RhoA rescues cardiac senescence by regulating Parkin-mediated mitophagy.

Heart failure is one of the leading causes of death worldwide. RhoA, a small GTPase, governs actin dynamics in various tissue and cell types, including cardiomyocytes; however, its involvement in cardiac function has not been fully elucidated. Here, we generated cardiomyocyte-specific RhoA conditional knockout (cKO) mice, which demonstrated a significantly shorter lifespan with left ventricular dilation and severely impaired ejection fraction. We found that the cardiac tissues of the cKO mice exhibited structural disorganization with fibrosis and also exhibited enhanced senescence compared with control mice. In addition, we show that cardiomyocyte mitochondria were structurally abnormal in the aged cKO hearts. Clearance of damaged mitochondria by mitophagy was remarkably inhibited in both cKO cardiomyocytes and RhoA-knockdown HL-1 cultured cardiomyocytes. In RhoA-depleted cardiomyocytes, we reveal that the expression of Parkin, an E3 ubiquitin ligase that plays a crucial role in mitophagy, was reduced, and expression of N-Myc, a negative regulator of Parkin, was increased. We further reveal that the RhoA-Rho kinase axis induced N-Myc phosphorylation, which led to N-Myc degradation and Parkin upregulation. Re-expression of Parkin in RhoA-depleted cardiomyocytes restored mitophagy, reduced mitochondrial damage, attenuated cardiomyocyte senescence, and rescued cardiac function both in vitro and in vivo. Finally, we found that patients with idiopathic dilated cardiomyopathy without causal mutations for dilated cardiomyopathy showed reduced cardiac expression of RhoA and Parkin. These results suggest that RhoA promotes Parkin-mediated mitophagy as an indispensable mechanism contributing to cardioprotection in the aging heart.

Impact of pureed and liquidised diets on health-related quality of life scores in older patients during postacute rehabilitation: A pilot study.

BACKGROUND: Consuming texture-modified diets was considered to negatively affect health-related quality of life (HR-QoL). However, the precise relationship between the levels of food texture consumed and HR-QoL remains uncertain. The present study aimed to determine the association between levels of food texture consumed and HR-QoL. METHODS: This cross-sectional study included 122 hospitalised patients aged ≥ 65 years (mean ± SD age 78.9 ± 7.9 years; 48.4% female) who required postacute rehabilitation. Consumed food texture levels were classified using the International Dysphagia Diet Standardization Initiative (IDDSI) framework 2.0. HR-QoL was evaluated using the five-level EuroQoL five-dimension (EQ-5D-5L). The association between food texture levels and HR-QoL was analysed using a multivariate linear regression model. RESULTS: The lower food texture levels consumed were associated with significantly lower EQ-5D-5L scores. Consumption of pureed (IDDSI Level 4) and liqudised (IDDSI Level 3) diets were associated with lower EQ-5D-5L scores among older patients undergoing postacute rehabilitation (regression coefficient -0.304; 95% confidence interval = -0.472 to -0.137 and regression coefficient, -0.444, 95% confidence interval = -0.676 to -0.213, respectivly). CONCLUSIONS: The findings suggest a link between the consumption of pureed and liquidised diets and lower HR-QoL scores in older hospitalised patients undergoing rehabilitation. To establish a clearer cause-and-effect relationship, future research should encompass multicentre and longitudinal studies, building upon the insights from the present study.

The effect of preoperative rehabilitation on the prevention of postoperative ileus in colorectal cancer patients.

