Korean J Parasitol 2021;59(1):67-75.
Published online February 19, 2021; DOI: https://doi.org/10.3347/kjp.2021.59.1.67

Abstract
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Legionella pneumophila is an opportunistic pathogen that survives and proliferates within protists such as Acanthamoeba spp. in environment. However, intracellular pathogenic endosymbiosis and its implications within Acanthamoeba spp. remain poorly understood. In this study, RNA sequencing analysis was used to investigate transcriptional changes in A. castellanii in response to L. pneumophila infection. Based on RNA sequencing data, we identified 1,211 upregulated genes and 1,131 downregulated genes in A. castellanii infected with L. pneumophila for 12 hr. After 24 hr, 1,321 upregulated genes and 1,379 downregulated genes were identified. Gene ontology (GO) analysis revealed that L. pneumophila endosymbiosis enhanced hydrolase activity, catalytic activity, and DNA binding while reducing oxidoreductase activity in the molecular function (MF) domain. In particular, multiple genes associated with the GO term ‘integral component of membrane’ were downregulated during endosymbiosis. The endosymbiont also induced differential expression of various methyltransferases and acetyltransferases in A. castellanii. Findings herein are may significantly contribute to understanding endosymbiosis of L. pneumophila within A. castellanii.

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Oxford Nanopore Technology-Based Identification of an Acanthamoeba castellanii Endosymbiosis in Microbial Keratitis
Sebastian Alexander Scharf, Lennart Friedrichs, Robert Bock, Maria Borrelli, Colin MacKenzie, Klaus Pfeffer, Birgit Henrich
Microorganisms.2024; 12(11): 2292. CrossRef
Transcription dynamics of heat-shock proteins (Hsps) and endosymbiont titres in response to thermal stress in whitefly, Bemisia tabaci (Asia-I)
Mritunjoy Barman, Snigdha Samanta, Bulbul Ahmed, Soumik Dey, Swati Chakraborty, M.G. Deeksha, Subham Dutta, Arunava Samanta, Jayanta Tarafdar, Deepayan Roy
Frontiers in Physiology.2023;[Epub] CrossRef
Proteomic analysis of Acanthamoeba castellanii response to Legionella pneumophila infection
Alban Hay, Steven Rolland, Clément Bernard, Yann Héchard, Romain Villéger, Ascel Samba-Louaka
FEMS Microbiology Letters.2023;[Epub] CrossRef
Comparative analysis of differentially expressed genes in Acanthamoeba after ingestion of Legionella pneumophila and Escherichia coli
Eun-Kyung Moon, Min-Jeong Kim, Hae-Ahm Lee, Fu-Shi Quan, Hyun-Hee Kong
Experimental Parasitology.2022; 232: 108188. CrossRef

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Original Article

Comparison of Proteins Secreted into Extracellular Space of Pathogenic and Non-pathogenic Acanthamoeba castellanii: Eun-Kyung Moon, Hyun-Seo Choi, So-Min Park, Hyun-Hee Kong, Fu-Shi Quan; Korean J Parasitol 2018;56(6):553-558.
Published online December 31, 2018; DOI: https://doi.org/10.3347/kjp.2018.56.6.553

Pathogenic Acanthamoeba spp. cause granulomatous amoebic encephalitis and keratitis. Acanthamoeba keratitis (AK) is a rare but serious ocular infection that can result in permanent visual impairment or blindness. However, pathogenic factors of AK remain unclear and treatment for AK is arduous. Expression levels of proteins secreted into extracellular space were compared between A. castellanii pathogenic (ACP) and non-pathogenic strains. Two-dimensional polyacrylamide gel electrophoresis revealed 123 differentially expressed proteins, including 34 increased proteins , 7 qualitative increased proteins, 65 decreased proteins, and 17 qualitative decreased proteins in ACP strain. Twenty protein spots with greater than 5-fold increase in ACP strain were analyzed by liquid chromatography triple quadrupole mass spectrometry. These proteins showed similarity each to inosine-uridine preferring nucleoside hydrolase, carboxylesterase, oxygen-dependent choline dehydrogenase, periplasmic-binding protein proteinases and hypothetical proteins. These proteins expressed higher in ACP may provide some information to understand pathogenicity of Acanthamoeba.

