Pyrenoid proteomics reveals independent evolution of the CO2-concentrating organelle in chlorarachniophytes.

Pyrenoids are microcompartments that are universally found in the photosynthetic plastids of various eukaryotic algae. They contain ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) and play a pivotal role in facilitating CO2 assimilation via CO2-concentrating mechanisms (CCMs). Recent investigations involving model algae have revealed that pyrenoid-associated proteins participate in pyrenoid biogenesis and CCMs. However, these organisms represent only a small part of algal lineages, which limits our comprehensive understanding of the diversity and evolution of pyrenoid-based CCMs. Here we report a pyrenoid proteome of the chlorarachniophyte alga Amorphochlora amoebiformis, which possesses complex plastids acquired through secondary endosymbiosis with green algae. Proteomic analysis using mass spectrometry resulted in the identification of 154 potential pyrenoid components. Subsequent localization experiments demonstrated the specific targeting of eight proteins to pyrenoids. These included a putative Rubisco-binding linker, carbonic anhydrase, membrane transporter, and uncharacterized GTPase proteins. Notably, most of these proteins were unique to this algal lineage. We suggest a plausible scenario in which pyrenoids in chlorarachniophytes have evolved independently, as their components are not inherited from green algal pyrenoids.

Encyclopedia of Family A DNA Polymerases Localized in Organelles: Evolutionary Contribution of Bacteria Including the Proto-Mitochondrion.

DNA polymerases synthesize DNA from deoxyribonucleotides in a semiconservative manner and serve as the core of DNA replication and repair machinery. In eukaryotic cells, there are 2 genome-containing organelles, mitochondria, and plastids, which were derived from an alphaproteobacterium and a cyanobacterium, respectively. Except for rare cases of genome-lacking mitochondria and plastids, both organelles must be served by nucleus-encoded DNA polymerases that localize and work in them to maintain their genomes. The evolution of organellar DNA polymerases has yet to be fully understood because of 2 unsettled issues. First, the diversity of organellar DNA polymerases has not been elucidated in the full spectrum of eukaryotes. Second, it is unclear when the DNA polymerases that were used originally in the endosymbiotic bacteria giving rise to mitochondria and plastids were discarded, as the organellar DNA polymerases known to date show no phylogenetic affinity to those of the extant alphaproteobacteria or cyanobacteria. In this study, we identified from diverse eukaryotes 134 family A DNA polymerase sequences, which were classified into 10 novel types, and explored their evolutionary origins. The subcellular localizations of selected DNA polymerases were further examined experimentally. The results presented here suggest that the diversity of organellar DNA polymerases has been shaped by multiple transfers of the PolI gene from phylogenetically broad bacteria, and their occurrence in eukaryotes was additionally impacted by secondary plastid endosymbioses. Finally, we propose that the last eukaryotic common ancestor may have possessed 2 mitochondrial DNA polymerases, POP, and a candidate of the direct descendant of the proto-mitochondrial DNA polymerase I, rdxPolA, identified in this study.

Nonrestorative Sleep and Type 2 Diabetes Incidence: the Aichi Workers' Cohort Study.

BACKGROUND: The term "nonrestorative sleep (NRS)" refers to unrefreshed feeling at wake-up and is a domain of poor sleep quality. Previous researches have demonstrated that NRS is linked to a number of diseases and adverse health outcomes, but less is known regarding the link between NRS and diabetes, particularly in Japanese. METHODS: We studied 3665 middle-aged male participants of the Aichi Workers' Cohort Study who were followed-up from 2002 to 2019. Cox proportional hazards models estimated hazard ratios (HRs) and 95% confidence intervals (CIs) of incident type 2 diabetes mellitus (T2DM) in relation to NRS adjusted for potential confounding variables. RESULTS: During a median follow-up of 14.6 years, 421 type 2 diabetes cases were identified. Participants with NRS had a higher crude incidence rate of T2DM (11.2/1,000 person-years), compared to participants without NRS (9.3/1,000 person-years). In the fully adjusted model, individuals who reported having NRS had a significantly higher risk of developing T2DM (HR: 1.36, 95% CI: 1.10-1.67). The association was observed only in participants under 50 years old (HR: 1.82, 95% CI: 1.36-2.43), not in the older (50 years or older) participants (P for interaction =0.025). In contrast, stratified analyses by the presence of shift work, obesity or sleep duration showed similar associations in all the strata. CONCLUSIONS: NRS was associated with higher risk of T2DM in middle-aged Japanese male workers independent of a variety of lifestyle factors and other sleep problems.

