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Aims The aims were to determine the effects of subinhibitory concentrations of eight cephem and carbapenem antibiotics on the biofilm formation of Acinetobacter baumannii cells and examine their effects on pre-established biofilms. Methods and Results Effects of antibiotics on biofilm formation were assayed using microtitre plates with polystyrene peg-lids. Cefmetazole, ceftriaxone, ceftazidime and cefpirome increased the biomass of pre-established biofilms on pegs in the range of their sub-minimum inhibitory concentrations (MICs), whereas none increased biofilm formation by planktonic cells. Carbapenems had a negative effect. The constituents of antibiotic-induced biofilms were analysed. Ceftriaxone or ceftazidime treatment markedly increased the matrix constituent amounts in the biofilms (carbohydrate, 2.7-fold; protein, 8.9–12.7-fold; lipid, 3.3–3.6-fold; DNA, 9.1–12.2-fold; outer membrane vesicles, 2.7–3.8-fold and viable cells, 6.8–10.1-fold). The antibiotic-enhanced biofilms had increased outer membrane protein A and were resistant to the anti-biofilm effect of azithromycin. Conclusions Some cephems increased the biomass of pre-established biofilms in the ranges of their sub-MICs. The antibiotic-enhanced biofilms possessed more virulent characteristics than normal biofilms. Significance and Impact of the Study Incomplete administration of certain cephems following biofilm-related Ac. baumannii infections could adversely cause exacerbated and chronic clinical results.

Ehlers–Danlos syndrome spondylodysplastic type 3 (EDSSPD3, OMIM 612350) is an inherited recessive connective tissues disease caused by loss of function of SLC39A13/ZIP13, a zinc transporter belonging to the Slc39a/ZIP family. Patients with EDSSPD3 suffer from impaired development of bone and connective tissues, and muscular hypotonia, or myopathy. However, whether ZIP13 participates in the early differentiation process of these cell types remains unclear. In this study, we investigated the role of ZIP13 in myogenic differentiation using murine myoblast cell line (C2C12) as well as human patient-derived induced pluripotent stem cells (iPSCs). We found that ZIP13 expression was upregulated by myogenic stimulation in C2C12 cells, and its knockdown disrupted myotubular differentiation. Myocytes differentiated from iPSCs of patients with EDSSPD3 (EDSSPD3-iPSCs) exhibited incomplete myogenic differentiation. Moreover, the phenotypic abnormalities of EDSSPD3-iPSC-derived myocytes were corrected by genomic editing of the pathogenic ZIP13 mutation, suggesting the indispensable role of ZIP13 in myogenic differentiation. These results clearly indicate that ZIP13 is required for proper myogenic differentiation and that the study of EDSSPD3-iPSCs may help shed light on the molecular basis underlying various clinical features caused by the loss of ZIP13.

Human-induced pluripotent stem cells (hiPSCs) are useful tools to examine human neuronal maturation processes. In this chapter, we describe the maturation of human neuronal precursor cells derived from hiPSCs by cellular communication network family member 2, also known as connective tissue growth factor. We describe the (1) preparation of feeder cells for undifferentiated culture of hiPSCs, (2) undifferentiated culture of hiPSCs, (3) induction of neuronal precursor cells from hiPSCs, (4) maturation of neuronal precursor cells from hiPSCs, (5) immunofluorescent staining of neuronal cells from hiPSCs, and (6) immunofluorescence analysis.

Low health literacy results in health inequity are linked with poor adherence to medical care. In the globalized Japanese context, the number of migrants with Japanese as a second language is increasing year after year. Since limited Japanese proficiency may pose a greater health risk, dentists are expected to manage cross-cultural communication and provide dental care to foreign patients. This study explored dentists’ experiences of treating patients with limited Japanese proficiencies. Semi-structured interviews were conducted with 11 community dentists and the qualitative data were analyzed through a thematic analysis approach. Their major challenges were classified into three themes—linguistic aspect (e.g., complicated explanation regarding root canal treatment), sociolinguistic aspect (e.g., communication with foreign residents with limited dental knowledge), and sociocultural aspect (e.g., cultural differences in their dental aesthetics and insurance treatment system). Several management strategies were employed, including linguistic accommodation, avoidance of complexities, use of various communication tools, and getting help from others. However, they were unsatisfied with their practice because they could not understand the patients’ psychosocial aspects due to incomplete communication. These findings provided insights into dentists’ practice in the globalized context.

