Factors related to hospital nurse intention to leave: Does striving for work-life balance and sense of coherence affect wishes to stay in the organization?

The aim of this study is to identify relationships between striving for work-life balance (S-WLB), sense of coherence (SOC), and intention to leave among hospital nurses. In August of 2017, we conducted a self-administered questionnaire survey to 2239 nurses at nine public hospitals. The questionnaire included demographic factors, work environmental factors, organizational factors, striving for worklife balance, a 13-item SOC, the Maslach Burnout Inventory, and intention to leave. We carried out a multiple regression analysis with intention to leave as the dependent variable and others as independent variables. We obtained valid responses from 1368 full-time employed nurses (61.1%). The mean age of respondents was 36.38 ± 10.18 years, and the length of clinical experience 13.4 ± 9.65 years. The mean total score of the intention to leave was 14.58 ± 5.09. As a result of the multiple regression analysis, the SWLB and the SOC were found to be statistically related to the intention to leave even after controlling for the confounding factor of burnout. In addition, the satisfaction of desire level for the actual working assignment at the workplace was a significant independent factor. In this study, the full-time nursing staff had a low intention to leave score when they had a high score in the SWLB and the SOC. It was also found that the level of satisfaction with appointment to a desired assignment is a significant factor in the intention to leave. In establishing an attractive workplace for nursing staff, it is important to create a workplace environment and supports to be able to properly evaluate workplace satisfaction after assignments are made while taking into account individual S-WLB and SOC functions

Bluetongue Virus VP1 Polymerase Activity In Vitro: Template Dependency, Dinucleotide Priming and Cap Dependency

BACKGROUND: Bluetongue virus (BTV) protein, VP1, is known to possess an intrinsic polymerase function, unlike rotavirus VP1, which requires the capsid protein VP2 for its catalytic activity. However, compared with the polymerases of other members of the Reoviridae family, BTV VP1 has not been characterized in detail. METHODS AND FINDINGS: Using an in vitro polymerase assay system, we demonstrated that BTV VP1 could synthesize the ten dsRNAs simultaneously from BTV core-derived ssRNA templates in a single in vitro reaction as well as genomic dsRNA segments from rotavirus core-derived ssRNA templates that possess no sequence similarity with BTV. In contrast, dsRNAs were not synthesized from non-viral ssRNA templates by VP1, unless they were fused with specific BTV sequences. Further, we showed that synthesis of dsRNAs from capped ssRNA templates was significantly higher than that from uncapped ssRNA templates and the addition of dinucleotides enhanced activity as long as the last base of the dinucleotide complemented the 3' -terminal nucleotide of the ssRNA template. CONCLUSIONS: We showed that the polymerase activity was stimulated by two different factors: cap structure, likely due to allosteric effect, and dinucleotides due to priming. Our results also suggested the possible presence of cis-acting elements shared by ssRNAs in the members of family Reoviridae

Development of reverse genetics for Ibaraki virus to produce viable VP6-tagged IBAV.

Ibaraki virus (IBAV) is a member of the epizootic hemorrhagic disease virus (EHDV) serogroup, which belongs to the Orbivirus genus of the Reoviridae family. Although EHDV, including IBAV, represents an ongoing threat to livestock in the world, molecular mechanisms of EHDV replication and pathogenesis have been unclear. The reverse genetics (RG) system is one of the strong tools to understand molecular mechanisms of virus replication. Here, we developed a RG system for IBAV to identify the nonessential region of a minor structural protein, VP6, by generating VP6-truncated IBAV. Moreover, several tags were inserted into the truncated region to produce VP6-tagged IBAV. We demonstrated that all VP6-tagged IBAV could replicate in BHK cells in the absence of any helper VP6 protein. Further, tagged-VP6 proteins were first assembled into puncta in cells infected with VP6-tagged IBAV. Our data suggests that, in order to initiate primary replication, IBAV VP6 is likely to accumulate in some parts of infected cells to assemble efficiently into the primary replication complex (subcore)

Interaction between a Unique Minor Protein and a Major Capsid Protein of Bluetongue Virus Controls Virus Infectivity.

