Parasites, Hosts and Diseases

"cytotoxicity"

Original Articles

In vitro anti-Trypanosoma cruzi activity of methanolic extract of Bidens pilosa and identification of active compounds by gas chromatography-mass spectrometry analysis: Gabriel Enrique Cázares-Jaramillo, Zinnia Judith Molina-Garza, Itza Eloisa Luna-Cruz, Luisa Yolanda Solís-Soto, José Luis Rosales-Encina, Lucio Galaviz-Silva; Parasites Hosts Dis 2023;61(4):405-417.
Published online November 28, 2023; DOI: https://doi.org/10.3347/PHD.23069

Abstract
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Chagas disease, caused by Trypanosoma cruzi parasite, is a significant but neglected tropical public health issue in Latin America due to the diversity of its genotypes and pathogenic profiles. This complexity is compounded by the adverse effects of current treatments, underscoring the need for new therapeutic options that employ medicinal plant extracts without negative side effects. Our research aimed to evaluate the trypanocidal activity of Bidens pilosa fractions against epimastigote and trypomastigote stages of T. cruzi, specifically targeting the Brener and Nuevo León strains—the latter isolated from Triatoma gerstaeckeri in General Terán, Nuevo León, México. We processed the plant’s aerial parts (stems, leaves, and flowers) to obtain a methanolic extract (Bp-mOH) and fractions with varying solvent polarities. These preparations inhibited more than 90% of growth at concentrations as low as 800 μg/ml for both parasite stages. The median lethal concentration (LC50) values for the Bp-mOH extract and its fractions were below 500 μg/ml. Tests for cytotoxicity using Artemia salina and Vero cells and hemolytic activity assays for the extract and its fractions yielded negative results. The methanol fraction (BPFC3MOH1) exhibited superior inhibitory activity. Its functional groups, identified as phenols, enols, alkaloids, carbohydrates, and proteins, include compounds such as 2-hydroxy-3-methylbenzaldehyde (50.9%), pentadecyl prop-2-enoate (22.1%), and linalool (15.4%). Eight compounds were identified, with a match confirmed by the National Institute of Standards and Technology (NIST-MS) software through mass spectrometry analysis.

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Inhibitory Activity of Compounds Obtained from Streptomyces Against Trypanosoma cruzi
Jorge Andrés Delgado-Garduño, Lucio Galaviz-Silva, Ma Guadalupe Rojas-Verde, Joel Horacio Elizondo-Luevano, Lidia Baylón-Pacheco, José Luis Rosales-Encina, Guadalupe Gutiérrez-Soto, Zinnia Judith Molina-Garza
Pathogens.2025; 14(7): 638. CrossRef
Nanostructured Polyurethane‐Collagen Hydrogels: Bioactive Crosslinked Networks for Enhanced Wound Healing
María I. León‐Campos, Jesús A. Claudio‐Rizo, Luis E. Cobos‐Puc, Denis A. Cabrera‐Munguía, Anilú Rubio‐Rios, Ernesto Oyervides‐Muñoz, Angélica Velázquez‐Arrellano
Journal of Biomedical Materials Research Part A.2025;[Epub] CrossRef
Assessment of Anticancer Properties of Argemone mexicana L. and Berberine: A Comparative Study
Joel H. Elizondo-Luevano, Ramiro Quintanilla-Licea, Imelda N. Monroy-García, Miroslava Kačániová, Uziel Castillo-Velázquez, Aldo F. Bazaldúa-Rodríguez, Lourdes M. Garza-Vega, Ángel D. Torres-Hernández, Abelardo Chávez-Montes
Plants.2024; 13(10): 1374. CrossRef
In Vitro Evaluation of Anti-Hemolytic and Cytotoxic Effects of Traditional Mexican Medicinal Plant Extracts on Human Erythrocytes and Cell Cultures
Joel H. Elizondo-Luevano, Ramiro Quintanilla-Licea, Sandra L. Castillo-Hernández, Eduardo Sánchez-García, Minerva Bautista-Villarreal, Georgia M. González-Meza, Marcela A. Gloria-Garza, Osvelia E. Rodríguez-Luis, Maciej Ireneusz Kluz, Miroslava Kačániová
Life.2024; 14(9): 1176. 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.

Citations

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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

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.

