Mesenchymal stem cells (MSC) have piqued worldwide interest for their extensive potential to treat a large array of clinical indications, their unique and controversial immunogenic and immune modulatory properties allowing ample discussions and debates for their possible applications. Emerging data demonstrating that the interaction of biomaterials and physical cues with MSC can guide their differentiation into specific cell lineages also provide new interesting insights for further MSC manipulation in different clinical applications. Moreover, recent discoveries of some regulatory molecules and signaling pathways in MSC niche that may regulate cell fate to distinct lineage herald breakthroughs in regenerative medicine. Although the advancement and success in the MSC field had led to an enormous increase in the amount of ongoing clinical trials, we still lack defined clinical therapeutic protocols. This review will explore the exciting opportunities offered by human and animal MSC, describing relevant biological properties of these cells in the light of the novel emerging evidence mentioned above while addressing the limitations and challenges MSC are still facing. Thanks to more than two decades of extensive research, scientific evidence confirms a promising potential for mesenchymal stem cell (MSC)-based therapeutic strategies. This is largely due to the advancement in the knowledge of their basic biological properties, which was translated into clinical opportunities. MSC are isolated from different tissue/organs of both humans and animals. Their exclusive properties such as: 1) multipotency, 2) ability to adhere to plastic surfaces, 3) expression of specific cell surface antigens [ multipot ability  expressiDue to the variety of clinical applications that MSC maybe engaged in, this review will focus and highlight some emerging evidence about the biological and physiological properties of both human and animal MSC. Promising animal models to monitor MSC in order to improve their homing and survival in translational medicine settings will be considered. MSC interaction with different cells of the human body including immune cells for immunomodulation and endothelial cells for angiogenesis and bone healing will be covered. Their interplay with biomaterials as a therapeutic option will also be adressed. Present challenges in their translation into clinical practice will be finally highlighted.MSC isolated from different tissue sources offer the basis for research studies focusing on their characterization to address potential therapeutic options. Animal-derived MSC provide an abundant resource to explore the specific properties of these cells to evaluate their potential functional benefits. In fact, the possibility to isolate and characterize specific biological functions of animal MSC both from adult and young tissues offers new possibilities for their therapeutic potential. Human bone marrow MSC were the first source discovered and the most frequently studied . They arAmong animal species, canine MSC (cMSC) can be considered a good model to evaluate potential biological mechanisms that could be exploited for their possible application in therapy. cMSC isolated from different tissue sources  demonstrated similarities to human MSC (hMSC), apart from minor differences . A discrRegarding the potentialities of growth factor for MSC biology during regenerative processes, data from Mellado-López and co-workers demonstrated that plasma rich in growth factors (PRGF) from platelet-rich plasma significantly improves cell survival of ASCs when are exposed to proapoptotic concentrations of hydrogen peroxide (used for mimicking oxidative stress as occurs at site of injury and/or transplantation area) by inducing phosphorylation of AKT . These fMore recently, other studies evidenced the role of the fibroblast growth factor-2 (FGF-2) in improving canine BM-periadipocyte cells (PAC) proliferation . In the Accordingly, spheroids tend to express more cartilaginous ECM because of their oxygen gradient in their microenvironment, which exposes cells in the central area to hypoxic conditions, therefore enhancing chondrogenesis  Table 1Table 1. Other sources of cMSC can be the ovaries and placenta. In particular, placental cMSC demonstrated adequate paracrine secretory factors  which facilitated neural networks formation when co-cultured in the presence of SH-SY5Y cells. Ovary-derived cMSC showed morphological kinetic properties comparable to adipose tissue-derived cMSC. The latter demonstrated high capacity to differentiate not only into adipo-, osteo- and chondrogenic tissue, but also towards neurogenic and endodermal lineages .In addition to canines, other animals can be used as sources for MSC for translational studies in regenerative medicine. Ovine species offered the setting for several preclinical studies relevant to human physiology ,20,21. IRecently, new therapeutic approaches, such as the use of ovine peripheral blood derived MSCs (PB-MSCs) in skin lesion, were tested. Results demonstrated a discrete efficacy in PB-MSCs showing a good skin re-epithelialization. The ability of these cells to speed up the formation of granulation tissue, stimulate neovascularization, and increase structural proteins and skin adnexa, suggests their future application for deeper lesions .Similarities between the human and porcine anatomy make the latter an attractive model for preclinical studies on MSC. Pig MSC (pMSC) characterized for their morphology revealed homogenous cell populations (size and granularity) coherently with flow cytometric results obtained from their hMSC counterparts . RecentlRecently, new and interesting results have been obtained by studies on secretome by porcine vascular wall–mesenchymal stem cells (pVW-MSCs), demonstrating a particular composition with high levels of IL-8, GM-CSF, IFN-γ and other immunomodulatory proteins: IL-6 IL-18 IL-4 IL-2 IL-10. Moreover, conditioned medium from unstimulated pVW-MSCs promoted in vitro endothelial angiogenesis, which manifested more strongly when the conditioned medium was from LPS .MSC isolated from some tissues of equine species showed good regenerative potential of soft tissues. This ability seems principally related to the angiogenesis stimulation by trophic factors secreted by equine MSC. In particular, studies revealed that VEGF plays a particular role through its paracrine bioactive factors IL-8, PDGF-AA, ET-1 and IGFBP2  Table 1Table 1. Recent innovative studies have been focused to apply new strategies for mapping the equine MSCs surface proteome by using biotin-enrichment and mass spectrometry (MS) analysis in consideration of limited availability of suitable antibodies of high quality. Results demonstrated for the first time in horse species the possibility to use this