Parasites, Hosts and Diseases

"Ho-Joon Shin"

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Understanding the pathogenicity of Naegleria fowleri in association with N. fowleri antigen-1 (Nfa1): Jong-Hyun Kim, Hae-Jin Sohn, Ho-Joon Shin, Stacy E. Walz, Suk-Yul Jung; Parasites Hosts Dis 2024;62(4):385-398.
Published online November 22, 2024; DOI: https://doi.org/10.3347/PHD.24025

Abstract
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Naegleria fowleri, a brain-eating amoeba, thrives in lakes and rivers with aquatic vegetation and causes primary amoebic meningoencephalitis (PAM) in humans. Most recently, it has become such a serious problem that N. fowleri was detected in tap water in Houston, USA. Several pathogenic factors are considered very important to destroy target cells in the brain. In particular, the food-cup where N. fowleri antigen-1 (Nfa1) is located, is strongly expressed in pseudopodia involved in the movement of N. fowleri, and is involved in phagocytosis by attaching to target cells. In this article, we reviewed the role of the Nfa1 protein and its associated pathogenicity. The nfa1 gene was cloned by cDNA library immunoscreening using infection serum and immune serum. Nfa1 protein is mainly distributed in pseudopodia important to movement and vacuoles. Moreover, heat shock protein 70, cathepsin-like proteare and Nf-actin are also associated with pseudopodia in which Nfa1 is localized. Interestingly, the amount of the nfa1 gene changed as N. fowleri trophozoites transformed into cysts. Polyclonal antiserum against Nfa1 showed a protective effect against cytotoxicity of approximately 19.7%. Nfa1-specific IgA antibodies prevent N. fowleri trophozoites from adhering to the nasal mucosa, delaying invasion. The nfa1-vaccinated mice showed significantly higher levels of Nfa1-specific antibody. The duration of anti-Nfa1 IgG in the vaccinated mice lasted 12 weeks, strongly suggesting that nfa1 is a significant pathogenic gene and that Nfa1 is a pathogenic protein. Several factors related to pseudopodia and locomotion have been linked to Nfa1. A clearer function of N. fowleri targeting nfa1 with other genes might enable target-based inhibition of N. fowleri pathogenicity.

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Computational exploration of natural inhibitors against toxin-associated proteins in Naegleria fowleri Karachi strain
Rabia Faizan, Muhammad Naveed, Inmaculada Bellido Estevez, Nimra Hanif, Arooj Arshad, Tariq Aziz, Abdulhakeem S. Alamri, Walaa F. Alsanie, Majid Alhomrani
Pathology - Research and Practice.2025; 274: 156184. CrossRef
A review of the mechanism, diagnosis, and treatment of Naegleria fowleri infection
Ling Dai, Xin-Ru Guo, Xu-Rui Chen, Ming-Hao Ma, Zi-Han Liu, Juan Lai, Jun Lu, Ming Feng, Xi-Xia Liu, Sheng-Hui Yang
Frontiers in Microbiology.2025;[Epub] CrossRef

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Original Articles

An Evaluation of a New Quantitative Point-of Care Diagnostic to Measure Glucose-6-phosphate Dehydrogenase Activity: Young Yil Bahk, Seong Kyu Ahn, Heung Jin Jeon, Byoung-Kuk Na, Sung-Keun Lee, Ho-Joon Shin; Korean J Parasitol 2022;60(4):281-288.
Published online August 24, 2022; DOI: https://doi.org/10.3347/kjp.2022.60.4.281

Abstract
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Malaria continues to be one of the most crucial infectious burdens in endemic areas worldwide, as well as for travelers visiting malaria transmission regions. It has been reported that 8-aminoquinolines are effective against the Plasmodium species, particularly primaquine, for anti-hypnozoite therapy in P. vivax malaria. However, primaquine causes acute hemolytic anemia in individuals with glucose-6-phosphate dehydrogenase (G6PD) deficiency. Therefore, G6PD deficiency testing should precede hypnozoite elimination with 8-aminoquinoline. Several point-of-care devices have been developed to detect G6PD deficiency. The aim of the present study was to evaluate the performance of a novel, quantitative G6PD diagnostics based on a metagenomic blue fluorescent protein (mBFP). We comparatively evaluated the sensitivity and specificity of the G6PD diagnostic modality with standard methods using 120 human whole blood samples. The G6PD deficiency was spectrophotometrically confirmed. The performance of the G6PD quantitative test kit was compared with that of a licensed control medical device, the G6PD strip. The G6PD quantitative test kit had a sensitivity of 95% (95% confidence interval (CI): 89.3-100%) and a specificity of 100% (95% CI: 94.3-100%). This study shows that the novel diagnostic G6PD quantitative test kit could be a cost-effective and time-efficient, and universally mandated screening tool for G6PD deficiency.

