Immunization with inactivated whole virus particle influenza virus vaccines improves the humoral response landscape in cynomolgus macaques.

Although antibody-inducing split virus vaccines (SV) are currently the most effective way to combat seasonal influenza, their efficacy can be modest, especially in immunologically-naïve individuals. We investigated immune responses towards inactivated whole influenza virus particle vaccine (WPV) formulations, predicated to be more immunogenic, in a non-human primate model, as an important step towards clinical testing in humans. Comprehensive analyses were used to capture 46 immune parameters to profile how WPV-induced responses differed to those elicited by antigenically-similar SV formulations. Naïve cynomolgus macaques vaccinated with either monovalent or quadrivalent WPV consistently induced stronger antibody responses and hemagglutination inhibition (HI) antibody titres against vaccine-matched viruses compared to SV formulations, while acute reactogenic effects were similar. Responses in WPV-primed animals were further increased by boosting with the same formulation, conversely to modest responses after priming and boosting with SV. 28-parameter multiplex bead array defined key antibody features and showed that while both WPV and SV induced elevated IgG responses against A/H1N1 nucleoprotein, only WPV increased IgG responses against A/H1N1 hemagglutinin (HA) and HA-Stem, and higher IgA responses to A/H1N1-HA after each vaccine dose. Antibodies to A/H1N1-HA and HA-Stem that could engage FcγR2a and FcγR3a were also present at higher levels after one dose of WPV compared to SV and remained elevated after the second dose. Furthermore, WPV-enhanced antibody responses were associated with higher frequencies of HA-specific B-cells and IFN-γ-producing CD4+ T-cell responses. Our data additionally demonstrate stronger boosting of HI titres by WPV following prior infection and support WPV administered as a priming dose irrespective of the follow up vaccine for the second dose. Our findings thus show that compared to SV vaccination, WPV-induced humoral responses are significantly increased in scope and magnitude, advocating WPV vaccination regimens for priming immunologically-naïve individuals and also in the event of a pandemic outbreak.

Association of oral frailty with medical expenditure in older Japanese adults: The study of late-stage older adults in Tottori (START Tottori).

OBJECTS: This study aimed to determine the association between annual medical expenses and oral frailty in later-stage older adults (aged ≥ 75 years). No studies have investigated the association between medical costs and oral frailty, which would elucidate the association between oral frailty and the deterioration of mental and overall physical function. MATERIALS AND METHODS: In this cross-sectional study, 2190 adults (860 men and 1330 women aged 75-94 years) covered by the Medical System for the Elderly and residing in Tottori Prefecture, Japan, between April 2016 and March 2019, were included. Participants were classified into three groups: healthy, pre-orally frail or orally frail, based on dental health screening findings. The medical and dental expenses over the years, number of days of consultations and comorbidities were obtained from the Japanese Health Insurance Claims Database. RESULTS: The number of days of medical and dental consultations and annual medical expenses for outpatient care differed among the three study groups. A significant association was observed between oral frailty and high annual expenses for outpatient medical and dental care. Oral frailty was associated with higher medical expenses in participants with poor masticatory function. Higher and lower dental expenses were associated with subjective poor masticatory function and subjective impairment of swallowing function respectively. CONCLUSION: Medical and dental expenses for orally frail older adults are high, indicating that oral frailty may be related to the occurrence and severity of diseases other than oral health issues. Future studies should examine the mechanism by which oral weakness affects physical and mental functions.

Serum levels of vitamin D and periodontal inflammation in community-dwelling older Japanese adults: The Otassha Study.

