The isolation and characterisation of protein partners for Cdc42Hs

Cdc42Hs is a member of the Rho family of GTPases that has a role in fundamental cellular processes such as the regulation of cell morphology, JNK activation, neurite outgrowth, cell polarity and cell transformation. To investigate how Cdc42Hs can control such diverse functions, a yeast two- hybrid screen was performed to identify novel Cdc42Hs-binding partners, which may be involved in mediating pathways underlying these processes. A screen of a human brain cDNA library with constitutively active Cdc42Hs bait resulted in the isolation of 358 putative targets. Seven positive clones (Cdc42Hs-Binding Protein 1-7) were characterised. CBP2, CBP6 and CBP3 were identified to be PAK, aPAK and ACK, respectively. CBP4 and CBP7 exhibit no similarity to known sequences. CBP1 and CBP5 showed identity to the hamster cDNA, encoding the insulin receptor tyrosine kinase p53/58 substrate. Full-length CBP5 cDNA is 83% identical to the hamster IRSp53 gene suggesting CBP5 is the human homologue of the hamster cDNA and was therefore renamed as insulin receptor tyrosine kinase substrate, IRS-58. Northern analysis of IRS-58 mRNA expression identified two transcripts, one of which was ubiquitously expressed, while the other is brain-enriched and possibly brain-specific. IRS-58 encodes four protein-protein interaction motifs; an SH3 domain, an SH3-binding domain, a WW-binding domain and, a novel Cdc42Hs-binding domain that exhibits similarity to, but is distinct from, the conserved p21 binding CRIB motif. Expression of IRS-58 in Swiss 3T3 fibroblasts results in the reorganisation of the actin cytoskeleton including a loss of stress fibres, cell retraction and the formation of F-actin rich clusters. Expression of IRS-58 in N1E-115 neuroblastoma cells induces the formation of thick, highly branched neurites that exhibit numerous projections along the length of the neurite. IRS-58-induced neurite formation is dependent on active Cdc42Hs and Rac as well as direct Cdc42Hs binding. Taken together these data suggest that IRS-58 is a novel effector for Cdc42Hs involved in remodelling of the F-actin cytoskeleton and in inducing morphological differentiation

Reversing T cell immunosenescence: why, who, and how

Immunosenescence is the term commonly used to describe the multifaceted phenomenon encompassing all changes occurring in the immune system during aging. It contributes to render older adults more prone to develop infectious disease and main age-related diseases. While age clearly imposes drastic changes in immune physiology, older adults have heterogeneous health and immune phenotypes. This confronts scientists and researcher to develop more age-specific interventions rather than simply adopting intervention regimes used in younger people and this in order to maintain immune protection in older adults. Thus, this review provides evidences of the central role played by cell-mediated immunity in the immunosenescence process and explores the means by which senescent state of the cell-mediated immune function could be identified and predicted using biomarkers. Furthermore considerations are given to recent advances made in the field of age-specific immune interventions that could contribute to maintain immune protection, to improve quality of life, and/or to promote healthy aging of the growing part of the populatio

Effectiveness of influenza vaccine in aging and older adults: comprehensive analysis of the evidence

Foremost amongst the diseases preventable by vaccination is influenza. Worldwide, influenza virus infection is associated with serious adverse events leading to hospitalization, debilitating complications, and death in elderly individuals. Immunization is considered to be the cornerstone for preventing these adverse health outcomes, and vaccination programs are timed to optimize protection during the annual influenza season. Trivalent inactivated influenza virus vaccines are believed to be both effective and cost-saving; however, in spite of widespread influenza vaccination programs, rates of hospitalization for acute respiratory illness and cardiovascular diseases have been increasing in this population during recent annual influenza seasons. From meta-analyses summarizing estimates of influenza vaccine effectiveness from available observational clinical studies, this review aims to examine how effective current influenza vaccine strategies are in the aging and older adult population and to analyze which are the most important biases that interfere with measurements of influenza vaccine effectiveness. Furthermore, consideration is given to strategies that should be adopted in order to optimize influenza vaccine effectiveness in the face of immune exhaustion

Recommendations for enterovirus diagnostics and characterisation within and beyond Europe.