method and thereby identify 1239 proteins including 19 CD markers.M) encapsulated in a three-dimensional alginate hydrogel scaffold differentiated into chondrocytes, opening new possibilities for clinical application in the treatment of joint injuries in horses [In particular, regarding chondrogenic differentiation,  demonstrn horses . Moreoven horses . In addin horses . This wan horses .A recent innovative method utilizing electroacupuncture (EA) has been tested for mobilization of horse MSC into peripheral blood. The obtained results demonstrated a good efficacy, and the immunomodulatory properties of MSC indicate this method as a valid alternative to surgical procedure .In the last five years a growing interest on feline species, confirmed by a discrete amount of studies, had emerged. Albeit the isolation and the characterization of feline BM-derived MSC started in 2002 , severalThe growing interest in veterinary MSC research is a mandate not only to reinforce our current knowledge of the biological properties of MSC for translational human applications but for creation of novel methodologies and therapeutic applications in veterinary medicine too. Indeed, the last decade had experienced a vast majority of animal MSC research investigating new insights into untraditional clinical therapy benefiting human and animal health .In addition, the possibility of in vivo cross-species administration of MSC in a variety of experimental models has been proven in these last ten years. Results from literature show in vivo cross-species administration of MSC in a variety of experimental models. In particular, among the tested animal species, pig MSC showed a good function in a different species, e.g., humans . Moreover, results from a recent paper by Daems et co-workers  aimed toAlbeit the benefits, limitations in the use of MSC from animal species still exist, some of which are: (i) high variability in differentiation ability and in surface marker expression, (ii) controversy regarding the safety procedures in xenogenic MSC transplantation, (iii) lack of adequate knowledge on the appropriate MSC source to be utilized to cure certain pathologies, (iv) dilemma of ethical concerns are currently being discussed that require the engagement of specialists outside the scientific community. Despite the advancement in the animal field of MSC, further investigations are needed to confirm the effective efficacy of animal MSC in regenerative medicine.+ CD1a− precursors into dermal/interstitial DC without affecting the generation of CD1a+ Langerhans cells through PGE2, IL-10 and downstream signaling via the JAK-STAT pathway [versus host disease (GvHD) and autoimmune diseases such as systemic lupus erythematosus [MSC have attracted much attention for their ability to regulate the immune system in vivo and in vitro. Their therapeutic potential is currently being investigated in various immunological disorders such as Crohn’s disease , graft-v pathway . TNF-α-s pathway . MSC hav pathway . However pathway ,66. Inde pathway . Bioacti pathway . The mic pathway . MSC decematosus . Howeverematosus . Inflammematosus . Whetherematosus . MSC exeematosus .In the majority of the completed clinical trials, recipients of MSC-based therapy demonstrated good tolerance and improved clinical symptoms. There remain challenges to the future development of MSC for immunomodulation and a need for improved quality control. Another limiting factor is that MSC for basic research and clinical applications are manufactured and developed as unique cell products by many different laboratories, often under different conditioned media. Immune modulatory effects of MSC are indeed altered by the different expansion media . Human pMSC enhance angiogenesis by phenotypically switching into the endothelial lineage and mainly exerting a paracrine action into the microenvironment . This isThe angiogenic fate of MSC can be also addressed by the degree of hypoxia levels and injury, the in-situ production of inflammatory molecules, the mechanical properties of the tissue , the hos+/CD31− MSC pool with VEGF/IGF, enhances endothelial differentiation, strengthening the importance of the endothelial contamination originally present in the stromal fraction [Additional and major drawbacks also originate from the need to define the stromal population by a panel of multiple markers excluding CD31, CD117 and CD34 ,100,101.fraction .A top priority should be to verify if MSC-derived new formed vessels are only temporarily functioning or they can foster the engraftment of circulating endothelial progenitor and display a specific endothelial metabolic asset including nitric oxide production, response to shear stress and uptake of LDL ,103. ResTheoretically, the precursors of MSC are the pericytes exerting a physiological and protective role on the vasculature . The supWe should also consider that the stromal tissue origin determines several biological properties of MSC ,128, incOne of the most significant modalities of the MSC-mediated paracrine mechanism is in the form of microvescicles and exosomes, known to control the physiological process of angiogenesis ,143,144.To date, the precise contribution of some miRNAs derived from extracellular vesicles of stromal origin has been individually established as reported for stem cells which are protected by exosomes-derived MSC through miR22 and Mecp2 pathways upon ischemia . miR675-The MSC-derived miRNA cargo is the product of a balanced mix of positive and negative endothelial modulators, representing an interesting tool to control or even to arrest angiogenesis. In fact, in rheumatoid arthritis, the delivery of the negative modulator miR150-5p by means of exosomes allows direct downregulation of MMP14 and VEGF, both mainly responsible for angiogenic/inflammatory-based clinical complications . BesidesMore recently, a role of exosomes has been evidence in relation to the particular involvement of MSCs in the tumor microenvironment and thus in the cancer development metastasis process. In fact, it has well shown that tumor migratory MSCs similar to cancer stem cells (CSCs) act in promoting tumor growing and progression . In partDifferently, considering the antitumor activity, MSC-derived exosomes seemed to inhibit prostate cancer via delivery of miR-145 by reducing the activity of Bcl-xL and stimulating apoptosis by acting on the caspase-3/7 pathway . More reThus, the modulation of the mesenchymal secretome is a current reality. Accordingly, packaged microparticles of soluble mediators released by MSC and coated with stromal membranes have being developed to synthetically mimic the angiogenic paracrine asset of MSC, providing the advantage to control the releasing dose and the stability over the time upon myocardial infarction . AlternaRecently, bioengineering