Among the Reoviridae family of double-stranded RNA viruses, only members of the Orbivirus genus possess a unique structural protein, termed VP6, within their particles. Bluetongue virus (BTV), an important livestock pathogen, is the prototype Orbivirus BTV VP6 is an ATP-dependent RNA helicase, and it is indispensable for virus replication. In the study described in this report, we investigated how VP6 might be recruited to the virus capsid and whether the BTV structural protein VP3, which forms the internal layer of the virus capsid core, is involved in VP6 recruitment. We first demonstrated that VP6 interacts with VP3 and colocalizes with VP3 during capsid assembly. A series of VP6 mutants was then generated, and in combination with immunoprecipitation and size exclusion chromatographic analyses, we demonstrated that VP6 directly interacts with VP3 via a specific region of the C-terminal portion of VP6. Finally, using our reverse genetics system, mutant VP6 proteins were introduced into the BTV genome and interactions between VP6 and VP3 were shown in a live cell system. We demonstrate that BTV strains possessing a mutant VP6 are replication deficient in wild-type BSR cells and fail to recruit the viral replicase complex into the virus particle core. Taken together, these data suggest that the interaction between VP3 and VP6 could be important in the packaging of the viral genome and early stages of particle formation.IMPORTANCE The orbivirus bluetongue virus (BTV) is the causative agent of bluetongue disease of livestock, often causing significant economic and agricultural impacts in the livestock industry. In the study described in this report, we identified the essential region and residues of the unique orbivirus capsid protein VP6 which are responsible for its interaction with other BTV proteins and its subsequent recruitment into the virus particle. The nature and mechanism of these interactions suggest that VP6 has a key role in packaging of the BTV genome into the virus particle. As such, this is a highly significant finding, as this new understanding of BTV assembly could be exploited to design novel vaccines and antivirals against bluetongue disease

Management of Volunteer Activities among Elderly Persons

Purpose: Volunteer activities in Japan may be expected to result in a variety of effects on health conditions and social contributions of the elderly depending on the degree of their activity participation. This study aims to identify differences in the health conditions and social contributions of the elderly in relation to the degree of volunteer activity participation, and determine what effective support can be provided for these elderly persons. Further, focusing on the differences in the level of activity of the elderly persons in volunteer activities, we discuss the effect of management through SWOT analysis from the perspectives suggested by the Balanced Scorecard (BSC), which is a performance management tool. Participants and Methods: The participants are elderly persons participating in volunteer activities, living in municipal housing owned by the Kumamoto prefecture in Japan. We classified the participants into an active group who contributed to the raising of funds for activities, and a non-active group. We conducted group interviews with the volunteers, organized interview data on the thoughts of the two groups according to the five perspectives of the balanced score card (BSC) management tool: (1) learning and development, (2) participants, (3) financial matters, (4) process of conducting activities, and (5) social contributions, and conducted a Strength-Weakness-OpportunityThreat (SWOT) analysis. In the SWOT analysis internal factors are distinguished into strengths and weaknesses, and external factors into opportunities and threats. Combining the internal and external factors, problems were extracted from the perspective of a positive strategy (S+O: strength + opportunity), a differentiation strategy (S+T: strength L threat), a stepwise strategy (W+O: weakness + opportunity), and a defensive strategy (W+T: weakness + threat). Then, we confirmed the main results by identifying the major factors in success to achieve salient aims, activity goals, and specific measures from the five perspectives provided by the BSC. Results: When comparing thestrengths of the two groups, the active group showed the following characteristics in the information collecting: more variety in the collected information (perspective of learning and development), ease of obtaining information of potential volunteers (perspective of participants), knowledge of how to raise funds (perspective of financial matters), and being physically more active and knowledgeable of the local community (perspective of social contributions).From the perspective of the process of conducting activities, both groups place importance on the relations among people. For weaknesses, and from the perspective of learning and development, the active group had difficulty in maintaining their motivation, and from the perspective of financial matters this group had difficulties in securing funds. From the perspective of social contributions, the non-active group depended on the active members. From the perspective of participants, both groups experienced a decline in physical function with age, and from the perspective of the process of activities there was a possibility that family or individual circumstances make it difficult to participate in all activities. In the SWOT analysis, combining the internal and external factors, we evaluated the directionality of volunteer activities from the five perspectives of the BSC. From the perspective of learning and development, regardless of the differences in the level of activity of the elderly persons in the volunteer activities, the volunteer members conducted a signature-collecting campaign to call for continuing volunteer activities for residents among those in their surroundings, and this made it possible to continue volunteer activities. From the perspective of the process of conducting activities, regardless of the differences in the level of activity of the elderly persons in the volunteer activities, the volunteer members became active in making remarks, and started to assume roles voluntarily. From the perspective of financial matters, the volunteer members were able to obtain donations for volunteer activities from neighboring establishments, as well as financial support from the social welfare council, by applying for subsidies for activities. From the perspective of participants, the number of new members increased through word of mouth spread by the existing membership. From the perspective of social cost, there was a remark that “I would like my schedule to be full of enjoyable events so that I become too busy to go to the hospital”, and in fact some members did reduce the frequency of hospital visitsDiscussion: Participants in this study are members of a volunteer group of elderly persons voluntarily formed in a regional city in Kumamoto prefecture where the elderly population accounts for 25.7% and is expected to increase. It appears that there is a self- and mutual- help relationship between active and non-active members where non-active members rely on the active members who assist the non-active members to be in harmony with the volunteer activities led by the active members, and that this relationship helps maintain independent lives. For the strengths, active members were able to collect more information from the perspective of learning and development, and contributed more through physical activity from the perspective of social contributions. It can be inferred that physically more active elderly persons are in a situation where they remain able to use their cognitive functions to collect information. For the weaknesses, the active members experienced loss of motivation. This may be due to the possibility that active members feel it a burden to assist non-active members. We wish to conduct further studies to understand how to maintain and improve motivation. The results of the SWOT analysis from the perspectives suggested by the BSC showed that there are differences in health conditions and social contributions of the elderly depending on the participation in volunteer activities, and what types of support from health services can be expected to be effective. This study evaluated the achievements of the volunteer activity goals of elderly persons by identifying the details of volunteer activities and quantifying the goals. The elderly members began to seek for new ideas to achieve better results when they were convinced that there were favorable results of their activities. This suggests that the PDCA cycle in volunteer activities may work by effecting empowerment in the process of BSC.