The post-translational acetylation of lysine residues is found in many nonhistone proteins and is involved in a wide range of biological processes. Recently, we showed that the nucleoprotein of the influenza A virus is acetylated by histone acetyltransferases (HATs), a phenomenon that affects viral transcription. Here, we report that the PA subunit of influenza A virus RNA-dependent RNA polymerase is acetylated by the HATs, P300/CREB-binding protein-associated factor (PCAF), and general control nonderepressible 5 (GCN5), resulting in accelerated endonuclease activity. Specifically, the full-length PA subunit expressed in cultured 293T cells was found to be strongly acetylated. Moreover, the partial recombinant protein of the PA N-terminal region containing the endonuclease domain was also acetylated by PCAF and GCN5 in vitro, which facilitated its endonuclease activity. Mass spectrometry analyses identified K19 as a candidate acetylation target in the PA N-terminal region. Notably, the substitution of the lysine residue at position 19 with glutamine, a mimic of the acetyl-lysine residue, enhanced its endonuclease activity in vitro; this point mutation also accelerated influenza A virus RNA-dependent RNA polymerase activity in the cell. Our findings suggest that PA acetylation is important for the regulation of the endonuclease and RNA polymerase activities of the influenza A virus.

In 2016, the Ministry of Health, Labour and Welfare established the Health Support Pharmacy Certification System. The certification requirements include a track record of counseling regarding the use of over-the-counter drugs (OTC). Therefore, pharmacists must increase their self-efficacy for counseling.To determine pharmacists' self-efficacy for OTC counseling and related factors.A web-based survey was conducted. Multivariate analysis was conducted to test the relationship among the mean scores of self-efficacy for OTC counselling for 25 symptoms, pharmacist attributes, years of work, psychosocial factor, job satisfaction, and level of trust from the community and patients.We received responses from 250 people. The overall self-efficacy was 5.8 (SD= 2.4) but varied depending on the symptoms. Self-efficacy was relatively high for allergic symptoms (6.9), cold/influenza (6.9), and constipation (7.1), but relatively low for contraceptive drugs (3.8), palpitation/shortness of breath (4.6), and abnormal taste/smell (4.2). In bivariate analysis, items related to self-efficacy included "age" (Spearman correlation= 0.276, P0.001), "academic background" (-0.208, P=0.001), "number of years of work" (0.267, P0.001), "level of trust from the community" (0.155, P=0.014), "level of trust from patients" (0.271, P0.001), "job satisfaction" (0.236, P0.001), "role clarity" (0.181, P=0.004), and "positive challenge at work" (0.271, P0.001). Multivariate analysis indicated that the number of years of work (Standardizing Coefficient: 0.22), trust from patients (0.13), and positive challenge at work (0.25) had a positive effect on self-efficacy.Years of work, recognition that they are trusted by patients, and positive challenge at work were important for the counseling self-efficacy of pharmacists. These results provide implications for pharmacy management and lifelong education strategies to promote self-efficacy in pharmacist counseling.

Background Prejudice and stigma against patients with depression may lead to adverse clinical consequences, such as delayed initiation and premature termination of therapy. However, available survey results on pharmacists' awareness showed a lack of confidence in dealing with patients with depression. Until recently, few studies addressed the relationship between pharmacists' perceptions of their social distance from and confidence in interacting with patients with depression. Objectives To determine pharmacists' social distance from patients with depression, and investigate the factors that influence and are influenced by it. Methods The target population of this survey study included 161 pharmacists employed at a drug chain who had participated in the Effects of Community Pharmacists' Medication Assistance for the Patients Taking Antidepressant study. The questionnaire included six questions on social distance, which were adopted from a previous nation-wide survey of mental illness stigma in Japan. Other questions related to the respondents' attributes, personal experience (living with depression, abusive behaviors inflicted by patients with depression), and confidence in interacting with patients with depression. Results Valid responses were obtained from 77 study participants. The mean total social distance score was 12.49. Greater total social distance scores were associated with greater difficulty in relationship building, information provision, and comprehension of condition. Pharmacists' personal experience of living with depression decreased their social distance from patients with depression, whereas their experience of being verbally or physically abused by patients with depression increased their social distance. Conclusion This study showed that community pharmacists in Japan had less social distance from patients with depression than the general population. A significant negative correlation was observed between community pharmacists' social distance from and their confidence in interacting with patients with depression.