approaches by dosing the amount of soluble angiogenic factors in transfected MSC have being a relevant strategy to empower MSC functionality . AlternaMechanobiology of stem cells is today a significant issue. Elucidating the interactions between biomaterials and cells has potential to be used for medical purposes. Indeed, the design of biomaterials to mimic natural scaffolds is a novel perspective for developing more efficient stem cell-based therapies in regenerative medicine. To this aim we need to understand that forces generated by adhesion to synthetic extracellular matrices affect stem cell gene expression profiles and ultimately their phenotype. In addition, a better comprehension of how interaction between stem cells and biomaterials, as well as compression or tension changes affects the cells, is also required.Cells are heterogeneous gels endowed with highly dynamic behaviour in terms of their elastic properties. In this respect, their response to a biomaterial can be monitored as their tensile elasticity, i.e., the Young’s modulus, which measures in pascal units (Pa) deformation when receiving a physical stress. A high Young’s modulus is consistent with a stiff behaviour, indicating reduced ability to deform. Undifferentiated hMSC display Young’s modulus values in the range of kPa magnitude order .As an example, when administered a soluble cocktail inducing an osteogenic phenotype commitment, hMSC show a pronounced decrease in their elastic modulus  AlthoughAll the above-mentioned mechanical stimuli are in fact activating surface receptors inducing intracellular signaling cascades driving specific molecular activation, such as RhoA and Rho-associated protein kinase (Rock) signaling , and conActive physiologic mechanical solicitations can be administered as differentiation cue via actuators—namely, bioreactor systems—that are used in vitro to prime cell-based 3D tissue constructs, potentially aiming at a following implant in vivo. Several configurations were designed for different kind of stimuli. Most diffused bioreactor systems are rotating wall systems, spinner flask systems, perfusion system, compression and strain systems . NowadayOther interesting aspects and new perspectives in the field of tissue regeneration and applicative use of MSC are strictly related to their potential to tackle the increase in bone diseases related with the extended life expectancy. Indeed, in many countries bone diseases are becoming a relevant cause of disability, even if they are acute, such as fractures, or chronic, such as osteoporosis and tumours. In any case these pathologies require treatments to enhance the healing and regenerative capacity of bone tissue. The common therapeutic strategies based on bone grafting  show some disadvantages. Autografts are limited by the bone volume that can be harvested from the iliac crest and present surgical risks such as bleeding, inflammation, infection, chronic pain, damage at the donor site and morbidity. Allografts also have some limitations, such as the lack of donors, high costs, the need for sterilization and the risk of infectious agent transmission or immune mediated tissue rejection.These limitations and disadvantages associated with auto- and allograft approaches indicate a clinical need for alternative therapeutic strategies aimed at bone healing and regeneration. Thus, new biomaterials and scaffolds, in association with MSC and growth factors, having requirements of biocompatibility, osteoinductive and osteoconductive properties, are investigated to improve bone repair ,196,197.hMSC have been found in several adult tissues, including the synovial membrane, the adipose tissue, the dental pulp tissue, or in perinatal tissues . All these cells are suited to therapeutic applications for bone regeneration, by means of two different ways of autologous cells transplantation: (1) cell therapies without expansion in culture (one-step procedure) and (2) cell therapies with ex vivo expansion. In the first case, cells are harvested during surgery. In 2010, Jagër et al., successfully treated more than 100 patients with local bone healing disorders using a biomaterial composite in association with BM aspiration concentrate . They obBone marrow contains MSC, hematopoietic stem cells, endothelial progenitor cells, other progenitor cells and growth factors, including bone morphogenetic proteins (BMP), platelet-derived growth factor (PDGF), transforming growth factor-β (TGF-β), vascular endothelial growth factor (VEGF), interleukin-8 (IL-8), and IL-1 receptor antagonist. It has been identified as an excellent source of cells and growth factors and it has been used with success for bone, cartilage and soft tissue healing ,202,203.We focused our attention on the use of BM-derived MSC in spinal fusion surgical procedures, which is the most definitive treatment performed to restore the structural stability of the spinal column in degenerative and oncological spine diseases. Currently, the gold standard stimulus to achieve spinal fusion is autologous bone (autograft), which is commonly harvested from the iliac crest or obtained from the surgical site . As autologous bone possesses osteogenic, osteoinductive and osteoconductive abilities; the success rate of spinal fusion with autograft is high (up to 95%). However, autograft material is limited in quantity and its quality varies depending on the patient. Moreover, significant morbidity is associated with bone harvesting, as previously described. Between many alternatives proposed to achieve spinal fusion, MSC are interesting because they can provide osteogenic properties allowing bone regeneration.We recently published a descriptive systematic literature review in order to understand if the use of MSC may represent a valid strategy to facilitate and accelerate bone regeneration and fusion during spine surgery . In thisSo far, few published clinical studies employ stem cells for spinal fusion ,205. MosDespite these results, some critical limitations exist, including the site of BM harvest, the choice of the optimal cell concentration, the methods to obtain the maximum number of cells, the delivery method (appropriate scaffold), the ideal manipulation procedure (one-step or ex vivo expansion procedure) and the best implantation technique. Thus, more in vitro and animal studies are useful to highlight these concerns and randomized controlled clinical trials are necessary to carefully evaluate the safety and efficacy of MSC use in spine surgery.In this scenario, our in vitro studies focused on the site of BM harvest, as the withdrawal from iliac crest during spine surgery leads to an increase in operative and rehabilitation time and to possible donor site morbidity. We previously observed that stromal populations from different