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PloS One 2023As the number of contact lens users increases, contact lens induced corneal infection is becoming more common. Acanthamoeba keratitis (AK) is a type of those which is...
As the number of contact lens users increases, contact lens induced corneal infection is becoming more common. Acanthamoeba keratitis (AK) is a type of those which is caused by Acanthamoeba species, and may cause severe ocular inflammation and visual loss. We evaluated whether Torreya nucifera (T. nucifera) extract has an anti-amoebic effect and studied its mechanism of action on Acanthamoeba lugdunensis (A. lugdunensis). Cell viability was tested using the alamarBlue™ method, and the cell death mechanism was confirmed using the Tali® Apoptosis Kit. The SYTOX® Green assay was performed to check the plasma membrane permeability. The JC-1 dye was used to measure the mitochondrial membrane potential. A CellTiter-Glo® Luminescent Assay was used to measure the adenosine-triphosphate (ATP) level. Morphological changes in the mitochondria were examined by transmission electron microscopy (TEM). Cystic changes and a decrease in cell viability after treatment with T. nucifera were observed. Both apoptotic and necrotic cells were found in the Tali® Apoptosis assay. There was no significant difference in plasma membrane permeability between the control and T. nucifera treated groups. The collapse of the mitochondrial membrane potential and reduced ATP level in A. lugdunensis was confirmed in the groups treated with T. nucifera. Structural damage to the mitochondria was observed on TEM in the groups treated with T. nucifera. T. nucifera showed an anti-amoebic effect on A. lugdunensis, by inducing the loss of mitochondrial membrane potential. Thus, it could be a future therapeutic agent for AK.
Topics: Humans; Acanthamoeba; Amebicides; Acanthamoeba Keratitis; Adenosine Triphosphate; Plant Extracts
PubMed: 36745609
DOI: 10.1371/journal.pone.0281141 -
Journal of Korean Medical Science Jan 2018This study aimed to evaluate the adhesion of Acanthamoeba trophozoites on cosmetic contact lenses (CLs) with and without CL care multipurpose solution (MPS) treatment.
BACKGROUND
This study aimed to evaluate the adhesion of Acanthamoeba trophozoites on cosmetic contact lenses (CLs) with and without CL care multipurpose solution (MPS) treatment.
METHODS
Acanthamoeba lugdunensis L3a trophozoites were inoculated onto disks trimmed from CLs: 1-day Acuvue moist, 1-day Acuvue define, Acuvue 2, and Acuvue 2 define. After 18-hour inoculation, the number of adherent trophozoites was counted under phase contrast microscopy. The effects of MPS, Opti-Free Express, soaking CLs for 6 hours, on Acanthamoeba adhesion were analyzed. Scanning electron microscopic examination was performed for assessment of Acanthamoeba attached on the lens surface.
RESULTS
Acanthamoeba trophozoites showed greater adhesion to cosmetic CL (P = 0.017 for 1-day CL and P = 0.009 for 2-week CL) although there was no significant difference between the types of cosmetic CL. On all lenses, the number of adherent Acanthamoeba was significantly reduced after treatment with MPS (P < 0.001 for 1-day Acuvue moist, P = 0.046 for 1-day Acuvue define, P < 0.001 for Acuvue 2, and P = 0.015 for Acuvue 2 define), but there was still significant difference between conventional and cosmetic CLs (P = 0.003 for 1-day CL and P < 0.001 for 2-week CL, respectively). More attachment of Acanthamoeba was observed on colored area and the acanthopodia of Acanthamoeba was placed on the rough surface of colored area.
CONCLUSION
Acanthamoeba showed a greater affinity for cosmetic CL and mostly attached on colored area. Although MPS that contained myristamidopropyl dimethylamine reduced the adhesion rate, there was a significant difference between conventional and cosmetic CLs.
Topics: Acanthamoeba; Contact Lenses; Disinfectants; Humans; Microscopy, Electron, Scanning; Trophozoites
PubMed: 29318793
DOI: 10.3346/jkms.2018.33.e26 -
The Korean Journal of Parasitology Aug 2022The high percentage of Vermamoeba was found in tap water in Korea. This study investigated whether Vermamoeba induced allergic airway inflammation in mice. We selected 2...