In Japan, there is a pressing need to improve community health care to cope with the rapid aging of the population. In this context, there have been private-sector-led approaches to enhance community dietary support by employing dietitians in pharmacies.To evaluate the effects of collaboration between dietitians and pharmacists working in pharmacies to support patients with type 2 diabetes.A single group pre- and post-comparative study was conducted on patients with type 2 diabetes mellitus. The intervention period was 6 months. During the intervention period, the dietitians provided dietary support to the patients after first providing them with medication guidance. The contents of these instructions were shared with the pharmacists. The contents of the instructions were recorded, and confirmed in monthly meetings with the principal investigator. The primary endpoint was the Hemoglobin A1c(HbA1c) level, and the secondary endpoints were high-density lipoprotein-cholesterol (HDL-c), low-density lipoprotein-cholesterol (LDL-c), Triglyceride (TG), degree of dietary self-management, degree of unbalanced diet and satisfaction with pharmacy services.Eight patients completed the intervention period. The first patient's intervention started in March 2021, and all patients' interventions were completed by December 2021. The primary endpoint, the mean (SD) HbA1c, was 7.26 (0.96) at baseline and decreased to 6.63 (0.79) after 6 months (p=0.028, r=0.72). Also, the HDL-c increased from 55.00 (14.81) to 63.14 (10.11) (p=0.110, r=0.51) and the Diabetes Mellitus Dietary Self Efficacy Scale score increased from 51.67 (8.31) to 60.17 (8.45) (p=0.025, r=0.79) and the patient satisfaction score increased 24.0 (4.0) to 26.1 (3.3) (p=0.161, r=0.51). Moderate decreases were also observed in LDL-c (p=0.235, r=0.47) and TG (p=0.368, r=0.37).Collaboration between dietitians and pharmacists working in pharmacies may improve the dietary habits and glycemic control of patients with type 2 diabetes. To verify this hypothesis more reliably, randomized controlled trials need to be conducted.

In Japan, there has been a private-sector initiative to register dietitians in pharmacies. There is not yet an adequate amount of data on the attitudes of dietitians in pharmacies regarding their work.To assess the attitudes of pharmacists and dietitians working in community pharmacies in Japan about their work, and particularly about demonstrating expertise and overall job satisfaction.We administered a web-based questionnaire to pharmacists and dietitians working in pharmacy branches with registered dietitians at two pharmacy chains that agreed to cooperate. We used our own 7-item questionnaire and compared the scores for each item between pharmacists and dietitians.A total of 61 persons (22 pharmacists and 39 dietitians) answered the questionnaire. The item with the highest mean score (standard deviation) for pharmacists was "I feel that I am helpful to patients," at 3.50 (0.74), and for dietitians it was "I have colleagues at my workplace whom I can talk to when I have a problem," at 3.51 (1.00). Conversely, the items with the lowest mean score (standard deviation) were "I am satisfied with my current job" for pharmacists, at 3.14 (0.83), and "I can grow sufficiently as a professional" for dietitians, at 2.41 (0.97). As a result of factor analysis, the seven-item question could be divided into two factors: "demonstrating expertise" and "overall job satisfaction." The factor-score for "demonstrating expertise" was significantly lower for dietitians than for pharmacists (p0.01), but there was no significant difference in overall job satisfaction between the two professions (p=0.36). Further research is needed to understand why dietitians find "demonstrating expertise" difficult in pharmacies.The attitudes of pharmacists and dietitians working in pharmacies in Japan toward their jobs were shown to consist of "demonstration of expertise" and "overall job satisfaction. Dietitians had significantly lower scores on "demonstration of expertise" than pharmacists.

Jiadifenolide has been reported to have neurotrophin-like activity in primary rat cortical neurons, and also possesses neurotrophic effects in neuronal precursor cells derived from human induced pluripotent stem cells (hiPSCs), as we have previously reported. However, the molecular mechanisms by which jiadifenolide exerts its neurotrophic effects in rat and human neurons are unknown. Thus, we aimed to investigate the molecular mechanisms and pathways by which jiadifenolide promotes neurotrophic effects. Here, we found that jiadifenolide activated cellular communication network factor (CCN) signaling pathways by up-regulating mRNA level expression of CCN genes in human neuronal cells. We also found that CCN2 (also known as connective tissue growth factor, CTGF) protein promotes neurotrophic effects through activation of the p44/42 mitogen-activated protein kinase signaling pathway. This is the first discovery which links neurotrophic activity with CCN signaling.