sources, although immunophenotypically similar, display distinct signatures of HOX and TALE homeobox genes . Our datAfterwards, we analyzed the strategies developed for cell-based therapies, encompassing in vitro expanded MSC and “one-step” procedures using BM in toto  or BMC. Specifically, using BMA appears as a promising technique, showing increased regenerative potential by the addition of marrow elements and the possibility to perform the entire procedure directly in the operating room ,218.Following collection of vertebral BMA samples, we analyzed the biological characteristics and HOX and TALE gene expression in vMSC derived from whole BMA and from density-gradient centrifugation of BMA, both cultured under the hypoxic stimulus, in order to identify the best cell isolation technique and condition for spinal surgery application  Figure .Our results showed that the in vitro expansion rate and CFU potency of vMSC derived from whole BMA under hypoxia were increased compared to all the other culture conditions analyzed.Although all culture conditions generate mature cells of all mesenchymal lineages when induced under osteogenic, adipogenic and chondrogenic differentiation, RT-PCR showed a significant upregulation of RUNX2 and ALPL in vMSCs derived from whole BMA under hypoxia in comparison to normoxia. A different behavior was observed in vMSCs derived from density gradient centrifugation where a lower expression of COL1A1 was observed in hypoxia in comparison to normoxia. This lower expression of COL1A1 under hypoxia indicated a decreased efficiency in the final osteogenic potential, previously observed ,221,222 In vMSCs derived from whole BMA, it was seen that hypoxia strongly up-regulated the expression of chondrogenic genes SOX9 and ACAN in comparison to normoxia. Similarly, vMSC derived from density gradient centrifugation highlighted an upregulation of ACAN. However, in vMSCs derived from whole BMA, contrary to osteogenic and chondrogenic gene expression, adipogenic genes, PPAR-γ and ADIPOQ, were up-regulated under normoxia. These results indicate that vMSCs derived from whole BMA under hypoxia differentiated onto the adipogenic lineage following exposure to specific inducing agents but with a lower efficiency in comparison with all the other culture methods and condition. Thus, we hypothesized that low oxygen tension in vMSCs derived from whole BMA reduced adipogenesis, as postulated by other authors, in a hypoxia inducible factor (HIF)-1 dependent manner.This is interesting for the improvement of spinal surgery where the hypoxic condition of the site would facilitate the differentiation of vMSC towards osteoblasts, inhibiting adipogenesis and favoring the initiation of proper bone regeneration .We also addressed the question of whether vMSCs isolated with different techniques might, following osteogenic induction, modulate the expression levels of HOX and/or TALE genes. We found that vMSCs derived from whole BMA up-regulated HOXB8 under hypoxia. HOXB8 is involved in the expansion of hematopoietic stem and early progenitor cells and it is also implicated in vertebral development, since its inactivation causes slight vertebral abnormalities while its increase can help by-pass the block of posterior elongation of axial tissues ,224,225.The results of this study showed for the first time that hypoxic preconditioning of vMSCs derived from whole BMA can enhance proliferation, morphology, osteogenesis and chondrogenesis, inhibit adipogenesis and up-regulate distinct level of HOX signatures. These aspects could open new perspectives for the improvement of spinal surgical procedures wherever the hypoxic nature of the site would facilitate the differentiation of vMSCs towards osteoblasts, giving the biological rationale for the use of vertebral bone marrow in spinal surgery.Because of the presence of megakaryocytes and platelets, BMA is prone to clotting, even after the addition of anticoagulants . When weAccording to our data ,218,219,We support the idea that stem cells have a big potential that can be exploited to treat many diseases and structural damages using cell therapy and regenerative medicine strategies. However, as previously observed, some limitations have to be overcome in order to use MSCs in clinical settings . Most coMoreover, the results of recent studies suggest that MSCs found in various niches of perinatal and adult tissues have a common source, primary mesenchyme. After migration from the mesenchyme during development, they then adapt to the designated niche. MSCs from each of the niches maintain certain common characteristics  while varying in others . This observation suggests that in clinical applications it is necessary to select the correct tissue for deriving MSCs in order to achieve the greatest outcome for the treatment of a particular disease. Following in vitro studies, we propose to use vertebral bone marrow as best source of MSCs for spinal fusion.In conclusion, based on our previous data, we are setting up a clinical trial in which vertebral bone marrow harvested from pedicles during the surgical procedure is re-implanted in the arthrodesis area through a “one-step” procedure, using an appropriate scaffold, without any additional surgical time or any involvement of other anatomical sites. In our opinion, the results of this trial could add new insights into the potential of MSCs in the clinical reality.The worldwide growing interest in MSC and their possible utilization in therapy has evolved to studies aimed to deepen the knowledge about MSC physiology, including issues such as their behaviour in response to autocrine and paracrine factors, their survival time, ability to migrate into organs and tissues and their donor-to-donor variability. All of these aspects certainly could complete the existing framework about MSC biology by providing support in the communication between basic scientists and clinicians for more useful applications in therapy. In fact, with the current review, we have tried to demonstrate data from basic science to clinical trials not only for a substential overview on the current status of human and animal MSC but highlighting the limitations and challenges MSC continue to face despite the advancement of the field.In the near future, opportunities offered by new synergies and disciplines might stimulate research at the interface between biomedical sciences, engineering and clinical application to enhance the translation of MSC from basic research into the clinics.The discovery of the possible use of MSC derivatives, such as conditioned medium, exosomes and extracellular vesicle, represents a promising aspect for future clinical applications where active MSC products might act as substitute mediators of local MSC action in regenerative medicine protocols.