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Performance of quantitative point-of-care tests to measure G6PD activity: An individual participant data meta-analysis
Arkasha Sadhewa, Ari Winasti Satyagraha, Mohammad Shafiul Alam, Wondimagegn Adissu, Anup Anvikar, Germana Bancone, Praveen K. Bharti, Vinod K. Bhutani, Santasabuj Das, Muzamil Mahdi Abdel Hamid, Mohammad Sharif Hossain, Nitika Nitika, Bernard A. Okech, Ly
PLOS Neglected Tropical Diseases.2025; 19(3): e0012864. CrossRef
Utilization of Glucose-6-Phosphate Dehydrogenase Test and the Prevalence of Enzyme Deficiency in Korea
Rihwa Choi, Wonseo Park, Gayoung Chun, Sang Gon Lee, Eun Hee Lee
Journal of Clinical Medicine.2023; 12(9): 3179. CrossRef

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Cytopathic Change and Inflammatory Response of Human Corneal Epithelial Cells Induced by Acanthamoeba castellanii Trophozoites and Cysts: Hae-Jin Sohn, Ga-Eun Seo, Jae-Ho Lee, A-Jeong Ham, Young-Hwan Oh, Heekyoung Kang, Ho-Joon Shin; Korean J Parasitol 2019;57(3):217-223.
Published online June 30, 2019; DOI: https://doi.org/10.3347/kjp.2019.57.3.217

Abstract
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Acanthamoeba castellanii has ubiquitous distribution and causes primary acanthamoebic keratitis (AK). AK is a common disease in contact lens wearers and results in permanent visual impairment or blindness. In this study, we observed the cytopathic effect, in vitro cytotoxicity, and secretion pattern of cytokines in human corneal epithelial cells (HCECs) induced by A. castellanii trophozoites and/or cysts. Morphological observation revealed that panked dendritic HCECs co-cultured with amoeba cysts had changed into round shape and gradually died. Such changes were more severe in co-culture with cyst than those of co-cultivation with trophozoites. In vitro cytotoxicity assay revealed the highest cytotoxicity to HCECs in the co-culture system with amoeba cysts. A. castellanii induced the expression of IL-1α, IL-6, IL-8, and CXCL1 in HCECs. Secreted levels of IL-1α, IL-6, and IL-8 in HCECs co-cultured with both trophozoites and cysts were increased at an early incubation time (3 and 6 hr). These results suggested that cytopathic changes and pro-inflammatory cytokines release of HCECs in response to A. castellanii, especially amoebic cysts, are an important mechanism for AK development.

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A Synthetic View on Acanthamoeba Keratitis Host Immune Response: Potential Factors Influencing the Development of Chronic Inflammation
Bianca Prado-Costa, Larissa Fagundes Pinto, Mariana Fernandes Fonseca, Denise de Freitas, Larissa Magalhães Alvarenga
Cornea.2025; 44(1): 118. CrossRef
In Vitro Efficacy of Miltefosine Against Clinical Isolates of Acanthamoeba spp. from Patients with Keratitis
Lakshminarayanan Gowtham, Savitri Sharma, Bhupesh Bagga
Seminars in Ophthalmology.2025; 40(8): 767. CrossRef
Diagnostic features of Acanthamoeba keratitis via in vivo confocal microscopy
Joanna Przybek-Skrzypecka, Malcolm Armstrong, Jennifer Kim, Andrew Walkden, Leon Au, Arun Brahma, Fiona Carley, Jaya Devi Chidambaram
Scientific Reports.2025;[Epub] CrossRef
Assessing Acanthamoeba cytotoxicity: comparison of common cell viability assays
Alvie Loufouma Mbouaka, Iwona Lesiak-Markowicz, Irene Heredero-Bermejo, Rounik Mazumdar, Julia Walochnik, Tania Martín-Pérez
Frontiers in Microbiology.2023;[Epub] CrossRef
Host cell-type and pathogen-specific immunomodulatory functions of macrophage migration inhibitory factor (MIF) in infectious keratitis
Swagata Ghosh, AH Humera Khathun, G.S. Athulya, P. Vignesh, L Mathan, Ninad Mudaraddi, Siddharth Narendran, Prajna Lalitha, N. Venkatesh Prajna
Experimental Eye Research.2023; 236: 109669. CrossRef
Aspergillus fumigatus-Stimulated Human Corneal Epithelial Cells Induce Pyroptosis of THP-1 Macrophages by Secreting TSLP
Qingshan Ji, Lisong Wang, Jiajia Liu, Yali Wu, Huayi Lv, Yuechun Wen, Lei Shi, Bin Qu, Nóra Szentmáry
Inflammation.2021; 44(2): 682. CrossRef
Corneal Changes in Acanthamoeba Keratitis at Various Levels of Severity: An In Vivo Confocal Microscopic Study
Zhenyu Wei, Kai Cao, Leying Wang, Christophe Baudouin, Antoine Labbé, Qingfeng Liang
Translational Vision Science & Technology.2021; 10(7): 10. CrossRef
Polymicrobial Keratitis: Risk Factors, Clinical Characteristics, Bacterial Profile, and Antimicrobial Resistance
Laura A. González-Dibildox, José A. Oyervidez-Alvarado, Kristian A. Vazquez-Romo, Nallely Ramos-Betancourt, Everardo Hernandez-Quintela, Francisco Beltran, Manuel Garza-Leon
Eye & Contact Lens: Science & Clinical Practice.2021; 47(8): 465. CrossRef

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Brief Communication

Monitoring of Noxious Protozoa for Management of Natural Water Resources: Young Yil Bahk, Pyo Yun Cho, Sung Kyu Ahn, Sangjung Park, Won Hwa Jheong, Yun-Kyu Park, Ho-Joon Shin, Sang-Seob Lee, Okjae Rhee, Tong-Soo Kim; Korean J Parasitol 2018;56(2):205-210.
Published online April 30, 2018; DOI: https://doi.org/10.3347/kjp.2018.56.2.205

