RNA-methylation-dependent RNA processing controls the speed of the circadian clock.

The eukaryotic biological clock involves a negative transcription-translation feedback loop in which clock genes regulate their own transcription and that of output genes of metabolic significance. While around 10% of the liver transcriptome is rhythmic, only about a fifth is driven by de novo transcription, indicating mRNA processing is a major circadian component. Here, we report that inhibition of transmethylation reactions elongates the circadian period. RNA sequencing then reveals methylation inhibition causes widespread changes in the transcription of the RNA processing machinery, associated with m(6)A-RNA methylation. We identify m(6)A sites on many clock gene transcripts and show that specific inhibition of m(6)A methylation by silencing of the m(6)A methylase Mettl3 is sufficient to elicit circadian period elongation and RNA processing delay. Analysis of the circadian nucleocytoplasmic distribution of clock genes Per2 and Arntl then revealed an uncoupling between steady-state pre-mRNA and cytoplasmic mRNA rhythms when m(6)A methylation is inhibited.

An intact pituitary vasopressin system is critical for building a robust circadian clock in the suprachiasmatic nucleus.

The circadian clock is a biological timekeeping system that oscillates with a circa-24-h period, reset by environmental timing cues, especially light, to the 24-h day-night cycle. In mammals, a "central" clock in the hypothalamic suprachiasmatic nucleus (SCN) synchronizes "peripheral" clocks throughout the body to regulate behavior, metabolism, and physiology. A key feature of the clock's oscillation is resistance to abrupt perturbations, but the mechanisms underlying such robustness are not well understood. Here, we probe clock robustness to unexpected photic perturbation by measuring the speed of reentrainment of the murine locomotor rhythm after an abrupt advance of the light-dark cycle. Using an intersectional genetic approach, we implicate a critical role for arginine vasopressin pathways, both central within the SCN and peripheral from the anterior pituitary.

Calcitonin receptors are ancient modulators for rhythms of preferential temperature in insects and body temperature in mammals.

Daily body temperature rhythm (BTR) is essential for maintaining homeostasis. BTR is regulated separately from locomotor activity rhythms, but its molecular basis is largely unknown. While mammals internally regulate BTR, ectotherms, including Drosophila, exhibit temperature preference rhythm (TPR) behavior to regulate BTR. Here, we demonstrate that the diuretic hormone 31 receptor (DH31R) mediates TPR during the active phase in Drosophila DH31R is expressed in clock cells, and its ligand, DH31, acts on clock cells to regulate TPR during the active phase. Surprisingly, the mouse homolog of DH31R, calcitonin receptor (Calcr), is expressed in the suprachiasmatic nucleus (SCN) and mediates body temperature fluctuations during the active phase in mice. Importantly, DH31R and Calcr are not required for coordinating locomotor activity rhythms. Our results represent the first molecular evidence that BTR is regulated distinctly from locomotor activity rhythms and show that DH31R/Calcr is an ancient specific mediator of BTR during the active phase in organisms ranging from ectotherms to endotherms.

Circadian protection against bacterial skin infection by epidermal CXCL14-mediated innate immunity.

The epidermis is the outermost layer of the skin and the body's primary barrier to external pathogens; however, the early epidermal immune response remains to be mechanistically understood. We show that the chemokine CXCL14, produced by epidermal keratinocytes, exhibits robust circadian fluctuations and initiates innate immunity. Clearance of the skin pathogen Staphylococcus aureus in nocturnal mice was associated with CXCL14 expression, which was high during subjective daytime and low at night. In contrast, in marmosets, a diurnal primate, circadian CXCL14 expression was reversed. Rhythmically expressed CXCL14 binds to S. aureus DNA and induces inflammatory cytokine production by activating Toll-like receptor (TLR)9-dependent innate pathways in dendritic cells and macrophages underneath the epidermis. CXCL14 also promoted phagocytosis by macrophages in a TLR9-independent manner. These data indicate that circadian production of the epidermal chemokine CXCL14 rhythmically suppresses skin bacterial proliferation in mammals by activating the innate immune system.