PURPOSE: Previous research suggests that the preoperative rehabilitation of colorectal cancer patients can reduce postoperative ileus. However, the evidence is insufficient and further research is warranted. This study aimed to investigate whether short-term preoperative rehabilitation, both on an outpatient and inpatient basis, can reduce the incidence of postoperative ileus after colorectal cancer surgery. METHODS: This was a retrospective cohort study that drew on data from multicenter electronic medical records. Patients with stage 1-3 colorectal cancer who underwent surgery and postoperative rehabilitation were included. The incidence of postoperative ileus was compared between patients who received short-term preoperative rehabilitation and those who did not. Propensity score adjustment using inverse probability weighting and subgroup analysis by type of surgery was performed. RESULTS: Four thousand seventy-six eligible patients (43.4% female; mean age 75.1 ± 10.9 years) were included; 1914 (47.0%) received short-term preoperative rehabilitation. The preoperative rehabilitation group had a significantly lower incidence of postoperative ileus than the no preoperative rehabilitation group (pre-adjustment: 5.5% vs. 9.9%, p < 0.001; post-adjustment: 5.2% vs. 9.0%, p < 0.001). Therefore, preoperative rehabilitation was significantly associated with a lower incidence of postoperative ileus (OR: 0.554, 95% CI: 0.415-0.739, p < 0.001). In an adjusted analysis of surgery type subgroups, the incidence of postoperative ileus was significantly lower in the preoperative rehabilitation group for all types of surgery. CONCLUSION: Our study showed that short-term preoperative rehabilitation for patients with stage 1-3 colorectal cancer, both with inpatients and outpatients, significantly reduces the incidence of postoperative ileus.

Early physical rehabilitation effectiveness in frail older patients hospitalized for community-acquired pneumonia: analysis of a nationwide database in Japan.

BACKGROUND: Frailty is a significant predictor of prognosis in older patients with community-acquired pneumonia (CAP). No effective therapy has been reported in frail patients with CAP, with frailty determined using the Hospital Frailty Risk Score (HFRS). AIMS: To investigate whether early physical rehabilitation intervention would effectively minimize adverse outcomes in frail older patients (determined using the HFRS) hospitalized for CAP. METHODS: This retrospective cohort analysis involved patients with CAP aged ≥ 65 years enrolled in the Japanese Diagnostic Procedure Combination Database between 2014 and 2020 and assessed as being frail. We compared 30-day mortality and readmission rates for patients who did and who did not receive physical rehabilitation within three days of admission and evaluated the association between outcomes and receiving early physical rehabilitation using Cox regression models and inverse probability weighting (IPW) for sensitivity analysis. RESULTS: The analysis involved 31,133 frail older patients hospitalized for CAP (mean age 84.3 ± 6.3 years; females, 49.1%), including 11,515 (37.0%) who received early physical rehabilitation. Cox regression analysis showed that early physical rehabilitation intervention was inversely associated with 30-day mortality and readmission rates. The IPW model also showed similar results. DISCUSSION: Early physical rehabilitation was associated with reduced risks of 30-day mortality, overall in-hospital mortality, and 30-day readmission rates in frail older patients with CAP. CONCLUSIONS: Early physical rehabilitation in frail older patients hospitalized for CAP may improve outcomes. This finding highlights the importance of simultaneously introducing the HFRS and early physical rehabilitation intervention into clinical practice for frail older patients with CAP.

Associations between oral health status and short-term functional outcomes in hospitalized patients aged over 65 years with acute ischemic stroke: a retrospective observational study.

OBJECTIVE: To assess the associations between oral health status and short-term functional outcomes in hospitalized patients aged over 65 years with acute ischemic stroke. MATERIALS AND METHODS: This retrospective observational analysis included older adult patients (age, ≥ 65 years) admitted for acute ischemic stroke. The oral health status at admission was evaluated using the Oral Health Assessment Tool (OHAT). Patients were categorized into the normal oral health (OHAT score, 0-2) or poor oral health (OHAT score, ≥ 3) group. Stroke severity, Functional Oral Intake Scale (FOIS), and medical history were compared. Multivariate analysis was used to determine the association between the OHAT score and modified Rankin Scale (mRS) score at discharge, FOIS score at discharge, and length of hospital stay. RESULTS: The study comprised 129 patients (mean age: 78.8 ± 7.7 years). The poor oral health group (n = 22) had a higher stroke severity and lower FOIS scores than the normal oral health group (n = 107). The poor oral health group exhibited significantly higher rates of moderate to severe disability at discharge (odds ratio = 9.18, 95% confidence interval [CI]: 1.74-48.30, P = 0.009), lower FOIS scores at discharge (ß = -0.96, 95% CI: -1.71 to -0.20, P = 0.014), and longer hospital stays (ß = 10.70, 95% CI: 0.80-20.61, P = 0.034) than the other group. CONCLUSION: In older patients with acute ischemic stroke, poor oral health status at admission was associated with worse short-term functional outcomes, including increased disability, dysphagia, and longer hospital stay. CLINICAL RELEVANCE: Assessing and addressing the oral health status of this population can potentially improve short-term functional outcomes and enhance comprehensive stroke care.