Citations

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Simultaneous quantitative determination of low-concentration ternary pesticide mixtures in wheat flour based on terahertz spectroscopy and BPNN
Qingxiao Ma, Yan Teng, Chun Li, Ling Jiang
Food Chemistry.2022; 377: 132030. CrossRef
Acanthamoeba keratitis: a review of biology, pathophysiology and epidemiology
Ana Gomes de Lacerda, Madalena Lira
Ophthalmic and Physiological Optics.2021; 41(1): 116. CrossRef
Differential Antimicrobial Efficacy of Multipurpose Solutions against Acanthamoeba Trophozoites
Rhonda Walters, Elise Miller, Allison Campolo, Manal M. Gabriel, Paul Shannon, Cindy McAnally, Monica Crary
Optometry and Vision Science.2021; 98(12): 1379. 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

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Original Articles

Pyruvate Protects Giardia Trophozoites from Cysteine-Ascorbate Deprived Medium Induced Cytotoxicity: Dibyendu Raj, Punam Chowdhury, Rituparna Sarkar, Yumiko Saito-Nakano, Keinosuke Okamoto, Shanta Dutta, Tomoyoshi Nozaki, Sandipan Ganguly; Korean J Parasitol 2018;56(1):1-9.
Published online February 28, 2018; DOI: https://doi.org/10.3347/kjp.2018.56.1.1

Giardia lamblia, an anaerobic, amitochondriate protozoan parasite causes parasitic infection giardiasis in children and young adults. It produces pyruvate, a major metabolic product for its fermentative metabolism. The current study was undertaken to explore the effects of pyruvate as a physiological antioxidant during oxidative stress in Giardia by cysteine-ascorbate deprivation and further investigation upon the hypothesis that oxidative stress due to metabolism was the reason behind the cytotoxicity. We have estimated intracellular reactive oxygen species generation due to cysteine-ascorbate deprivation in Giardia. In the present study, we have examined the effects of extracellular addition of pyruvate, during oxidative stress generated from cysteine-ascorbate deprivation in culture media on DNA damage in Giardia. The intracellular pyruvate concentrations at several time points were measured in the trophozoites during stress. Trophozoites viability under cysteine-ascorbate deprived (CAD) medium in presence and absence of extracellular pyruvate has also been measured. The exogenous addition of a physiologically relevant concentration of pyruvate to trophozoites suspension was shown to attenuate the rate of ROS generation. We have demonstrated that Giardia protects itself from destructive consequences of ROS by maintaining the intracellular pyruvate concentration. Pyruvate recovers Giardia trophozoites from oxidative stress by decreasing the number of DNA breaks that might favor DNA repair.

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A myeloid leukemia factor homolog is involved in tolerance to stresses and stress-induced protein metabolism in Giardia lamblia
Jui-Hsuan Wu, Jen-Chi Lee, Chun-Che Ho, Pei-Wei Chiu, Chin-Hung Sun
Biology Direct.2023;[Epub] CrossRef
Escherichia coli mediated resistance of Entamoeba histolytica to oxidative stress is triggered by oxaloacetate
Yana Shaulov, Chikako Shimokawa, Meirav Trebicz-Geffen, Shruti Nagaraja, Karen Methling, Michael Lalk, Lea Weiss-Cerem, Ayelet T. Lamm, Hajime Hisaeda, Serge Ankri, William A. Petri
PLOS Pathogens.2018; 14(10): e1007295. 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
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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.