The high percentage of Vermamoeba was found in tap water in Korea. This study investigated whether Vermamoeba induced allergic airway inflammation in mice. We selected 2 free-living amoebas (FLAs) isolated from tap water, which included Korean FLA 5 (KFA5; Vermamoeba vermiformis) and 21 (an homolog of Acanthamoeba lugdunensis KA/ E2). We axenically cultured KFA5 and KFA21. We applied approximately 1 × 106 to mice's nasal passages 6 times and investigated their pathogenicity. The airway resistance value was significantly increased after KFA5 and KFA21 treatments. The eosinophil recruitment and goblet cell hyperplasia were concomitantly observed in bronchial alveolar lavage (BAL) fluid and lung tissue in mice infected with KFA5 and KFA21. These infections also activated the Th2-related interleukin 25, thymic stromal lymphopoietin, and thymus and activation-regulated chemokines gene expression in mouse lung epithelial cells. The CD4+ interleukin 4+ cell population was increased in the lung, and the secretion of Th2-, Th17-, and Th1-associated cytokines were upregulated during KFA5 and KFA21 infection in the spleen, lung-draining lymph nodes, and BAL fluid. The pathogenicity (allergenicity) of KFA5 and KFA21 might not have drastically changed during the long-term in vitro culture. Our results suggested that Vermamoeba could elicit allergic airway inflammation and may be an airway allergen.
Topics: Acanthamoeba; Amoeba; Animals; Bronchoalveolar Lavage Fluid; Eosinophils; Inflammation; Mice; Water
PubMed: 36041484
DOI: 10.3347/kjp.2022.60.4.229 -
Korean Journal of Ophthalmology : KJO Jun 1997We applied ribosomal RNA gene (rDNA) polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) to identify Acanthamoeba isolates from contact lens...
We applied ribosomal RNA gene (rDNA) polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) to identify Acanthamoeba isolates from contact lens paraphernalia, and characterized these on the basis of mitochondrial DNA (mtDNA) RFLP and isoenzyme analysis. The 22 Acanthamoeba strains used as reference strains were obtained from the American Type Culture Collection. Twenty-eight isolates were classified into six ribogroups, as follows: Acanthamoeba ribogroup (AcRG) 1 consisted of 18 isolates; AcRG 2, of three, AcRG 3, of three; AcRG 4, of two; AcRG 5, of one, and AcRG 6, of one. AcRG 1, which was the most frequently isolated type, was identified as A. lugdunensis, and AcRG 2 as A. hatchetti. AcRG 4 was identified as A. triangularis, while AcRG 3 and AcRG 5 were closely related to A. triangularis. AcRG 6 was identified as A. castellanii. The mtDNA RFLP patterns and zymograms for five isoenzymes of the isolates belonging to a ribogroup were identical to one another. The mtDNA digestion phenotype and zymogram for acid phosphatase (AcP) of AcRG 1 were identical to those of A. lugdunensis L3a and KA/E2, the type strain and corneal isolates from a Korean keratitis patient, respectively. The mtDNA digestion phenotype and zymogram for AcP of AcRG 6 were identical to those of A. castellanii Castellani and KA/E3, the type strain and another corneal isolate found in Korea, respectively. The mtDNA RFLP and zymogram for AcP of AcRG 2 were very similar to those of A. hatchetti BH-2 and Chang, respectively the type strain and a pathogen. The mtDNA RFLP and zymogram for AcP of AcRG 4 were similar to those of A. triangularis SH621, the type strain. The mtDNA RFLP patterns of AcRG 3 and 5 were unique. These results showed that the riboprints, mtDNA RFLP and zymograms of 22 of 28 Acanthamoeba isolates were the same as or very similar to those of the clinical isolates, which can probably be regarded as keratopathogens. More attention should be paid to the prevention of contamination by Acanthamoeba and to the disinfection of contact lens paraphernalia.
Topics: Acanthamoeba; Acanthamoeba Keratitis; Animals; Contact Lenses; DNA, Mitochondrial; DNA, Protozoan; Electrophoresis, Agar Gel; Equipment Contamination; Humans; Isoelectric Focusing; Isoenzymes; Polymerase Chain Reaction; Polymorphism, Restriction Fragment Length; RNA, Protozoan; RNA, Ribosomal
PubMed: 9283153
DOI: 10.3341/kjo.1997.11.1.39 -
The Korean Journal of Parasitology Apr 2020Free-living amoebas (FLAs) can cause severe disease in humans and animals when they become infected. However, there are no accurate survey reports on the prevalence of...