Association of poor oral health status and faecal incontinence in patients with dysphagia: A cross-sectional analysis from the Sarcopenic Dysphagia Database.

BACKGROUND: Poor oral health status may alter oral and gut microbiota. Previous studies have shown that poor oral health can exacerbate gut inflammation. Therefore, poor oral health status may be related to faecal incontinence via changes in the gut. OBJECTIVE: To investigate the association between poor oral health status and faecal incontinence in inpatients with dysphagia. METHODS: This multicentre cross-sectional study included 423 patients (mean age 79.8 ± 11.5 years, 48.2% female) with dysphagia. Oral health status was assessed at each facility using the Oral Health Assessment Tool (OHAT) or the Revised Oral Assessment Guide (ROAG). Poor oral health status was defined as an OHAT score of ≥3 or a ROAG score of ≥13. A multivariate logistic model was used to analyse the association between poor oral health status and faecal incontinence. RESULTS: A total of 351 (83.0%) patients had poor oral health and 97 (22.7%) had faecal incontinence. Patients with poor oral health status had a higher proportion of faecal incontinence than those with normal oral health status (25.4% vs. 11.1%, p = .009). A multivariate logistic model revealed an association between faecal incontinence and poor oral health status (adjusted odds ratio = 2.501, 95% confidence interval = 1.065-5.873, p = .035). CONCLUSIONS: Poor oral health status assessed by OHAT or ROAG in inpatients with dysphagia may adversely affect faecal incontinence. Further studies are needed to determine the causal relationship between poor oral health status and faecal incontinence.

Cardio- and reno-protective effects of dipeptidyl peptidase III in diabetic mice.

Diabetes mellitus (DM) causes injury to tissues and organs, including to the heart and kidney, resulting in increased morbidity and mortality. Thus, novel potential therapeutics are continuously required to minimize DM-related organ damage. We have previously shown that dipeptidyl peptidase III (DPPIII) has beneficial roles in a hypertensive mouse model, but it is unknown whether DPPIII has any effects on DM. In this study, we found that intravenous administration of recombinant DPPIII in diabetic db/db mice for 8 weeks suppressed the DM-induced cardiac diastolic dysfunctions and renal injury without alteration of the blood glucose level. This treatment inhibited inflammatory cell infiltration and fibrosis in the heart and blocked the increase in albuminuria by attenuating the disruption of the glomerular microvasculature and inhibiting the effacement of podocyte foot processes in the kidney. The beneficial role of DPPIII was, at least in part, mediated by the cleavage of a cytotoxic peptide, named Peptide 2, which was increased in db/db mice compared with normal mice. This peptide consisted of nine amino acids, was a digested fragment of complement component 3 (C3), and had an anaphylatoxin-like effect determined by the Miles assay and chemoattractant analysis. The effect was dependent on its interaction with the C3a receptor and protein kinase C-mediated RhoA activation downstream of the receptor in endothelial cells. In conclusion, DPPIII plays a protective role in the heart and kidney in a DM animal model through cleavage of a peptide that is a part of C3.

Pituitary luteinizing hormone synthesis starts in aromatase (cyp19a1b)-positive cells expressing esr1 and esr2b at the onset of puberty in Takifugu rubripes (fugu).