AIM: To evaluate the association between vitamin D status and periodontal inflammation as determined by the periodontal inflamed surface area (PISA) in community-dwelling older adults. MATERIALS AND METHODS: This cross-sectional study included 467 Japanese adults (mean age = 73.1 years) who underwent full-mouth periodontal examinations and measurements of serum levels of 25-hydroxyvitamin D (25(OH)D). We used linear regression and restricted cubic spline models to analyse the association between exposure (serum 25(OH)D) and outcome (PISA). RESULTS: The linear regression model showed that, after adjusting for potential confounders, participants in the lowest quartile of serum 25(OH)D had 41.0 mm2 more PISA (95% confidence interval [CI]: 4.6-77.5) than the reference group (the highest quartile of serum 25(OH)D). The spline model showed that the association between serum 25(OH)D and PISA was non-linear and restricted to the low 25(OH)D range. PISA initially sharply decreased as serum 25(OH)D increased, and then the decreasing trend slowed and plateaued. The inflection point with the minimum PISA value was a serum 25(OH)D level of 27.1 ng/mL, above which there was no decreasing trend in PISA with increasing serum 25(OH)D levels. CONCLUSIONS: Low vitamin D status had an L-shaped association with periodontal inflammation in this cohort of Japanese adults.

Predictors of early and late mortality after the treatment for early gastric cancers.

OBJECTIVES: Although many patients with early gastric cancers (EGCs) die of non-gastric cancer-related causes, the association of the risk categories of lymph node metastasis (LNM) with all-cause mortality remains unclear. We aimed to clarify the predictors of early and late mortality, separately. METHODS: Patients with endoscopic resection or gastrectomy for EGCs between 2003 and 2017 were retrospectively enrolled. We analyzed predictors for early and late mortality, including risk categories of LNM, treatment method, and nine non-cancer-related indices, separately, with a cut-off value of 3 years. RESULTS: We enrolled 1439 patients with a median follow-up period of 79 months. The 5-year overall survival rate was 86.8%. In the multivariate Cox analysis, the most important predictors for early and late mortality were age ≥85 years (hazard ratio [HR] 2.88 and 4.54, respectively) and Eastern Cooperative Oncology Group Performance Status ≥2 (HR 3.00 and 4.19, respectively). Charlson comorbidity index ≥2 (HR 2.76 and 1.99, respectively), American Society of Anesthesiologists Physical Status ≥3 (HR 2.35 and 1.79, respectively), and C-reactive protein/albumin ratio ≥0.028 (HR 2.30 and 1.58, respectively) were also predictors for both early and late mortality. Male (HR 2.26), intermediate- (HR 2.12)/high-risk (HR 1.85) of LNM in eCura system, and sarcopenia evaluated by the psoas muscle mass index (HR 1.70) were predictors for early mortality. CONCLUSION: The combined assessment of multiple predictors might help to predict early and/or late mortality in patients with EGCs. The eCura system was associated with early mortality.

Association between dry mouth and physical frailty among community-dwelling older adults in Japan: The Otassha Study.

OBJECTIVE: To elucidate the relationship between physical frailty and dry mouth among community-dwelling older adults in Japan. BACKGROUND: Oral hypofunction is associated with physical frailty in older adults. Dry mouth, subjectively evaluated as xerostomia and objectively measured as hyposalivation, is highly prevalent in older adults. However, few reports have used both measures of dry mouth and investigated their association with physical frailty. MATERIALS AND METHODS: This cross-sectional study included 643 individuals (mean age, 73.9 years) from the Otassha Study in 2019. Xerostomia was assessed using the 5-item Summated Xerostomia Inventory (SXI). Hyposalivation was assessed by measuring the amount of resting saliva using the modified cotton roll method. Frailty phenotypes were defined by weakness, slowness, weight loss, low physical activity level and exhaustion components, and served as the outcome variable. Participants with three or more components were considered physically frail, and those with one or two components were considered pre-frail. Ordinal logistic regression analyses were performed to evaluate the associations between physical frailty and xerostomia and hyposalivation. RESULTS: Among the participants, 4.5% and 49.8% had physical frailty and pre-frailty, respectively. In ordinal logistic regression analysis adjusted for demographic and health characteristics, the SXI score was associated with physical frailty status (adjusted odds ratio for a one-point increase in SXI, 1.12; 95% confidence interval, 1.06-1.19). However, no difference was observed between the amount of resting saliva and severity of physical frailty. CONCLUSION: Xerostomia is associated with physical frailty among community-dwelling older adults in Japan. CLINICAL TRIAL REGISTRATION: Not applicable.

Oral hypofunction and malnutrition among community-dwelling older adults: Evidence from the Otassha study.