Enteroviruses (EV) can cause severe neurological and respiratory infections, and occasionally lead to devastating outbreaks as previously demonstrated with EV-A71 and EV-D68 in Europe. However, these infections are still often underdiagnosed and EV typing data is not currently collected at European level. In order to improve EV diagnostics, collate data on severe EV infections and monitor the circulation of EV types, we have established European non-polio enterovirus network (ENPEN). First task of this cross-border network has been to ensure prompt and adequate diagnosis of these infections in Europe, and hence we present recommendations for non-polio EV detection and typing based on the consensus view of this multidisciplinary team including experts from over 20 European countries. We recommend that respiratory and stool samples in addition to cerebrospinal fluid (CSF) and blood samples are submitted for EV testing from patients with suspected neurological infections. This is vital since viruses like EV-D68 are rarely detectable in CSF or stool samples. Furthermore, reverse transcriptase PCR (RT-PCR) targeting the 5'noncoding regions (5'NCR) should be used for diagnosis of EVs due to their sensitivity, specificity and short turnaround time. Sequencing of the VP1 capsid protein gene is recommended for EV typing; EV typing cannot be based on the 5'NCR sequences due to frequent recombination events and should not rely on virus isolation. Effective and standardized laboratory diagnostics and characterisation of circulating virus strains are the first step towards effective and continuous surveillance activities, which in turn will be used to provide better estimation on EV disease burden

Reducing the environmental impact of surgery on a global scale: systematic review and co-prioritization with healthcare workers in 132 countries

Abstract Background Healthcare cannot achieve net-zero carbon without addressing operating theatres. The aim of this study was to prioritize feasible interventions to reduce the environmental impact of operating theatres. Methods This study adopted a four-phase Delphi consensus co-prioritization methodology. In phase 1, a systematic review of published interventions and global consultation of perioperative healthcare professionals were used to longlist interventions. In phase 2, iterative thematic analysis consolidated comparable interventions into a shortlist. In phase 3, the shortlist was co-prioritized based on patient and clinician views on acceptability, feasibility, and safety. In phase 4, ranked lists of interventions were presented by their relevance to high-income countries and low–middle-income countries. Results In phase 1, 43 interventions were identified, which had low uptake in practice according to 3042 professionals globally. In phase 2, a shortlist of 15 intervention domains was generated. In phase 3, interventions were deemed acceptable for more than 90 per cent of patients except for reducing general anaesthesia (84 per cent) and re-sterilization of ‘single-use’ consumables (86 per cent). In phase 4, the top three shortlisted interventions for high-income countries were: introducing recycling; reducing use of anaesthetic gases; and appropriate clinical waste processing. In phase 4, the top three shortlisted interventions for low–middle-income countries were: introducing reusable surgical devices; reducing use of consumables; and reducing the use of general anaesthesia. Conclusion This is a step toward environmentally sustainable operating environments with actionable interventions applicable to both high– and low–middle–income countries

Immunotherapy of immunosenesence; who, how and when?

Major changes in social welfare, economic conditions and medical knowledge over the last 60 years have pro-duced a demographic shift in the population. More individuals are living longer, and in a decade there will be more people over 65 than infants under 5 for the first time in history. Taking the analysis beyond mere numbers reveals that older indi-viduals are now physically more active than their forebears and travel more widely. This provides a greater opportunity for encountering infectious agents which could present a considerable problem. Older individuals are more susceptible to infection and do not respond as well as younger people to vaccination because of an age related decline in immunity, a state which has been termed immunosenesence. This decline is not uniform and some older individuals show a greater de-cline in their immune response than others. In this review we have sought to consider who are the ‘at risk' individuals, how they might best be treated and when

Vaccine effectiveness in older individuals: what has been learned from the influenza-vaccine experience

Vaccination policies in most high-income countries attempt to reduce the adverse impact of influenza targeting people aged at least 60 years. However, while it is widely believed that the current immunization strategy saves many lives, influenza infection still remains a severe burden in aged individuals leading to a wide debate on the exact magnitude of the benefit of vaccination in this population. The first aim of the present review is to examine how effective current influenza-vaccine strategies are in aged adults, by analysing which are the most important factors modulating the interpretation of study results in this population. Furthermore, consideration will be given to how immune factors influence the measurement of vaccine efficacy/effectiveness, where advancing age leads to deleterious changes in the adaptive immune system, resulting in less than optimal responses to infectious agents and vaccination. Finally this review concludes with possible strategies to improve the ability of the senescent immune system to respond to vaccination