Citations

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Amoebicidal and cysticidal in vitro activity of cationic dendritic molecules against Acanthamoeba polyphaga and Acanthamoeba griffini
Cristina Verdú-Expósito, Tania Martín-Pérez, Jorge Pérez-Serrano, Javier Sanchez-Nieves, Francisco Javier de la Mata, Irene Heredero-Bermejo
Parasitology Research.2024;[Epub] CrossRef
Tannic Acid-Modified Silver Nanoparticles in Conjunction with Contact Lens Solutions Are Useful for Progress against the Adhesion of Acanthamoeba spp. to Contact Lenses
Marcin Padzik, Lidia Chomicz, Julita Bluszcz, Karolina Maleszewska, Jaroslaw Grobelny, David Bruce Conn, Edyta B. Hendiger
Microorganisms.2022; 10(6): 1076. CrossRef
In vitro effects of multi-purpose contact lens disinfecting solutions towards survivability of Acanthamoeba genotype T4 in Malaysia
Rosnani Hanim Mohd Hussain, Wan Nur Afiqah, Mohamed Kamel Abdul Ghani, Naveed Ahmed Khan, Ruqaiyyah Siddiqui, Tengku Shahrul Anuar
Saudi Journal of Biological Sciences.2021; 28(4): 2352. CrossRef
Development of anti-acanthamoebic approaches
Mohammad Ridwane Mungroo, Tommy Tong, Naveed Ahmed Khan, Tengku Shahrul Anuar, Sutherland K. Maciver, Ruqaiyyah Siddiqui
International Microbiology.2021; 24(3): 363. CrossRef
BCLA CLEAR - Contact lens wettability, cleaning, disinfection and interactions with tears
Mark Willcox, Nancy Keir, Vinod Maseedupally, Simin Masoudi, Alison McDermott, Rabia Mobeen, Christine Purslow, Jacinto Santodomingo-Rubido, Silvia Tavazzi, Fabrizio Zeri, Lyndon Jones
Contact Lens and Anterior Eye.2021; 44(2): 157. CrossRef
In Vitro Evaluation of the Combination of Melaleuca alternifolia (Tea Tree) Oil and Dimethyl Sulfoxide (DMSO) against Trophozoites and Cysts of Acanthamoeba Strains. Oxygen Consumption Rate (OCR) Assay as a Method for Drug Screening
Tania Martín-Pérez, Irene Heredero-Bermejo, Cristina Verdú-Expósito, Jorge Pérez-Serrano
Pathogens.2021; 10(4): 491. CrossRef
Combination of tert-butyl hydroperoxide with vorinostat induces cell death of Acanthamoeba through cell cycle arrest
Hae-Ahm Lee, Eun-Kyung Moon, Fu-Shi Quan
Experimental Parasitology.2020; 210: 107833. CrossRef
Anti-staphylococcal Effect of a Nephrite-containing Contact Lens Storage Case
Su Hwan Park, Seung Uk Lee, Yoon Kyung Kim, Hak Sun Yu, Sung Hee Park, Jung Hyo Ahn, Su Jin Kim, Jong Hoon Shin, Ji Eun Lee
Journal of the Korean Ophthalmological Society.2020; 61(8): 868. CrossRef
Synthesis and in vitro activity of new biguanide-containing dendrimers on pathogenic isolates of Acanthamoeba polyphaga and Acanthamoeba griffini
T. Martín-Pérez, T. Lozano-Cruz, A. Criado-Fornelio, P. Ortega, R. Gómez, F. J. de la Mata, J. Pérez-Serrano
Parasitology Research.2019; 118(6): 1953. CrossRef
Chloroquine as a possible disinfection adjunct of disinfection solutions against Acanthamoeba
Eun-Kyung Moon, Seungeun Lee, Fu-Shi Quan, Hyun-Hee Kong
Experimental Parasitology.2018; 188: 102. CrossRef
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
Journal of the Korean Ophthalmological Society.2017; 58(5): 509. CrossRef
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
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
Seung-Mok Lee, Da-In Lee, Sung-Hee Park, Hak-Sun Yu, Ji-Eun Lee, Jong-Soo Lee
Cornea.2017; 36(12): 1538. CrossRef

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In Vitro Scolicidal Effects of Salvadora persica Root Extract against Protoscolices of Echinococcus granulosus: Abdel-Azeem S. Abdel-Baki, Esam Almalki, Lamjed Mansour, Saleh Al-Quarishy; Korean J Parasitol 2016;54(1):61-66.
Published online February 26, 2016; DOI: https://doi.org/10.3347/kjp.2016.54.1.61

Abstract
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It has been known that Arak, Salvadora persica, has a number of medicinal properties. We tried to investigate in vitro scolicidal effect of root extracts of this plant against protoscolices from hydatid cysts of Echinococcus granulosus. Protoscolices were aseptically collected from sheep livers containing hydatid cysts. S. persica root extract was used in 10, 30, and 50 mg/ml concentration for 10, 20, and 30 min. The viability of protoscolices was ascertained by 0.1% eosin staining. Scolicidal activity of S. persica extract at a concentration of 10 mg/ml was 36.3%, 50.3%, and 70.8% after 10, 20, and 30 min of exposure, respectively. The scolicidal effect of this extract at a concentration of 30 mg/ml was 52.9%, 86.7%, and 100% after 10, 20, and 30 min of exposure, respectively. S. persica extract at a concentration of 50 mg/ml, meanwhile, killed 81.4%, 100%, and 100% of protoscolices after 10, 20, and 30 min, respectively. Also, the cytotoxic potential of S. persica was assessed on human liver cells (HepG2) using trypan blue exclusion test. No cytotoxic effect was observed on HepG2 cell line. The present study confirmed for the first time that the ethanolic extract of S. persica has high scolicidal power in vitro. However, in vivo effect of this material remains to be studied for treatment of echinococcosis in humans and herbivorous animals.

