Behavioral and psychological symptoms of dementia (BPSD) and delirium superimposed on dementia (DSD) can lead to severe complications if they are not accurately identified and managed. Effective dementia care therefore requires clear differentiation, systematic assessment, and appropriate nursing interventions. This study aimed to develop VRDementia: BPSD/DSD, a virtual reality simulation program, and to qualitatively examine its validity and usability as a development-based pilot study. Methods: Using the ADDIE model (Analysis, Design, Development, Implementation, Evaluation), the program was systematically developed. During the analysis phase, qualitative interviews and literature reviews identified educational needs among nurses in long-term care hospitals. Key challenges included distinguishing agitation/aggression (BPSD) from hyperactive DSD, and depression (BPSD) from hypoactive DSD. Based on these findings, four case-based scenarios were created. Content validity and usability were qualitatively evaluated through semi-structured interviews with five experienced nurses (≥5 years of clinical experience). Results: The program consists of four sessions addressing agitation/aggression and depression (BPSD), and hyperactivity and hypoactivity (DSD). Nurses practice symptom assessment, therapeutic communication, physician reporting, and nursing interventions. The simulation is accessible via head-mounted display (HMD), mobile devices, and PC (including laptops). Qualitative feedback indicated that participants perceived the program as useful and applicable for dementia care education, including its potential use in interdisciplinary training contexts. Conclusion: VRDementia: BPSD/DSD is a valid, practical educational tool that improves nurses’ competence in distinguishing and managing BPSD and DSD. This program may contribute to higher quality dementia care in clinical settings.

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.

Background: and Purpose: Recognizing cognitive decline patterns in mild cognitive impairment (MCI) is crucial for early screening and preventive interventions. However, studies on the trajectory of individual cognitive functions in MCI are limited. Thus, the purpose of this study was to identify subtypes and stages of cognitive decline in MCI using a machine learning method. Methods: A total of 944 subjects consisting of those who were cognitively normal and those with MCI were enrolled. Fifteen neuropsychological tasks were used in the analysis.The optimal number of subtypes was determined based on the cross-validation information criterion. Fifteen stages of cognitive trajectory were estimated for each subtype. Results: The following three subtypes were identified: amnestic-verbal subtype, dysexecutive subtype, and amnestic-visual subtype. Of 723 (76.6%) subjects who had reached stage 1 at least, amnestic-verbal subtype accounted for the most (n=340, 47.0%), followed by dysexecutive subtype (n=253, 35.0%) and amnestic-visual subtype (n=130, 18%). The amnestic-verbal subtype had significantly more males (amnestic-verbal: 41.8%, dysexecutive: 31.2%, and amnestic-visual: 28.5%), younger subjects (amnestic-verbal: 72.01 years, dysexecutive: 74.43 years, and amnestic-visual: 75.06 years), higher educational years (amnestic-verbal: 11.06 years, dysexecutive: 9.53 years, and amnestic-visual: 9.79 years), lower Clinical Dementia Rating sum of boxes (amnestic-verbal: 1.40, dysexecutive: 1.61, and amnestic-visual: 1.71), and lower Korean-Instrumental Activities of Daily Living score (amnestic-verbal: 0.20, dysexecutive: 0.27, and amnestic-visual: 0.26). Conclusions Three types of MCIs were extracted using SuStaIn. Pathways of MCI deterioration could be different. The amnestic type could be bisected based on whether episodic verbal or visual memory is degraded first.

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.

Background: and Purpose: Recognizing cognitive decline patterns in mild cognitive impairment (MCI) is crucial for early screening and preventive interventions. However, studies on the trajectory of individual cognitive functions in MCI are limited. Thus, the purpose of this study was to identify subtypes and stages of cognitive decline in MCI using a machine learning method. Methods: A total of 944 subjects consisting of those who were cognitively normal and those with MCI were enrolled. Fifteen neuropsychological tasks were used in the analysis.The optimal number of subtypes was determined based on the cross-validation information criterion. Fifteen stages of cognitive trajectory were estimated for each subtype. Results: The following three subtypes were identified: amnestic-verbal subtype, dysexecutive subtype, and amnestic-visual subtype. Of 723 (76.6%) subjects who had reached stage 1 at least, amnestic-verbal subtype accounted for the most (n=340, 47.0%), followed by dysexecutive subtype (n=253, 35.0%) and amnestic-visual subtype (n=130, 18%). The amnestic-verbal subtype had significantly more males (amnestic-verbal: 41.8%, dysexecutive: 31.2%, and amnestic-visual: 28.5%), younger subjects (amnestic-verbal: 72.01 years, dysexecutive: 74.43 years, and amnestic-visual: 75.06 years), higher educational years (amnestic-verbal: 11.06 years, dysexecutive: 9.53 years, and amnestic-visual: 9.79 years), lower Clinical Dementia Rating sum of boxes (amnestic-verbal: 1.40, dysexecutive: 1.61, and amnestic-visual: 1.71), and lower Korean-Instrumental Activities of Daily Living score (amnestic-verbal: 0.20, dysexecutive: 0.27, and amnestic-visual: 0.26). Conclusions Three types of MCIs were extracted using SuStaIn. Pathways of MCI deterioration could be different. The amnestic type could be bisected based on whether episodic verbal or visual memory is degraded first.