Abstract
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Waterborne parasitic protozoa, particularly Giardia lamblia and Cryptosporidium spp., are common causes of diarrhea and gastroenteritis worldwide. The most frequently identified source of infestation is water, and exposure involves either drinking water or recreation in swimming pools or natural bodies of water. In practice, studies on Cryptosporidium oocysts and Giardia cysts in surface water are challenging owing to the low concentrations of these microorganisms because of dilution. In this study, a 3-year monitoring of Cryptosporidium parvum, Giardia lamblia, and Naegleria fowleri was conducted from August 2014 to June 2016 at 5 surface water sites including 2 lakes, 1 river, and 2 water intake plants. A total of 50 water samples of 40 L were examined. Cryptosporidium oocysts were detected in 22% of samples and Giardia cysts in 32%. Water at the 5 sampling sites was all contaminated with Cryptosporidium oocysts (0-36/L), Giardia cysts (0-39/L), or both. The geometric mean concentrations of Cryptosporidium and Giardia were 1.14 oocysts/L and 4.62 cysts/L, respectively. Thus, effective monitoring plans must take into account the spatial and temporal parameters of contamination because they affect the prevalence and distribution of these protozoan cysts in local water resources.

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Cryptosporidium and cryptosporidiosis: An update of Asian perspectives in humans, water and food, 2015–2025
Shahira Abdelaziz Ali Ahmed, Sonia Boughattas, Mohammad Reza Mahmoudi, Huma Khan, Simuzar Mamedova, Ardra Namboodiri, Frederick R. Masangkay, Panagiotis Karanis
Current Research in Parasitology & Vector-Borne Diseases.2025; 8: 100311. CrossRef
Recent progress of functionalized nanomaterials-based biosensing for monitoring of food- and water-borne pathogens
Piyush Dey, Mohd Jamshaiya Raza, Alisha Khera, Akanksha Sharma, Akhil Khajuria, Ankur Pandey, Chandra Mouli Pandey, Rakesh Kumar Sharma, Gurpal Singh, Ravi Pratap Barnwal
Environmental Nanotechnology, Monitoring & Management.2024; 21: 100914. CrossRef
Performance of sewage treatment technologies for the removal of Cryptosporidium sp. and Giardia sp.: Toward water circularity
Pilar Suarez, José Luis Alonso, Gloria Gómez, Gladys Vidal
Journal of Environmental Management.2022; 324: 116320. CrossRef
Incorporating Microbial Species Interaction in Management of Freshwater Toxic Cyanobacteria: A Systems Science Challenge
Aabir Banerji, Kasey Benesh
Ecologies.2022; 3(4): 570. CrossRef

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Original Articles

Development of Monoclonal Antibodies for Diagnosis of Plasmodium vivax: Nguyen Thi Phuong Linh, Hyun Park, Jinyoung Lee, Dong-Xu Liu, Ga-Eun Seo, Hae-Jin Sohn, Jin-Hee Han, Eun-Taek Han, Ho-Joon Shin, Seon-Ju Yeo; Korean J Parasitol 2017;55(6):623-630.
Published online December 31, 2017; DOI: https://doi.org/10.3347/kjp.2017.55.6.623

Abstract
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Plasmodium lactate dehydrogenase (pLDH) is a strong target antigen for the determination of infection with Plasmodium species specifically. However, a more effective antibody is needed because of the low sensitivity of the current antibody in many immunological diagnostic assays. In this study, recombinant Plasmodium vivax LDH (PvLDH) was experimentally constructed and expressed as a native antigen to develop an effective P. vivax-specific monoclonal antibody (mAb). Two mAbs (2CF5 and 1G10) were tested using ELISA and immunofluorescence assays (IFA), as both demonstrated reactivity against pLDH antigen. Of the 2 antibodies, 2CF5 was not able to detect P. falciparum, suggesting that it might possess P. vivax-specificity. The detection limit for a pair of 2 mAbs-linked sandwich ELISA was 31.3 ng/ml of the recombinant antigen. The P. vivax-specific performance of mAbs-linked ELISA was confirmed by in vitro-cultured P. falciparum and P. vivax-infected patient blood samples. In conclusion, the 2 new antibodies possessed the potential to detect P. vivax and will be useful in immunoassay.

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Diagnostic Methods for Non-Falciparum Malaria
Alba Marina Gimenez, Rodolfo F. Marques, Matías Regiart, Daniel Youssef Bargieri
Frontiers in Cellular and Infection Microbiology.2021;[Epub] CrossRef
Plasmodium falciparum Parasitemia and Band Sensitivity of the SD Bioline Malaria Ag P.f/Pan Rapid Diagnostic Test in Madagascar
Rajeev K. Mehlotra, Rosalind E. Howes, Estee Y. Cramer, Riley E. Tedrow, Tovonahary A. Rakotomanga, Stéphanie Ramboarina, Arsène C. Ratsimbasoa, Peter A. Zimmerman
The American Journal of Tropical Medicine and Hygiene.2019; 100(5): 1196. CrossRef

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Efficient Liquid Media for Encystation of Pathogenic Free-Living Amoebae: Hae-Jin Sohn, Heekyoung Kang, Ga-Eun Seo, Jong-Hyun Kim, Suk-Yul Jung, Ho-Joon Shin; Korean J Parasitol 2017;55(3):233-238.
Published online June 30, 2017; DOI: https://doi.org/10.3347/kjp.2017.55.3.233