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Brief Communications

Effects of Mannose on Pathogenesis of Acanthamoeba castellanii: Kyung-Tae Yoo, Suk-Yul Jung; Korean J Parasitol 2012;50(4):365-369.
Published online November 26, 2012; DOI: https://doi.org/10.3347/kjp.2012.50.4.365

Abstract
PDF

Acanthamoeba spp. are single-celled protozoan organisms that are widely distributed in the environment. In this study, to understand functional roles of a mannose-binding protein (MBP), Acanthamoeba castellanii was treated with methyl-alpha-D-mannopyranoside (mannose), and adhesion and cytotoxicity of the amoeba were analyzed. In addition, to understand the association of MBP for amoeba phagocytosis, phagocytosis assay was analyzed using non-pathogenic bacterium, Escherichia coli K12. Amoebae treated with mannose for 20 cycles exhibited larger vacuoles occupying the most area of the amoebic cytoplasm in comparison with the control group amoebae and glucose-treated amoebae. Mannose-selected amoebae exhibited lower levels of binding to Chinese hamster ovary (CHO) cells. Exogenous mannose inhibited >50% inhibition of amoebae (control group) binding to CHO cells. Moreover, exogenous mannose inhibited amoebae (i.e., man-treated) binding to CHO cells by <15%. Mannose-selected amoebae exhibited significantly decreased cytotoxicity to CHO cells compared with the control group amoebae, 25.1% vs 92.1%. In phagocytic assay, mannose-selected amoebae exhibited significant decreases in bacterial uptake in comparison with the control group, 0.019% vs 0.03% (P<0.05). Taken together, it is suggested that mannose-selected A. castellanii trophozoites should be severely damaged and do not well interact with a target cell via a lectin of MBP.

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Muhammad Ilyas, Fiona Stapleton, Mark D. P. Willcox, Fiona Henriquez, Hari Kumar Peguda, Binod Rayamajhee, Tasbiha Zahid, Constantinos Petsoglou, Nicole A. Carnt
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Yuehua Wang, Linzhe Jiang, Yitong Zhao, Xiaohong Ju, Le Wang, Liang Jin, Ryan D. Fine, Mingguang Li
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Acanthamoeba-mediated cytopathic effect correlates with MBP and AhLBP mRNA expression
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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
PDF

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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Fowlerstefin, a cysteine protease inhibitor of Naegleria fowleri, induces inflammatory responses in BV-2 microglial cells in vitro
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Original Articles

Phospholipase Activities in Clinical and Environmental Isolates of Acanthamoeba: Abdul Matin, Suk-Yul Jung; Korean J Parasitol 2011;49(1):1-8.
Published online March 18, 2011; DOI: https://doi.org/10.3347/kjp.2011.49.1.1

Abstract
PDF

The pathogenesis and pathophysiology of Acanthamoeba infections remain incompletely understood. Phos-pholipases are known to cleave phospholipids, suggesting their possible involvement in the host cell plasma membrane disruption leading to host cell penetration and lysis. The aims of the present study were to determine phospholipase activities in Acanthamoeba and to determine their roles in the pathogenesis of Acanthamoeba. Using an encephalitis isolate (T1 genotype), a keratitis isolate (T4 genotype), and an environmental isolate (T7 genotype), we demonstrated that Acanthamoeba exhibited phospholipase A₂ (PLA₂) and phospholipase D (PLD) activities in a spectrophotometry-based assay. Interestingly, the encephalitis isolates of Acanthamoeba exhibited higher phospholipase activities as compared with the keratitis isolates, but the environmental isolates exhibited the highest phospholipase activities. Moreover, Acanthamoeba isolates exhibited higher PLD activities compared with the PLA₂. Acanthamoeba exhibited optimal phospholipase activities at 37℃ and at neutral pH indicating their physiological relevance. The functional role of phospholipases was determined by in vitro assays using human brain microvascular endothelial cells (HBMEC), which constitute the blood-brain barrier. We observed that a PLD-specific inhibitor, i.e., compound 48/80, partially inhibited Acanthamoeba encephalitis isolate cytotoxicity of the host cells, while PLA₂-specific inhibitor, i.e., cytidine 5'-diphosphocholine, had no effect on parasite-mediated HBMEC cytotoxicity. Overall, the T7 exhibited higher phospholipase activities as compared to the T4. In contract, the T7 exhibited minimal binding to, or cytotoxicity of, HBMEC.

