Acanthamoeba is an opportunistic pathogen responsible for granulomatous amoebic encephalitis and amoebic keratitis. Despite its clinical significance, effective treatments remain challenging due to a limited understanding of its pathogenic mechanism. This study developed a genetic manipulation system in Acanthamoeba to facilitate gene function and drug screening studies. We applied the Cre/loxP system to integrate the gene encoding the tdTomato fluorescent protein into the genome of Acanthamoeba castellanii via homologous recombination. The polyubiquitin gene and its untranslated regions were identified and verified, after which the tdTomato gene was cloned between the untranslated regions of the polyubiquitin gene. The construct was then introduced into the Acanthamoeba genome using a modified pLPBLP vector containing loxP sites. Cre recombinase was utilized to remove the neomycin resistance cassette flanked by loxP sites, and genetically modified cells were selected by clonal dilution. The integration of the tdTomato gene, confirmed through PCR and fluorescence microscopy, showed stable expression in both trophozoites and cysts without the need for antibiotic selection. We demonstrated the feasibility of antibiotic-free reporter gene expression in Acanthamoeba. The system provides a valuable tool for functional genomics, allowing us to explore gene functions in Acanthamoeba and develop reliable drug screening models. Furthermore, the ability to express genes without the continuous use of selection markers opens up new possibilities for studying the pathobiology of this pathogen and advancing the development of novel therapeutic strategies against Acanthamoeba infections.

Acanthamoeba is an opportunistic pathogen responsible for granulomatous amoebic encephalitis and amoebic keratitis. Despite its clinical significance, effective treatments remain challenging due to a limited understanding of its pathogenic mechanism. This study developed a genetic manipulation system in Acanthamoeba to facilitate gene function and drug screening studies. We applied the Cre/loxP system to integrate the gene encoding the tdTomato fluorescent protein into the genome of Acanthamoeba castellanii via homologous recombination. The polyubiquitin gene and its untranslated regions were identified and verified, after which the tdTomato gene was cloned between the untranslated regions of the polyubiquitin gene. The construct was then introduced into the Acanthamoeba genome using a modified pLPBLP vector containing loxP sites. Cre recombinase was utilized to remove the neomycin resistance cassette flanked by loxP sites, and genetically modified cells were selected by clonal dilution. The integration of the tdTomato gene, confirmed through PCR and fluorescence microscopy, showed stable expression in both trophozoites and cysts without the need for antibiotic selection. We demonstrated the feasibility of antibiotic-free reporter gene expression in Acanthamoeba. The system provides a valuable tool for functional genomics, allowing us to explore gene functions in Acanthamoeba and develop reliable drug screening models. Furthermore, the ability to express genes without the continuous use of selection markers opens up new possibilities for studying the pathobiology of this pathogen and advancing the development of novel therapeutic strategies against Acanthamoeba infections.

Acanthamoeba is an opportunistic pathogen responsible for granulomatous amoebic encephalitis and amoebic keratitis. Despite its clinical significance, effective treatments remain challenging due to a limited understanding of its pathogenic mechanism. This study developed a genetic manipulation system in Acanthamoeba to facilitate gene function and drug screening studies. We applied the Cre/loxP system to integrate the gene encoding the tdTomato fluorescent protein into the genome of Acanthamoeba castellanii via homologous recombination. The polyubiquitin gene and its untranslated regions were identified and verified, after which the tdTomato gene was cloned between the untranslated regions of the polyubiquitin gene. The construct was then introduced into the Acanthamoeba genome using a modified pLPBLP vector containing loxP sites. Cre recombinase was utilized to remove the neomycin resistance cassette flanked by loxP sites, and genetically modified cells were selected by clonal dilution. The integration of the tdTomato gene, confirmed through PCR and fluorescence microscopy, showed stable expression in both trophozoites and cysts without the need for antibiotic selection. We demonstrated the feasibility of antibiotic-free reporter gene expression in Acanthamoeba. The system provides a valuable tool for functional genomics, allowing us to explore gene functions in Acanthamoeba and develop reliable drug screening models. Furthermore, the ability to express genes without the continuous use of selection markers opens up new possibilities for studying the pathobiology of this pathogen and advancing the development of novel therapeutic strategies against Acanthamoeba infections.