Differential Localization of Structural and Non-Structural Proteins during the Bluetongue Virus Replication Cycle.

Members of the Reoviridae family assemble virus factories within the cytoplasm of infected cells to replicate and assemble virus particles. Bluetongue virus (BTV) forms virus inclusion bodies (VIBs) that are aggregates of viral RNA, certain viral proteins, and host factors, and have been shown to be sites of the initial assembly of transcriptionally active virus-like particles. This study sought to characterize the formation, composition, and ultrastructure of VIBs, particularly in relation to virus replication. In this study we have utilized various microscopic techniques, including structured illumination microscopy, and virological assays to show for the first time that the outer capsid protein VP5, which is essential for virus maturation, is also associated with VIBs. The addition of VP5 to assembled virus cores exiting VIBs is required to arrest transcriptionally active core particles, facilitating virus maturation. Furthermore, we observed a time-dependent association of the glycosylated non-structural protein 3 (NS3) with VIBs, and report on the importance of the two polybasic motifs within NS3 that facilitate virus trafficking and egress from infected cells at the plasma membrane. Thus, the presence of VP5 and the dynamic nature of NS3 association with VIBs that we report here provide novel insight into these previously less well-characterized processes

Structure based modification of Bluetongue virus helicase protein VP6 to produce a viable VP6-truncated BTV.

Bluetongue virus core protein VP6 is an ATP hydrolysis dependent RNA helicase. However, despite much study, the precise role of VP6 within the viral capsid and its structure remain unclear. To investigate the requirement of VP6 in BTV replication, we initiated a structural and biological study. Multinuclear nuclear magnetic resonance spectra were assigned on his-tagged full-length VP6 (329 amino acid residues) as well as several truncated VP6 variants. The analysis revealed a large structured domain with two large loop regions that exhibit significant conformational exchange. One of the loops (amino acid position 34-130) could be removed without affecting the overall fold of the protein. Moreover, using a BTV reverse genetics system, it was possible to demonstrate that the VP6-truncated BTV was viable in BHK cells in the absence of any helper VP6 protein, suggesting that a large portion of this loop region is not absolutely required for BTV replication

Oligomerization of Hepatitis C Virus Core Protein is Crucial for Interaction with the Cytoplasmic Domain of E1 Envelope Protein

Hepatitis C virus (HCV) contains two membrane-associated envelope glycoproteins, E1 and E2, which assemble as a heterodimer in the endoplasmic reticulum (ER). In this study, predictive algorithms and genetic analyses of deletion mutants and glycosylation site variants of the E1 glycoprotein were used to suggest that the glycoprotein can adopt two topologies in the ER membrane: the conventional type I membrane topology and a polytopic topology in which the protein spans the ER membrane twice with an intervening cytoplasmic loop (amino acid residues 288 to 360). We also demonstrate that the E1 glycoprotein is able to associate with the HCV core protein, but only upon oligomerization of the core protein in the presence of tRNA to form capsid-like structures. Yeast two-hybrid and immunoprecipitation analyses reveal that oligomerization of the core protein is promoted by amino acid residues 72 to 91 in the core. Furthermore, the association between the E1 glycoprotein and the assembled core can be recapitulated using a fusion protein containing the putative cytoplasmic loop of the E1 glycoprotein. This fusion protein is also able to compete with the intact E1 glycoprotein for binding to the core. Mutagenesis of the cytoplasmic loop of E1 was used to define a region of four amino acids (residues 312 to 315) that is important for interaction with the assembled HCV core. Taken together, our studies suggest that interaction between the self-oligomerized HCV core and the E1 glycoprotein is mediated through the cytoplasmic loop present in a polytopic form of the E1 glycoprotein