Nipah virus induces two inclusion body populations: Identification of novel inclusions at the plasma membrane.

Formation of cytoplasmic inclusion bodies (IBs) is a hallmark of infections with non-segmented negative-strand RNA viruses (order Mononegavirales). We show here that Nipah virus (NiV), a bat-derived highly pathogenic member of the Paramyxoviridae family, differs from mononegaviruses of the Rhabdo-, Filo- and Pneumoviridae families by forming two types of IBs with distinct localizations, formation kinetics, and protein compositions. IBs in the perinuclear region form rapidly upon expression of the nucleocapsid proteins. These IBperi are highly mobile and associate with the aggresome marker y-tubulin. IBperi can recruit unrelated overexpressed cytosolic proteins but do not contain the viral matrix (M) protein. Additionally, NiV forms an as yet undescribed IB population at the plasma membrane (IBPM) that is y-tubulin-negative but contains the M protein. Infection studies with recombinant NiV revealed that IBPM require the M protein for their formation, and most likely represent sites of NiV assembly and budding. The identification of this novel type of plasma membrane-associated IBs not only provides new insights into NiV biology and may open new avenues to develop novel antiviral approaches to treat these highly pathogenic viruses, it also provides a basis for a more detailed characterization of IBs and their role in virus assembly and replication in infections with other Mononegavirales.

Influenza virus-mediated suppression of bronchial Chitinase-3-like 1 secretion promotes secondary pneumococcal infection.

Infections of the lung are among the leading causes of death worldwide. Despite the preactivation of innate defense programs during viral infection, secondary bacterial infection substantially elevates morbidity and mortality rates. Particularly problematic are co-infections with influenza A virus (IAV) and the major bacterial pathogen Streptococcus pneumoniae. However, the molecular processes underlying the severe course of such co-infections are not fully understood. Previously, the absence of secreted glycoprotein Chitinase-3-like 1 (CHI3L1) was shown to increase pneumococcal replication in mice. We therefore hypothesized that an IAV preinfection decreases CHI3L1 levels to promote pneumococcal infection. Indeed, in an air-liquid interface model of primary human bronchial epithelial cells (hBECs), IAV preinfection interfered with apical but not basolateral CHI3L1 release. Confocal time-lapse microscopy revealed that the gradual loss of apical CHI3L1 localization during co-infection with influenza and S. pneumoniae coincided with the disappearance of goblet as well as ciliated cells and increased S. pneumoniae replication. Importantly, extracellular restoration of CHI3L1 levels using recombinant protein significantly reduced bacterial load in influenza preinfected bronchial models. Thus, recombinant CHI3L1 may provide a novel therapeutic means to lower morbidity and mortality associated with post-influenza pneumococcal infections.

Replication of a Nipah Virus Encoding a Nuclear-Retained Matrix Protein.

Nipah virus (NiV) matrix protein (NiV M) plays a major role in virus assembly. It undergoes nuclear transit before accumulating at the plasma membrane and recruiting nucleocapsids to the budding sites. Because nuclear NiV M cannot be detected in all cell types, we wondered whether it can reach the cell surface by bypassing the nucleus. Using an M mutant with a defective nuclear export signal (MNESmut), however, we revealed that the nuclear import of M is ubiquitous, because MNESmut was retained in the nuclei of all cell types tested. Because a functional nuclear transit is a general prerequisite for M surface transport, we wanted to characterize the effect of nuclear-retained M protein in a full viral context and generated a recombinant NiV-MNESmut. Mutant NiV-MNESmut caused increased cell-cell fusion and produced lower virus titers. As expected for an assembly defective NiV, perinuclear inclusions (IBperi) were formed, but inclusions at the plasma membrane (IBPM), which probably represent the viral assembly platforms, were not found. It is interesting to note that the transport-defective MNESmut was recruited to IBperi. This probably prevents overaccumulation of nonfunctional M proteins in the cytoplasm and nuclei of NiV-infected cells and thus provides first evidence that IBperi are functionally relevant aggresome-like compartments.

Institutional review board training when patients and community members are engaged as researchers.