OBJECTIVE: This study investigated the association between oral hypofunction and malnutrition among community-dwelling older adults. BACKGROUND: Oral hypofunction, a 7-component phenotype of the clinical features of oral health, has been described in Japan. Data about its association with nutritional status are limited. MATERIALS AND METHODS: This cross-sectional study enrolled 715 adults (mean age, 73.5 years; range, 65-91 years) from the Otassha Study. Oral hypofunction was defined as the presence of ≥3 of the following 7 components: poor oral hygiene; oral dryness; low occlusal force; low articulatory oral motor skill; low tongue pressure; low masticatory performance; and compromised swallowing function. The Mini Nutritional Assessment® -Short Form (MNA® -SF) was used to assess the nutritional status. Malnutrition was defined as having an MNA® -SF score of seven or less. The association between oral hypofunction and nutritional status was evaluated using ordinal logistic regression analyses. RESULTS: The prevalence rates of oral hypofunction and malnutrition were 42.7% and 4.1%, respectively. Multivariable Poisson regression analyses revealed that oral hypofunction was associated with malnutrition. The adjusted relative risk of malnutrition in the study participants with oral hypofunction was 3.00 with a 95% confidence interval of 1.29-6.98. CONCLUSION: Community-dwelling older adults whose oral functions were compromised in multiple aspects had poor nutritional status.

Pneumonia incidence and oral health management by dental hygienists in long-term care facilities: A 1-year prospective multicentre cohort study.

OBJECTIVE: To investigate the association between oral health management (OHM) by dental hygienists and the occurrence of pneumonia, and determine the effectiveness of OHM in pneumonia prevention. BACKGROUND: In long-term care facilities in Japan, the need for professional OHM is increasing with an increase in the number of severely debilitated residents. MATERIALS AND METHODS: A 1-year prospective multicentre cohort study was conducted using data from 504 residents (63 men; mean age: 87.4 ± 7.8 years) in Japanese long-term care facilities. Basic information, medical history, willingness to engage in oral hygiene behaviour, need for OHM and oral conditions were investigated at baseline. In addition, information on the occurrence of pneumonia was collected using a follow-up survey after one year. A Poisson regression analysis with robust standard errors was conducted, with pneumonia as the dependent variable, and factors associated with OHM and pneumonia occurrence as explanatory variables. RESULTS: Overall, 349 (69.2%) residents required OHM by dental hygienists during that year of follow-up. Of those, 238 (68.2%) were provided with OHM, and 18 (7.5%) developed pneumonia. Among the 111 patients (31.8%) who were not provided with OHM, 21 (18.9%) developed pneumonia. The OHM group had lower pneumonia rates than the non-OHM group (prevalence rate ratio: 0.374; 95% CI: 0.210-0.665). CONCLUSION: Oral health management by dental hygienists was associated with a lower incidence of pneumonia among residents of long-term care facilities, underlining the importance of professional OHM for such individuals. It is recommended that OHM be practised routinely in long-term care facilities.

Immune profiling of influenza-specific B- and T-cell responses in macaques using flow cytometry-based assays.

Influenza remains a significant global public health burden, despite substantial annual vaccination efforts against circulating virus strains. As a result, novel vaccine approaches are needed to generate long-lasting and universal broadly cross-reactive immunity against distinct influenza virus strains and subtypes. Several new vaccine candidates are currently under development and/or in clinical trials. The successful development of new vaccines requires testing in animal models, other than mice, which capture the complexity of the human immune system. Importantly, following vaccination or challenge, the assessment of adaptive immunity at the antigen-specific level is particularly informative. In this study, using peripheral blood mononuclear cells (PBMCs) from cynomolgus macaques, we describe detection methods and in-depth analyses of influenza virus-specific B cells by recombinant hemagglutinin probes and flow cytometry, as well as the detection of influenza virus-specific CD8+ and CD4+ T cells by stimulation with live influenza A virus and intracellular cytokine staining. We highlight the potential of these assays to be used with PBMCs from other macaque species, including rhesus macaques, pigtail macaques and African green monkeys. We also demonstrate the use of a human cytometric bead array kit in detecting inflammatory cytokines and chemokines from cynomolgus macaques to assess cytokine/chemokine milieu. Overall, the detection of influenza virus-specific B and T cells, together with inflammatory responses, as described in our study, provides useful insights for evaluating novel influenza vaccines. Our data deciphering immune responses toward influenza viruses can be also adapted to understanding immunity to other infections or vaccination approaches in macaque models.