Novel antiviral agents for influenza, which poses a substantial threat to humans, are required. Cyclobakuchiols A and B have been isolated from Psoralea glandulosa, and cyclobakuchiol C has been isolated from P. corylifolia. The structural differences between cyclobakuchiol A and C arise due to the oxidation state of isopropyl group, and these compounds can be derived from (+)-(S)-bakuchiol, a phenolic isoprenoid compound present in P. corylifolia seeds. We previously reported that bakuchiol induces enantiospecific anti-influenza A virus activity involving nuclear factor erythroid 2-related factor 2 (Nrf2) activation. However, it remains unclear whether cyclobakuchiols A–C induce anti-influenza A virus activity. In this study, cyclobakuchiols A, B, and C along with cyclobakuchiol D, a new artificial compound derived from cyclobakuchiol B, were synthesized and examined for their anti-influenza A virus activities using Madin-Darby canine kidney cells. As a result, cyclobakuchiols A–D were found to inhibit influenza A viral infection, growth, and the reduction of expression of viral mRNAs and proteins in influenza A virus-infected cells. Additionally, these compounds markedly reduced the mRNA expression of the host cell influenza A virus-induced immune response genes, interferon-β and myxovirus-resistant protein 1. In addition, cyclobakuchiols A–D upregulated the mRNA levels of NAD(P)H quinone oxidoreductase 1, an Nrf2-induced gene, in influenza A virus-infected cells. Notably, cyclobakuchiols A, B, and C, but not D, induced the Nrf2 activation pathway. These findings demonstrate that cyclobakuchiols have anti-influenza viral activity involving host cell oxidative stress response. In addition, our results suggest that the suitably spatial configuration between oxidized isopropyl group and phenol moiety in the structure of cyclobakuchiols is required for their effect.

Human influenza virus infections occur annually worldwide and are associated with high morbidity and mortality. Hence, development of novel anti-influenza drugs is urgently required. Rice Power® extract developed by the Yushin Brewer Co. Ltd. is a novel aqueous extract of rice obtained via saccharization and fermentation with various microorganisms, such as Aspergillus oryzae, yeast [such as Saccharomyces cerevisiae], and lactic acid bacteria, possessing various biological and pharmacological properties. In our previous experimental screening with thirty types of Rice Power® extracts, we observed that the 30th Rice Power® (Y30) extract promoted the survival of influenza A virus-infected Madin-Darby canine kidney (MDCK) cells. Therefore, to identify compounds for the development of novel anti-influenza drugs, we aimed to investigate whether the Y30 extract exhibits anti-influenza A virus activity. In the present study, we demonstrated that the Y30 extract strongly promoted the survival of influenza A H1N1 Puerto Rico 8/34 (A/PR/8/34), California 7/09, or H3N2 Aichi 2/68 (A/Aichi/2/68) viruses-infected MDCK cells and inhibited A/PR/8/34 or A/Aichi/2/68 viruses infection and growth in the co-treatment and pre-infection experiments. The pre-treatment of Y30 extract on MDCK cells did not induce anti-influenza activity in the cell. The Y30 extract did not significantly affect influenza A virus hemagglutination, and neuraminidase and RNA-dependent RNA polymerase activities. Interestingly, the electron microscopy experiment revealed that the Y30 extract disrupts the integrity of influenza A virus particles by permeabilizing the viral membrane envelope, suggesting that Y30 extract has a direct virucidal effect against influenza A virus. Furthermore, we observed that compared to the ethyl acetate (EtOAc) extract, the water extract of Y30 extract considerably promoted the survival of cells infected with A/PR/8/34 virus. These results indicated that more anti-influenza components were present in the water extract of Y30 extract than in the EtOAc extract. Our results highlight the potential of a rice extract fermented with A. oryzae and S. cerevisiae as an anti-influenza medicine and a drug source for the development of anti-influenza compounds.

Recently, opportunistic nosocomial infections caused by Acinetobacter baumannii have become increasingly prevalent worldwide. The pathogen often establishes biofilms that adhere to medical devices, causing chronic infections refractory to antimicrobial therapy. Clinical reports have indicated that some macrolide antibiotics are effective against chronic biofilm-related infections. In this study, we examined the direct anti-biofilm effects of seven macrolides (azithromycin, clarithromycin, erythromycin, josamycin, spiramycin, fidaxomicin, and ivermectin) on A. baumannii using a simple and newly established in vitro assay system for the swift and serial spectrophotometric determinations of two biofilm-amount indexes of viability and biomass. These macrolides were found to possess direct anti-biofilm effects exerting specific anti-biofilm effects not exclusively depending on their bacteriostatic/bactericidal effects. The anti-biofilm effect of azithromycin was found to be the strongest, while those of fidaxomicin and ivermectin were weak and limited. These results provide insights into possible adjunctive chemotherapy with macrolides for A. baumannii infection. Common five macrolides also interfered with the Agrobacterium tumefaciens NTL(pCF218) (pCF372) bioassay system of N-acyl homoserine lactones, providing insights into sample preparation for the bioassay, and putatively suggesting the actions of macrolides as remote signals in bacterial quorum sensing systems.