Abstract
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Pathogenic Naegleria fowleri, Acanthamoeba castellanii, and Acanthamoeba polyphaga, are distributed worldwide. They are causative agents of primary amoebic meningoencephalitis or acanthamoebic keratitis in humans, respectively. Trophozoites encyst in unfavorable environments, such as exhausted food supply and desiccation. Until recently, the method of N. fowleri encystation used solid non-nutrient agar medium supplemented with heat-inactivated Escherichia coli; however, for the amoebic encystment of Acanthamoeba spp., a defined, slightly modified liquid media is used. In this study, in order to generate pure N. fowleri cysts, a liquid encystment medium (buffer 1) modified from Page’s amoeba saline was applied for encystation of N. fowleri. N. fowleri cysts were well induced after 24 hr with the above defined liquid encystment medium (buffer 1). This was confirmed by observation of a high expression of differential mRNA of nfa1 and actin genes in trophozoites. Thus, this liquid medium can replace the earlier non-nutrient agar medium for obtaining pure N. fowleri cysts. In addition, for cyst formation of Acanthamoeba spp., buffer 2 (adjusted to pH 9.0) was the more efficient medium. To summarize, these liquid encystment media may be useful for further studies which require axenic and pure amoebic cysts.

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Development of a novel encystment medium: Enhancing diagnostic potential of Acanthamoeba spp.
Julalak Chuprom, Suthinee Sangkanu, Watcharapong Mitsuwan, Rachasak Boonhok, Alok K. Paul, Sonia M. Rodrigues Oliveira, Maria L. Pereira, Tajudeen O. Jimoh, Mohammed Rahmatullah, Polrat Wilairatana, Christophe Wiart, Ajoy K. Verma, Veeranoot Nissapatorn
Veterinary World.2025; : 110. CrossRef
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Xiangyu Ren, Chen Liu, Yang Yue, Shuo Sun, Lang Zhao, Chengyuan Tao, Beibei Wang, Wu Xiong, Zongzhuan Shen, Rong Li, Qirong Shen
Microbiological Research.2025; 295: 128109. CrossRef
Cloning of nf-profilin and intercellular interaction with nf-actin in Naegleria fowleri cysts
Hae-Jin Sohn, A-Jeong Ham, A-Young Park, Jeong-Heon Lee, Sun Park, Ho-Joon Shin, Jong-Hyun Kim
Scientific Reports.2025;[Epub] CrossRef
Ouabain, ATPase inhibitor, potentially enhances the effect of polyhexamethylene biguanide on Acanthamoeba castellanii
Kuang-Yi Shih, Yao-Tsung Chang, Yu-Jen Wang, Jian-Ming Huang
International Journal for Parasitology: Drugs and Drug Resistance.2024; 25: 100550. CrossRef
Understanding the pathogenicity of Naegleria fowleri in association with N. fowleri antigen-1 (Nfa1)
Jong-Hyun Kim, Hae-Jin Sohn, Ho-Joon Shin, Stacy E. Walz, Suk-Yul Jung
Parasites, Hosts and Diseases.2024; 62(4): 385. CrossRef
De Novo Transcriptome Profiling of Naegleria fowleri Trophozoites and Cysts via RNA Sequencing
Hae-Jin Sohn, Jong-Hyun Kim, Kyongmin Kim, Sun Park, Ho-Joon Shin
Pathogens.2023; 12(2): 174. CrossRef
An International External Quality Assessment Scheme to Assess the Diagnostic Performance of Polymerase Chain Reaction Detection of Acanthamoeba Keratitis
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Brian Shing, Mina Balen, William Fenical, Anjan Debnath, Wei-Hua Chen
Microbiology Spectrum.2022;[Epub] CrossRef
Curcumin effect on Acanthamoeba triangularis encystation under nutrient starvation
Rachasak Boonhok, Suthinee Sangkanu, Suganya Phumjan, Ramita Jongboonjua, Nawarat Sangnopparat, Pattamaporn Kwankaew, Aman Tedasen, Chooi Ling Lim, Maria de Lourdes Pereira, Mohammed Rahmatullah, Polrat Wilairatana, Christophe Wiart, Karma G. Dolma, Alok
PeerJ.2022; 10: e13657. CrossRef
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Noor Akbar, Wendy E. Kaman, Maarten Sarink, Kamran Nazmi, Floris J. Bikker, Naveed Ahmed Khan, Ruqaiyyah Siddiqui
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Combinatorial treatment with β-glucanase enzyme and chlorhexidine induces cysticidal effects in Acanthamoeba cyst
Nurhidayana Mohd Rased, Syed Ahmad Tajudin Tuan Johari, Hazlina Ahamad Zakeri, Nyuk Ling Ma, Siti Aisyah Razali, Fatimah Hashim
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The Acanthamoeba SBDS, a cytoskeleton-associated gene, is highly expressed during phagocytosis and encystation
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RETRACTED ARTICLE: Establishment of an Acanthamoeba keratitis mouse model confirmed by amoebic DNA amplification
Heekyoung Kang, Hae-Jin Sohn, A-Young Park, A-Jeong Ham, Jeong-Heon Lee, Young-Hwan Oh, Yong-Joon Chwae, Kyongmin Kim, Sun Park, Hongseok Yang, Suk-Yul Jung, Jong-Hyun Kim, Ho-Joon Shin
Scientific Reports.2021;[Epub] CrossRef
A Novel Cysteine Protease Inhibitor of Naegleria fowleri That Is Specifically Expressed during Encystation and at Mature Cysts
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Pathogens.2021; 10(4): 388. CrossRef
Inhibitory Effect of Host Ocular Microenvironmental Factors on Chlorhexidine Digluconate Activity
Chun-Hsien Chen, Yu-Jen Wang, Jian-Ming Huang, Fu-Chin Huang, Wei-Chen Lin
Antimicrobial Agents and Chemotherapy.2021;[Epub] CrossRef
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Biomolecules.2021; 11(9): 1320. CrossRef
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Pharmaceuticals.2021; 14(12): 1294. CrossRef
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Chun-Hsien Chen, Chao-Li Huang, Ming-Shan He, Fu-Chin Huang, Wei-Chen Lin
International Journal of Antimicrobial Agents.2020; 55(2): 105823. CrossRef
Molecular detection of free-living amoebae from Namhangang (southern Han River) in Korea
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Scientific Reports.2020;[Epub] CrossRef
Fowlerstefin, a cysteine protease inhibitor of Naegleria fowleri, induces inflammatory responses in BV-2 microglial cells in vitro
Thị Lam Thái, Jung-Mi Kang, Hương Giang Lê, Jinyoung Lee, Won Gi Yoo, Ho-Joon Shin, Woon-Mok Sohn, Byoung-Kuk Na
Parasites & Vectors.2020;[Epub] CrossRef
The Antifungal Drug Isavuconazole Is both Amebicidal and Cysticidal against Acanthamoeba castellanii
Brian Shing, Seema Singh, Larissa M. Podust, James H. McKerrow, Anjan Debnath
Antimicrobial Agents and Chemotherapy.2020;[Epub] CrossRef
Effects of Acanthamoeba castellanii on the dissolved oxygen and the microbial community under the experimental aquatic model
Chih-Ming Tsai, Jenn-Wei Chen, Wei-Chen Lin
Experimental Parasitology.2020; 218: 107985. CrossRef
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Rasha Maal-Bared, Brent Dixon, Diana Axelsson-Olsson
Experimental Parasitology.2019; 199: 104. CrossRef
Cellular characterization of actin gene concerned with contact‐dependent mechanisms in Naegleria fowleri
Hae‐Jin Sohn, Kyoung‐Ju Song, Heekyoung Kang, A‐Jeong Ham, Jae‐Ho Lee, Yong‐Joon Chwae, Kyongmin Kim, Sun Park, Jong‐Hyun Kim, Ho‐Joon Shin
Parasite Immunology.2019;[Epub] CrossRef
Cytopathic Change and Inflammatory Response of Human Corneal Epithelial Cells Induced by Acanthamoeba castellanii Trophozoites and Cysts
Hae-Jin Sohn, Ga-Eun Seo, Jae-Ho Lee, A-Jeong Ham, Young-Hwan Oh, Heekyoung Kang, Ho-Joon Shin
The Korean Journal of Parasitology.2019; 57(3): 217. CrossRef
Production and characterization of monoclonal antibodies against cathepsin B and cathepsin B-Like proteins of Naegleria fowleri
Gi-Sang Seong, Hae-Jin Sohn, Heekyoung Kang, Ga-Eun Seo, Jong-Hyun Kim, Ho-Joon Shin
Experimental Parasitology.2017; 183: 171. CrossRef