Acanthamoeba is an opportunistic pathogen responsible for granulomatous amoebic encephalitis and amoebic keratitis. Despite its clinical significance, effective treatments remain challenging due to a limited understanding of its pathogenic mechanism. This study developed a genetic manipulation system in Acanthamoeba to facilitate gene function and drug screening studies. We applied the Cre/loxP system to integrate the gene encoding the tdTomato fluorescent protein into the genome of Acanthamoeba castellanii via homologous recombination. The polyubiquitin gene and its untranslated regions were identified and verified, after which the tdTomato gene was cloned between the untranslated regions of the polyubiquitin gene. The construct was then introduced into the Acanthamoeba genome using a modified pLPBLP vector containing loxP sites. Cre recombinase was utilized to remove the neomycin resistance cassette flanked by loxP sites, and genetically modified cells were selected by clonal dilution. The integration of the tdTomato gene, confirmed through PCR and fluorescence microscopy, showed stable expression in both trophozoites and cysts without the need for antibiotic selection. We demonstrated the feasibility of antibiotic-free reporter gene expression in Acanthamoeba. The system provides a valuable tool for functional genomics, allowing us to explore gene functions in Acanthamoeba and develop reliable drug screening models. Furthermore, the ability to express genes without the continuous use of selection markers opens up new possibilities for studying the pathobiology of this pathogen and advancing the development of novel therapeutic strategies against Acanthamoeba infections.

Acanthamoeba is an opportunistic pathogen responsible for granulomatous amoebic encephalitis and amoebic keratitis. Despite its clinical significance, effective treatments remain challenging due to a limited understanding of its pathogenic mechanism. This study developed a genetic manipulation system in Acanthamoeba to facilitate gene function and drug screening studies. We applied the Cre/loxP system to integrate the gene encoding the tdTomato fluorescent protein into the genome of Acanthamoeba castellanii via homologous recombination. The polyubiquitin gene and its untranslated regions were identified and verified, after which the tdTomato gene was cloned between the untranslated regions of the polyubiquitin gene. The construct was then introduced into the Acanthamoeba genome using a modified pLPBLP vector containing loxP sites. Cre recombinase was utilized to remove the neomycin resistance cassette flanked by loxP sites, and genetically modified cells were selected by clonal dilution. The integration of the tdTomato gene, confirmed through PCR and fluorescence microscopy, showed stable expression in both trophozoites and cysts without the need for antibiotic selection. We demonstrated the feasibility of antibiotic-free reporter gene expression in Acanthamoeba. The system provides a valuable tool for functional genomics, allowing us to explore gene functions in Acanthamoeba and develop reliable drug screening models. Furthermore, the ability to express genes without the continuous use of selection markers opens up new possibilities for studying the pathobiology of this pathogen and advancing the development of novel therapeutic strategies against Acanthamoeba infections.

Purpose: : This study aimed to understand the current state of patient-centered care for people with disabilities by reviewing the key definitions, elements, and aspects of previously published care models/programs that borrow a patient-centered approach using a scoping review methodology. Methods : Following Arksey and O’Malley’s five-step methodology, we conducted a keyword search in EMBASE, Web of Science, and Pubmed for articles published from 2000 to January 2022, and a total of 30 articles were selected from 1,305 initially extracted articles. Results : A total of 13 studies developed models of care for people with disabilities based on the concept of patient-centeredness, 11 studies developed and evaluated care programs with patient- centered components, and 6 studies prospectively evaluated the effectiveness of patient-centered programs in clinical trials. The studies were generally based on multidisciplinary teams and emphasized two-way communication, collaborative relationships, multidimensional assessment, and measurement of patient needs. However, elements of patient-centeredness varied across studies, and methodologies for measuring needs and evaluating effectiveness were inconsistent. Conclusion : Based on this study, there is a need for systematic research to identify integrated and desirable patient-centeredness elements in healthcare models/programs for people with disabilities. This will enable the development of more patient-centered and friendly healthcare services for people with disabilities.

Purpose: : Health literacy, or the ability to find, understand, and use health information, is a key focus in South Korea’s ‘5th National Health Promotion Comprehensive Plan.’ However, despite South Korea’s high internet penetration, research on digital health information use among people with developmental disabilities is limited. This study assesses the current state of digital health literacy in this population. Methods: : A survey was conducted on the people with disabilities and people without disabilities. General digital health information was measured using an 8-item scale by Skinner (2006), and COVID-19-related health information was assessed with a 10-item tool developed by the research team. A total of 365 responses were analyzed, comprising people with physical disabilities (189), developmental disabilities (82), and those without disabilities (94). Data analysis was performed using SPSS version 28, including MANOVA and descriptive statistics. Results: : eHealth scores for individuals with developmental disabilities (24.6 ± 9.8) were significantly lower than those with physical disabilities (32.6 ± 7.9) and those without disabilities (33.9 ± 9.1). Similarly, Covid-eHealth scores were lower for individuals with developmental disabilities (24.6 ± 8.4) compared to those with physical disabilities (29.5 ± 6.8) and those without disabilities (31.3 ± 7.7). The differences in both cases were statistically significant (p < 0.001). Conclusion : This study represents the first comprehensive examination of digital health literacy among people with developmental disabilities in South Korea. The findings indicate that while this group has lower digital health literacy, appropriate education and support can improve their skills. This underscores the need for policies and support systems to ensure they can access and effectively use digital health information, especially as healthcare becomes more digital.