In Japanese pharmaceutical education, the Model Core Curriculum was revised in 2013 to train pharmacists who can appropriately evaluate literature and use evidence-based medicine (EBM). However, in the investigation of EBM education at pharmaceutical universities in 2015, it was found that literature evaluation was hardly performed in the education of undergraduate students. One of the reason is the lack of EBM lecturers at each universities. Therefore, we adopted team-based learning (TBL) to educate more than 50 undergraduate students on the practical evaluation of literatures and the understanding of EBM concepts. The learning outcomes of this strategy were evaluated using the scores of individual tests before and after the class. As a result, the mean scores on the post-test significantly improved from 4.34 to 6.42 out of 10 total points (p

Histone acetylation plays crucial roles in transcriptional regulation and chromatin organization. Viral RNA of the influenza virus interacts with its nucleoprotein (NP), whose function corresponds to that of eukaryotic histones. NP regulates viral replication and has been shown to undergo acetylation by the cAMP-response element (CRE)–binding protein (CBP) from the host. However, whether NP is the target of other host acetyltransferases is unknown. Here, we show that influenza virus NP undergoes acetylation by the two host acetyltransferases GCN5 and P300/CBP-associated factor (PCAF) and that this modification affects viral polymerase activities. Western blot analysis with anti–acetyl-lysine antibody on cultured A549 human lung adenocarcinoma epithelial cells infected with different influenza virus strains indicated acetylation of the viral NP. A series of biochemical analyses disclosed that the host lysine acetyltransferases GCN5 and PCAF acetylate NP in vitro. MS experiments identified three lysine residues as acetylation targets in the host cells and suggested that Lys-31 and Lys-90 are acetylated by PCAF and GCN5, respectively. RNAi-mediated silencing of GCN5 and PCAF did not change acetylation levels of NP. However, interestingly, viral polymerase activities were increased by the PCAF silencing and were decreased by the GCN5 silencing, suggesting that acetylation of the Lys-31 and Lys-90 residues has opposing effects on viral replication. Our findings suggest that epigenetic control of NP via acetylation by host acetyltransferases contributes to regulation of polymerase activity in the influenza A virus.

The objective of this survey is investigating pharmacists' recognition of and belief in the importance of doing PRE-AVOID using the drug profile book (DPB). The main survey items were: 1. Awareness and self-confidence concerning PRE-AVOID using DPB, 2.Status of educating patients of the merits of the DPB, 3. Status of educating patients of the utilities of the DPB, 4. Status of educating patients of the roles of the DPB, 5. Guidance provided on methods of DPB usage, and 6. Interactions with patients. A multiple regression analysis was performed using "Awareness and self-confidence concerning PRE-AVOID using DPB" as a dependent variable. A regression equation including three items ("Educating the roles", "Blood test", and "Not using multiple DPBs at same time") was derived for "Self-confidence in PRE-AVOID work" (R2=0.20). The item most affecting "Self-confidence in PRE-AVOID work" was "Educating the roles" (R=0.28). A regression equation with "Awareness of PRE-AVOID work" as a dependent variable was not derived. The results of the study suggest that "Educating the roles" or "Not using multiple DPBs at same time" for patients, and asking the results of "Blood test" in a medication consultation may increase pharmacists belief in doing PRE-AVOID.

Objectives To develop a semi-quantitative instrument to assess pharmacists' confidence in medication counseling for patients with depression, The Pharmacists' Confidence scale about Medication Consultation for Depressive patients (PCMCD), and investigated its validity. Methods Following discussions with practicing pharmacists, we developed a 12-item questionnaire to assess pharmacists' confidence in medication counseling for patients with depression. We launched web-based cross-sectional survey during November and December 2018 to 77 pharmacists employed at drug chain stores in Kansai area. Factor analysis was performed to evaluate the configuration concept validity. The least-squares method was used for factor extraction, and the resulting factors were subjected to direct oblimin rotation, with a factor loading cut-off of 0.4. To assess internal consistency, Cronbach's alpha values were calculated for each of the extracted factors (subscales). A multiple regression analysis was performed using simultaneous forced entry, with the scores obtained for each subscale as dependent variables and responder attributes as independent variables in order to investigate the factors associated with each subscale. Results During the factor analysis procedure, four questions were excluded by the cut-off rule. Eventually, a model with three subscales was identified, with a cumulative sum of squared loadings being 61.9%. The subscales were termed "relationship building," "comprehension of condition," and "information provision" based on the nature of the questions relevant for each of them. The Cronbach's alpha values for these subscales were 0.92, 0.73, and 0.72, respectively. The average inter-item correlation was 0.378. In addition, multiple regression analysis revealed that there were significant correlations between pharmacist career and both relationship building and information provision. Conclusions The PCMCD model demonstrated a satisfactory construct validity and internal consistency. This model will provide an excellent tool for assessing pharmacists' confidence in depression care.