Factors influencing interprofessional collaboration in long-term care from a multidisciplinary perspective: a case study approach.

Systematic assessments of interprofessional collaboration barriers and enablers in long-term care settings are critical for delivering person-centered healthcare. However, research on factors influencing interprofessional collaboration in long-term care settings is limited. For this study, 65 healthcare professionals across multiple facilities experienced in long-term care in Japan participated in online focus group discussions and individual interviews to discuss cases. The qualitative data were analyzed using qualitative content analysis. Seven themes emerged: coordination, the need for care manager training, hierarchy among healthcare professionals, specialization but not the mind-set of overspecialization, casual conversations, electronic group communication tools, and excessive fear of personal information protection. These findings highlight the need to develop coordinator roles and for interprofessional education on the proper approach to personal information protection laws. Furthermore, daily casual conversations, the use of online platforms, and the prevention of patients being left behind due to overspecialization are required.

Challenges faced by older people with dementia during the COVID-19 pandemic as perceived by professionals: a qualitative study with interviews.

BACKGROUND: Previous studies have highlighted a decline in the mental health of older adults over the course of the coronavirus disease 2019 (COVID-19) pandemic. Few studies have determined the possible causes of behavioural and psychological symptoms of dementia during COVID-19 in a comprehensive manner. We aimed to identify the challenges faced by older adults with dementia during the COVID-19 pandemic. METHODS: This study adopted a qualitative approach to understanding the perceptions of healthcare professionals, such as regarding the negative effects of COVID-19 on the mental health of people with dementia. Between January and March 2022, the authors conducted individual in-depth interviews on how COVID-19 affected the stress levels, care, and self-determination of people with dementia. Qualitative data from the individual interviews were data cleansed to ensure the clarity and readability of the transcripts. The qualitative data were then analyzed by inductive manual coding using a qualitative content analysis approach. The grouping process involved reading and comparing individual labels to cluster similar labels into categories and inductively formulate themes. RESULTS: Qualitative analysis extracted 61 different semantic units that were duplicated. Seven categories were inductively extracted using a grouping process. These were further integrated to extract the following four themes: fear of personal protective equipment (PPE), loneliness, dissatisfaction with behavioural restrictions and limitations of video calls, and family interference with service use. DISCUSSION: People with dementia often faced mental distress during the pandemic owing to preventive measures against COVID-19, and a lack of awareness and understanding of such preventive measures worsened their distress. They experienced a severe sense of social isolation and loneliness. Findings also indicated that families tended to ignore the needs of people with dementia and their decisions and opinions regarding healthcare service use.

Publisher Correction: Genetic tool development in marine protists: emerging model organisms for experimental cell biology.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Genetic tool development in marine protists: emerging model organisms for experimental cell biology.

Diverse microbial ecosystems underpin life in the sea. Among these microbes are many unicellular eukaryotes that span the diversity of the eukaryotic tree of life. However, genetic tractability has been limited to a few species, which do not represent eukaryotic diversity or environmentally relevant taxa. Here, we report on the development of genetic tools in a range of protists primarily from marine environments. We present evidence for foreign DNA delivery and expression in 13 species never before transformed and for advancement of tools for eight other species, as well as potential reasons for why transformation of yet another 17 species tested was not achieved. Our resource in genetic manipulation will provide insights into the ancestral eukaryotic lifeforms, general eukaryote cell biology, protein diversification and the evolution of cellular pathways.

New Parabasalia symbionts Snyderella spp. and Daimonympha gen. nov. from South American Rugitermes termites and the parallel evolution of a cell with a rotating "head".