Discriminant Accuracy of Standing Balance Tests for the Level of Gait Dependency in Hospitalized Patients with Alzheimer's Disease.

INTRODUCTION: When determining the level of gait independence in patients with Alzheimer's disease (AD), detailed functional assessment is difficult in some patients. The previous literature has suggested simple standing balance tests for patients with AD due to their ease of implementation in clinical practice and relevance to gait. However, their usefulness for discriminating the level of gait independence remains unclear. This study aimed to investigate the discrimination accuracy of a simple standing balance test in the level of gait independence among hospitalized patients with AD. METHODS: This cross-sectional study was a post hoc analysis of a study conducted on 63 inpatients with AD in a single hospital. Participants were divided into three groups according to their level of gait independence: independent, modified independent (independent, walking with walking aids), and dependent groups (supervision). Gait independence was determined using the Functional Independence Measure. Four standing balance tests were used - closed-leg, semi-tandem, tandem, and one-leg standings - and the discrimination accuracy of each test was calculated by receiver operating characteristic analysis. RESULTS: One-leg standing was best at discriminating between the independent and modified independent groups (positive predictive value = 80.0%, negative predictive value = 94.1%). Tandem standing was best at discriminating between the modified independent and dependent groups (positive predictive value = 74.1%, negative predictive value = 93.3%). CONCLUSION: A simple standing balance test may assist in the determining level of gait independence in patients with AD when it is difficult to perform a mobility assessment.

Combining Physical and Cognitive Functions to Discriminate Level of Gait Independence in Hospitalized Patients with Alzheimer's Disease.

INTRODUCTION: Both physical and cognitive functions are required to be assessed to determine the level of gait independence in patients with Alzheimer's disease (AD); nonetheless, a method to achieve this assessment has not been established. This study aimed to investigate the accuracy of an assessment method that combined muscle strength, balance ability, and cognitive function parameters in discriminating the level of gait independence in a real-world setting in hospitalized patients with AD. METHODS: In this cross-sectional study, 63 patients with AD (mean age: 86.1 ± 5.8 years) were classified into three groups according to their gait level: independent, modified independent (independent walking with walking aids), and dependent groups. Discrimination accuracy was calculated for single items of muscle strength, balance ability, and cognitive function tests and for combinations of each. RESULTS: The combined accuracy of muscle strength, balance ability, and cognitive function had a positive predictive value of 100.0% and a negative predictive value of 67.7% between the independent and modified independent groups. The positive and negative predictive values were 100.0% and 72.4%, respectively, between the modified independent and dependent groups. CONCLUSION: This study emphasizes the importance of assessing the level of gait independence in a real-world setting in patients with AD from the perspective of both physical and cognitive functions and proposes a novel method for discriminating an optimal state.

Factors Associated with Long-term Medication Adherence in Patients Who Participated in a Short-term Group Psychoeducation Program for Bipolar Disorder.

Improving adherence to medication and quality of life is a challenge in treating bipolar disorder. Therefore, psychoeducation plays an important role. This study examined factors associated with long-term medication adherence in patients with bipolar disorder who participated in a short-term psychoeducation program. Additionally, the relationships between medication adherence and medication attitudes and quality of life (QOL) were assessed. Multiple regression analysis was conducted on 67 inpatients and outpatients using medication adherence (Brief Evaluation for Medication Influences and Beliefs [BEMIB] score) as the dependent variable and clinical and demographic variables before and after the program as explanatory variables, one year after completion of the program. Pearson's correlation coefficients were calculated for associations between patients' BEMIB score and medication attitudes (Drug Attitude Inventory-10 [DAI-10]) and QOL (World Health Organization QOL-26 [WHOQOL-26] score) before and after the program and one year after program completion. The results showed that the CSQ-8 J (Client Satisfaction Questionnaire-8 Japanese) and DAI-10 scores immediately after the program were significantly related to the BEMIB score one year after program completion. Both the BEMIB and DAI-10 showed significant positive correlations with several items of the WHOQOL-26, both after the program and one year after completion of the program. The results suggest that medication attitudes acquired through psychoeducation and program satisfaction impact long-term medication adherence. The study also indicates that medication attitudes and medication adherence after a psychoeducation program are associated with QOL. Thus, patients' subjective views after a psychoeducation program can play an important role in long-term medication adherence and QOL.