With an ageing world population, preservation of older adults’ health and quality of life (QoL) is paramount. Due to lower levels of physical functionality, older adults are particularly susceptible to local environment influences, especially those living alone and lacking family support. Using generalised additive mixed models, we examined associations and confounder-adjusted associations between objectively-measured neighbourhood attributes and QoL domains in 909 Hong Kong Chinese elderly community dwellers. Most examined neighbourhood attributes were not associated with QoL in the whole sample. Neighbourhood residential and entertainment density was curvilinearly and/or linearly related to specific QoL domains. Number of parks was negatively associated with social QoL and having well-treed parks with higher levels of social QoL. Older adults living alone in neighbourhoods with poor access to destinations and few activities in parks showed lower environmental and/or social QoL than their counterparts. Neighbourhood built environment characteristics do not seem to impact Hong Kong older adults’ physical and psychological QoL. Medium-to-high density, well-ordered neighbourhoods with optimal mixes of well-treed public open spaces and services meeting their daily needs may significantly contribute to social and environmental QoL in this population and appear particularly important to those living alone. According to the World Health Organisation (WHO), quality of life (QoL) refers to people’s perceptions of their status in life in relation to their goals, standards and concerns, and within the context of their culture and value systems. This concept spans a wide range of aspects of life, including and beyond mere physical and mental health defined as the presence and severity of diseases . The breWith extended life expectancy, an increasing proportion of the world population is likely to experience poorer QoL due to age-related chronic health problems and declines in physical functioning and mobility . It is, Several studies have found associations between physical attributes of the neighbourhood environment and QoL in older adults. For example, various aspects of neighbourhood safety and quality were positively associated with health-related QoL ,10. HighMost of the studies examining physical environmental correlates of QoL used self-report measures of environmental attributes . Among tSocial-ecological models posit that health and well-being outcomes are shaped by the interaction of multiple levels of influences, including environmental, social and individual factors . In the Our study aimed to examine the associations between objectively-measured neighbourhood physical environmental attributes and QoL domains in Hong Kong older community dwellers and estimate the moderating effects of neighbourhood environmental attributes on the associations of living arrangements with QoL. We hypothesised that: (1) objective measures of availability of/access to destinations, public open spaces/parks and pedestrian-friendly infrastructure would be positively associated with QoL domains; (2) adverse environmental attributes, such as pollution and traffic-related hazards, would be negatively associated with QoL domains; (3) better access to/availability of destinations and lower levels of adverse environmental attributes would mitigate the negative effect of living alone on QoL.This study used data from the Active Lifestyle and the Environment in Chinese Seniors (ALECS) project, an observational investigation of associations between neighbourhood environment, physical activity, depressive symptoms and QoL in Hong Kong Chinese elderly community dwellers. The ALECS project used a two-stage sampling method to recruit participants from 124 tertiary planning units (TPUs) stratified by high/low socio-economic status (SES) and walkability to maximise the variation in environmental characteristics. TPUs are the smallest administrative area units with census data in Hong Kong. TPU-level SES was defined using Census data on median household income. TPUs were classified into high and low walkable based on a walkability index consisting of the sum of TPU-level z-scores for net residential density, intersection density and land-use mix derived using GIS data. Details about study design and neighbourhood selection have been reported elsewhere . Ethics p = 0.018), more likely to be men (p = 0.010) and reporting lower environmental QoL (p = 0.002) than their counterparts.Participants were recruited from the Elderly Health Centres (EHCs) and elderly community centres located in the pre-selected TPUs. The EHCs were established by the Department of Health of the Hong Kong Special Administrative Region to provide healthcare services to persons aged 65+ years and are distributed across all 18 Hong Kong districts. Although EHC clients are representative of the general Hong Kong elderly population in terms of age and SES, they tend to be more health-conscious than their counterparts . To examDue to restricted access to residential addresses and other contact details mandated by the Hong Kong Personal Data (Privacy) Ordinance , potentiExtant GIS data and newly-collected data from environmental audits were used to objectively assess neighbourhood environmental attributes. Extant GIS data were sourced from the Census and Statistics, Lands, and Planning Departments of the Hong Kong Special Administrative Region. To quantify relevant neighbourhood attributes using extant GIS data, participant residential buffers, representing individual neighbourhood boundaries, were created by geocoding the residential addresses of each participant and then tracing from the participants’ residences through the unique street networks in all directions for 400 and 800 m, which are considered to be walkable distances and appropriate geographical scales for older adults living in high density environments ,23. GIS-Environmental audits were employed to quantify neighbourhood attributes that were not assessable via extant GIS data  or for which the archival GIS database was outdated or incomplete. Crow-fly buffers of 400 m centred at participant residences were used as areas for environmental audits. We used 400 m crow-fly buffers rather than 400 m and 800 m street-network buffers for environmental audits for two reasons. First, study resources were insufficient to conduct environmental audits of all 400 m and 800 m street-network buffers  used for the computation of neighbourhood attributes based on extant GIS data. Second, previous audits conducted in Hong Kong indicated that a 400 m crow-fly distance corresponded to a street network distance from 400 to ~900 m ,35, implSingle and multiple EAST-HK items assessed the presence of particular environmental attributes  in each QoL was measured via interviewer-administration of the WHOQOL-BREF, the abbreviated QoL questionnaire developed by the WHO. The Hong Kong Chinese version of the WHOQOL-BREF  containsCovariates included in all analyses were age (years), gender , educational attainment , marital status , living arrangement , housing type , availability of car in the household (yes vs. no), area-level SES (high vs. low), type of recruitment centres (EHC vs. elderly community centre) and the number of diagnosed health problems. The latter was obtained using information from a clinical health-problems checklist obtained from the EHC  or, for those recruited at elderly community centres, from the participants at the interview.Descriptive statistics were computed for all variables. Generalised additive mixed models (GAMMs) with Gaussian variance and identity link functions were used to estimate the confounder-adjusted associations between objectively-assessed physical neighbourhood attributes and QoL domains. GAMMs can model outcomes with various distributional assumptions, spatially correlated data and curvilinear relationships of unknown form .p < 0.05) were