Citations

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The gene expression and proteomic profiling of Acanthamoeba isolates
Chayan Sharma, Sumeeta Khurana, Alka Bhatia, Amit Arora, Amit Gupta
Experimental Parasitology.2023; 255: 108630. CrossRef
Detection of Acanthamoeba spp. using carboxylesterase antibody and its usage for diagnosing Acanthamoeba-keratitis
Min-Jeong Kim, Ki-Back Chu, Hae-Ahm Lee, Fu-Shi Quan, Hyun-Hee Kong, Eun-Kyung Moon, Ashok Kumar
PLOS ONE.2022; 17(1): e0262223. CrossRef
Characterization of a Peptide Antibody Specific to the Adenylyl Cyclase-Associated Protein of Acanthamoeba castellanii
Min-Jeong Kim, Hae-Ahm Lee, Fu-Shi Quan, Hyun-Hee Kong, Eun-Kyung Moon
The Korean Journal of Parasitology.2022; 60(1): 7. CrossRef
Specific Detection of Acanthamoeba species using Polyclonal Peptide Antibody Targeting the Periplasmic Binding Protein of A. castellanii
Min-Jeong Kim, Fu-Shi Quan, Hyun-Hee Kong, Jong-Hyun Kim, Eun-Kyung Moon
The Korean Journal of Parasitology.2022; 60(2): 143. CrossRef
Host-Parasite Interaction of Atlantic salmon (Salmo salar) and the Ectoparasite Neoparamoeba perurans in Amoebic Gill Disease
Natasha A. Botwright, Amin R. Mohamed, Joel Slinger, Paula C. Lima, James W. Wynne
Frontiers in Immunology.2021;[Epub] CrossRef
Proteomic analysis of the skeletal muscles from dysferlinopathy patients
Young-Chul Choi, Ji-Man Hong, Kee Duk Park, Ha Young Shin, Seung Min Kim, Hyung Jun Park
Journal of Clinical Neuroscience.2020; 71: 186. CrossRef
Production of a polyclonal antibody against inosine-uridine preferring nucleoside hydrolase of Acanthamoeba castellanii and its access to diagnosis of Acanthamoeba keratitis
So-Min Park, Hae-Ahm Lee, Ki-Back Chu, Fu-Shi Quan, Su-Jung Kim, Eun-Kyung Moon, Paulo Lee Ho
PLOS ONE.2020; 15(9): e0239867. CrossRef
Quantitative proteomic analysis and functional characterization of Acanthamoeba castellanii exosome-like vesicles
Wei-Chen Lin, Chia-Yun Tsai, Jian-Ming Huang, Shang-Rung Wu, Lichieh Julie Chu, Kuo-Yang Huang
Parasites & Vectors.2019;[Epub] CrossRef

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Brief Communication

Effect of 2, 6-Dichlorobenzonitrile on Amoebicidal Activity of Multipurpose Contact Lens Disinfecting Solutions: Eun-Kyung Moon, Seungeun Lee, Fu-Shi Quan, Hyun-Hee Kong; Korean J Parasitol 2018;56(5):491-494.
Published online October 31, 2018; DOI: https://doi.org/10.3347/kjp.2018.56.5.491

Multipurpose contact lens disinfecting solutions (MPDS) are widely used to cleanse and disinfect microorganisms. However, disinfection efficacy of these MPDS against Acanthamoeba cyst remain insufficient. 2, 6-dichlorobenzonitrile (DCB), a cellulose synthesis inhibitor, is capable of increasing the amoebical effect against Acanthamoeba by inhibiting its encystation. In this study, we investigated the possibility of DCB as a disinfecting agent to improve the amoebicidal activity of MPDS against Acanthamoeba cyst. Eight commercial MPDS (from a to h) were assessed, all of which displayed insufficient amoebicidal activity against the mature cysts. Solution e, f, and h showed strong amoebicidal effect on the immature cysts. Amoebicidal efficacy against mature cysts remained inadequate even when the 8 MPDS were combined with 100 μM DCB. However, 4 kinds of MPDS (solution d, e, f, and h) including 100 μM DCB demonstrated strong amoebicidal activity against the immature cysts. The amoebicidal activity of solution d was increased by addition of DCB. Cytotoxicity was absent in human corneal epithelial cells treated with either DCB or mixture of DCB with MPDS. These results suggested that DCB can enhance the amoebicical activity of MPDS against Acanthamoeba immature cyst in vitro.

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Original Articles

Influenza M1 Virus-Like Particles Consisting of Toxoplasma gondii Rhoptry Protein 4: Su-Hwa Lee, Dong-Hun Lee, Ying Piao, Eun-Kyung Moon, Fu-Shi Quan; Korean J Parasitol 2017;55(2):143-148.
Published online April 30, 2017; DOI: https://doi.org/10.3347/kjp.2017.55.2.143

Abstract
PDF

Toxoplasma gondii infections occur throughout the world, and efforts are needed to develop various vaccine candidates expressing recombinant protein antigens. In this study, influenza matrix protein (M1) virus-like particles (VLPs) consisting of T. gondii rhoptry antigen 4 (ROP4 protein) were generated using baculovirus (rBV) expression system. Recombinant ROP4 protein with influenza M1 were cloned and expressed in rBV. SF9 insect cells were coinfected with recombinant rBVs expressing T. gondii ROP4 and influenza M1. As the results, influenza M1 VLPs showed spherical shapes, and T. gondii ROP4 protein exhibited as spikes on VLP surface under transmission electron microscopy (TEM). The M1 VLPs resemble virions in morphology and size. We found that M1 VLPs reacted with antibody from T. gondii-infected mice by western blot and ELISA. This study demonstrated that T. gondii ROP4 protein can be expressed on the surface of influenza M1 VLPs and the M1 VLPs containing T. gondii ROP4 reacted with T. gondii-infected sera, indicating the possibility that M1 VLPs could be used as a coating antigen for diagnostic and/or vaccine candidate against T. gondii infection.