IgA tetramerization improves target breadth but not peak potency of functionality of anti-influenza virus broadly neutralizing antibody.

Mucosal immunoglobulins comprise mainly secretory IgA antibodies (SIgAs), which are the major contributor to pathogen-specific immune responses in mucosal tissues. These SIgAs are highly heterogeneous in terms of their quaternary structure. A recent report shows that the polymerization status of SIgA defines their functionality in the human upper respiratory mucosa. Higher order polymerization of SIgA (i.e., tetramers) leads to a marked increase in neutralizing activity against influenza viruses. However, the precise molecular mechanisms underlying the effects of SIgA polymerization remain elusive. Here, we developed a method for generating recombinant tetrameric monoclonal SIgAs. We then compared the anti-viral activities of these tetrameric SIgAs, which possessed variable regions identical to that of a broadly neutralizing anti-influenza antibody F045-092 against influenza A viruses, with that of monomeric IgG or IgA. The tetrameric SIgA showed anti-viral inhibitory activity superior to that of other forms only when the antibody exhibits low-affinity binding to the target. By contrast, SIgA tetramerization did not substantially modify anti-viral activity against targets with high-affinity binding. Taken together, the data suggest that tetramerization of SIgA improved target breadth, but not peak potency of antiviral functions of the broadly neutralizing anti-influenza antibody. This phenomenon presumably represents one of the mechanisms by which SIgAs present in human respiratory mucosa prevent infection by antigen-drifted influenza viruses. Understanding the mechanisms involved in cross neutralization of viruses by SIgAs might facilitate the development of vaccine strategies against viral infection of mucosal tissues.

Interrelationships among whole-body skeletal muscle mass, masseter muscle mass, oral function, and dentition status in older Japanese adults.

BACKGROUND: Generalized loss of skeletal muscle mass (SMM) may modulate or otherwise affect the loss of masseter muscle mass and be responsible for low masseter muscle performance and strength (i.e., low oral function). Moreover, dentition status can affect oral function independent of the muscle state. This cross-sectional study aimed to simultaneously investigate the relationships among whole-body SMM, masseter muscle mass, oral function (masseter muscle performance and strength), and dentition status in 1349 Japanese adults (mean age = 73.6 years). METHODS: We determined the estimated masseter muscle mass (e-MMM) based on morphological measurements of the masseter muscle. Masseter muscle performance was assessed via masticatory performance evaluation scores using gum, and strength was assessed as the maximal occlusal force. Dentition status was assessed as the number of functional teeth. SMM was measured by bioelectrical impedance analysis. Structural equation modeling stratified by sex was employed to investigate associations among SMM, e-MMM, gum score, occlusal force, and number of functional teeth. RESULTS: The direct path from SMM to e-MMM was statistically significant, as was the direct path from e-MMM to oral function (gum score and maximum occlusal force) for both sexes. We additionally confirmed that SMM indirectly affected gum score and maximum occlusal force via e-MMM (men; standardized coefficient [95% CI] = 3.64 [1.31 to 5.96] for maximum occlusal force and 0.01 [0.01 to 0.02] for gum score, women; 2.01 [0.38 to 3.81] for maximum occlusal force and 0.01 [0.002 to 0.01] for gum score). The number of functional teeth had direct effects on e-MMM, gum score, and maximum occlusal force. CONCLUSIONS: Low SMM was significantly indirectly associated with poor oral function through a low masseter muscle mass, and dentition status was independently associated with oral function.

[The oral function of community-dwelling older adults complaining of an oral function deterioration: An examination using the oral function-related items of the Questionnaire for Latter-stage Elderly People].