Factors Associated with Caregiver Burden and Depressive States among Family Caregivers of Patients Admitted due to Behavioral and Psychological Symptoms of Dementia.

INTRODUCTION: Among those caring for people with dementia (PwD) at home, more than 60% feel a caregiver burden (CB), and one in three are depressed. Reducing feelings of burden and depressive states in caregiving families will improve the living environment for PwD. However, very few studies have focused on effective methods and reducing feelings of burden and depressive states of caregivers. Thus, using data from a previous study, we aimed to determine the factors associated with perceived CB and depressive states experienced by caregivers for PwD with behavioral and psychological symptoms of dementia (BPSD) at home. METHODS: We performed single regression analysis on 285 participants' data to determine the association between each item and the Zarit Caregiver Burden Interview and Center for Epidemiologic Studies-Depression Scale scores. We performed multiple regression analysis with variables considered in the single regression analysis as independent variables. RESULTS: Severity of BPSD, caregivers' subjective health status (SHS), time of caregiving, and depressive states were related to CB, and caregivers' SHS and CB were related to depressive states. CONCLUSIONS: Similar to previous studies, we identified an association between family caregivers' perceived CB and BPSD in PwD. Additionally, we found that caregivers' SHS is commonly associated with both perceived CB and depressive states. This is a new finding. Our results suggest that interventions focusing on family caregivers' health status can help not only to reduce family caregivers' CB and depressive states but also stabilize patients' symptoms and provide home-based care for a longer time.

CNOT1 regulates circadian behaviour through Per2 mRNA decay in a deadenylation-dependent manner.

Circadian clocks are an endogenous internal timekeeping mechanism that drives the rhythmic expression of genes, controlling the 24 h oscillatory pattern in behaviour and physiology. It has been recently shown that post-transcriptional mechanisms are essential for controlling rhythmic gene expression. Controlling the stability of mRNA through poly(A) tail length modulation is one such mechanism. In this study, we show that Cnot1, encoding the scaffold protein of the CCR4-NOT deadenylase complex, is highly expressed in the suprachiasmatic nucleus, the master timekeeper. CNOT1 deficiency in mice results in circadian period lengthening and alterations in the mRNA and protein expression patterns of various clock genes, mainly Per2. Per2 mRNA exhibited a longer poly(A) tail and increased mRNA stability in Cnot1+/- mice. CNOT1 is recruited to Per2 mRNA through BRF1 (ZFP36L1), which itself oscillates in antiphase with Per2 mRNA. Upon Brf1 knockdown, Per2 mRNA is stabilized leading to increased PER2 expression levels. This suggests that CNOT1 plays a role in tuning and regulating the mammalian circadian clock.

Nmu/Nms/Gpr176 Triple-Deficient Mice Show Enhanced Light-Resetting of Circadian Locomotor Activity.

The suprachiasmatic nucleus (SCN) is the master circadian clock in mammals and is properly entrained by environmental light cycle. However, the molecular mechanism(s) determining the magnitude of phase shift by light is still not fully understood. The orphan G-protein-coupled receptor Gpr176 is enriched in the SCN, controls the pace (period) of the circadian rhythm in behavior but is not apparently involved in the light entrainment; Gpr176-/- animals display a shortened circadian period in constant darkness but their phase-resetting responses to light are normal. Here, we performed microarray analysis and identified enhanced mRNA expression of neuromedin U (Nmu) and neuromedin S (Nms) in the SCN of Gpr176-/- mice. By generating C57BL/6J-backcrossed Nmu/Nms/Gpr176 triple knockout mice, we noted that the mutant mice had a greater magnitude of phase shift in response to early subjective night light than wildtype mice, while Nmu/Nms double knockout mice as well as Gpr176 knockout mice are normal in the phase shifts induced by light. At the molecular level, Nmu-/-Nms-/-Gpr176-/- mice had a reduced induction of Per1 and cFos mRNA expression in the SCN by light and mildly upregulated circadian expression of Per2, Prok2, Rgs16, and Rasl11b. These expressional changes may underlie the phenotype of the Nmu/Nms/Gpr176 knockout mice. Our data argue that there is a mechanism requiring Nmu, Nms, and Gpr176 for the proper modulation of light-induced phase shift in mice. Simultaneous modulation of Nmu/Nms/Gpr176 may provide a potential target option for modulating the circadian clock.