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Karolina Sołowińska, Lucyna Holec-Gąsior
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Protective immunity induced by CpG ODN‐adjuvanted virus‐like particles containing Toxoplasma gondii proteins
Hae‐Ji Kang, Ki‐Back Chu, Min‐Ju Kim, Su‐Hwa Lee, Hyunwoo Park, Hui Jin, Eun‐Kyung Moon, Fu‐Shi Quan
Parasite Immunology.2021;[Epub] CrossRef
Detection of Toxoplasma gondii Infections using Virus-Like Particles Displaying T. gondii ROP4 Antigen
Min-Ju Kim, Jie Mao, Hae-Ji Kang, Ki-Back Chu, Fu-Shi Quan
The Korean Journal of Parasitology.2021; 59(6): 565. CrossRef
Virus-like particle vaccine displaying Toxoplasma gondii apical membrane antigen 1 induces protection against T. gondii ME49 infection in mice
Min-Ju Kim, Su-Hwa Lee, Hae-Ji Kang, Ki-Back Chu, Hyunwoo Park, Hui Jin, Eun-Kyung Moon, Sung Soo Kim, Fu-Shi Quan
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Toxoplasma gondii virus‐like particle vaccination alleviates inflammatory response in the brain upon T gondii infection
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Parasite Immunology.2020;[Epub] CrossRef
Previous Infection with Plasmodium berghei Confers Resistance to Toxoplasma gondii Infection in Mice
Dong-Hun Lee, Ki-Back Chu, Hae-Ji Kang, Su-Hwa Lee, Fu-Shi Quan
The Korean Journal of Parasitology.2019; 57(2): 93. CrossRef
Virus-Like Particles Expressing Toxoplasma gondii Rhoptry Protein 18 Induces Better Protection Than Rhoptry Protein 4 against T. gondii Infection
Hae-Ji Kang, Su-Hwa Lee, Ki-Back Chu, Dong-Hun Lee, Fu-Shi Quan
The Korean Journal of Parasitology.2018; 56(5): 429. CrossRef

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DNA Methylation of Gene Expression in Acanthamoeba castellanii Encystation: Eun-Kyung Moon, Yeonchul Hong, Hae-Ahm Lee, Fu-Shi Quan, Hyun-Hee Kong; Korean J Parasitol 2017;55(2):115-120.
Published online April 30, 2017; DOI: https://doi.org/10.3347/kjp.2017.55.2.115

Abstract
PDF

Encystation mediating cyst specific cysteine proteinase (CSCP) of Acanthamoeba castellanii is expressed remarkably during encystation. However, the molecular mechanism involved in the regulation of CSCP gene expression remains unclear. In this study, we focused on epigenetic regulation of gene expression during encystation of Acanthamoeba. To evaluate methylation as a potential mechanism involved in the regulation of CSCP expression, we first investigated the correlation between promoter methylation status of CSCP gene and its expression. A 2,878 bp of promoter sequence of CSCP gene was amplified by PCR. Three CpG islands (island 1-3) were detected in this sequence using bioinformatics tools. Methylation of CpG island in trophozoites and cysts was measured by bisulfite sequence PCR. CSCP promoter methylation of CpG island 1 (1,633 bp) was found in 8.2% of trophozoites and 7.3% of cysts. Methylation of CpG island 2 (625 bp) was observed in 4.2% of trophozoites and 5.8% of cysts. Methylation of CpG island 3 (367 bp) in trophozoites and cysts was both 3.6%. These results suggest that DNA methylation system is present in CSCP gene expression of Acanthamoeba. In addition, the expression of encystation mediating CSCP is correlated with promoter CpG island 1 hypomethylation.

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The Korean Journal of Parasitology.2019; 57(3): 217. CrossRef

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Identification and Characterization of Protein Arginine Methyltransferase 1 in Acanthamoeba castellanii: Eun-Kyung Moon, Hyun-Hee Kong, Yeonchul Hong, Hae-Ahm Lee, Fu-Shi Quan; Korean J Parasitol 2017;55(2):109-114.
Published online April 30, 2017; DOI: https://doi.org/10.3347/kjp.2017.55.2.109

Abstract
PDF

Protein arginine methyltransferase (PRMT) is an important epigenetic regulator in eukaryotic cells. During encystation, an essential process for Acanthamoeba survival, the expression of a lot of genes involved in the encystation process has to be regulated in order to be induced or inhibited. However, the regulation mechanism of these genes is yet unknown. In this study, the full-length 1,059 bp cDNA sequence of Acanthamoeba castellanii PRMT1 (AcPRMT1) was cloned for the first time. The AcPRMT1 protein comprised of 352 amino acids with a SAM-dependent methyltransferase PRMT-type domain. The expression level of AcPRMT1 was highly increased during encystation of A. castellanii. The EGFPAcPRMT1 fusion protein was distributed over the cytoplasm, but it was mainly localized in the nucleus of Acanthamoeba. Knock down of AcPRMT1 by synthetic siRNA with a complementary sequence failed to form mature cysts. These findings suggested that AcPRMT1 plays a critical role in the regulation of encystation of A. castellanii. The target gene of AcPRMT1 regulation and the detailed mechanisms need to be investigated by further studies.