Free-living amoebas (FLAs) can cause severe disease in humans and animals when they become infected. However, there are no accurate survey reports on the prevalence of FLAs in Korea. In this study, we collected 163 tap water samples from buildings, apartments, and restrooms of highway service areas in 7 Korean provinces with high population density. All these buildings and facilities have water storage tanks in common. The survey was separated into categories of buildings, apartments, and highway service areas. Five hundred milliliters of tap water from each building was collected and filtered with 0.2 µm pore filter paper. The filters were incubated in agar plates with heated E. coli at 25°C. After axenization, genomic DNA was collected from each FLA, and species classification was performed using partial 18S-rDNA PCR-sequencing analysis. We found that 12.9% of tap water from buildings with storage tanks in Korea was contaminated with FLAs. The highway service areas had the highest contamination rate at 33.3%. All of the FLAs, except one, were genetically similar to Vermamoeba vermiformis (Hartmannella vermiformis). The remaining FLA (KFA21) was very similar to Acanthamoeba lugdunensis (KA/E26). Although cases of human infection by V. vermiformis are very rare, we must pay attention to the fact that one-third of tap water supplies in highway service areas have been contaminated.
Topics: Amoeba; Republic of Korea; Sequence Analysis, DNA; Water Microbiology; Water Supply
PubMed: 32418389
DOI: 10.3347/kjp.2020.58.2.191 -
The Korean Journal of Parasitology Jun 1998Subgenus classification of Acanthamoeba remains uncertain. Twenty-three reference strains of Acanthamoeba including 18 (neo)type-strains were subjected for...
Subgenus classification of Acanthamoeba remains uncertain. Twenty-three reference strains of Acanthamoeba including 18 (neo)type-strains were subjected for classification at the subgenus level by riboprinting. PCR/RFLP analysis of 18S rRNA gene (rDNA). On the dendrogram reconstructed on the basis of riboprint analyses, two type-strains (A. astronyxis and A. tubiashi) of morphological group 1 diverged early from the other strains and were quite distinct from each other. Four type-strains of morphological group 3, A. culbertsoni, A. palestinensis, A. healyi were considered taxonomically valid, but A. pustulosa was regarded as an invalid synonym of A. palestinensis. Strains of morphological group 2 were classified into 6 subgroups. Among them, A. griffini which has an intron in its 18S rDNA was the most divergent from the remaining strains. Acanthamoeba castellanii Castellani, A. quina Vil3, A. lugdunensis L3a, A. polyphaga Jones, A. triangularis SH621, and A. castellanii Ma strains belonged to a subgroup, A. castellanii complex. However, A. quina and A. lugdunensis were regarded as synonyms of A. castellanii. The Chang strain could be regarded as A. hatchetti. Acanthamoeba mauritaniensis, A. divionensis, A. paradivionensis could be considered as synonyms of A. rhysodes. Neff strain was regarded as A. polyphaga rather than as A. castellanii. It is likely that riboprinting can be applied for rapid identification of Acanthamoeba isolated from the clinical specimens and environments.
Topics: Acanthamoeba; Animals; DNA, Protozoan; Polymerase Chain Reaction; Polymorphism, Restriction Fragment Length; RNA, Protozoan; RNA, Ribosomal, 18S
PubMed: 9637824
DOI: 10.3347/kjp.1998.36.2.69 -
The Korean Journal of Parasitology Dec 2003In order to evaluate the possible roles of secretory proteases in the pathogenesis of amoebic keratitis, we purified and characterized a serine protease secreted by...
In order to evaluate the possible roles of secretory proteases in the pathogenesis of amoebic keratitis, we purified and characterized a serine protease secreted by Acanthamoeba lugdunensis KA/E2, isolated from a Korean keratitis patient. The ammonium sulfate-precipitated culture supernatant of the isolate was purified by sequential chromatography on CM-Sepharose, Sephacryl S-200, and mono Q-anion exchange column. The purified 33 kDa protease had a pH optimum of 8.5 and a temperature optimum of 55 degrees C. Phenylmethylsulfonylfluoride and 4-(2- Aminoethyl)-benzenesulfonyl-fluoride, both serine protease specific inhibitors, inhibited almost completely the activity of the 33 kDa protease whereas other classes of inhibitors did not affect its activity. The 33 kDa enzyme degraded various extracellular matrix proteins and serum proteins. Our results strongly suggest that the 33 kDa serine protease secreted from this keratopathogenic Acanthamoeba play important roles in the pathogenesis of amoebic keratitis, such as in corneal tissue invasion, immune evasion and nutrient uptake.