Midwifery scale to support shared decision-making for unplanned pregnancies: A cross-sectional study.

Midwives significantly support women with unplanned pregnancies-promoting a shared perspective on the decision-making process. This study aimed to develop a scale to support midwives to self-assess their practice of this vital role. Following the derivation of scale items and pilot testing, the final version of the scale was administered to 531 midwives to establish internal consistency and construct criterion-related validity. Through exploratory factor analysis, 35 items with a five-factor structure were retained to form the midwifery practice self-assessment scale to promote shared decision-making in women with unplanned pregnancies. These factors illustrate midwives' general aptitude and competencies in understanding environmental factors, collaborating with significant others and the interprofessional group, forming rapport and problem sharing, focusing on consultation content, and promoting autonomous decision-making. There were high and low scores on the scales after attendance of the workshops to support the decision-making of women with unplanned pregnancies. The reliability analysis showed acceptable Cronbach's alpha values for the five factors, from 0.85 to 0.87. The scale was demonstrated to be a reliable and valid measure that would help improve the quality of midwives' practice.

Association Between Subjective Voice Assessment and Psychological Distress After Thyroidectomy.

PURPOSE: This study aimed to determine the association between postoperative subjective voice function and psychological distress in patients without laryngeal nerve injury after thyroidectomy. DESIGN: A prospective cohort study. METHODS: We investigated the factors associated with subjective voice function in patients who underwent thyroidectomy without laryngeal nerve injury between October 2018 and July 2020. The Voice Handicap Index was used to assess subjective voice function, the GRBAS (grade, roughness, breathiness, asthenia, strain) scale to assess objective voice, and the Hospital Anxiety and Depression Scale to assess psychological distress. FINDINGS: Among 39 patients who underwent thyroidectomy, 32 had no postoperative laryngeal nerve injury. Postoperative Voice Handicap Index was significantly associated with Hospital Anxiety and Depression Scale-Anxiety score after surgery (rs = 0.448, P = .010). CONCLUSIONS: In this study, an association was observed between subjective voice function and anxiety following surgery. The finding suggested that nurses and medical practitioners need to consider postoperative anxiety when evaluating patients' voice function after thyroidectomy.

Fetal Leydig cells dedifferentiate and serve as adult Leydig stem cells.

Previous studies have established that fetal Leydig cells (FLCs) and adult Leydig cells (ALCs) show distinct functional characteristics. However, the lineage relationship between FLCs and ALCs has not been clarified yet. Here, we reveal that a subset of FLCs dedifferentiate at fetal stages to give rise to ALCs at the pubertal stage. Moreover, the dedifferentiated cells contribute to the peritubular myoid cell and vascular pericyte populations in the neonatal testis, and these non-steroidogenic cells serve as potential ALC stem cells. We generated FLC lineage-specific Nr5a1 (Ad4BP/SF-1) gene-disrupted mice and mice lacking the fetal Leydig enhancer (FLE) of the Nr5a1 gene. Phenotypes of these mice support the conclusion that most of the ALCs arise from dedifferentiated FLCs, and that the FLE of the Nr5a1 gene is essential for both initial FLC differentiation and pubertal ALC redifferentiation.

The influence of coping types on post-traumatic growth in patients with primary breast cancer.