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PRMT5 Inhibitor EPZ015666 Decreases the Viability and Encystment of Entamoeba invadens
Rigoberto Ortiz-Hernández, Elmer Joel Millán-Casarrubias, Jeni Bolaños, Susana Munguía-Robledo, Carlos Vázquez-Calzada, Elisa Azuara-Licéaga, Jesús Valdés, Mario Alberto Rodríguez
Molecules.2024; 30(1): 62. CrossRef
Comparative analysis of differentially expressed genes in Acanthamoeba after ingestion of Legionella pneumophila and Escherichia coli
Eun-Kyung Moon, Min-Jeong Kim, Hae-Ahm Lee, Fu-Shi Quan, Hyun-Hee Kong
Experimental Parasitology.2022; 232: 108188. CrossRef
Drug Discovery against Acanthamoeba Infections: Present Knowledge and Unmet Needs
Hany M. Elsheikha, Ruqaiyyah Siddiqui, Naveed Ahmed Khan
Pathogens.2020; 9(5): 405. CrossRef
Free-living amoebae and squatters in the wild: ecological and molecular features
Ascel Samba-Louaka, Vincent Delafont, Marie-Hélène Rodier, Estelle Cateau, Yann Héchard
FEMS Microbiology Reviews.2019; 43(4): 415. CrossRef

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Efficacy of Korean Multipurpose Contact Lens Disinfecting Solutions against Acanthamoeba castellanii: Eun-Kyung Moon, Hye-Ryun Park, Fu-Shi Quan, Hyun-Hee Kong; Korean J Parasitol 2016;54(6):697-702.
Published online December 31, 2016; DOI: https://doi.org/10.3347/kjp.2016.54.6.697

Abstract
PDF

Acanthamoeba keratitis has been increasing in recent years. Main risk factors are contact lens wear and their cleaning solutions. Most contact lens wearers use multipurpose disinfecting solutions (MPDS) for cleansing and disinfecting microorganisms because of its convenience. We determined amoebicidal effects of MPDS made in Korea and their cytotoxicity on human corneal epithelium cells. Fifteen commercial MPDS (A to O) were tested for their amoebicidal effects on Acanthamoeba castellanii trophozoites and cysts by using a most probable number (MPN) technique. Among them, 7 kinds of MPDS showed little or no amoebicidal effects for 24 hr exposure. Solutions A, B, G, H, L, and O showed positive amoebicidal effects, and solutions M and N killed almost all trophozoites and cysts after 24 hr exposure. However, 50%-N solution showed 56% cytotoxicity on human corneal epithelial cells within 4 hr exposure, and 50%-O solution also showed 62% cytotoxicity on human cells within 4 hr exposure. Solution A did not show any cytotoxicity on human cells. These results revealed that most MPDS made in Korea were ineffective to kill Acanthamoeba. The solutions having amoebicidal activity also showed high levels of cytotoxicity on human corneal epithelial cells. New formulations for improved MPDS that are amoebicidal but safe for host cells are needed to prevent Acanthamoeba keratitis.

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Tannic Acid-Modified Silver Nanoparticles in Conjunction with Contact Lens Solutions Are Useful for Progress against the Adhesion of Acanthamoeba spp. to Contact Lenses
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In vitro effects of multi-purpose contact lens disinfecting solutions towards survivability of Acanthamoeba genotype T4 in Malaysia
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Development of anti-acanthamoebic approaches
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BCLA CLEAR - Contact lens wettability, cleaning, disinfection and interactions with tears
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Tania Martín-Pérez, Irene Heredero-Bermejo, Cristina Verdú-Expósito, Jorge Pérez-Serrano
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Combination of tert-butyl hydroperoxide with vorinostat induces cell death of Acanthamoeba through cell cycle arrest
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T. Martín-Pérez, T. Lozano-Cruz, A. Criado-Fornelio, P. Ortega, R. Gómez, F. J. de la Mata, J. Pérez-Serrano
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Effect of 2, 6-Dichlorobenzonitrile on Amoebicidal Activity of Multipurpose Contact Lens Disinfecting Solutions
Eun-Kyung Moon, Seungeun Lee, Fu-Shi Quan, Hyun-Hee Kong
The Korean Journal of Parasitology.2018; 56(5): 491. CrossRef
Amoebicidal Effect of Nephrite-containing Contact Lens Storage Case
Jae Woo Jung, Jong Heon Lee, Sung Hee Park, Hak Sun Yu, Yoon Kyung Kim, Ji-Eun Lee
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Efficient Liquid Media for Encystation of Pathogenic Free-Living Amoebae
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The Korean Journal of Parasitology.2017; 55(3): 233. CrossRef
Effect of Multipurpose Solution Combined With Autophagy Inhibitors on Adhesion of Acanthamoeba trophozoites to Silicone Hydrogel Contact Lenses
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Cornea.2017; 36(12): 1538. CrossRef

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Identification of Protein Arginine Methyltransferase 5 as a Regulator for Encystation of Acanthamoeba: Eun-Kyung Moon, Yeonchul Hong, Dong-Il Chung, Youn-Kyoung Goo, Hyun-Hee Kong; Korean J Parasitol 2016;54(2):133-138.
Published online April 30, 2016; DOI: https://doi.org/10.3347/kjp.2016.54.2.133