Most Parabasalia are symbionts in the hindgut of "lower" (non-Termitidae) termites, where they widely vary in morphology and degree of morphological complexity. Large and complex cells in the class Cristamonadea evolved by replicating a fundamental unit, the karyomastigont, in various ways. We describe here four new species of Calonymphidae (Cristamonadea) from Rugitermes hosts, assigned to the genus Snyderella based on diagnostic features (including the karyomastigont pattern) and molecular phylogeny. We also report a new genus of Calonymphidae, Daimonympha, from Rugitermes laticollis. Daimonympha's morphology does not match that of any known Parabasalia, and its SSU rRNA gene sequence corroborates this distinction. Daimonympha does however share a puzzling feature with a few previously described, but distantly related, Cristamonadea: a rapid, smooth, and continuous rotation of the anterior end of the cell, including the many karyomastigont nuclei. The function of this rotatory movement, the cellular mechanisms enabling it, and the way the cell deals with the consequent cell membrane shear, are all unknown. "Rotating wheel" structures are famously rare in biology, with prokaryotic flagella being the main exception; these mysterious spinning cells found only among Parabasalia are another, far less understood, example.

Work and Family Conflicts, Depression, and "Ikigai": A Mediation Analysis in a Cross-cultural Study Between Japanese and Egyptian Civil Workers.

BACKGROUND: Total work-family conflicts (TWFCs) could associate with mental health, and having ikigai (a purpose of life) may mediate this association. METHODS: In a cross-cultural study of 4,792 Japanese Aichi Workers' Cohort study participants and 3,109 Egyptian civil workers, the Midlife Development in the United States (MIDUS) questionnaire measured TWFCs and the Center for Epidemiological Studies Depression (CES-D) 11-item scale measured depression. We used logistic regression models to estimate odds ratios (ORs) of having depression and a high-ikigai across levels of TWFCs (low, moderate, and high), and the PROCESS macro of Hayes to test the mediation effect. RESULTS: The prevalence of high TWFCs, depression, and having a high ikigai were 17.9%, 39.4%, and 70.1% in Japanese women, 10.5%, 26.8%, and 70.1% in Japanese men, 23.7%, 58.2%, and 24.7% in Egyptian women, and 19.1%, 38.9%, and 36.9% in Egyptian men, respectively. Compared with participants with low TWFCs, the multivariable ORs of depression in Japanese women and men with high TWFCs were 4.11 (95% confidence interval [CI], 2.99-5.65) and 5.42 (95% CI, 4.18-7.02), and those in Egyptian women and men were 4.43 (95% CI, 3.30-5.95) and 4.79 (95% CI, 3.53-6.48), respectively. The respective ORs of having a high-ikigai were 0.46 (95% CI, 0.33-0.64) and 0.40 (95% CI, 0.31-0.52) in Japanese women and men and were 0.34 (95% CI, 0.24-0.48) and 0.28 (95% CI, 0.20-0.39) in Egyptian women and men. No interaction between TWFCs and country was observed for the associations with depression or ikigai. Ikigai mediated (up to 18%) the associations between the TWFCs and depression, especially in Egyptian civil workers. CONCLUSION: TWFCs were associated with depression, and having low ikigai mediated these associations in Japanese and Egyptian civil workers.

Relationship Between Fasting Blood Glucose Levels in Middle Age and Cognitive Function in Later Life: The Aichi Workers' Cohort Study.

BACKGROUND: There is limited evidence regarding the relationship between Diabetes mellitus (DM) in middle age and mild cognitive impairment after a follow-up. Therefore, we investigated the relationship between fasting blood glucose (FBG) levels in middle age and cognitive function assessed using the Japanese version of the Montreal Cognitive Assessment (MoCA-J) in later life, following over 15 years of follow-up in the Aichi Workers' Cohort Study in Japan. METHODS: Participants were 253 former local government employees aged 60-79 years in 2018 who participated in a baseline survey conducted in 2002. Using baseline FBG levels and self-reported history, participants were classified into the normal, impaired fasting glucose (IFG) and, and DM groups. Total MoCA-J score ranges from 0 to 30, and cognitive impairment was defined as MoCA-J score ≤25 in this study. A general linear model was used to estimate the mean MoCA-J scores in the FBG groups, adjusted for age, sex, educational year, smoking status, alcohol consumption, physical activity, body mass index, systolic blood pressure, total cholesterol, and estimated glomerular filtration rate. RESULTS: The mean MoCA-J score in the total population was 25.0, and the prevalence of MoCA-J score ≤25 was 49.0%. Multivariable-adjusted total MoCA-J scores were 25.2, 24.8, and 23.4 in the normal, IFG, and DM groups, respectively. The odds ratio of MoCA-J score ≤25 in the DM group was 3.29. CONCLUSION: FBG level in middle age was negatively associated with total MoCA-J scores assessed later in life, independent of confounding variables.