BACKGROUND: The physical and mental impacts of breast cancer diagnosis on women are substantial. Several studies have investigated the negative mental health effects of breast cancer. However, in recent years, there has also been growing interest in post-traumatic growth, a positive response to stressful events. Considering positive psychology focuses on such virtues, proactive coping theory was chosen as a theoretical guide. This study investigates how breast cancer patients' post-traumatic growth is associated with proactive coping and mental well-being. METHODS: A self-administered questionnaire survey was conducted with 80 breast cancer patients aged 20-70 years attending an outpatient clinic. The survey was conducted using the Posttraumatic Growth Inventory-Japanese version (PTGI-J), Proactive Coping Inventory-Japanese version and the Japanese version of the General Health Questionnaire. Single regression and multiple regression analyses with PTGI-J as the dependent variable were performed. RESULTS: The multiple regression analysis extracted proactive coping (P = 0.006), emotional support seeking (P = 0.004) and avoidance coping (P = 0.001) as factors associated with post-traumatic growth in breast cancer patients. CONCLUSIONS: These results suggest that using proactive coping for conflicts caused by a breast cancer diagnosis and temporary avoidant coping for daily stresses during the treatment process may enhance post-traumatic growth while preventing deterioration in mental well-being. Additionally, seeking emotional support is important for post-traumatic growth.

Factors associated with facilitating advance care planning based on the theory of planned behaviour.

BACKGROUND: In Japan, educational support for advance care planning is still limited, and the factors facilitating advance care planning practice have not been identified. The aim of this study is to identify the factors associated with advance care planning practice in Japan using the theory of planned behaviour. METHODS: We conducted a cross-sectional study through a nationwide online questionnaire survey with 446 Japanese residents aged 20 years and older. After categorizing participants into two groups based on who performed advance care planning, comparisons were made (univariate analysis) regarding attitudes towards advance care planning, willingness to perform advance care planning, subjective norms, sense of control in performing advance care planning, presence of and relationship with a family doctor, experience of seeing or using a ventilator, and degree of self-determination (Autonomy Preference Index) as well as sociodemographic data. In addition, a logistic regression analysis was performed using the factors with significant differences in the univariate analysis as independent variables and the existence of the implementation of advance care planning as a dependent variable. RESULTS: The group with advance care planning consisted of 106 participants (23.8%), and the group without advance care planning comprised 340 participants (76.2%). Through logistic regression analysis, age (odds ratio: 1.020), subjective norms (odds ratio: 3.276) and experience with mechanical ventilation (odds ratio: 1.997) were extracted as significant factors influencing advance care planning implementation. CONCLUSIONS: Advance care planning may be facilitated by providing support not only to the patients but also to their family members to be positively disposed towards advance care planning and by offering comprehensive education regarding using a ventilator.

Changes in motor function and quality of life after surgery in patients with pancreatic cancer.

OBJECTIVE: To determine the changes in motor function and health-related quality of life after pancreatectomy and identify factors influencing postoperative physical functioning in health-related quality of life. METHODS: This single-centre, prospective, observational study measured 6-min walking distance, grip strength, knee extension strength and health-related quality of life variables in patients with pancreatic cancer, before and after surgery. Paired t- and Wilcoxon signed-rank tests were used to compare pre- and postoperative motor function and health-related quality-of-life variables. Factors associated with postoperative physical functioning scores of health-related quality of life were assessed using multiple regression analysis. RESULTS: Fifty-nine individuals were enrolled. Motor function values decreased significantly postoperatively, including 6-min walking distance (mean ± standard deviation: 402.5 ± 95.4 vs. 497.7 ± 80.4 m, p < 0.001), knee extensor strength (0.42 ± 0.10 vs. 0.47 ± 0.10 kgf/kg, p < 0.001) and grip strength (22.0 ± 8.9 vs. 24.5 ± 9.2 kg, p = 0.001). Multiple regression analysis showed significant association between 6-min walking distance change and postoperative physical functioning scores of health-related quality of life (p = 0.036). CONCLUSION: The results suggest that motor function decreases postoperatively, and a decrease in 6-min walking distance after surgery is associated with postoperative physical functioning in patients with pancreatic cancer.

CK1δ/ε protein kinase primes the PER2 circadian phosphoswitch.