Unlike mammals, teleost fish have high aromatase activity (AA) in the pituitary. However, the cells responsible for oestradiol synthesis and the local physiological roles of this hormone remain unclear. Hence, we investigated the effects of age and development on steroidogenic activity, mRNA expression, and cyp19a1b localization in the pituitary gland of the Japanese pufferfish Takifugu rubripes. Under aquaculture conditions, AA was highest after puberty, and the mRNA expression levels of cyp19a1b and the oestrogen receptors esr1 and 2b and the level of serum testosterone (T) were significantly increased after puberty compared with the other developmental stages in male and female pufferfish. Immunohistochemistry using multiple antibodies and in situ hybridization analysis revealed that Cyp19a1b colocalizes with luteinizing hormone (LH) in pituitary cells. Furthermore, Esr1 was localized in the nuclei of all hormone-producing cells, whereas Esr2b was localized only in the nuclei of Cyp19- and LH-positive cells. The administration of an aromatizable androgen (T) or oestrogen (E2) to reproductively inactive females induced LH synthesis in vivo. We prepared spheroids from pituitary cells to investigate the role of local E2 in LH synthesis in vitro. Immunohistochemical analysis of spheroids showed that T-induced LH synthesis could be blocked by an aromatase inhibitor and/or an ER antagonist but not an AR antagonist. Taken together, these findings suggest that LH synthesis is initiated in cyp19a1b-, esr1-, and esr2b-expressing cells at the onset of puberty under the control of steroidal feedback, and both feedback and local oestrogen may be involved in controlling LH synthesis in these cells.

High SARC-F score predicts poor survival of patients with cancer receiving palliative care.

PURPOSE: A high score determined by SARC-F, a simple screening questionnaire for sarcopenia, has been reportedly associated with worse medical outcomes. However, information regarding whether high SARC-F scores are associated with a poor prognosis in patients with advanced cancer remains limited. We clarified whether a SARC-F score ≥ 4 predicts poor prognosis in patients with cancer receiving palliative care. METHODS: We conducted a retrospective cohort study of patients with cancer who received palliative care at a university hospital between May 2019 and April 2020. Patient characteristics including age, sex, height, weight, cancer type, serum albumin level, C-reactive protein level, presence of edema, Eastern Cooperative Oncology Group-Performance Status (ECOG-PS), SARC-F score, history of anticancer therapy, and clinical outcomes were collected from electronic medical records. RESULTS: Of 304 patients, 188 had a SARC-F score < 4, and 116 patients had a SARC-F score ≥ 4. The overall survival of patients with a SARC-F score ≥ 4 was 40 days (95% CI 29-47), which was significantly worse than 121 days (95% CI 95-156) for patients with a SARC-F score < 4 (p < 0.001). SARC-F score ≥ 4 (hazard ratio: HR 1.56), edema (HR 1.94), head and neck cancer (HR 0.51), C-reactive protein (HR 1.05), ECOG-PS ≥ 3 (HR 1.47), and radiotherapy (HR 0.52) were associated with overall survival. The ability to climb stairs was a SARC-F sub-item significantly associated with mortality (HR 1.59). CONCLUSION: The SARC-F questionnaire is a useful predictor of prognosis for patients with cancer receiving palliative care because a SARC-F ≥ 4 score predicts worse overall survival.

Development of sandwich enzyme-linked immunosorbent assays for chub mackerel Scomber japonicus gonadotropins and regulation of their secretion in female reproduction.

The pituitary gonadotropins (Gths), follicle-stimulating hormone (Fsh) and luteinizing hormone (Lh), play critical roles in regulating gonadal development and sexual maturation in vertebrates. We developed non-competitive enzyme-linked immunosorbent assays (ELISAs) to measure Fsh and Lh in chub mackerel Scomber japonicus, which is a commercially important scombrid species. Mouse monoclonal antibodies specific for Fsh and Lh, and a rabbit polyclonal antibody against both Gths were produced by immunization with hormones purified from chub mackerel pituitaries. These monoclonal and polyclonal antibodies were used as capture and detection antibodies in the developed sandwich ELISAs. The ELISAs were reproducible, sensitive, and specific for chub mackerel Fsh and Lh. Parallelism between the standard curve and serial dilutions of chub mackerel serum and pituitary extract was observed for both Fsh and Lh ELISAs. Comparison between vitellogenic and immature females revealed that Fsh is secreted during vitellogenesis and Lh is barely released during immaturity. After gonadotropin-releasing hormone analog (GnRHa) injection, vitellogenic females showed increases in serum Lh, whereas serum levels of Fsh did not vary. Moreover, the serum steroid profiles revealed that estradiol-17ß was continuously produced after GnRHa treatment, whereas 17,20ß-dihydroxy-4-pregnen-3-one secretion was transiently induced. These results indicate that, in vitellogenic females, GnRHa stimulates the release of Lh, but not Fsh, which results in acceleration of vitellogenesis and induction of oocyte maturation via steroid production.