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Brief Communication

Naegleria fowleri Lysate Induces Strong Cytopathic Effects and Pro-inflammatory Cytokine Release in Rat Microglial Cells: Yang-Jin Lee, Chang-Eun Park, Jong-Hyun Kim, Hae-Jin Sohn, Jinyoung Lee, Suk-Yul Jung, Ho-Joon Shin; Korean J Parasitol 2011;49(3):285-290.
Published online September 30, 2011; DOI: https://doi.org/10.3347/kjp.2011.49.3.285

Abstract
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Naegleria fowleri, a ubiquitous free-living ameba, causes fatal primary amebic meningoencephalitis in humans. N. fowleri trophozoites are known to induce cytopathic changes upon contact with microglial cells, including necrotic and apoptotic cell death and pro-inflammatory cytokine release. In this study, we treated rat microglial cells with amebic lysate to probe contact-independent mechanisms for cytotoxicity, determining through a combination of light microscopy and scanning and transmission electron microscopy whether N. fowleri lysate could effect on both necrosis and apoptosis on microglia in a time- as well as dose-dependent fashion. A ⁵¹Cr release assay demonstrated pronounced lysate induction of cytotoxicity (71.5%) toward microglial cells by 24 hr after its addition to cultures. In an assay of pro-inflammatory cytokine release, microglial cells treated with N. fowleri lysate produced TNF-α, IL-6, and IL-1β, though generation of the former 2 cytokines was reduced with time, and that of the last increased throughout the experimental period. In summary, N. fowleri lysate exerted strong cytopathic effects on microglial cells, and elicited pro-inflammatory cytokine release as a primary immune response.