Multisite phosphorylation of the PERIOD 2 (PER2) protein is the key step that determines the period of the mammalian circadian clock. Previous studies concluded that an unidentified kinase is required to prime PER2 for subsequent phosphorylation by casein kinase 1 (CK1), an essential clock component that is conserved from algae to humans. These subsequent phosphorylations stabilize PER2, delay its degradation, and lengthen the period of the circadian clock. Here, we perform a comprehensive biochemical and biophysical analysis of mouse PER2 (mPER2) priming phosphorylation and demonstrate, surprisingly, that CK1δ/ε is indeed the priming kinase. We find that both CK1ε and a recently characterized CK1δ2 splice variant more efficiently prime mPER2 for downstream phosphorylation in cells than the well-studied splice variant CK1δ1. While CK1 phosphorylation of PER2 was previously shown to be robust to changes in the cellular environment, our phosphoswitch mathematical model of circadian rhythms shows that the CK1 carboxyl-terminal tail can allow the period of the clock to be sensitive to cellular signaling. These studies implicate the extreme carboxyl terminus of CK1 as a key regulator of circadian timing.

Two Ck1δ transcripts regulated by m6A methylation code for two antagonistic kinases in the control of the circadian clock.

The N6-methylation of internal adenosines (m6A) in mRNA has been quantified and localized throughout the transcriptome. However, the physiological significance of m6A in most highly methylated mRNAs is unknown. It was demonstrated previously that the circadian clock, based on transcription-translation negative feedback loops, is sensitive to the general inhibition of m6A. Here, we show that the Casein Kinase 1 Delta mRNA (Ck1δ), coding for a critical kinase in the control of circadian rhythms, cellular growth, and survival, is negatively regulated by m6A. Inhibition of Ck1δ mRNA methylation leads to increased translation of two alternatively spliced CK1δ isoforms, CK1δ1 and CK1δ2, uncharacterized until now. The expression ratio between these isoforms is tissue-specific, CK1δ1 and CK1δ2 have different kinase activities, and they cooperate in the phosphorylation of the circadian clock protein PER2. While CK1δ1 accelerates the circadian clock by promoting the decay of PER2 proteins, CK1δ2 slows it down by stabilizing PER2 via increased phosphorylation at a key residue on PER2 protein. These observations challenge the previously established model of PER2 phosphorylation and, given the multiple functions and targets of CK1δ, the existence of two isoforms calls for a re-evaluation of past research when CK1δ1 and CK1δ2 were simply CK1δ.

Correction to: Voice rehabilitation for laryngeal cancer after radiotherapy: a systematic review and meta-analysis.

In the original publication of the article, the reference 14 was published incorrectly. The correct reference is given below.

Effects of a Psychoeducation Program for People with Schizophrenia Aimed at Increasing Subjective Well-Being and the Factors Influencing Those Effects: a Preliminary Study.

There have been no intervention studies of psychoeducation programs for schizophrenia that focus on improving subjective well-being or studies to determine the factors influencing such effects. This study aimed to examine the effects of a psychoeducation program combining traditional psychoeducation with a focus on providing knowledge and information and a new intervention to raise patients' subjective well-being and to clarity the factors affecting the program's efficacy. Subjects were 117 patients who participated in a psychoeducation program for schizophrenia between 2012 and 2018. In addition to comparing subjective well-being (Subjective Well-being under Neuroleptic Drug Treatment Short Form, Japanese version, SWNS-J) and attitudes towards drugs (Drug Attitude Inventory-10, DAI-10) before and after the program, basic information such as psychiatric symptoms was surveyed. The factors influencing the effects of the program were assessed with multiple regression analysis. Scores for SWNS-J subscales and total SWNS-J score increased significantly after the program. Higher total scores on subjective well-being after the program were significantly associated with having less severe negative symptoms and higher total subjective well-being before the program, and with more positive attitudes toward drugs after the program. These results suggest that intervention with a focus on improving subjective well-being can be an effective part of psychoeducation programs for patients with schizophrenia. The results also suggest that the level of improvement in subjective well-being gained from the program may be higher in patients with milder negative symptoms and patients with a better understanding of drug treatment.