probed using Johnson–Neyman procedures, whereby we estimated the region of significance of moderators, i.e., the range of values of the environmental attributes for which the effects of living arrangements  on QoL domain scores were statistically significant.First, a set of GAMMs estimated the associations of all covariates and living arrangements with QoL domain scores. Another set of main-effect GAMMs estimated the dose-response relationships of single environmental variables with QoL domain scores. Curvilinear relationships of environmental variables with the outcomes were assessed with thin-plate smoothing spline terms in GAMMs. Thin-plate splines are a type of smoothing splines that are used to model and visualize complex curvilinear relationships between continuous exposure and outcome variables. Their advantage over other smoothing spline methods is that they do not require any a priori knowledge of the functional form of the relationship of interest . Smooth p-value < 0.10 were entered in multiple-environmental-variable GAMMs adjusted for all covariates. We adopted a probability level of 0.10 as a criterion for the preliminary selection of environmental attributes and interaction terms to be included in multiple-environmental-variable models in order to avoid missing potentially important explanatory variables due to negative confounding. Environmental attributes that were strongly correlated were combined into composite variables as appropriate. Only those environmental attributes and interaction terms that showed a significant independent effect on the outcomes (p < 0.05) were retained in the final multiple-environmental-variable models. Following the recommendations of statistical theorists, no adjustments for multiple testing were performed because the analyses were confirmatory and the outcomes and environmental exposures were correlated [All single environmental attributes and interaction terms with rrelated ,41. All rrelated ,43.Associations of QoL domains with socio-demographics and health conditions are shown in 2  and showing a positive trend  modified the negative effects of living alone on environmental QoL in the expected direction. Older adults who had poor access to retail, eateries, health-related services, places for socialising, civic/institutional destinations and good-quality parks reported lower levels of environmental QoL if living alone than if living with others. These findings suggest that these neighbourhood attributes are particularly important to older residents who live alone, enabling self-reliance in performing activities of daily living. Although to our knowledge no other study has examined neighbourhood characteristics as moderators of living arrangements-QoL associations, a recent survey of Dutch elderly reported that perceived access to facilities was positively related to environmental QoL, providing further support for the importance of neighbourhood destination accessibility to older residents’ satisfaction with the environment .2 (corresponding to the samples’ 25th and 87th percentiles of density) and a nil or slightly negative association thereafter. As residential density is necessary to support local commercial and public services, it is a proxy measure of overall destination accessibility. Medium-to-high density neighbourhoods are generally characterised by mixed-use developments with a large range of complementary destinations that offer opportunities for utilitarian walking and social activities [Whilst no significant positive associations between environmental QoL and access to single destination categories were found in the whole sample, we observed a curvilinear relationship with residential density suggestive of a positive association for values ranging from ~9000 to 23,000 dwellings/kmtivities ,46. Howetivities , which mtivities .2 (40th percentile) and a negative effect for values ranging from 5 to 13 destinations/km2 (89th percentile)  . A similParks and other public open spaces, where older residents can engage in physical and social activities , are anoIt is interesting that older adults having access to a wide range of activities in local parks had higher social QoL if living alone than living with others. Parks providing different activities are likely to be more frequently visited by a larger number of residents, which increases the opportunity for older adults living alone to develop and maintain social relations with likeminded neighbours. In contrast, those living with others may commit themselves to spending more time with household members within their typically overcrowded apartment, which would result in heightened levels of stress and, thus, negative social perceptions ,54.Another interesting moderating effect of park accessibility on the associations between living arrangements and QoL was found in relation physical QoL. Compared to those living with others, older people living alone reported lower physical QoL when residing in neighbourhoods intersected by larger park areas. In Hong Kong, these types of neighbourhoods are located in areas adjacent to country parks or natural reserves, which are often typified by steep terrain  and haveSigns of crime/disorder moderated the associations between living arrangements and psychological QoL. Compared to those living with others, older people living alone reported higher psychological QoL when residing in neighbourhoods with more signs of crime/disorder. However, it should be noted that the levels of signs of crime/disorder in our study were very low (on average 0.3 out of 100 points), which is typical of Hong Kong residential neighbourhoods . Some siTwo non-destination aspects of the environment that emerged as correlates of QoL domains in the whole sample were the presence of litter and/or decay and street intersection density. Whilst the former characteristic was, as anticipated, negatively associated with environmental QoL, the latter showed somewhat unexpected negative associations with environmental and social QoL. In Hong Kong, areas with a lot of street intersections  are also ultra-dense, filled with high-rise buildings, heavily trafficked and, hence, polluted and noisy , which mFinally, the small number of significant associations between the examined neighbourhood characteristics and QoL in the whole sample of older adults requires an explanation. Studies on factors influencing Hong Kong older adults’ physical activity, a lifestyle behaviour beneficial to QoL, identified a substantial number of neighbourhood physical attributes associated with walking within the neighbourhood ,60 but vThere are several strengths and limitations to our study. We assessed a large range of neighbourhood physical environmental attributes using objective methods that allowed us to control for a major potential source of reserve causality common in this research field—namely, positive affective states and better health status (high QoL) causing older adults to perceive their environment more favourably. This being a cross-sectional observational study, we cannot rule out an effect of neighbourhood self-selection bias on the findings. However, concerns about neighbourhood self-selection are mitigated by the study sampling design ensuring a balanced representation of high and low walkable neighbourhoods across SES strata . AdditioMedium-to-high density, well-ordered neighbourhoods with an optimal mix of well-treed public open spaces and services catering for older adults’ daily needs may significantly contribute to the social and environmental QoL of an ageing population in Hong Kong. Easy access to a variety of daily destinations and activities in local parks appears to be particularly important to the social and environmental QoL of older people living alone who are self-reliant in performing activities of daily living. The extent to which these findings are generalisable to other Asian mega-cities needs to be examined in future studies.