AIM: The oral function-related items of the newly developed "Questionnaire for Latter-stage Elderly People" are based on two items of the Kihon checklist assessing the masticatory and swallowing functions. In this study, we estimated the questionnaire's application rate and investigated the oral function of the respondents. METHODS: Included were 699 older adults (274 men/425 women, average age, 73.4±6.6 years old). Those who responded positively to the related items were considered to have decreased swallowing and masticatory functions. The specific oral function was assessed based on the number of present and functional teeth; oral hygiene; oral moisture; occlusal force; oral diadochokinesis /pa/, /ta/, /ka/; tongue pressure; mixing ability; shearing ability; and Eating Assessment Tool (EAT)-10 (reference, score ≥3). RESULTS: The rates of decreased masticatory and swallowing functions were 21.5% and 26.6%, respectively, while 7.4% of participants had both. Those with a decreased masticatory function showed fewer present teeth; a lower occlusal force, oral diadochokinesis /pa/, mixing ability, shearing ability, and higher EAT-10 scores.Those with a decreased swallowing function only had higher EAT-10 scores. In the early- and latter-stage elderly, the decreased masticatory function rates were 15.6% and 29.4%, respectively, the decreased swallowing function rates were 27.8% and 25.0%, respectively, and the rates of both decreased masticatory and swallowing functions were 6.0% and 9.5%, respectively. CONCLUSION: The older adults who responded positively to the related questionnaire items, especially for items related to masticatory function, had a decreased oral function in multiple respects. The items related to the oral function in the Questionnaire for Latter-stage Elderly People are considered useful for identifying older adults with a decreased oral function.

[Relationship between the rate of a decreased oral function and the nutrient intake in community-dwelling older persons: An examination using oral function-related items in a questionnaire for latter-stage elderly people].

AIM: To determine the rate of a decreased oral function using questions from the Kihon checklist corresponding to the Questionnaire for Latter-stage Elderly People and to clarify nutrient intake in older persons. METHODS: This study targeted 511 older people (217 men, 294 women, average age 73.1±5.6 years old). Their oral function was evaluated using questions on the masticatory function and swallowing function from the Kihon checklist, corresponding to questions on the oral function in the Questionnaire for Latter-stage Elderly People. Participants who had at least one symptom measured were defined as the applicable group (AG). In addition, to evaluate the nutrient intake of the participants, interviews were conducted using the Food Frequency Questionnaire Based on Food Groups. RESULTS: The rate of inclusion in the AG was 32.9% for the total sample, 28.2% for early-stage elderly people, and 40.1% for latter-stage elderly people. The AG rates did not differ significantly between men and women. For latter-stage elderly people, the protein-energy ratio and intakes of total energy, protein, pantothenic acid, folic acid, vitamin B6, niacin, vitamin K, copper, zinc, phosphorus, magnesium, potassium, and total dietary fiber were significantly lower in the AG than in the non-AG. CONCLUSION: The evaluation of placement in the AG through questions on the oral function from the Kihon checklist corresponding to the Questionnaire for Latter-stage Elderly People demonstrated that the rate of a decreased oral function was higher in latter-stage elderly people than in early-stage elderly people. In addition, the latter-stage elderly people in the AG had a lower nutrient intake.

Association between anorexia and hyposalivation in community-dwelling older adults in Japan: a 6-year longitudinal study.

BACKGROUND: Hyposalivation is associated with the nutritional status. Anorexia of ageing, defined as an age-related decrease in appetite and food intake, presents even in healthy adults and is considered an independent predictor of malnutrition, frailty, and mortality. However, the relationship between anorexia and hyposalivation of ageing is unclear. Thus, the present longitudinal study aimed to investigate the incidence of hyposalivation and its relationship with anorexia in community-dwelling older people in Japan. METHODS: The study population comprised 220 individuals (80 men and 140 women) aged 65-86 years at baseline. The participants underwent comprehensive health check-ups, including dental examinations and anthropometry, and face-to-face interviews in 2013 and 2019. Hyposalivation was determined on the basis of the unstimulated salivary flow rate measured using the modified cotton roll method. Anorexia was defined as a score of ≤29 in the Japanese version of the Council on Nutrition Appetite Questionnaire. Logistic regression analyses were used to test whether the presence of anorexia at baseline was an independent predictor of hyposalivation. RESULTS: Hyposalivation developed at a rate of 19.5% during the 6-year observation period. Anorexia was observed in 95 (43.2%) participants at baseline. After adjusting for potential confounding factors, anorexia (adjusted odds ratio [AOR], 2.65; 95% confidence interval [CI], 1.26-5.57) and polypharmacy (AOR, 3.29; CI, 1.06-10.19) were significant predictors of hyposalivation. CONCLUSION: Loss of appetite is independently correlated with and a risk factor for hyposalivation in older adults. Anorexia of ageing may have negative effects on the salivary flow rate in such settings. Salivation should be a standard feature in clinical assessments of the older adults.