Good death for people living with dementia: a qualitative study.

BACKGROUND: Ensuring a good death is one of the primary objectives of palliative care and end-of-life care. There is insufficient evidence regarding what defines a good death for people living with dementia. Obtaining an understanding of what constitutes a good death could help improve dementia care. This study aimed to explore how multiple stakeholders perceive a good death for people living with dementia. METHODS: This qualitative study was carried out across six prefectures in Japan. Enrollment of participants took place within dementia outpatient clinics, hospitals, daycare centers, and community centers. A total of thirty-three in-depth interviews with people living with dementia, physicians, and nurses were conducted. Six focus group discussions were performed with family caregivers and care workers. Verbatim transcripts of the interviews were prepared, and inductive content analysis was used to examine the data. FINDINGS: Regarding the perception of a good death, the following themes were derived: (1) painless death; (2) dying in a preferred environment; (3) family's coping with loss; (4) maintaining regular life; (5) living with respect; and (6) preparation for death. All these themes are interrelated. Participants viewed a good death as a process rather than a single event. CONCLUSION: This study identifies crucial components of a good death for people living with dementia. The findings could be used to improve dementia care.

[The current role of advance care planning for people with dementia - A narrative review].

Advance care planning (ACP) for people with dementia, as with other diseases, is a necessary process to realize medical treatment and care in the final stage of a person's life. On the other hand, dementia, a disease that is expected to make it difficult for people to make decisions on their own in the future, has a long course, and is characterized by uncertainty regarding the course of the disease, which may also be a limiting factor in the implementation of ACP for people with dementia. On the other hand, the uncertainties may also be a reason for implementing ACP. This paper reviews reports on ACP initiatives for people with dementia from many countries and presents their characteristics, cultural and customary influences, effects, facilitating and inhibiting factors, and recommendations for implementation, with the aim of promoting future ACP initiatives for people with dementia. The aim of the study was to promote future ACP initiatives for people with dementia.

Everyday wishes of older people living with dementia in care planning: a qualitative study.

BACKGROUND: The dementia care policy in Japan emphasizes the views of people living with dementia in care planning. An exploration of the everyday wishes of older people living with dementia can help clarify their priorities and assist in improving dementia care. This study aimed to explore the everyday wishes of older people living with dementia in Japan. METHODS: This qualitative study was conducted in Aichi prefecture in Japan. Older people with mild to moderate dementia were considered for inclusion. Participants were recruited from a dementia outpatient clinic. In-depth interviews were conducted with 36 participants in the same dementia outpatient clinic from January to October 2019. Audio-recorded interviews were transcribed verbatim. Inductive content analysis was carried out to analyze the data. FINDINGS: Participants expressed their everyday wishes within five themes (desire of being connected, freedom to decide, involvement in activities, status quo, and self-reliance). Older people living with dementia loved the connection with their family and wanted to have an enjoyable life by engaging in several activities without others' interference. They desired to maintain the status quo and not be a burden to others. CONCLUSIONS: This study provides evidence on the everyday wishes of people living with dementia. Identified wishes are mostly on emotional aspects of their daily lives. The findings of our study might help provide care for the people living with dementia considering their wishes. Further exploration, including people with severe dementia, is needed.

Contemporary Issues and Practicalities in Completing Advance Care Planning for Patients With Severe COPD Living Alone: A Qualitative Study.