Diet-induced inflammation is associated with sarcopenia and muscle strength in older adults who visit a frailty clinic.

BACKGROUND: Diet-induced inflammation may be associated with sarcopenia; however, few reports have examined this relationship. AIM: To examine the association between the dietary inflammatory index (DII) and sarcopenia in older adults who visited a frailty clinic in Japan. METHODS: This cross-sectional study used outpatient data from the Frailty Registry Study. The DII is an index of diet-induced inflammation, and a dietary assessment was performed using a brief self-administered diet history questionnaire to calculate the DII score. We classified DII scores by quartiles (Q1-Q4), and sarcopenia was diagnosed according to the Asian Working Group for Sarcopenia 2019 consensus. Logistic regression analyses for sarcopenia were performed. Age, sex, comorbidities, and physical activity were entered as confounding factors (Model 1) and Models 2, 3, and 4 with BMI, protein intake, and energy intake added to Model 1. RESULTS: We included 304 patients in the analysis (mean age, 77.6 ± 6.3 years; female, 67.4%). The prevalence of sarcopenia was 14.5%. Logistic regression analyses showed that DII scores were significantly associated with sarcopenia in Model 1 and 2 (Model 1, reference: Q1, Q4: OR 3.10, P = 0.020; Model 2, Q4: OR 3.40, P = 0,022) but not in Model 3 and 4. DISCUSSION: Diet-induced inflammation is associated with a higher likelihood of sarcopenia; however, this association disappeared after confounding for protein and energy intake. CONCLUSIONS: The results demonstrated that dietary protein and energy parameters were the main drivers for muscle health in medical patients.

Food appearance affects reward-related brain activity in healthy adults: a functional magnetic resonance imaging study.

A good appearance of food increases appetite. A new food product called iEat® resembles the appearance and softness of familiar foods. Previous studies have reported that iEat® foods increase appetite. However, the neuronal substrates underlying the increase in appetite following the observation of iEat® foods remain unknown. In the present study, the brain activity of 20 healthy adults during the visual presentation of iEat® and pureed foods and non-food objects was examined using functional magnetic resonance imaging. Compared with pureed foods and non-food objects, iEat® foods showed significantly greater activation in regions of the brain reward system, such as the amygdala, ventral striatum and orbital frontal cortex. In addition, individual differences in the activity of the left amygdala were positively correlated with subjective appetite ratings. These results suggest that the good appearance of foods, such as iEat® foods, may be useful for stimulating the appetite of patients with poor appetite.

Undernutrition at Admission Suppresses Post-Stroke Recovery of Trunk Function.

OBJECTIVES: Malnutrition and decreased trunk function have a negative influence on the activities of daily living (ADL) prognosis after stroke. However, the relationship between malnutrition and improvement in trunk function has not been clarified. We aimed to examine the influence of malnutrition on the improvement in trunk function in stroke patients. MATERIALS AND METHODS: This retrospective, observational study was conducted with cerebral infarction patients aged ≥ 65 years with stroke. The study period was from May 2018 to September 2020. Patients were divided into malnutrition and intact groups according to the Global Leadership Initiative on Malnutrition (GLIM) criteria. The primary outcome was the change in the Functional Assessment for Control of Trunk (FACT) score (FACT score at discharge - FACT score at admission). RESULTS: A total of 183 participants (mean age, 79.7 ± 7.5 years; males, 56.3%) were included. A total of 79 (43%) and 104 (57%) patients were divided into the malnutrition and intact groups, respectively. The malnutrition group had a lower FACT score at admission (7.7 ± 7.3 vs. 11.9 ± 6.3, P < 0.001) and a lower FACT score at discharge (8.0 ± 7.7 vs. 13.3 ± 6.1, P < 0.001) than the intact group. After adjustment for the confounding factors, malnutrition was associated with a smaller change in the FACT score (coefficient = -1.871, 95% CI = -3.401 to -0.340, P = 0.017). CONCLUSIONS: Malnutrition has a negative influence on the recovery of trunk function in post-stroke patients. This finding should be verified through additional prospective studies.