Association among kidney function, frailty, and oral function in patients with chronic kidney disease: a cross-sectional study.

BACKGROUND: Chronic kidney disease (CKD) involves many factors that can cause frailty and oral hypofunction. We aimed to investigate the prevalence of frailty and oral hypofunction and to examine the associations among kidney function, frailty, and oral function in adults with CKD in Japan. METHODS: This cross-sectional study was conducted at two institutions. The participants included 109 patients with CKD stages 3-5 who visited outpatient clinics or were admitted for inpatient treatment. Frailty was evaluated using the Japanese version of the Cardiovascular Health Study frailty criteria. Oral function was evaluated by assessing oral motor skills [oral diadochokinesis (ODK) rate], masticatory ability, and the repetitive saliva swallowing test. The estimated glomerular filtration rate (eGFR) was used to indicate kidney function. We examined the associations among kidney function, frailty, and oral function using binomial logistic regression analysis. RESULTS: In total, 31 participants (28.4%) were classified as being frail. Univariate analysis showed that age, body mass index, eGFR, and haemoglobin level were significantly associated with frailty. ODK and swallowing function were significantly associated with frailty. Multivariate analysis revealed that frailty was significantly associated with eGFR [odds ratio (OR) 0.96, 95% confidence interval (CI) 0.92-1.00, p = 0.048] and ODK rate (OR 0.68, CI 0.47-0.98, p = 0.038). However, no significant association was found between CKD severity and masticatory or swallowing function. CONCLUSION: We found a high prevalence of frailty in patients with CKD and a significant association between frailty and oral motor skills, affecting the swallowing function of patients with nondialysis CKD. The high prevalence of frailty among patients with CKD suggests that routine assessment of frailty is necessary to prevent the development of severe complications. In addition, oral and kidney function should be carefully evaluated, and oral health education and interventions should be performed for patients with CKD.

Development of a simple method to measure masseter muscle mass.

OBJECTIVE: To develop a simple method to estimate masseter muscle mass. BACKGROUND: The masseter muscle is important for optimal oral function. A decrease in the masseter muscle mass may affect the oral and physical function of the whole body; therefore, it is an important entity to measure. However, to date, no easy measurement method has been developed for the same. Herein, we devised a simple method to estimate masseter muscle mass. In addition, we compared our method with magnetic resonance imaging (MRI) and skeletal muscle mass (SMM) to verify its validity. MATERIALS AND METHODS: We recorded the age, sex, masseter muscle mass (by MRI), number of retained natural teeth, number of functional teeth, length and thickness of masseter muscle, and skeletal muscle mass in 53 community-dwelling elderly people (21 men and 32 women, average age 72.9 ± 4.5 years). The estimated masseter muscle mass was calculated by multiplying the length, width and thickness of the muscle. RESULTS: The correlation coefficient between masseter muscle mass and estimated masseter muscle mass was r = .903; thus, a strong correlation was recognised. The correlation coefficient between the estimated masseter muscle mass and SMM was r = .279, considering age and sex. CONCLUSION: We obtained substantial results using our method for estimating the masseter muscle mass and verified its validity by comparing it with masseter muscle mass measured by MRI and SMM. We believe that our proposed technique is simple and useful for estimating masseter muscle mass.

pH-Responsive Polyion Complex Vesicle with Polyphosphobetaine Shells.