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Providing an in vitro depiction of microglial cells challenged with immunostimulatory extracellular vesicles of Naegleria fowleri
Lissette Retana Moreira, Alberto Cornet-Gomez, M. Rosario Sepulveda, Silvia Molina-Castro, Johan Alvarado-Ocampo, Frida Chaves Monge, Mariana Jara Rojas, Antonio Osuna, Elizabeth Abrahams Sandí
Frontiers in Microbiology.2024;[Epub] CrossRef
Transcriptomic profiling of “brain-eating amoeba” Naegleria fowleri infection in mice: the host and the protozoa perspectives
Vincent Guerlais, Nina Allouch, E. Ashley Moseman, Alicja W. Wojciechowska, Jakub W. Wojciechowski, Isabel Marcelino
Frontiers in Cellular and Infection Microbiology.2024;[Epub] CrossRef
Pro-inflammatory cytokine responses to Naegleria fowleri infection
Ching-wen Chen, E. Ashley Moseman
Frontiers in Tropical Diseases.2023;[Epub] CrossRef
Naegleria fowleri Extracellular Vesicles Induce Proinflammatory Immune Responses in BV-2 Microglial Cells
Hương Giang Lê, Jung-Mi Kang, Tuấn Cường Võ, Won Gi Yoo, Byoung-Kuk Na
International Journal of Molecular Sciences.2023; 24(17): 13623. CrossRef
Three encephalitis-causing amoebae and their distinct interactions with the host
Maarten J. Sarink, Nadia L. van der Meijs, Kristin Denzer, Leo Koenderman, Aloysius G.M. Tielens, Jaap J. van Hellemond
Trends in Parasitology.2022; 38(3): 230. CrossRef
Extracellular Vesicles from Naegleria fowleri Induce IL-8 Response in THP-1 Macrophage
Sakaorat Lertjuthaporn, Jinjuta Somkird, Kittima Lekmanee, Anyapat Atipimonpat, Kasama Sukapirom, Hathai Sawasdipokin, Supathra Tiewcharoen, Kovit Pattanapanyasat, Ladawan Khowawisetsut
Pathogens.2022; 11(6): 632. CrossRef
Naegleria fowleri Cathepsin B Induces a Pro-Inflammatory Immune Response in BV-2 Microglial Cells via NF-κB and AP-1 Dependent-MAPK Signaling Pathway
Hương Giang Lê, Jung-Mi Kang, Tuấn Cường Võ, Byoung-Kuk Na
International Journal of Molecular Sciences.2022; 23(15): 8388. CrossRef
Parasite infections, neuroinflammation, and potential contributions of gut microbiota
Jérémy Alloo, Ines Leleu, Corinne Grangette, Sylviane Pied
Frontiers in Immunology.2022;[Epub] CrossRef
Fowlerstefin, a cysteine protease inhibitor of Naegleria fowleri, induces inflammatory responses in BV-2 microglial cells in vitro
Thị Lam Thái, Jung-Mi Kang, Hương Giang Lê, Jinyoung Lee, Won Gi Yoo, Ho-Joon Shin, Woon-Mok Sohn, Byoung-Kuk Na
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Original Article

Contact-Independent Cell Death of Human Microglial Cells due to Pathogenic Naegleria fowleri Trophozoites: Jong-Hyun Kim, Daesik Kim, Ho-Joon Shin; Korean J Parasitol 2008;46(4):217-221.
Published online December 20, 2008; DOI: https://doi.org/10.3347/kjp.2008.46.4.217

Abstract
PDF

Free-living Naegleria fowleri leads to a fatal infection known as primary amebic meningoencephalitis in humans. Previously, the target cell death could be induced by phagocytic activity of N. fowleri as a contact-dependent mechanism. However, in this study we investigated the target cell death under a non-contact system using a tissue-culture insert. The human microglial cells, U87MG cells, co-cultured with N. fowleri trophozoites for 30 min in a non-contact system showed morphological changes such as the cell membrane destruction and a reduction in the number. By fluorescence-activated cell sorter (FACS) analysis, U87MG cells co-cultured with N. fowleri trophozoites in a non-contact system showed a significant increasse of apoptotic cells (16%) in comparison with that of the control or N. fowleri lysate. When U87MG cells were co-cultured with N. fowleri trophozoites in a non-contact system for 30 min, 2 hr, and 4 hr, the cytotoxicity of amebae against target cells was 40.5, 44.2, and 45.6%, respectively. By contrast, the cytotoxicity of non-pathogenic N. gruberi trophozoites was 10.2, 12.4, and 13.2%, respectively. These results suggest that the molecules released from N. fowleri in a contact-independent manner as well as phagocytosis in a contact-dependent manner may induce the host cell death.

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Brief Communication

Seroprevalence of toxoplasmosis in Korean pregnant women: Kyoung-Ju Song, Jong-Chul Shin, Ho-Joon Shin, Ho-Woo Nam; Korean J Parasitol 2005;43(2):69-71.
Published online June 20, 2005; DOI: https://doi.org/10.3347/kjp.2005.43.2.69

Abstract
PDF

This study was performed in order to evaluate the sero-epidemiological status of toxoplasmosis in pregnant Korean women. Among 5,175 sera and 750 amniotic fluid samples obtained from pregnant women, 41 serum samples (0.79%) and 10 (1.33%) amniotic fluid samples tested positive for IgG antibodies by ELISA. Fifty one cases showing a score more than 0.25 on ELISA were tested for PCR reaction against the SAG1 gene. Only one case of the 51 ELISA positive cases exhibited a positive reaction on all tests. This case had a history of acute nephropyelitis during early pregnancy, but fortunately, had delivered a phenotypically healthy baby. In this study, the seroprevalence of toxoplasmosis in pregnant women was found to be comparatively low, consistent with previous reports from Korea. However our trials, performed with a variety of diagnostic tools, were considered to be useful for the precise diagnosis of congenital toxoplasmosis.