Background: and Purpose: Recognizing cognitive decline patterns in mild cognitive impairment (MCI) is crucial for early screening and preventive interventions. However, studies on the trajectory of individual cognitive functions in MCI are limited. Thus, the purpose of this study was to identify subtypes and stages of cognitive decline in MCI using a machine learning method. Methods: A total of 944 subjects consisting of those who were cognitively normal and those with MCI were enrolled. Fifteen neuropsychological tasks were used in the analysis.The optimal number of subtypes was determined based on the cross-validation information criterion. Fifteen stages of cognitive trajectory were estimated for each subtype. Results: The following three subtypes were identified: amnestic-verbal subtype, dysexecutive subtype, and amnestic-visual subtype. Of 723 (76.6%) subjects who had reached stage 1 at least, amnestic-verbal subtype accounted for the most (n=340, 47.0%), followed by dysexecutive subtype (n=253, 35.0%) and amnestic-visual subtype (n=130, 18%). The amnestic-verbal subtype had significantly more males (amnestic-verbal: 41.8%, dysexecutive: 31.2%, and amnestic-visual: 28.5%), younger subjects (amnestic-verbal: 72.01 years, dysexecutive: 74.43 years, and amnestic-visual: 75.06 years), higher educational years (amnestic-verbal: 11.06 years, dysexecutive: 9.53 years, and amnestic-visual: 9.79 years), lower Clinical Dementia Rating sum of boxes (amnestic-verbal: 1.40, dysexecutive: 1.61, and amnestic-visual: 1.71), and lower Korean-Instrumental Activities of Daily Living score (amnestic-verbal: 0.20, dysexecutive: 0.27, and amnestic-visual: 0.26). Conclusions Three types of MCIs were extracted using SuStaIn. Pathways of MCI deterioration could be different. The amnestic type could be bisected based on whether episodic verbal or visual memory is degraded first.

Background: and Purpose: Recognizing cognitive decline patterns in mild cognitive impairment (MCI) is crucial for early screening and preventive interventions. However, studies on the trajectory of individual cognitive functions in MCI are limited. Thus, the purpose of this study was to identify subtypes and stages of cognitive decline in MCI using a machine learning method. Methods: A total of 944 subjects consisting of those who were cognitively normal and those with MCI were enrolled. Fifteen neuropsychological tasks were used in the analysis.The optimal number of subtypes was determined based on the cross-validation information criterion. Fifteen stages of cognitive trajectory were estimated for each subtype. Results: The following three subtypes were identified: amnestic-verbal subtype, dysexecutive subtype, and amnestic-visual subtype. Of 723 (76.6%) subjects who had reached stage 1 at least, amnestic-verbal subtype accounted for the most (n=340, 47.0%), followed by dysexecutive subtype (n=253, 35.0%) and amnestic-visual subtype (n=130, 18%). The amnestic-verbal subtype had significantly more males (amnestic-verbal: 41.8%, dysexecutive: 31.2%, and amnestic-visual: 28.5%), younger subjects (amnestic-verbal: 72.01 years, dysexecutive: 74.43 years, and amnestic-visual: 75.06 years), higher educational years (amnestic-verbal: 11.06 years, dysexecutive: 9.53 years, and amnestic-visual: 9.79 years), lower Clinical Dementia Rating sum of boxes (amnestic-verbal: 1.40, dysexecutive: 1.61, and amnestic-visual: 1.71), and lower Korean-Instrumental Activities of Daily Living score (amnestic-verbal: 0.20, dysexecutive: 0.27, and amnestic-visual: 0.26). Conclusions Three types of MCIs were extracted using SuStaIn. Pathways of MCI deterioration could be different. The amnestic type could be bisected based on whether episodic verbal or visual memory is degraded first.

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.