Exosomes and microvesicles are two major categories of extracellular vesicles (EVs) released by almost all cell types and are highly abundant in biological fluids. Both the molecular composition of EVs and their release are thought to be strictly regulated by external stimuli. Multiple studies have consistently demonstrated that EVs transfer proteins, lipids and RNA between various cell types, thus mediating intercellular communication, and signaling. Importantly, small non-coding RNAs within EVs are thought to be major contributors to the molecular events occurring in the recipient cell. Furthermore, RNA cargo in exosomes and microvesicles could hold tremendous potential as non-invasive biomarkers for multiple disorders, including pathologies of the immune system. This mini-review is aimed to provide the state-of-the-art in the EVs-associated RNA transcriptome field, as well as the comprehensive analysis of previous studies characterizing RNA content within EVs released by various cells using next-generation sequencing. Finally, we highlight the technical challenges associated with obtaining pure EVs and deep sequencing of the EV-associated RNAs. The “Extracellular Vesicles (EVs)” is a general term used to describe various types of spheroid structures, encircled by a lipid membrane bilayer, which are secreted by mammalian cells either passively or upon certain stimuli . Since tThere are three distinct types of EVs (as classified by their origin and biogenesis)—apoptotic bodies (ABs), microvesicles , and exosomes . The ABsImportantly, multiple research reports have demonstrated that various RNA species  entrapped within EVs can be transferred from donor to acceptor cells and interfere in gene expression in the latter , 14. ThiThe techniques widely used so far for isolating EVs include ultracentrifugation, density gradient flotation, ultrafiltration, chromatography, polymer-based precipitation, and immunoprecipitation . DiffereWhile each of the above-mentioned approaches harnesses certain differences in biophysical or molecular properties of EVs , neither method can recover a pure material and allows only an enrichment for certain subpopulations of EVs in a sample , 25. As Finally, certain cell types  exhibit a membrane polarity . TherefoThe presence of mRNAs, miRNAs, and lncRNAs within exosomes and MVs have been consistently shown with microarrays and RT-qPCR techniques in multiple early , 30–36 aIn a pioneering work, Nolte-'t Hoen et al. characterized small RNA content in EVs released by the immune cells in culture using deep sequencing. Interestingly, the majority of total RNA isolated from EVs consisted of small RNAs (<200 nt), with minor amounts of 18S and 28S rRNA. Those short RNA fragments were primarily mapped to protein-coding regions and genomic repeats including SINE, LINE, and LTR sequences . On the The sequencing of total  RNAs in the EVs was reported by Jenjaroenpun et al.  and MiraR = 0.91), MVs and exosomes (R = 0.86), as well as MVs and parental cells (R = 0.86). While a less strong correlation was found between ABs and exosomes (R = 0.79) and exosomes and cells (R = 0.75). Despite the fact that EVs subsets were different only to a minor degree from the aspect of their miRNA cargo, a significant number of miRNAs were detected only in exosomes and were absent in both ABs and MVs, supporting the concept of specific RNA loading into exosomes. It has to be mentioned that other ncRNA species were not only significantly more abundant as compared to miRNA but also selectively enriched in different EVs subtypes released by melanoma cells, which adds another level of complexity to investigating extracellular vesicle RNA cargo and its function  8 signaling, which induces cytokine secretion, presumably by mimicking viral RNA . The TLRMassive parallel sequencing has enabled characterization of the whole spectrum of nucleic acids in a given sample, and was consistently applied to demonstrate the presence of the complex RNA cargo within EVs populations released by various cells. Interestingly, the intravesicular miRNAs (which were well-documented previously using microarray and qPCR-based methods) represented only a very marginal proportion as compared to other RNA species including various small non-coding RNAs, lncRNAs, and mRNA fragments. A putative biological impact of the EVs-associated transcriptomes remains to be validated; however, multiple studies indicated that, at least, exosomal miRNA could mediate communication among various cell types including the immune cells. In addition, EVs-encapsulated miRNAs have been shown to serve as highly specific biomarkers for various pathological conditions and correlate with the presence of malignant tumors. Indeed, exosomes carrying a tumor-specific miRNA repertoire have been consistently detected in the venous blood of cancer patients and mouse models. The collective finding that non-miRNA species are in fact much more abundant in the isolated EVs populations, suggests that they could serve as even more promising non-invasive biomarkers for cancer and/or other disorders.ATo and ATu conceived the study. ATu prepared the tables and figures and wrote the manuscript. ATo and OD participated in final manuscript design, and provided experts' opinion on the content and critical revision.ATu is affiliated with the company SciBerg e.Kfm. ATo is affiliated with the company SRC BioClinicum. The remaining author declares that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


The Wheat@URGI portal has been developed to provide the international community of researchers and breeders with access to the bread wheat reference genome sequence produced by the International Wheat Genome Sequencing Consortium. Genome browsers, BLAST, and InterMine tools have been established for in-depth exploration of the genome sequence together with additional linked datasets including physical maps, sequence variations, gene expression, and genetic and phenomic data from other international collaborative projects already stored in the GnpIS information system. The portal provides enhanced search and browser features that will facilitate the deployment of the latest genomics resources in wheat improvement.The online version of this article (10.1186/s13059-018-1491-4) contains supplementary material, which is available to authorized users. The International Wheat Genome Sequencing Consortium (IWGSC)  is an inTo ensure that wheat breeding and research programs can make the most of this extensive genomic resource, the IWGSC endorsed the establishment of a data repository at URGI (Unité de Recherche Génomique Info/research unit in genomics and bioinformatics) from INRA  to develop databases and browsers with relevant links to public data available worldwide. The IWGSC data repository is thus hosted by URGI to support public and private parties in data management as well as analysis and usage of the sequence data. Wheat functional genomics , genetic, and phenomic data have increased concurrently, requiring the development of additional tools and resources to integrate different data for biologists and