Early Wearing of Knee-Ankle-Foot Orthosis Improves Functional Prognosis in Patients after Stroke.

OBJECTIVES: Knee-ankle-foot orthosis (KAFO) is sometimes used for gait training in stroke patients. The impact of the time of wearing KAFO on activities of daily living (ADL) recovery has not been clarified. This study aimed to examine the relationship between the days from onset to KAFO wearing and functional prognosis in patients after stroke. MATERIALS AND METHODS: This retrospective, observational study was conducted with stroke patients who were prescribed a KAFO. Patients were divided into early and delayed groups according to the median days from onset to KAFO wearing. Baseline characteristics were evaluated at the initiation of KAFO wearing. The primary outcome was the Functional Independence Measure (FIM) gain, which was scored by the nurse at baseline and discharge. RESULTS: 112 participants (mean age 67.9 ± 14.0 years, 51.8% male) were included. The time period measure from day of onset to KAFO wearing for the early group was significantly shorter than the delayed group (35.8 ± 6.6 days vs. 73.5 ± 28.9 days). The early group had a higher FIM at discharge (84.9 ± 28.0% vs. 65.1 ± 29.0%, P < 0.001) and higher FIM gain (36.9 ± 19.8% vs. 26.8 ± 22.3, P = 0.013) than did the delayed group. Multiple regression analysis showed that the early group was associated with FIM gain (coefficient = 8.607, P = 0.032). CONCLUSIONS: Early wearing of KAFO, irrespective of the difference in ADL at the time of KAFO wearing, may have a positive impact on the improvement of ADL in patients after stroke.

Poor oral health and anorexia in older rehabilitation patients.

OBJECTIVE: To investigate the association between anorexia and comprehensive oral health status in older inpatients. BACKGROUND: Anorexia in older inpatients is a major concern, but whether it is associated with oral problems is currently unclear. METHODS: This cross-sectional study included 160 participants (42.5% men) aged ≥65 years (mean age 78.6 ± 7.9) who had been admitted to a rehabilitation hospital. A score of ≤14 on the Simplified Nutritional Appetite Questionnaire for Japanese Elderly indicated anorexia. A score of ≥3 on the Oral Health Assessment Tool (OHAT) indicated poor oral health. Malnutrition was diagnosed according to the criteria set out by the Global Leadership Initiative on Malnutrition. Multivariate logistic regression was used to investigate the association between poor oral health and anorexia and, additionally, which subcategory of the OHAT (represented by a score ≥1) was associated with anorexia. RESULTS: Anorexia and poor oral health status were observed in 86 (53.8%) and 85 (53.1%), respectively. Poor oral health was associated with anorexia after adjusting for potential confounders (adjusted odds ratio [AOR] 2.7; 95% confidence interval [CI]: 1.3-5.9). Additionally, poor status of dentures (AOR 2.6; 95% CI: 1.2-5.7) and poor oral cleanliness (AOR 3.0; 95% CI: 1.4-6.4) were independently associated with anorexia. CONCLUSIONS: Poor oral health was associated with anorexia in older inpatients. Detection of poor oral health status using a comprehensive oral health assessment may be useful for anorexic patients. Early detection for poor oral health using comprehensive oral health assessments and oral care and prosthetic treatment may be useful for anorexic patients.

Cell-to-cell contact-mediated regulation of tumor behavior in the tumor microenvironment.