When a bioactive molecule is taken into cells by endocytosis, it is sometimes unable to escape from the lysosomes, resulting in inefficient drug release. We prepared pH-responsive polyion complex (PIC) vesicles that collapse under acidic conditions such as those inside a lysosome. Furthermore, under acidic conditions, cationic polymer was released from the PIC vesicles to break the lysosome membranes. Diblock copolymers (P20M167 and P20A190) consisting of water-soluble zwitterionic poly(2-methacryloyloxyethyl phosphorylcholine) (PMPC) block and cationic or anionic blocks were synthesized via reversible addition-fragmentation chain transfer (RAFT) radical polymerization. Poly(3-(methacrylamidopropyl) trimethylammonium chloride) (PMAPTAC) and poly(sodium 6-acrylamidohexanoate) (PAaH) were used as the cationic and anionic blocks, respectively. The pendant hexanoate groups in the PAaH block are ionized in basic water and in phosphate buffered saline (PBS), while the hexanoate groups are protonated in acidic water. In basic water, PIC vesicles were formed from a charge neutralized mixture of oppositely charged diblock copolymers. At the interface of PIC vesicle and water exists biocompatible PMPC shells. Under acidic conditions, the PIC vesicles collapsed, because the charge balance shifted due to protonation of the PAaH block. After collapse of the PIC vesicles, P20A190 formed micelles composed of protonated PAaH core and PMPC shells, while P20M167 was released as unimers. PIC vesicles can encapsulate hydrophilic nonionic guest molecules into their hollow core. Under acidic conditions, the PIC vesicles can release the guest molecules and P20M167. The cationic P20M167 can break the lysosome membrane to efficiently release the guest molecules from the lysosomes to the cytoplasm.

Qualitative analysis of the vocabulary used in work logs of a preventive programme for elderly oral function and nutrition.

BACKGROUND: In Japan, day care services for elders include programmes aimed at improving nutrition and oral and motor functions. Few studies have qualitatively assessed these interventions. OBJECTIVE: To qualitatively search for the characteristic words used in the work logs of a preventive programme on oral function and nutrition for elders by intervention period and intervention type． METHODS: We included 83 participants (81.3 ± 8.2 years) from four day care services in Japan and divided them into the following groups randomly: those who received oral function intervention only, nutritional intervention only and those who received combined oral function plus nutritional intervention. The interventions were conducted twice per month for 24 months. Data from handwritten work logs were entered into a computer as text files. Monitoring of frequently appearing words, co-occurrence analysis and cross-tabulation by intervention period and intervention types was conducted using text mining analysis. RESULTS: Correspondence analysis revealed that the words used during 1-6 months and 7-12 months were similar in participants' subjective content, and those used in objective content in 13-18 months and 19-24 months were similar. These results indicate that subjective improvements increased after 13 months, and it was maintained within 24 months. The combined intervention type is ideal for oral and nutrition problems. CONCLUSION: Because this text mining approach revealed the changes in the words used and could be used to monitor any subjective improvement, this approach may help evaluate the effects of preventive care.

Reversible bacterial immobilization based on the salt-dependent adhesion of the bacterionanofiber protein AtaA.

BACKGROUND: Immobilization of microbial cells is an important strategy for the efficient use of whole-cell catalysts because it simplifies product separation, enables the cell concentration to be increased, stabilizes enzymatic activity, and permits repeated or continuous biocatalyst use. However, conventional immobilization methods have practical limitations, such as limited mass transfer in the inner part of a gel, gel fragility, cell leakage from the support matrix, and adverse effects on cell viability and catalytic activity. We previously showed a new method for bacterial cell immobilization using AtaA, a member of the trimeric autotransporter adhesin family found in Acinetobacter sp. Tol 5. This approach is expected to solve the drawbacks of conventional immobilization methods. However, similar to all other immobilization methods, the use of support materials increases the cost of bioprocesses and subsequent waste materials. RESULTS: We found that the stickiness of the AtaA molecule isolated from Tol 5 cells is drastically diminished at ionic strengths lower than 10 mM and that it cannot adhere in deionized water, which also inhibits cell adhesion mediated by AtaA. Cells immobilized on well plates and polyurethane foam in a salt solution were detached in deionized water by rinsing and shaking, respectively. The detached cells regained their adhesiveness in a salt solution and could rapidly be re-immobilized. The cells expressing the ataA gene maintained their adhesiveness throughout four repeated immobilization and detachment cycles and could be repeatedly immobilized to polyurethane foam by a 10-min shake in a flask. We also demonstrated that both bacterial cells and a support used in a reaction could be reused for a different type of reaction after detachment of the initially immobilized cells from the support and a subsequent immobilization step. CONCLUSIONS: We invented a unique reversible immobilization method based on the salt-dependent adhesion of the AtaA molecule that allows us to reuse bacterial cells and supports by a simple manipulation involving a deionized water wash. This mitigates problems caused by the use of support materials and greatly helps to enhance the efficiency and productivity of microbial production processes.