Topics: Acanthamoeba; Acanthamoeba Keratitis; Animals; Cornea; Humans; Hydrogen-Ion Concentration; Korea; Serine Endopeptidases; Substrate Specificity; Temperature; Virulence Factors
PubMed: 14699259
DOI: 10.3347/kjp.2003.41.4.189 -
Molecules (Basel, Switzerland) Mar 2019On its own, rosmarinic acid possesses multiple biological activities such as anti-inflammatory, antimicrobial, cardioprotective and antitumor properties, and these are...
On its own, rosmarinic acid possesses multiple biological activities such as anti-inflammatory, antimicrobial, cardioprotective and antitumor properties, and these are the consequence of its ROS scavenging and inhibitory effect on inflammation. In this study, two quaternary phosphonium salts of rosmarinic acid were prepared for the purpose of increasing its penetration into biological systems with the aim of improving its antimicrobial, antifungal, antiprotozoal and antitumor activity. The synthetized molecules, the triphenylphosphonium and tricyclohexylphosphonium salts of rosmarinic acid, exhibited significantly stronger inhibitory effects on the growth of HCT116 cells with IC values of 7.28 or 8.13 μM in comparison to the initial substance, rosmarinic acid (>300 μM). For the synthesized derivatives, we detected a greater than three-fold increase of activity against and a greater than eight-fold increase of activity against in comparison to rosmarinic acid. Furthermore, we recorded significantly higher antimicrobial activity of the synthetized derivatives when compared to rosmarinic acid itself. Both synthetized quaternary phosphonium salts of rosmarinic acid appear to be promising antitumor and antimicrobial agents, as well as impressive molecules for further research.
Topics: Acanthamoeba; Anti-Bacterial Agents; Anti-Infective Agents; Antifungal Agents; Antiprotozoal Agents; Cinnamates; Depsides; HCT116 Cells; Humans; Microbial Sensitivity Tests; Organophosphorus Compounds; Rosmarinic Acid
PubMed: 30893808
DOI: 10.3390/molecules24061078 -
PloS One 2023[This corrects the article DOI: 10.1371/journal.pone.0281141.].
[This corrects the article DOI: 10.1371/journal.pone.0281141.].
PubMed: 37922291
DOI: 10.1371/journal.pone.0294194 -
The Korean Journal of Parasitology Sep 1999We investigated the value of mitochondrial small subunit rRNA gene (mt SSU rDNA) PCR-RFLP as a taxonomic tool for Acanthamoeba isolates with close inter-relationships....
We investigated the value of mitochondrial small subunit rRNA gene (mt SSU rDNA) PCR-RFLP as a taxonomic tool for Acanthamoeba isolates with close inter-relationships. Twenty-five isolates representing 20 species were included in the analysis. As in nuclear 18S rDNA analysis, two type strains (A. astronyxis and A. tubiashi) of morphological group 1 diverged earliest from the other strains, but the divergence between them was less than in 18S riboprinting. Acanthamoeba griffini of morphological group 2 branched between pathogenic (A. culbertsoni A-1 and A. healyi OC-3A) and nonpathogenic (A. palestinensis Reich, A. pustulosa GE-3a, A. royreba Oak Ridge, and A lenticulata PD2S) strains of morphological group 3. Among the remaining isolates of morphological group 2, the Chang strain had the identical mitochondrial riboprints as the type strain of A. hatchetti. AA2 and AA1, the type strains of A. divionensis and A. paradivionensis, respectively, had the identical riboprints as A. quina Vil3 and A. castellanii Ma. Although the branching orders of A. castellanii Neff, A. polyphaga P23, A. triangularis SH621, and A. lugdunensis L3a were different from those in 18S riboprinting analysis, the results obtained from this study generally coincided well with those from 18S riboprinting. Mitochondrial riboprinting may have an advantage over nuclear 18S rDNA riboprinting because the mt SSU rDNAs do not seem to have introns that are found in the 18S genes of Acanthamoeba and that distort phylogenetic analyses.
Topics: Acanthamoeba; Animals; DNA, Mitochondrial; DNA, Protozoan; DNA, Ribosomal; Phylogeny; Polymerase Chain Reaction; Polymorphism, Restriction Fragment Length
PubMed: 10507226
DOI: 10.3347/kjp.1999.37.3.181