Abstract
PDF

Encystation is an essential process for Acanthamoeba survival under nutrient-limiting conditions and exposure to drugs. The expression of several genes has been observed to increase or decrease during encystation. Epigenetic processes involved in regulation of gene expression have been shown to play a role in several pathogenic parasites. In the present study, we identified the protein arginine methyltransferase 5 (PRMT5), a known epigenetic regulator, in Acanthamoeba castellanii. PRMT5 of A. castellanii (AcPRMT5) contained domains found in S-adenosylmethionine-dependent methyltransferases and in PRMT5 arginine-N-methyltransferase. Expression levels of AcPRMT5 were increased during encystation of A. castellanii. The EGFP-PRMT5 fusion protein was mainly localized in the nucleus of trophozoites. A. castellanii transfected with siRNA designed against AcPRMT5 failed to form mature cysts. The findings of this study lead to a better understanding of epigenetic mechanisms behind the regulation of encystation in cyst-forming pathogenic protozoa.

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Encystment and Excystment Processes in Acanthamoeba castellanii: An Emphasis on Cellulose Involvement
Mathew Choaji, Ascel Samba-Louaka, Zineb Fechtali-Moute, Willy Aucher, Sébastien Pomel
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Oxidase enzyme genes are differentially expressed during Acanthamoeba castellanii encystment
Christian Q. Scheckhuber, Rebeca Damián Ferrara, Jesús Gómez-Montalvo, Sutherland K. Maciver, Alvaro de Obeso Fernández del Valle
Parasitology Research.2024;[Epub] CrossRef
Acanthamoeba keratitis: new hopes for potential interventions for a curable but often refractory disease
Bader Saleem Alawfi, Naveed Ahmed Khan, David Lloyd, Ruqaiyyah Siddiqui
Expert Review of Ophthalmology.2024; 19(4): 271. CrossRef
Biological characteristics and pathogenicity of Acanthamoeba
Yuehua Wang, Linzhe Jiang, Yitong Zhao, Xiaohong Ju, Le Wang, Liang Jin, Ryan D. Fine, Mingguang Li
Frontiers in Microbiology.2023;[Epub] CrossRef
Comparative analysis of differentially expressed genes in Acanthamoeba after ingestion of Legionella pneumophila and Escherichia coli
Eun-Kyung Moon, Min-Jeong Kim, Hae-Ahm Lee, Fu-Shi Quan, Hyun-Hee Kong
Experimental Parasitology.2022; 232: 108188. CrossRef
Stimulation of Acanthamoeba castellanii excystment by enzyme treatment and consequences on trophozoite growth
Zineb Fechtali-Moute, Philippe M. Loiseau, Sébastien Pomel
Frontiers in Cell and Developmental Biology.2022;[Epub] CrossRef
Free-living amoebae and squatters in the wild: ecological and molecular features
Ascel Samba-Louaka, Vincent Delafont, Marie-Hélène Rodier, Estelle Cateau, Yann Héchard
FEMS Microbiology Reviews.2019; 43(4): 415. CrossRef
Encystation: the most prevalent and underinvestigated differentiation pathway of eukaryotes
Pauline Schaap, Christina Schilde
Microbiology.2018; 164(5): 727. CrossRef
Identification and Characterization of Protein Arginine Methyltransferase 1 in Acanthamoeba castellanii
Eun-Kyung Moon, Hyun-Hee Kong, Yeonchul Hong, Hae-Ahm Lee, Fu-Shi Quan
The Korean Journal of Parasitology.2017; 55(2): 109. CrossRef
DNA Methylation of Gene Expression in Acanthamoeba castellanii Encystation
Eun-Kyung Moon, Yeonchul Hong, Hae-Ahm Lee, Fu-Shi Quan, Hyun-Hee Kong
The Korean Journal of Parasitology.2017; 55(2): 115. CrossRef

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10 Web of Science
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Down-Regulation of Cellulose Synthase Inhibits the Formation of Endocysts in Acanthamoeba: Eun-Kyung Moon, Yeonchul Hong, Dong-Il Chung, Youn-Kyoung Goo, Hyun-Hee Kong; Korean J Parasitol 2014;52(2):131-135.
Published online April 18, 2014; DOI: https://doi.org/10.3347/kjp.2014.52.2.131

Abstract
PDF

Acanthamoeba cysts are resistant to unfavorable physiological conditions and various disinfectants. Acanthamoeba cysts have 2 walls containing various sugar moieties, and in particular, one third of the inner wall is composed of cellulose. In this study, it has been shown that down-regulation of cellulose synthase by small interfering RNA (siRNA) significantly inhibits the formation of mature Acanthamoeba castellanii cysts. Calcofluor white staining and transmission electron microscopy revealed that siRNA transfected amoeba failed to form an inner wall during encystation and thus are likely to be more vulnerable. In addition, the expression of xylose isomerase, which is involved in cyst wall formation, was not altered in cellulose synthase down-regulated amoeba, indicating that cellulose synthase is a crucial factor for inner wall formation by Acanthamoeba during encystation.