Health care professionals working with older people living alone with chronic obstructive pulmonary disease (COPD) to complete advance care planning (ACP) often encounter the double burden of social isolation and acute exacerbations in this planning. The study explored clinicians' perceptions regarding factors influencing the completion of ACP for older people with COPD living alone. Individual interviews were conducted with 18 health care professionals using the video meeting platform in 2020. A semi-structured interview guide included: (a) behavior and lifestyle related to decision-making, (b) desired place to die, and (c) facilitators and barriers to autonomy in patients with severe COPD who live alone. Five main themes were identified: information sharing among team members, patient readiness, desired place of death, economic constraints, and care at the time of and after death. Partly due to the heterogeneity and complexity of clinical courses and treatment responses of COPD, a wide range of social issues of a person's life were related to practicality in the completion of ACP for older people with COPD living alone. Social work knowledge and skills such as in-depth interviewing, outreach finance and welfare support, and holistic perspective play an essential role in completing ACP for COPD patients living alone.

Characterization of a novel type of carbonic anhydrase that acts without metal cofactors.

BACKGROUND: Carbonic anhydrases (CAs) are universal metalloenzymes that catalyze the reversible conversion of carbon dioxide (CO2) and bicarbonate (HCO3-). They are involved in various biological processes, including pH control, respiration, and photosynthesis. To date, eight evolutionarily unrelated classes of CA families (α, ß, γ, δ, ζ, η, θ, and ι) have been identified. All are characterized by an active site accommodating the binding of a metal cofactor, which is assumed to play a central role in catalysis. This feature is thought to be the result of convergent evolution. RESULTS: Here, we report that a previously uncharacterized protein group, named "COG4337," constitutes metal-independent CAs from the newly discovered ι-class. Genes coding for COG4337 proteins are found in various bacteria and photosynthetic eukaryotic algae. Biochemical assays demonstrated that recombinant COG4337 proteins from a cyanobacterium (Anabaena sp. PCC7120) and a chlorarachniophyte alga (Bigelowiella natans) accelerated CO2 hydration. Unexpectedly, these proteins exhibited their activity under metal-free conditions. Based on X-ray crystallography and point mutation analysis, we identified a metal-free active site within the cone-shaped α+ß barrel structure. Furthermore, subcellular localization experiments revealed that COG4337 proteins are targeted into plastids and mitochondria of B. natans, implicating their involvement in CO2 metabolism in these organelles. CONCLUSIONS: COG4337 proteins shared a short sequence motif and overall structure with ι-class CAs, whereas they were characterized by metal independence, unlike any known CAs. Therefore, COG4337 proteins could be treated as a variant type of ι-class CAs. Our findings suggested that this novel type of ι-CAs can function even in metal-poor environments (e.g., the open ocean) without competition with other metalloproteins for trace metals. Considering the widespread prevalence of ι-CAs across microalgae, this class of CAs may play a role in the global carbon cycle.

Diabetes management in people living with dementia in Japan: a qualitative exploration.

BACKGROUND: Dementia and diabetes are highly prevalent among the older society in Japan. People living with dementia (PLWD) often face hurdles in managing diabetes. This study explored the perspectives of healthcare providers in Japan regarding diabetes management in PLWD. METHODS: We conducted a qualitative study using in-depth interviews as a data collection method. A total of 15 physicians and nurses were interviewed. A qualitative content analysis of the codes was performed to generate the themes. RESULTS: The major themes focused on the management of medications/therapeutic regimen, difficulties of continuing health care, emotional aspects of PLWD for adherence to lifestyle modification, and varying direction and degree of family support for diabetes care. CONCLUSION: PLWD in Japan face challenges in medication management, food restriction, and lifestyle modification. Policies to engage home visit care workers in medication management, consideration of the emotional aspect of PLWD, and utilisation of social support might help in the proper management of diabetes in PLWD.

Improving the quality of community primary palliative care in COPD: A qualitative study of health-care providers.

Primary palliative care is essential for the continuity of care in severe COPD. This study aimed to identify essential factors and aspects to enhance the quality of primary palliative care for adults with severe COPD living in the community. Interviews with medical professionals from six institutions located in two major metropolitan areas in Japan were conducted, and these interviews were analyzed by using a qualitative content analysis approach. Results indicate that effective collaborative communication among team members, long-term care insurance system and related services, and palliative care techniques were the primary themes.