Tumor growth and progression are complex processes mediated by mutual interactions between cancer cells and their surrounding stroma that include diverse cell types and acellular components, which form the tumor microenvironment. In this environment, direct intercellular communications play important roles in the regulation of the biological behaviors of tumors. However, the underlying molecular mechanisms are insufficiently defined. We used an in vitro coculture system to identify genes that were specifically expressed at higher levels in cancer cells associated with stromal cells. Major examples included epithelial membrane protein 1 (EMP1) and stomatin, which positively and negatively regulate tumor progression, respectively. EMP1 promotes tumor cell migration and metastasis via activation of the small GTPase Rac1, while stomatin strongly suppresses cell proliferation and induces apoptosis of cancer cells via inhibition of Akt signaling. Here we highlight important aspects of EMP1, stomatin, and their family members in cancer biology. Furthermore, we consider the molecules that participate in intercellular communications and signaling transduction between cancer cells and stromal cells, which may affect the phenotypes of cancer cells in the tumor microenvironment.

Identification of transmembrane protein 168 mutation in familial Brugada syndrome.

Brugada syndrome (BrS) is an inherited channelopathy responsible for almost 20% of sudden cardiac deaths in patients with nonstructural cardiac diseases. Approximately 70% of BrS patients, the causative gene mutation(s) remains unknown. In this study, we used whole exome sequencing to investigate candidate mutations in a family clinically diagnosed with BrS. A heterozygous 1616G>A substitution (R539Q mutation) was identified in the transmembrane protein 168 (TMEM168) gene of symptomatic individuals. Similar to endogenous TMEM168, both TMEM168 wild-type (WT) and mutant proteins that were ectopically induced in HL-1 cells showed nuclear membrane localization. A significant decrease in Na+ current and Nav 1.5 protein expression was observed in HL-1 cardiomyocytes expressing mutant TMEM168. Ventricular tachyarrhythmias and conduction disorders were induced in the heterozygous Tmem168 1616G>A knock-in mice by pharmacological stimulation, but not in WT mice. Na+ current was reduced in ventricular cardiomyocytes isolated from the Tmem168 knock-in heart, and Nav 1.5 expression was also impaired. This impairment was dependent on increased Nedd4-2 binding to Nav 1.5 and subsequent ubiquitination. Collectively, our results show an association between the TMEM168 1616G>A mutation and arrhythmogenesis in a family with BrS.

Mutant KCNJ3 and KCNJ5 Potassium Channels as Novel Molecular Targets in Bradyarrhythmias and Atrial Fibrillation.

BACKGROUND: Bradyarrhythmia is a common clinical manifestation. Although the majority of cases are acquired, genetic analysis of families with bradyarrhythmia has identified a growing number of causative gene mutations. Because the only ultimate treatment for symptomatic bradyarrhythmia has been invasive surgical implantation of a pacemaker, the discovery of novel therapeutic molecular targets is necessary to improve prognosis and quality of life. METHODS: We investigated a family containing 7 individuals with autosomal dominant bradyarrhythmias of sinus node dysfunction, atrial fibrillation with slow ventricular response, and atrioventricular block. To identify the causative mutation, we conducted the family-based whole exome sequencing and genome-wide linkage analysis. We characterized the mutation-related mechanisms based on the pathophysiology in vitro. After generating a transgenic animal model to confirm the human phenotypes of bradyarrhythmia, we also evaluated the efficacy of a newly identified molecular-targeted compound to upregulate heart rate in bradyarrhythmias by using the animal model. RESULTS: We identified one heterozygous mutation, KCNJ3 c.247A>C, p.N83H, as a novel cause of hereditary bradyarrhythmias in this family. KCNJ3 encodes the inwardly rectifying potassium channel Kir3.1, which combines with Kir3.4 (encoded by KCNJ5) to form the acetylcholine-activated potassium channel ( IKACh channel) with specific expression in the atrium. An additional study using a genome cohort of 2185 patients with sporadic atrial fibrillation revealed another 5 rare mutations in KCNJ3 and KCNJ5, suggesting the relevance of both genes to these arrhythmias. Cellular electrophysiological studies revealed that the KCNJ3 p.N83H mutation caused a gain of IKACh channel function by increasing the basal current, even in the absence of m2 muscarinic receptor stimulation. We generated transgenic zebrafish expressing mutant human KCNJ3 in the atrium specifically. It is interesting to note that the selective IKACh channel blocker NIP-151 repressed the increased current and improved bradyarrhythmia phenotypes in the mutant zebrafish. CONCLUSIONS: The IKACh channel is associated with the pathophysiology of bradyarrhythmia and atrial fibrillation, and the mutant IKACh channel ( KCNJ3 p.N83H) can be effectively inhibited by NIP-151, a selective IKACh channel blocker. Thus, the IKACh channel might be considered to be a suitable pharmacological target for patients who have bradyarrhythmia with a gain-of-function mutation in the IKACh channel.