Although jiadifenolide has been reported to neurotrophin-like activity in primary cultured rat cortical neurons, it is unknown on that of activity in human neurons. Thus, we aimed to assess neurotrophin-like activity by jiadifenolide in human neuronal cells. We analyzed neuronal precursor cells derived from human induced pluripotent stem cells for microtuble-associated-protein-2 expression by immunofluorescence and western blot, following jiadifenolide treatment. Jiadifenolide promoted dendrite outgrowth, facilitated growth, and prevented death in neuronal cells derived from human induced pluripotent stem cells. Interestingly, jiadifenolide also increased postsynaptic density-95 protein expression suggesting that jiadifenolide promotes neuronal maturation and post-synaptic formation. We demonstrate for the first time that jiadifenolide exhibits neurotrophic effects on human neuronal precursor cells.

Background: Community pharmacists should be involved in diabetes care, while there has been less evidence about whether a brief lifestyle intervention is effective for diabetes care in community pharmacies. Objectives: To examine the effects of brief lifestyle intervention on glycemic control in patients with type 2 diabetes mellitus (T2D) by using a coaching style, provided by community pharmacists. Methods: A prospective, cluster-randomized, controlled trial was conducted in 50 groups of community pharmacies in Japan. In all, 132 patients with T2D (age, 20 - 75 years, ≥8.0% of hemoglobin A1c (HbA1c)) were assigned to the intervention group (n = 90) or the usual care group (n = 42). The intervention group (IG) underwent brief lifestyle coaching for self-care of T2D for 6 months. The standard care group (CG) received usual care by pharmacists and was given a general newsletter. The primary outcome was changes in HbA1c levels. Results: After 6 months, the IG had significantly improved HbA1c (IG: -0.6 ± 0.9 vs. CG: -0.2% ± 0.9%; p = 0.021 using the last observation carried forward analysis). Although the number of drugs reduced from 2.3 ± 0.8 to 2.0 ± 1.2 in the IG, the number increased from 2.3 ± 1.1 to 2.5 ± 1.1 in the CG (-0.2 ± 0.9 in IG vs. 0.2 ± 0.6 in CG; p = 0.023). Conclusions: The brief lifestyle intervention by community pharmacists improved glycemic control in patients with T2D. Community pharmacists may more positively participate as lifestyle coaches for diabetes care.

In Japanese pharmacies, Drug Profile Books (DPBs), which are a type of Personal Health Record (PHR), are incorporated in order to prevent duplicate medication and drug interactions in outpatients (patients) through the uniform management of drug administration information. In this study, we tried to clarify the effect on patient safety of brief interventions via DPBs by pharmacists. The study design was a randomized controlled trial on pharmacies as clusters. 65 pharmacies agreed to participate in the study (intervention group (IG): 33; control group (CG): 32). The primary outcomes were: rate of inquiry occurrence, rate of prescription change, and rates of duplicate medications & drug interactions. 56 pharmacies (IG: 29; CG: 27) completed the study. There was a higher tendency for prescription changes in the IG compared to the CG (IG: 0.03%; CG: 0.02%; P = 0.08). In addition, the rate of duplicate medications & drug interactions accounting for the inquiries was significantly higher in the IG than in the CG (IG: 89.2%; CG: 71.9%; P = 0.01). This implied that brief interventions by pharmacists using DPBs had an effect in raising patient safety.

Objectives To demonstrate how the confidence, attitude and job satisfaction of pharmacists who provide interventions and lifestyle support services to patients with hypertension are altered. Methods This study was a before-and-after study that used data from 50 pharmacists who participated in the Community Pharmacists Assist for Blood Pressure (COMPASS-BP) study, which was an interventional study that was conducted in Japan to evaluate the effects of lifestyle support services provided to hypertension patients. The pharmacists received 4 h of training before the study was initiated to learn how to quickly provide patients with guidance about diet and exercise and tobacco and alcohol consumption. During the interventional period, each pharmacist provided concise lifestyle guidance to 1–3 hypertension patients. The pharmacists met with the patients a total of 5 times. The parameters that were assessed were the pharmacists’ attitudes towards and confidence in their lifestyle support services, as well as their level of job satisfaction. We verified the scores of each parameter assessed before and after the study, as well as changes in the relationships among the different parameters. Key findings Significant increases from baseline to the end of the study were observed for Confidence (P < 0.01), Job satisfaction, including ‘Knowledge’ (P < 0.01), ‘Patient counselling management’ (P = 0.04) and ‘Sense of personal growth’ (P < 0.01). Additionally, path analysis results revealed that after the study, the causal relationship between ‘Knowledge’ and ‘Attitude’ disappeared, and a new causal relationship appeared between ‘Patient counselling and management’ and ‘Attitude’. Conclusions Providing lifestyle guidance improved pharmacists’ confidence and attitude. Also, ‘Attitude’ tended to improve as the ‘Patient counselling and management’ score increased.