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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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Parasites & Vectors.2020;[Epub] 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
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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
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Infection and Immunity.2016; 84(9): 2422. CrossRef
Novel cathepsin B and cathepsin B-like cysteine protease of Naegleria fowleri excretory–secretory proteins and their biochemical properties
Jinyoung Lee, Jong-Hyun Kim, Hae-Jin Sohn, Hee-Jong Yang, Byoung-Kuk Na, Yong-Joon Chwae, Sun Park, Kyongmin Kim, Ho-Joon Shin
Parasitology Research.2014; 113(8): 2765. CrossRef
Naegleria fowleriLysate 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
The Korean Journal of Parasitology.2011; 49(3): 285. CrossRef
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Immunodominant antigens in Naegleria fowleri excretory–secretory proteins were potential pathogenic factors
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Parasitology Research.2009; 105(6): 1675. CrossRef

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Brief Communication

Effect of Iron on Adherence and Cytotoxicity of Entamoeba histolytica to CHO Cell Monolayers: Jongweon Lee, Soon-Jung Park, Tai-Soon Yong; Korean J Parasitol 2008;46(1):37-40.
Published online March 20, 2008; DOI: https://doi.org/10.3347/kjp.2008.46.1.37

Abstract
PDF

Iron is an essential element for almost all living organisms. The possible role of iron for growth, adherence and cytotoxicity of Entamoeba histolytica was evaluated in this study. The absence of iron from TYI-S-33 medium stopped amebic growth in vitro. However, iron concentrations in the culture media of 21.4-285.6 ?M did not affect the growth of the amebae. Although growth was not retarded at these concentrations, the adhesive abilities of E. histolytica and their cytotoxicities to CHO cell monolayer were correlated with iron concentration. Amebic adhesion to CHO cell monolayers was significantly reduced by low-iron (24.6 ± 2.1%) compared with 62.7 ± 2.8 and 63.1 ± 1.4% of amebae grown in a normal-iron and high-iron media, respectively. E. histolytica cultured in the normal- and high-iron media destroyed 69.1 ± 4.3% and 72.6 ± 5.7% of cultured CHO cell monolayers, but amebae grown in the low-iron medium showed a significantly reduced level of cytotoxicity to CHO cells (2.8 ± 0.2%). Addition of divalent cations other than iron to amebic trophozoites grown in the low-iron medium failed to restore levels of the cytotoxicity. However, when E. histolytica grown in low-iron medium were transferred to normal-iron medium, the amebae showed completely restored cytotoxicity within 7 days. The result suggests that iron is an important factor in the adherence and cytotoxicity of E. histolytica to CHO cell monolayer.

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Original Article

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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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 Article

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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Experimental infection of T4 Acanthamoeba genotype determines the pathogenic potential
Daniella de Sousa Mendes Moreira Alves, Aline Silva Moraes, Luciano Moreira Alves, Rodrigo Gurgel-Gonçalves, Ruy de Souza Lino Junior, César Augusto Cuba-Cuba, Marina Clare Vinaud
Parasitology Research.2016; 115(9): 3435. CrossRef
Acanthamoeba royreba: Morphological features and in vitro cytopathic effect
Arturo González-Robles, Lizbeth Salazar-Villatoro, Maritza Omaña-Molina, Jacob Lorenzo-Morales, Adolfo Martínez-Palomo
Experimental Parasitology.2013; 133(4): 369. CrossRef
Pathogenic free-living amoebae in Korea
Ho-Joon Shin, Kyung-il Im
The Korean Journal of Parasitology.2004; 42(3): 93. CrossRef
Acanthamoeba sohi, n. sp., a pathogenic Korean isolate YM-4 from a freshwater fish
Kyung-il Im, Ho-Joon Shin
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Ho-Joon Shin, Myung-Soo Cho, Suk-Yul Jung, Hyung-Il Kim, Sun Park, Jang-Hoon Seo, Jung-Chil Yoo, Kyung-Il Im
Clinical Diagnostic Laboratory Immunology.2001; 8(4): 837. CrossRef
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
The Korean Journal of Parasitology.2000; 38(2): 99. CrossRef
Apoptosis of Primary-Culture Rat Microglial Cells Induced by PathogenicAcanthamoebaspp
Ho-Joon Shin, Myung-Soo Cho, Hyung-Il Kim, Millina Lee, Sun Park, Seonghyang Sohn, Kyung-Il Im
Clinical Diagnostic Laboratory Immunology.2000; 7(3): 510. CrossRef