Impact of Multiple Texture-Modified Diets on Oral Intake and Nutritional Status in Older Patients with Pneumonia: A Retrospective Cohort Study.

Texture-modified diets (TMD) is often used in clinical practices for the treatment and prevention of pneumonia. However, it is unclear how stages of TMD affect the swallowing ability and nutritional status in patients with pneumonia. This study aimed to investigate the relationship between the various stages of TMD and swallowing ability and nutritional status in older inpatients with pneumonia. In this retrospective cohort study, data for patients aged ≥ 65 years with pneumonia were obtained from the Japan Rehabilitation Nutrition Database. We performed coarsened exact matching with Mini Nutritional Assessment Short Form (MNA-SF) on admission. Ultimately, 218 patients (mean age 82.9 ± 9.8 years) were included and divided into two groups based on the stages of TMD in the facility: multiple TMD (M-TMD) group (stages of TMD ≥ 6) and control group (stages of TMD < 6). The main outcome was the rate of improvement in the Food Intake Level Scale (FILS) and the maintenance or improvement in the MNA-SF score. We used the within-hospital correction with generalized estimation equations that are commonly used to analyze clustered data while correcting for confounding factors by clustering. Multivariate multiple logistic analysis showed that M-TMD was independently associated with FILS improvement rate and the maintenance or improvement in the MNA-SF score (odds ratio [OR] 3.252; 95% confidence interval [CI] 1.602-6.601; p = 0.001 and OR 1.873; 95% CI 1.054-3.330; p = 0.032, respectively). M-TMD in the facility was associated with the maintenance or improvement in swallowing ability and the nutritional status of patients with pneumonia.

The sbGnRH-GTH system in the female short mackerel, Rastrelliger brachysoma (Bleeker, 1851), during breeding season: implications for low gamete production in captive broodstock.

The short mackerel (Rastrelliger brachysoma) is one of the most economically important fish in Thailand; it is also a prime candidate for mariculture but unfortunately is plagued by reproductive problems that cause low production of gametes in captivity. An understanding of how the brain, pituitary, and gonad axis (BPG) from the neuroendocrine system are involved in the reproductive activity of wild and captive R. brachysoma should help clarify the situation. In this study, we investigated changes in the sea bream gonadotropin-releasing hormone (sbGnRH)-gonadotropin (GTH) system in the female short mackerel, Rastrelliger brachysoma (Bleeker, 1851), during breeding season. sbGnRH-immunoreactive (ir) cell bodies were detected in the nucleus preopticus-periventricularis including nucleus periventricularis (NPT), nucleus preopticus (Np), and nucleus lateralis tuberis (NLT). Additionally, the sbGnRH-ir fibers protruded into the proximal par distalis (PPD) where GTH (FSH and LH) cells were detected. The number of sbGnRH-ir cell bodies and GTH cells and level of LH mRNA were significantly higher in the breeding season than those in the non-breeding season. Moreover, the number of sbGnRH-ir cell bodies and GTH cells and levels of sbGnRH and GTH (FSH and LH) mRNA were significantly higher in the wild fish than those in the cultured broodstock. It is suggested that the wild fish tended to have better reproductive system than hatchery fishes. This could be related to the endocrinological dysfunction and the reproductive failure in the hatchery condition. Moreover, the changes of all of the hormonal level could potentially be applied to R. brachysoma aquaculture.