Adenovirus vectors (Adv) elicit innate immune responses via several pattern-recognition receptors. Although it has been suggested that various Adv-induced mechanisms play important roles in the induction of innate immunity in vitro , the impacts of these mechanisms in vivo remain unclear. Viral nucleic acids elicit innate immune responses through the recognition of cytosolic nucleic acid sensors and transduce intracellular signals to TANK-binding kinase (TBK) 1. In this study, to determine the impacts of viral nucleic acids on innate immune responses in vivo , we administered transgene-expressing Adv to Tbk1 -deficient mice. The systemic Adv administration failed to induce type I interferons (type I IFNs) in the spleen, but not the liver, of Tbk1 -deficient mice, resulting in the increase of transgene-expressing cells in the spleen, but not the liver. Moreover, Adv failed to induce type I IFNs in the bone-marrow-derived dendritic cells, but not the mouse embryonic fibroblasts, from Tbk1 -deficient mice in vitro . These results support the idea that Adv elicit innate immunity in immune cells and non-immune cells in a TBK1-dependent and TBK1-independent manner, respectively.

The COMmunity Pharamcists ASSist for Blood Pressure (COMPASS-BP) study aimed to assess the effectiveness of lifestyle support programs administered in community pharmacies on hypertension control. This open-label, two-armed parallel group, cluster-randomized controlled trial included 73 pharmacies (clusters) in Japan randomized to a control or intervention group. Eligible hypertensive patients (n = 125), aged 20-75 years, received the intervention (n = 64) or the control treatment (n = 61), as dictated by their pharmacy randomization. Patients in the intervention group received brochures and healthy lifestyle advice from pharmacists using motivational interviewing methods during pharmacy visits over a 12-week period, with their usual pharmacy care. Conversely, the control group just received usual care. The main outcome measure was a change in morning systolic blood pressure (SBP) from baseline to week 12. The intervention group exhibited a decrease in morning SBP that was 6.0 mmHg greater than that of the control group (95% confidence interval [CI]: -11.0 to -0.9, p = 0.021). In a mixed-effect model for repeated measures analysis, the intergroup difference in morning SBP decrease was -4.5 mmHg (95% CI: -8.5 to -0.6, p = 0.024). Our findings indicate that implementation of a lifestyle advice program in pharmacies is feasible and may lead to reduced blood pressure.

Background: The PB2 subunit of RNA polymerase of influenza virus has a novel site for acetyl-CoA interaction, which contains a valine/arginine/glycine (VRG) motif. Results: A mutation in the VRG site inhibited acetyl-CoA interaction, RNA polymerase activity, and viral replication. Conclusion: Acetyl-CoA interaction at the VRG site is potentially essential for viral replication. Significance: The VRG site might be a novel target for drugs and vaccines., The PA, PB1, and PB2 subunits, components of the RNA-dependent RNA polymerase of influenza A virus, are essential for viral transcription and replication. The PB2 subunit binds to the host RNA cap (7-methylguanosine triphosphate (m7GTP)) and supports the endonuclease activity of PA to “snatch” the cap from host pre-mRNAs. However, the structure of PB2 is not fully understood, and the functional sites remain unknown. In this study, we describe a novel Val/Arg/Gly (VRG) site in the PB2 cap-binding domain, which is involved in interaction with acetyl-CoA found in eukaryotic histone acetyltransferases (HATs). In vitro experiments revealed that the recombinant PB2 cap-binding domain that includes the VRG site interacts with acetyl-CoA; moreover, it was found that this interaction could be blocked by CoA and various HAT inhibitors. Interestingly, m7GTP also inhibited this interaction, suggesting that the same active pocket is capable of interacting with acetyl-CoA and m7GTP. To elucidate the importance of the VRG site on PB2 function and viral replication, we constructed a PB2 recombinant protein and recombinant viruses including several patterns of amino acid mutations in the VRG site. Substitutions of the valine and arginine residues or of all 3 residues of the VRG site to alanine significantly reduced the binding ability of PB2 to acetyl-CoA and its RNA polymerase activity. Recombinant viruses containing the same mutations could not be replicated in cultured cells. These results indicate that the PB2 VRG sequence is a functional site that is essential for acetyl-CoA interaction, RNA polymerase activity, and viral replication.