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The role of the Acanthamoeba castellanii Sir2-like protein in the growth and encystation of Acanthamoeba
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Identification of Atg8 Isoform in Encysting Acanthamoeba: Eun-Kyung Moon, Yeonchul Hong, Dong-Il Chung, Hyun-Hee Kong; Korean J Parasitol 2013;51(5):497-502.
Published online October 31, 2013; DOI: https://doi.org/10.3347/kjp.2013.51.5.497

Abstract
PDF

Autophagy-related protein 8 (Atg8) is an essential component of autophagy formation and encystment of cyst-forming parasites, and some protozoa, such as, Acanthamoeba, Entamoeba, and Dictyostelium, have been reported to possess a type of Atg8. In this study, an isoform of Atg8 was identified and characterized in Acanthamoeba castellanii (AcAtg8b). AcAtg8b protein was found to encode 132 amino acids and to be longer than AcAtg8 protein, which encoded 117 amino acids. Real-time PCR analysis showed high expression levels of AcAtg8b and AcAtg8 during encystation. Fluorescence microscopy demonstrated that AcAtg8b is involved in the formation of the autophagosomal membrane. Chemically synthesized siRNA against AcAtg8b reduced the encystation efficiency of Acanthamoeba, confirming that AcAtg8b, like AcAtg8, is an essential component of cyst formation in Acanthamoeba. Our findings suggest that Acanthamoeba has doubled the number of Atg8 gene copies to ensure the successful encystation for survival when 1 copy is lost. These 2 types of Atg8 identified in Acanthamoeba provide important information regarding autophagy formation, encystation mechanism, and survival of primitive, cyst-forming protozoan parasites.

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Brief Communication

Short-Cut Pathway to Synthesize Cellulose of Encysting Acanthamoeba: Eun-Kyung Moon, Hyun-Hee Kong; Korean J Parasitol 2012;50(4):361-364.
Published online November 26, 2012; DOI: https://doi.org/10.3347/kjp.2012.50.4.361

Abstract
PDF

The mature cyst of Acanthamoeba is highly resistant to various antibiotics and therapeutic agents. Cyst wall of Acanthamoeba are composed of cellulose, acid-resistant proteins, lipids, and unidentified materials. Because cellulose is one of the primary components of the inner cyst wall, cellulose synthesis is essential to the process of cyst formation in Acanthamoeba. In this study, we hypothesized the key and short-step process in synthesis of cellulose from glycogen in encysting Acanthamoeba castellanii, and confirmed it by comparing the expression pattern of enzymes involving glycogenolysis and cellulose synthesis. The genes of 3 enzymes, glycogen phosphorylase, UDP-glucose pyrophosphorylase, and cellulose synthase, which are involved in the cellulose synthesis, were expressed high at the 1st and 2nd day of encystation. However, the phosphoglucomutase that facilitates the interconversion of glucose 1-phosphate and glucose 6-phosphate expressed low during encystation. This report identified the short-cut pathway of cellulose synthesis required for construction of the cyst wall during the encystation process in Acanthamoeba. This study provides important information to understand cyst wall formation in encysting Acanthamoeba.

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Original Articles

Microarray Analysis of Differentially Expressed Genes between Cysts and Trophozoites of Acanthamoeba castellanii: Eun-Kyung Moon, Ying-Hua Xuan, Dong-Il Chung, Yeonchul Hong, Hyun-Hee Kong; Korean J Parasitol 2011;49(4):341-347.
Published online December 16, 2011; DOI: https://doi.org/10.3347/kjp.2011.49.4.341

Abstract
PDF

Acanthamoeba infection is difficult to treat because of the resistance property of Acanthamoeba cyst against the host immune system, diverse antibiotics, and therapeutic agents. To identify encystation mediating factors of Acanthamoeba, we compared the transcription profile between cysts and trophozoites using microarray analysis. The DNA chip was composed of 12,544 genes based on expressed sequence tag (EST) from an Acanthamoeba ESTs database (DB) constructed in our laboratory, genetic information of Acanthamoeba from TBest DB, and all of Acanthamoeba related genes registered in the NCBI. Microarray analysis indicated that 701 genes showed higher expression than 2 folds in cysts than in trophozoites, and 859 genes were less expressed in cysts than in trophozoites. The results of real-time PCR analysis of randomly selected 9 genes of which expression was increased during cyst formation were coincided well with the microarray results. Eukaryotic orthologous groups (KOG) analysis showed an increment in T article (signal transduction mechanisms) and O article (posttranslational modification, protein turnover, and chaperones) whereas significant decrement of C article (energy production and conversion) during cyst formation. Especially, cystein proteinases showed high expression changes (282 folds) with significant increases in real-time PCR, suggesting a pivotal role of this proteinase in the cyst formation of Acanthamoeba. The present study provides important clues for the identification and characterization of encystation mediating factors of Acanthamoeba.