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Mini Review

Pathogenic free-living amoebae in Korea: Ho-Joon Shin, Kyung-il Im; Korean J Parasitol 2004;42(3):93-119.
Published online September 20, 2004; DOI: https://doi.org/10.3347/kjp.2004.42.3.93

Abstract
PDF

Acanthamoeba and Naegleria are widely distributed in fresh water, soil and dust throughout the world, and cause meningoencephalitis or keratoconjunctivitis in humans and other mammals. Korean isolates, namely, Naegleria sp. YM-1 and Acanthamoeba sp. YM-2, YM-3, YM-4, YM-5, YM-6 and YM-7, were collected from sewage, water puddles, a storage reservoir, the gills of a fresh water fish, and by corneal washing. These isolates were categorized into three groups based on the mortalities of infected mice namely, highly virulent (YM-4), moderately virulent (YM-2, YM-5 and YM-7) and nonpathogenic (YM-3). In addition, a new species of Acanthamoeba was isolated from a freshwater fish in Korea and tentatively named Korean isolate YM-4. The morphologic characters of its cysts were similar to those of A. culbertsoni and A. royreba, which were previously designated as Acanthamoeba group III. Based on experimentally infected mouse mortality, Acanthamoeba YM-4 was highly virulent. The isoenzymes profile of Acanthamoeba YM-4 was similar to that of A. royreba. Moreover, an anti-Acanthamoeba YM-4 monoclonal antibody reacted only with Acanthamoeba YM-4, and not with A. culbertsoni. Random amplified polymorphic DNA marker analysis and RFLP analysis of mitochondrial DNA and of a 18S small subunit ribosomal RNA, placed Acanthamoeba YM-4 in a separate cluster based on phylogenic distances. Thus Acanthamoeba YM-4 was identified as a new species, and assigned Acanthamoeba sohi. Up to the year 2002 in Korea, two clinical cases were found to be infected with Acanthamoeba spp. These patients died of meningoencephalitis. In addition, one case of Acanthamoeba pneumonia with an immunodeficient status was reported and Acanthamoeba was detected in several cases of chronic relapsing corneal ulcer, chronic conjunctivitis, and keratitis.

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Original Articles

Decreasing effect of an anti-Nfa1 polyclonal antibody on the in vitro cytotoxicity of pathogenic Naegleria fowleri: Seok-Ryoul Jeong, Su-Yeon Kang, Sang-Chul Lee, Kyoung-Ju Song, Kyung-il Im, Ho-Joon Shin; Korean J Parasitol 2004;42(1):35-40.
Published online March 20, 2004; DOI: https://doi.org/10.3347/kjp.2004.42.1.35

Abstract
PDF

The nfa1 gene was cloned from a cDNA library of pathogenic Naegleria fowleri by immunoscreening; it consisted of 360 bp and produced a 13.1 kDa recombinant protein (rNfa1) that showed the pseudopodia-specific localization by immunocytochemistry in the previous study. Based on the idea that the pseudopodia-specific Nfa1 protein mentioned above seems to be involved in the pathogenicity of N. fowleri, we observed the effect of an anti-Nfa1 antibody on the proliferation of N. fowleri trophozoites and the cytotoxicity of N. fowleri trophozoites on the target cells. The proliferation of N. fowleri trophozoites was inhibited after being treated with an anti-Nfa1 polyclonal antibody in a dose-dependent manner for 48 hrs. By a light microscope, CHO cells co-cultured with N. fowleri trophozoites (group I) for 48 hrs showed severe morphological destruction. On the contrary, CHO cells co-cultured with N. fowleri trophozoites and anti-Nfa1 polyclonal antibody (1:100 dilution) (group II) showed less destruction. In the LDH release assay results, group I showed 50.6% cytotoxicity, and group II showed 39.3%. Consequently, addition of an anti-Nfa1 polyclonal antibody produced a decreasing effect of in vitro cytotoxicity of N. fowleri in a dosedependent manner.

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Acanthamoeba sohi, n. sp., a pathogenic Korean isolate YM-4 from a freshwater fish: Kyung-il Im, Ho-Joon Shin; Korean J Parasitol 2003;41(4):181-188.
Published online December 20, 2003; DOI: https://doi.org/10.3347/kjp.2003.41.4.181

Abstract
PDF

A new species of Acanthamoeba was isolated from a freshwater fish in Korea and tentatively named Acanthamoeba sp. YM-4 (Korean isolate YM-4). The trophozoites were 11.0-23.0 ?m in length and had hyaline filamentous projections. Cysts were similar to those of A. culbertsoni and A. royreba, which were previously designated as Acanthamoeba group III. Acanthamoeba YM-4 can survive at 40℃, and its generation time was 19.6 hr, which was longer than that of A. culbertsoni. In terms of the in vitro cytotoxicity of lysates, Acanthamoeba YM-4 was weaker than A. culbertsoni, but stronger than A. polyphaga. On the basis of the mortality of experimentally infected mice, Acanthamoeba YM-4 was found to be highly virulent. The isoenzymes profile of Acanthamoeba YM-4 was similar to that of A. royreba. An anti-Acanthamoeba YM-4 monoclonal antibody, McAY7, was found to react only with Acanthamoeba YM-4, and not with A. culbertsoni. Random amplified polymorphic DNA marker analysis and RFLP analysis of mitochondrial DNA and of 18S small subunit ribosomal RNA, placed Acanthamoeba YM-4 in a separate cluster on the basis of phylogenetic distances. Thus the Acanthamoeba Korean isolate YM-4 was identified as a new species, and assigned as Acanthamoeba sohi.