Awa-ban tea is a uniquely flavored, pickled and anaerobically fermented tea found only in the Tokushima prefecture in the Shikoku region of Japan. Diabetes is a major health problem worldwide. Here, we report that Awa-ban tea suppressed the increase in blood glucose levels after oral administration of maltose, sucrose, or glucose in mice (n = 9–54). Awa-ban tea suppressed the area under the curve (AUC) of blood glucose by 72–83%. Thus, we propose that Awa-ban tea can be used as an alternative functional beverage for diabetes prevention.

This paper proposes a two-state Markov model to determine the storage size of a photovoltaic system. The system is usually connected to a distribution grid, but once a large disaster occurs, it becomes the main source of supply. Daily solar radiation changes due to the weather, and it could become lower than a specified supply level for a sequence of days. Transition probabilities are determined from meteorological data on the solar radiation. The Markov model shows that its frequency exponentially decreases as the sequence becomes long. One or two days of storage is adequate to reduce the number of insufficient supply days to an acceptable level, and the value of adding the storage exponentially decreases, too. Some simulations and experiments are executed to verify these facts.

Few of the vaccines in current use can induce antigen- (Ag-) specific immunity in both mucosal and systemic compartments. Hence, the development of vaccines that realize both mucosal and systemic protection against various pathogens is a high priority in global health. Recently, it has been reported that intramuscular (i.m.) vaccination of an adenovirus vector (Adv) can induce Ag-specific cytotoxic T lymphocytes (CTLs) in both systemic and gut mucosal compartments. We previously revealed that type I IFN signaling is required for the induction of gut mucosal CTLs, not systemic CTLs. However, the molecular mechanism via type I IFN signaling is largely unknown. Here, we report that type I IFN signaling following i.m. Adv vaccination is required for the expression of type I IFN in the inguinal lymph nodes (iLNs), which are the draining lymph nodes of the administration site. We also showed that the type I IFN signaling is indispensable for the early activation of CTLs in iLNs. These data suggested that type I IFN signaling has an important role in the translation of systemic innate immune response into mucosal adaptive immunity by amplifying the innate immune signaling and activating CTLs in the iLN.

The varicella-zoster virus (VZV) Oka vaccine strain (vOka) is a highly immunogenic and safe live vaccine that has long been used worldwide. Because its genome is large, making it suitable for inserting foreign genes, vOka is considered a candidate vector for novel polyvalent vaccines. Previously, a recombinant vOka, rvOka-HN, that expresses mumps virus (MuV) hemagglutinin-neuraminidase (HN) was generated by the present team. rvOka-HN induces production of neutralizing antibodies against MuV in guinea pigs. MuV also expresses fusion (F) protein, which is important for inducing neutralizing antibodies, in its viral envelope. To induce a more robust immune response against MuV than that obtained with rvOka-HN, here an rvOka expressing both HN and F (rvOka-HN-F) was generated. However, co-expression of HN and F caused the infected cells to form syncytia, which reduced virus titers. To reduce the amount of cell fusion, an rvOka expressing HN and a mutant F, F(S195Y) were generated. Almost no syncytia formed among the rvOka-HN-F(S195Y)-infected cells and the growth of rvOka-HN-F(S195Y) was similar to that of the original vOka clone. Moreover, replacement of serine 195 with tyrosine had no effect on the immunogenicity of F in mice and guinea pigs. Although obvious augmentation of neutralizing antibody production was not observed after adding F protein to vOka-HN, the anti-F antibodies did have neutralizing activity. These data suggest that F protein contributes to induction of immune protection against MuV. Therefore this recombinant virus is a promising candidate vaccine for polyvalent protection against both VZV and MuV.

This survey aimed to examine how patient-pharmacist communication using the drug profile book relates patient's behavior regarding its use. Among patients who visited one of the five pharmacies during the 4 months between July and October of 2013, 245 patients who had been prescribed antihypertensives were asked to complete a questionnaire. Items included patient attributes, whether the patient thought the drug profile book was useful to them ("sense of utility"), whether the patient has ever been questioned by a pharmacist while showing the drug profile book ("experience of being questioned by a pharmacist while showing the drug profile book"), and whether the patient has ever shown the drug profile book to the physician ("experience of showing the drug profile book to the physician"). In addition, pharmacists counted the frequency of patients bringing the drug profile book, and if so, the frequency of the sticker affix during the last 5 visits. 34.3% of responding patients answered that they had the "experience of being questioned while showing the drug profile book". Response rates of "frequency of bringing the drug profile book", "sense of utility", and "experience of showing the drug profile book to the physician" in the group with "experience of being questioned while showing the drug profile book" were significantly higher than those in the group without such experience. This survey indicated that experience of being questioned by a pharmacist while showing the drug profile book related patient's behavior regarding its use.