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Essential Role for an M17 Leucine Aminopeptidase in Encystation of Acanthamoeba castellanii
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The Korean Journal of Parasitology.2014; 52(2): 131. CrossRef
Microarray and KOG analysis of Acanthamoeba healyi genes up-regulated by mouse-brain passage
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Proteomic profiling of the infective trophozoite stage of Acanthamoeba polyphaga
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Cysteine Protease Inhibitor (AcStefin) Is Required for Complete Cyst Formation of Acanthamoeba
Jung-Yub Lee, Su-Min Song, Eun-Kyung Moon, Yu-Ran Lee, Bijay Kumar Jha, Dinzouna-Boutamba Sylvatrie Danne, Hee-Jae Cha, Hak Sun Yu, Hyun-Hee Kong, Dong-Il Chung, Yeonchul Hong
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Cysteine protease involving in autophagosomal degradation of mitochondria during encystation of Acanthamoeba
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Short-Cut Pathway to Synthesize Cellulose of Encysting Acanthamoeba
Eun-Kyung Moon, Hyun-Hee Kong
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Protein kinase C signaling molecules regulate encystation of Acanthamoeba
Eun-Kyung Moon, Dong-Il Chung, Yeonchul Hong, Hyun-Hee Kong
Experimental Parasitology.2012; 132(4): 524. CrossRef

9,832 View
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Expressed Sequence Tag Analysis of the Erythrocytic Stage of Plasmodium berghei: Ji-Woong Seok, Yong-Seok Lee, Eun-Kyung Moon, Jung-Yub Lee, Bijay Kumar Jha, Hyun-Hee Kong, Dong-Il Chung, Yeonchul Hong; Korean J Parasitol 2011;49(3):221-228.
Published online September 30, 2011; DOI: https://doi.org/10.3347/kjp.2011.49.3.221

Abstract
PDF

Rodent malaria parasites, such as Plasmodium berghei, are practical and useful model organisms for human malaria research because of their analogies to the human malaria in terms of structure, physiology, and life cycle. Exploiting the available genetic sequence information, we constructed a cDNA library from the erythrocytic stages of P. berghei and analyzed the expressed sequence tag (EST). A total of 10,040 ESTs were generated and assembled into 2,462 clusters. These EST clusters were compared against public protein databases and 48 putative new transcripts, most of which were hypothetical proteins with unknown function, were identified. Genes encoding ribosomal or membrane proteins and purine nucleotide phosphorylases were highly abundant clusters in P. berghei. Protein domain analyses and the Gene Ontology functional categorization revealed translation/protein folding, metabolism, protein degradation, and multiple family of variant antigens to be mainly prevalent. The presently-collected ESTs and its bioinformatic analysis will be useful resources to identify for drug target and vaccine candidates and validate gene predictions of P. berghei.

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Atg3-Mediated Lipidation of Atg8 Is Involved in Encystation of Acanthamoeba: Eun-Kyung Moon, Dong-Il Chung, Yeonchul Hong, Hyun-Hee Kong; Korean J Parasitol 2011;49(2):103-108.
Published online June 14, 2011; DOI: https://doi.org/10.3347/kjp.2011.49.2.103

Abstract
PDF

Autophagy is a catabolic process involved in the degradation of a cell's own components for cell growth, development, homeostasis, and the recycling of cellular products. Autophagosome is an essential component in the protozoan parasite during differentiation and encystation. The present study identified and characterized autophagy-related protein (Atg) 3, a member of Atg8 conjugation system, in Acanthamoeba castellanii (AcAtg3). AcAtg3 encoding a 304 amino acid protein showed high similarity with the catalytic cysteine site of other E2 like enzymes of ubiquitin system. Predicted 3D structure of AcAtg3 revealed a hammer-like shape, which is the characteristic structure of E2-like enzymes. The expression level of AcAtg3 did not increase during encystation. However, the formation of mature cysts was significantly reduced in Atg3-siRNA transfected cells in which the production of Atg8-phosphatidylethanolamine conjugate was inhibited. Fluorescent microscopic analysis revealed that dispersed AcAtg3-EGFP fusion protein gathered around autophagosomal membranes during encystation. These results provide important information for understanding autophagic machinery through the lipidation reaction mediated by Atg3 in Acanthamoeba.

Citations

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Encystment and Excystment Processes in Acanthamoeba castellanii: An Emphasis on Cellulose Involvement
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Pathogens.2025; 14(3): 268. CrossRef
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Construction of EST Database for Comparative Gene Studies of Acanthamoeba: Eun-Kyung Moon, Joung-Ok Kim, Ying-Hua Xuan, Young-Sun Yun, Se Won Kang, Yong Seok Lee, Tae-In Ahn, Yeon-Chul Hong, Dong-Il Chung, Hyun-Hee Kong; Korean J Parasitol 2009;47(2):103-107.
Published online May 26, 2009; DOI: https://doi.org/10.3347/kjp.2009.47.2.103

Abstract
PDF

The genus Acanthamoeba can cause severe infections such as granulomatous amebic encephalitis and amebic keratitis in humans. However, little genomic information of Acanthamoeba has been reported. Here, we constructed Acanthamoeba expressed sequence tags (EST) database (Acanthamoeba EST DB) derived from our 4 kinds of Acanthamoeba cDNA library. The Acanthamoeba EST DB contains 3,897 EST generated from amebae under various conditions of long term in vitro culture, mouse brain passage, or encystation, and downloaded data of Acanthamoeba from National Center for Biotechnology Information (NCBI) and Taxonomically Broad EST Database (TBestDB). The almost reported cDNA/genomic sequences of Acanthamoeba provide stand alone BLAST system with nucleotide (BLAST NT) and amino acid (BLAST AA) sequence database. In BLAST results, each gene links for the significant information including sequence data, gene orthology annotations, relevant references, and a BlastX result. This is the first attempt for construction of Acanthamoeba database with genes expressed in diverse conditions. These data were integrated into a database (http://www.amoeba.or.kr).