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Brief Communication

In vitro cytotoxicity of Acanthamoeba spp. isolated from contact lens containers in Korea by crystal violet staining and LDH release assay: Ho-Joon Shin, Myung-Soo Cho, Suk-Yul Jung, Hyung-Il Kim, Kyung-il Im; Korean J Parasitol 2000;38(2):99-102.
Published online June 30, 2000; DOI: https://doi.org/10.3347/kjp.2000.38.2.99

Abstract
PDF

In order to observe the cytotoxicity of Acanthamoeba spp., which were isolated from contact lens containers as ethiological agents for the probable amoebic keratitis in Korea, the crystal violet staining method and LDH release assay were carried out. In the crystal violet staining method, among eight contact lens container isolates, isolate 3 (Acanthamoeba KA/LS5) showed 83.6% and 81.8% of cytotoxicity, and isolate 7 (Acanthamoeba KA/LS37) showed 28.2% and 25.1% of cytotoxicity, in 1 mg/ml and 0.5 mg/ml lysate treatments, respectively. Acanthamoeba culbertsoni and A. healyi showed 84.0% and 82.8% of cytotoxicity. Similar results were observed in A. castellanii and A. hatchetti which showed 83.6% and 75.5% of cytotoxicity. Acanthamoeba royreba and A. polyphaga showed 9.0% and 1.7% of cytotoxicity. In the LDH release assay, isolate 3 (20.4%) showed higher cytotoxicity than other isolates in 1 mg/ml lysate treatment. The results provide that at least isolate 3 has the cytotoxic effect against CHO cells and seems to be the pathogenic strain.

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Original Articles

Isoenzyme patterns and phylogenetic relationships in Acanthamoeba spp. isolated from contact lens containers in Korea: Ho-Joon Shin, Myung-Soo Cho, Han-Jip Kim, Kyung-Il Im; Korean J Parasitol 1999;37(4):229-236.
Published online December 31, 1999; DOI: https://doi.org/10.3347/kjp.1999.37.4.229

Abstract
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In order to refer to the basic information regarding the identification of isolates obtained from a contact lens container in Korea, the isoelectric focusing gel electrophoresis was employed to compare the isoenzyme band patterns among Acanthamoeba spp. including eight isolates and the simple pairwise dissimilarity analysis was carried out. For an alkaline phosphate development, isolate 7 and Acanthamoeba polyphaga showed homologous band patterns, and isolates 1, 2, and 3 showed the same patterns. For lactate dehydrogenase, similar patterns were observed in isolates 2 and 3. Isolates 3 and 5 showed homologous band patterns for malate dehydrogenase and glucose phosphate isomerase. For hexokinase, isolates 4, 7, and A. hatchetti showed the same band patterns. In others, a considerable number of interstrain polymorphisms was observed in nine isoenzyme band patterns. In Acanthamoeba group II, genetic distances among isolates 1, 2, 3, 4, and 5 ranged from 0.104 to 0.200. In comparison to A. castellanii, A. hatchetti, and A. polyphaga, genetic distances of isolates 7 and 8 were 0.254 and 0.219, respectively. In Acanthamoeba group III, including A. culbertsoni, A. healyi, and A. royreba, isolate 6 had genetic distances which ranged from 0.314 to 0.336. Finally, when comparing to the six reference Acanthamoeba, it was possible to classify isolates 1, 2, 3, 4, and 5, as genetically close-related species and as independent species group. Furthermore, isolates 6, 7 and 8 were identified as independent species as well.

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Diagnosis of Acanthamoeba keratitis
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The Korean Journal of Parasitology.2007; 45(1): 11. CrossRef
Pathogenic free-living amoebae in Korea
Ho-Joon Shin, Kyung-il Im
The Korean Journal of Parasitology.2004; 42(3): 93. CrossRef

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Pathogenicity of Korean isolates of Acanthamoeba by observing the experimental infection and zymodemes of five isoenzymes: Kyung-Il Im, Ho-Joon Shin, Dong Whan Seo, Soung-Hoo Jeon, Tae-Eun Kim; Korean J Parasitol 1999;37(2):85-92.
Published online June 30, 1999; DOI: https://doi.org/10.3347/kjp.1999.37.2.85

Abstract
PDF

To determine the pathogenicity of Acanthamoeba spp. isolated in Korea and to develop a isoenzymatic maker, the mortality rate of infected mice, in vitro cytotoxicity against target cells and isoenzyme band patterns were observed. Five isolates of Acanthamoeba spp. (YM-2, YM-3, YM-4, YM-5, and YM-7) were used in this study as well as three reference Acanthamoeba spp. (A. culbertsoni, A. hatchetti, and A. royreba). According to the mortality rate of infected mice, Korean isolates could be categorized into three groups: high virulent (YM-4), low virulent (YM-2, YM-5, YM-7) and the nonpathogenic group (YM-3). In addition, the virulence of Acanthamoeba spp. was enhanced by brain passage in mice. In the cytotoxicity assay against chinese hamster ovary cells, especially, the cytotoxicity of brain-passaged amoebae was relatively higher than the long-term cultivated ones. The zymodeme patterns of glucose-6-phosphate dehydrogenase (G6PD), malate dehydrogenase (MDH), hexokinase (HK), glutamate oxaloacetate transaminase (GOT) and malic enzyme (ME) of Acanthamoeba spp. were different among each isolate, and also between long-term cultured amoebae and brain passaged ones. In spites of the polymorphic zymodemes, a slow band of G6PD and HK, and an intermediate band of MDH were only observed in pathogenic Acanthamoeba spp., which should be used as isoenzymatic makers.

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