Background: Patient engagement efforts often rely on a participatory research approach, which means engaging patients and community members in all aspects of research. As research team members, they require familiarity with the principles of human subject protection, privacy, and institutional review boards (IRB). However, the time required for individual IRB training may be a barrier to engaging community members in participatory research. As more community members participated in research, the State Networks of Colorado Practices and Partners (SNOCAP) was faced with finding a balance between including community members as part of the research team and the significant time commitment and institutional requirements for human subjects research oversight. Objective: Design and implement a community training on human subject protection in research. Methods: The SNOCAP team worked with the leadership from the Colorado Multi-Institutional Review Board (COMIRB) to develop a training programme that included the ethical principles and guidelines for the protection of human subjects. Results: The final training programme was based on the core principles of the Belmont Report: respect for persons, beneficence and justice. Privacy was taught using the Health Insurance Portability and Accountability Act (HIPAA) national guidelines. Conclusions: The members of the High Plains Research Network Community Advisory Council were fully engaged in developing the training programme, as well as in the training itself. They were committed to the principles and guidelines for protecting the rights and welfare of human subjects. Patients and community members have become a critical part of our research team. They understand the principles of human subjects protection and privacy and incorporate these principles into their research activities.

Formation of high-order oligomers is required for functional bioactivity of an African bat henipavirus surface glycoprotein.

Hendra virus (HeV) and Nipah virus (NiV) are highly pathogenic henipaviruses originating from fruit bats in Australia and Asia that can cause severe infections in livestock and humans. In recent years, also African bat henipaviruses were identified at the nucleic acid level. To assess their potential to replicate in non-bat species, several studies were performed to characterize the two surface glycoproteins required for virus entry and spread by cell-cell fusion. It has been shown that surface expression and fusion-helper function of the receptor-binding G protein of Kumasi virus (KV), the prototypic Ghanaian bat henipavirus, is reduced compared to other non-African henipavirus G proteins. Immunostainings and pulse-chase analysis revealed a delayed export of KV G from the ER. As defects in oligomerization of viral glycoproteins can be responsible for limited surface transport thereby restricting the bioactivity, we analyzed the oligomerization pattern of KV G. In contrast to HeV and NiV whose G proteins are known to be expressed at a dimer-tetramer ratio of 1:1, KV G almost exclusively formed stable tetramers or higher oligomers. KV G also showed less stringent requirements for defined stalk cysteines to form dimers and tetramers. Interestingly, any changes in the oligomeric forms negatively affected the fusion-helper activity although surface expression and receptor binding was unchanged. This clearly indicates that the formation of mostly higher oligomeric KV G forms is not a deficiency responsible for ER retention, but is rather a basic structural feature essential for the bioactivity of this African bat henipavirus glycoprotein.

How do rural patients benefit from the patient-centred medical home? A card study in the High Plains Research Network.

Context The patient-centred medical home (PCMH) has become a dominant model for improving the quality and cost of primary care. Geographic isolation, small populations, privacy concerns and staffing requirements may limit implementation of the PCMH in clinical practice. Objective To determine the primary care provider perceived benefit of PCMH for patients in rural Colorado. Design, setting and participants The High Plains Research Network (HPRN) is a community and practice-based research network spanning 30 000 square miles in 16 counties in eastern Colorado. The HPRN consists of 58 practices, 120 primary care clinicians and 145 000 residents. Main outcome measures Providers' perceived benefit of PCMH for individual patients. Results Seventy-eight providers in 37 practices saw 1093 patients and completed 1016 surveys. There was wide variation among the provider-perceived benefits of PCMH elements ranging from 9% for group visits to 64% for electronic prescribing. Provider-perceived benefit was higher for patients with a chronic medical condition. Conclusions Rural primary care providers perceived patient benefit for numerous elements of the PCMH. There is need to consider what PCMH elements may be required in practice and what components might be optional. Our findings reveal that rural practices share PCMH aspirations including commitment to quality, safety, outcomes, cost reduction, and patient and provider satisfaction. These findings support the need for ongoing conversation about how to best provide a locally relevant medical home.

Translating the medical home into patient-centred language.

Background The patient-centred medical home (PCMH) is a healthcare delivery model that aims to make health care more effective and affordable and to curb the rise in episodic care resulting from increasing costs and sub-specialisation of health care. Although the PCMH model has been implemented in many different healthcare settings, little is known about the PCMH in rural or underserved settings. Further, less is known about patients' understanding of the PCMH and its effect on their care. Aims The goal of this project was to ascertain the patient perspective of the PCMH and develop meaningful language around the PCMH to help inform and promote patients' participation with the PCMH. Method The High Plains Research Network Community Advisory Council (CAC) is comprised of a diverse group of individuals from rural eastern Colorado. The CAC and its academic partners started this project by receiving a comprehensive education on the PCMH. Using a community-based participatory research approach, the CAC translated technical medical jargon on the PCMH into a core message that the 'Medical Home is Relationship'. Results The PCMH should focus on the relationship of the patient with their personal physician. Medical home activities should be used to support and strengthen this relationship. Conclusion The findings serve as a reminder of the crucial elements of the PCMH that make it truly patient centred and the importance of engaging local patients in developing and implementing the medical home.