Purpose: : This study aimed to understand the current state of patient-centered care for people with disabilities by reviewing the key definitions, elements, and aspects of previously published care models/programs that borrow a patient-centered approach using a scoping review methodology. Methods : Following Arksey and O’Malley’s five-step methodology, we conducted a keyword search in EMBASE, Web of Science, and Pubmed for articles published from 2000 to January 2022, and a total of 30 articles were selected from 1,305 initially extracted articles. Results : A total of 13 studies developed models of care for people with disabilities based on the concept of patient-centeredness, 11 studies developed and evaluated care programs with patient- centered components, and 6 studies prospectively evaluated the effectiveness of patient-centered programs in clinical trials. The studies were generally based on multidisciplinary teams and emphasized two-way communication, collaborative relationships, multidimensional assessment, and measurement of patient needs. However, elements of patient-centeredness varied across studies, and methodologies for measuring needs and evaluating effectiveness were inconsistent. Conclusion : Based on this study, there is a need for systematic research to identify integrated and desirable patient-centeredness elements in healthcare models/programs for people with disabilities. This will enable the development of more patient-centered and friendly healthcare services for people with disabilities.

Purpose: : Health literacy, or the ability to find, understand, and use health information, is a key focus in South Korea’s ‘5th National Health Promotion Comprehensive Plan.’ However, despite South Korea’s high internet penetration, research on digital health information use among people with developmental disabilities is limited. This study assesses the current state of digital health literacy in this population. Methods: : A survey was conducted on the people with disabilities and people without disabilities. General digital health information was measured using an 8-item scale by Skinner (2006), and COVID-19-related health information was assessed with a 10-item tool developed by the research team. A total of 365 responses were analyzed, comprising people with physical disabilities (189), developmental disabilities (82), and those without disabilities (94). Data analysis was performed using SPSS version 28, including MANOVA and descriptive statistics. Results: : eHealth scores for individuals with developmental disabilities (24.6 ± 9.8) were significantly lower than those with physical disabilities (32.6 ± 7.9) and those without disabilities (33.9 ± 9.1). Similarly, Covid-eHealth scores were lower for individuals with developmental disabilities (24.6 ± 8.4) compared to those with physical disabilities (29.5 ± 6.8) and those without disabilities (31.3 ± 7.7). The differences in both cases were statistically significant (p < 0.001). Conclusion : This study represents the first comprehensive examination of digital health literacy among people with developmental disabilities in South Korea. The findings indicate that while this group has lower digital health literacy, appropriate education and support can improve their skills. This underscores the need for policies and support systems to ensure they can access and effectively use digital health information, especially as healthcare becomes more digital.

Purpose: : This study aimed to understand the current state of patient-centered care for people with disabilities by reviewing the key definitions, elements, and aspects of previously published care models/programs that borrow a patient-centered approach using a scoping review methodology. Methods : Following Arksey and O’Malley’s five-step methodology, we conducted a keyword search in EMBASE, Web of Science, and Pubmed for articles published from 2000 to January 2022, and a total of 30 articles were selected from 1,305 initially extracted articles. Results : A total of 13 studies developed models of care for people with disabilities based on the concept of patient-centeredness, 11 studies developed and evaluated care programs with patient- centered components, and 6 studies prospectively evaluated the effectiveness of patient-centered programs in clinical trials. The studies were generally based on multidisciplinary teams and emphasized two-way communication, collaborative relationships, multidimensional assessment, and measurement of patient needs. However, elements of patient-centeredness varied across studies, and methodologies for measuring needs and evaluating effectiveness were inconsistent. Conclusion : Based on this study, there is a need for systematic research to identify integrated and desirable patient-centeredness elements in healthcare models/programs for people with disabilities. This will enable the development of more patient-centered and friendly healthcare services for people with disabilities.