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Brief Communication

Differentially expressed genes of Acanthamoeba castellanii during encystation: Eun-Kyung Moon, Dong-Il Chung, Yeon-Chul Hong, Hyun-Hee Kong; Korean J Parasitol 2007;45(4):283-285.
Published online December 20, 2007; DOI: https://doi.org/10.3347/kjp.2007.45.4.283

Abstract
PDF

To examine the expressed gene profile during encystation of Acanthamoeba castellanii Castellani, we used differentially expressed gene (DGE) screening by RT-PCR with 20 sets of random primers. From this analysis, we found that approximately 16 genes showed upregulation during encystation. We chose 6 genes, which had relatively higher expression levels, for further investigation. Based on homology search in database, DEG2 showed 55% of similarity with xylose isomerase, DEG9 showed 37% of similarity with Na P-type ATPase, and DEG14 showed 77% of similarity with subtilisin-like serine proteinase. DEG3 and DEG26 were identified as hypothetical proteins and DEG25 exhibited no significant similarity to any known protein. Encystation of Acanthamoeba has been suggested to be a process to resist adverse environmental or nutritional conditions. Further characterization studies of these genes may provide us with more information on the encystation mechanism of Acanthamoeba.

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Original Articles

Molecular and biochemical characterization of a novel actin bundling protein in Acanthamoeba: Joanna It-itan Alafag, Eun-Kyung Moon, Yeon-Chul Hong, Dong-Il Chung, Hyun-Hee Kong; Korean J Parasitol 2006;44(4):331-341.
Published online December 20, 2006; DOI: https://doi.org/10.3347/kjp.2006.44.4.331

Abstract
PDF

Actin binding proteins play key roles in cell structure and movement particularly as regulators of the assembly, stability and localization of actin filaments in the cytoplasm. In the present study, a cDNA clone encoding an actin bundling protein named as AhABP was isolated from Acanthamoeba healyi, a causative agent of granulomatous amebic encephalitis. This clone exhibited high similarity with genes of Physarum polycephalum and Dictyostelium discoideum, which encode actin bundling proteins. Domain search analysis revealed the presence of essential conserved regions, i.e., an active actin binding site and 2 putative calcium binding EF-hands. Transfected amoeba cells demonstrated that AhABP is primarily localized in phagocytic cups, peripheral edges, pseudopods, and in cortical cytoplasm where actins are most abundant. Moreover, AhABP after the deletion of essential regions formed ellipsoidal inclusions within transfected cells. High-speed co-sedimentation assays revealed that AhABP directly interacted with actin in the presence of up to 10 ?M of calcium. Under the electron microscope, thick parallel bundles were formed by full length AhABP, in contrast to the thin actin bundles formed by constructs with deletion sites. In the light of these results, we conclude that AhABP is a novel actin bundling protein that is importantly associated with actin filaments in the cytoplasm.

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Hongshuai Li, Yuan Wang, Mengchu Ma, Lihong Hu, Xinxin Zhang, Lingbiao Xin, Wei Zhang, Xiaoming Sun, Yuanyuan Ren, Xinting Wang, Jie Yang
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Parasitology Research.2013; 112(3): 1125. CrossRef
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Abdul Mannan Baig, Junaid Iqbal, Naveed Ahmed Khan
Antimicrobial Agents and Chemotherapy.2013; 57(8): 3561. CrossRef

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Evaluation of taxonomic validity of four species of Acanthamoeba: A. divionensis, A. paradivionensis, A. mauritaniensis, and A. rhysodes, inferred from molecular analyses: Hua Liu, Eun-Kyung Moon, Hak-Sun Yu, Hae-Jin Jeong, Yeon-Chul Hong, Hyun-Hee Kong, Dong-Il Chung; Korean J Parasitol 2005;43(1):7-13.
Published online March 20, 2005; DOI: https://doi.org/10.3347/kjp.2005.43.1.7

Abstract
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The taxonomy of Acanthamoeba spp., an amphizoic amoeba which causes granulomatous amoebic encephalitis and chronic amoebic keratitis, has been revised many times. The taxonomic validity of some species has yet to be assessed. In this paper, we analyzed the morphological characteristics, nuclear 18s rDNA and mitochondrial 16s rDNA sequences and the Mt DNA RFLP of the type strains of four Acanthamoeba species, which had been previously designated as A. divionensis, A. parasidionensis, A. mauritaniensis, and A. rhysodes. The four isolates revealed characteristic group II morphology. They exhibited 18S rDNA sequence differences of 0.2-1.1% with each other, but more than 2% difference from the other compared reference strains. Four isolates formed a different clade from that of A. castellanii Castellani and the other strains in morphological group II on the phylogenetic tree. In light of these results, A. paradivionensis, A. divionensis, and A. mauritaniensis should be regarded as synonyms for A. rhysodes.

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