Factors affecting masticatory function of community-dwelling older people: Investigation of the differences in the relevant factors for subjective and objective assessment.

OBJECTIVE: This study aimed to examine relevant factors for subjective and objective assessment of masticatory functions and elucidate any differences between the two methods. BACKGROUND: Previous studies have reported that the results of subjective and objective assessment of masticatory function in older people do not necessarily agree. MATERIALS AND METHODS: This study included 487 community-dwelling Japanese older people (205 male and 282 female; mean age 74.1±6.3 years) who participated in a comprehensive geriatric health examination. Basic information (gender and age), higher level of competence in daily living, depression, subjective masticatory function (SMF) and objective masticatory function (OMF) assessments, cognitive function, skeletal muscle mass, handgrip strength, gait speed and oral status (number of remaining and functional teeth, mouth dryness and occlusal force) were recorded. RESULTS: Multiple logistic regression analysis showed that depression (odds ratio [OR]: 1.181, 95% confidence interval [CI]: 1.094-1.275), mouth dryness (OR: 2.037, CI: 1.212-3.423) and occlusal force (OR: 0.997, CI: 0.996-0.999) were significantly associated with SMF, whereas higher level of competence in daily living (OR: 0.730 CI: 0.586-0.910), skeletal muscle mass (OR: 0.521 CI: 0.283-0.960), number of functional teeth (OR: 0.862 CI: 0.775-0.959), number of remaining teeth (OR: 0.868 CI: 0.810-0.930) and occlusal force (OR: 0.994, CI: 0.991-0.998) were associated with OMF. CONCLUSION: Subjective masticatory functionSMF and OMF were associated with different factors, suggesting that both mental and physical factors should be taken into consideration when treating decreased masticatory function.

Oral function and nutritional status among the elderly with facial and oral tactile hypersensitivity who are under long-term care.

Objective　In oral health care, a refusal action can cause difficulties, and tactile hypersensitivity (TH) may be a contributing factor. People with TH of the face and mouth clench their jaws. Despite numerous reports on TH among children with disabilities, reports on TH in the elderly who are under long-term care are scarce. The purpose of this study was to investigate TH among the elderly who are under long-term care.Methods　We selected 80 residents (8 men and 72 women; mean age: 91.1±6.2y) in a Tokyo nursing home and investigated the presence of TH in them. We also obtained patients' (1) basic information (sex, age, stage of long-term care needs (SCN), and the degree of independent living (IL)); (2) oral information (swallowing status, choking tendency, and intraoral residue); and (3) nutritional information (serum albumin (Alb) levels and body mass index (BMI)). We assessed the face (the forehead, cheek, and perioral area) and the intraoral environment (buccal mucosa, buccal cavity, and palate) as testing sites for TH, using the tester's palm and forefinger. We confirmed the presence of TH when the tested areas reacted partially or fully by shuddering. We classified the subjects into 2 groups based on the presence or absence of TH and analyzed our results using a chi-square test and Mann-Whitney U test. This investigation was approved by the Dentistry Ethics Screening Committee, Tokyo Medical and Dental University.Results　A total of 18 residents were diagnosed with TH (22.5%). Significant differences in SCN, IL, choking tendency, intraoral residue, swallowing status, serum Alb levels, and BMI (P<0.05) were reported between this group and the non-TH group.Conclusion　We demonstrated that residents with TH were in an advanced SCN and had a lower IL score and a decreased swallowing and nutritional status. Therefore, oral health care, promotion of nutritional status, and meal support are particularly important for the elderly with TH.