Job satisfaction among physicians providing health care to the elderly in Japan: a qualitative study.

BACKGROUND: Physicians' job satisfaction plays an important role in service delivery. Japan is a country with a higher number of elderly people and their medical care is a crucial issue. To date, no study has been conducted on the job satisfaction of geriatricians serving the elderly in Japan. This study aims to explore the job satisfaction and motivation of physicians providing health care to the elderly working at different hospitals in Japan. METHODS: In-depth interview of 23 geriatric physicians was conducted and data were coded and analysed. Qualitative content analysis was conducted to identify the response themes. RESULTS: Three major themes emerged from the analysis and they are: satisfaction as a primary care provider, career development, and suitable communication with patients. Geriatric physicians enjoy their work with inter-professional collaboration and are satisfied to provide primary health care to elderly people. Geriatrics is a very promising career to them and they enjoy the communication and empathetic doctor-patient relationship in their service. CONCLUSIONS: Japanese physicians providing health care to the elderly are enjoying their work due to a suitable environment, multi-professional collaboration, work-life balance, and patient-doctor relationship.

Discrimination against the elderly in health-care services: a systematic review.

Ageism in health-care services is a cardinal public health concern in geriatric health. It is important to know the types and extent of discrimination experienced by elderly individuals while seeking health care. This systematic review aimed to explore the available research to identify discrimination of elderly individuals in health-care services. MEDLINE through PubMed, Web of Science, PsycInfo, and Cochrane Database were searched by using a comprehensive search strategy in September 2020 with no limitation in the year of publication and types of publication. Articles published in English focusing on disparities in health-care service among the elderly were included. Two independent authors screened, extracted, and assessed the quality of data using the Joanna Briggs Institute Critical Appraisal Tool. A narrative synthesis was conducted followed by qualitative content analysis to identify themes from the extracted outcomes of the included studies. Of 3117 articles retrieved from the initial search, 21 articles were included. A total of 611 608 participants from 38 countries in four geographic regions were reported. Articles were published between 2003 and 2020. Most of the studies (n = 19) were cross-sectional in design, and the remaining two studies were qualitative. Major themes of discrimination were age-related discrimination, racial discrimination, gender discrimination, wealth-related discrimination, and technology-related discrimination. This systematic review identified the major perspectives on disparities faced by the elderly in accessing health services. There is a huge research gap on this issue. Specific strategies should be incorporated to address the varying types of discrimination experienced by elderly individuals.

[A systematic review of distress due to pneumonia experienced by older people with end-stage dementia].

AIM: Many older people with end-stage dementia die of pneumonia, but the details of their distress have been unclear. The present study clarified the distress experienced by such individuals due to pneumonia. METHODS: We searched for literature corresponding to 5 clinical questions (CQs) concerning including "What distress is experienced by older people with end-stage dementia due to pneumonia (CQ1) ?" using a search formula with the Web version of the Central Medical Magazine, MEDLINE (STN)/EMBASE (STN), Cochrane Library, and extracted 604 articles. Forty-two articles corresponding to 5 CQs were extracted by primary screening using abstracts, and 17 articles were adopted after the secondary screening. A total of six articles corresponding to CQ1 were ultimately analyzed. RESULTS: Older people with end-stage dementia who died of pneumonia reported more discomfort and dyspnea than those who died of dehydration. Their main symptoms were respiratory symptoms, such as cough, sputum, and dyspnea, a fever and somnolence. Distress such as dyspnea and discomfort among older people with end-stage dementia recovering from pneumonia peaked on the day of the diagnosis. Furthermore, in older people with end-stage dementia dying of pneumonia, distresses began approximately one week before death and peaked on the day before death. A few days before death, several distressing symptoms, such as pain, dyspnea and restlessness/agitation, were often observed at the same time. CONCLUSIONS: If older people with end-stage dementia contract pneumonia and die from it, attentive palliative care is needed to manage their severe discomfort, dyspnea and other sources of distress.