breeders. To manage this escalation of data, URGI has built this data repository for the wheat community with the following specific aims: (1) to store resources for which no public archive exists ; (2) to enable pre-publication access to specific datasets ; and 3) to enable rapid release of integrated resources upon publication. The repository has been designed in accordance with the “FAIR” principles  to enabl combinedHere we describe the data and tools currently available through the Wheat@URGI portal , the priThe data hosted by the Wheat@URGI portal are available through flat files stored in the IWGSC data repository and through the GnpIS information system . GnpIS eTriticum aestivum L.) accession Chinese Spring. The IWGSC data hosted in the Wheat@URGI portal within the IWGSC data repository are shown in Table Through its concerted efforts to achieve a high-quality, functionally annotated reference wheat genome sequence, the IWGSC has developed a variety of resources for the bread wheat  fingerprinting (HICF)  or WholeThe IWGSC wheat genome sequence assemblies available for download, BLAST , and disTriticum durum cv. Cappelli, Triticum durum cv. Strongfield, Triticum durum cv. Svevo, Triticum monococcum, Triticum urartu, Aegilops speltoides, Aegilops sharonensis [In addition to the bread wheat sequence, the IWGSC also assembled seven diploid and tetraploid wheat-related species: ronensis . DownloaRNA-Seq expression data are available as read counts and transcripts per kilobase million (TPM) for the IWGSC RefSeq v1.1 annotation. It is a transcriptome atlas developed from 850 RNA-Seq datasets representing a diverse range of tissues, development stages, and environmental conditions .These data consist of downloadable variant call format (VCF) files from genotyping by sequencing (GBS) and whole exome capture experiments of 62 diverse wheat lines  and of tTriticum aestivum TGACv1 [Aegilops tauschii [As well as IWGSC resources, URGI also hosts other open access wheat sequence data to facilitate research into the wheat gene pool. Sequence assemblies available for download and BLAST include the bread wheat whole genome sequence assembly m TGACv1  and the tauschii .In addition to sequence data, the Wheat@URGI portal hosts, within GnpIS-coreDB, several sets of genetic and phenomic wheat data  that havGenetic information corresponds to genetically mapped markers, quantitative trait loci (QTLs), genetic resources (germplasms), and genetic studies (genome-wide association studies (GWASs)). Genomic information consists of variations from SNP discovery experiments, genotyping, comparative genomics (synteny), and expression data . Phenomic data are available as whole trials including phenotypic and environmental observations recorded using variables from ontologies with Minimum Information About a Plant Phenotyping Experiment (MIAPPE)  complianGermplasm data were mainly provided by the French small grain cereals genebank maintained by INRA at Clermont-Ferrand  but alsoData can be easily accessed through the Wheat@URGI portal  using  and the variables (i.e. observed traits), described with dedicated ontologies, are other important key data for genetic studies as they allow linking phenotype data to genetic associations and QTLs through traits. The genetic resources stored in GnpIS-coreDB displays the unambiguous identification of the accession used ) and a rich set of associated data following the Multi-Crop Passport Descriptors  standarTo complete these already rich integrated datasets, a gene-centric data warehouse, the WheatMine, has been set up using the well-established InterMine tool . The genFigure Finally, to facilitate data search and access to this wealth of data, we developed a data discovery tool, which, similar to a google search, allows the user to enter keywords or terms to find all the matching information in the various data warehouses. The results are presented in a table with details on the matches  and a direct link to the feature .Figure The Wheat@URGI portal hosts and gives access to essential, high-quality wheat data from the IWGSC, European, and international projects. Furthermore, its added value is that it integrates different data types altogether  and provides dedicated tools to explore them.As new wheat resources such as GWASs, genomic selection, and pan-genome data are generated in the frame of ongoing projects, GnpIS will allow their management and integration with other data already available in the information system, linking new upcoming data to this central IWGSC genomic resource.At a wider scale, an expert working group (EWG) of the international Wheat Initiative has built an international wheat information system, called WheatIS, with the aim of providing a single-access web-based system to all available wheat data resources and bioinformatics tools . The WheFigure Data integration is fundamental for researchers and breeders who want to use genomic information to improve wheat varieties. However, the diversity of data types and the concomitant lack of data harmonisation and standards hamper cross-referencing and meta-analysis. A joint action between the WheatIS EWG and a group of linked data scientists created the Wheat Data Interoperability Working Group under the Research Data Alliance (RDA) umbrella  to help Additional file 1:Supplementary data on software technologies and usage metrics. (PDF 541 kb)


Ethnic/racial minorities are under-represented in blood donor populations in most developed countries. This is of particular concern where minorities differ from a country's majority population in terms of blood or tissue typing, especially where type matching is required for effective management of rare disorders such as sickle-cell disease that require multiple transfusions. This systematic review assessed the effectiveness of interventions to increase blood donation among ethnic/racial minority populations in developed countries. We searched MEDLINE, EMBASE, CINAHL, and ProQuest on 20 March 2017 with no date restrictions and supplemented this with searches on Google Scholar, blood collection agency websites, reference lists of included studies, and a forward search of citations of included studies. We included intervention studies designed to increase recruitment and/or retention of adult, ethnic/racial minority blood donors in developed countries. The review identified eight studies reported in nine publications. Six were conducted in the USA with African Americans. Four studies reported on multifaceted, community-based interventions; three reported on one-off information and educational video interventions, presented face-to-face, or delivered via post or e-mail. The level of evidence for efficacy was low, and the majority of studies were assessed as having some risk of bias related to one or more methodological issues. All eight studies reported positive outcomes in blood donation and/or intention to donate. Seven trials found that the intervention increased presentation for donation, and three found an increase in the percentage of new donors from the ethnic minority targeted. The review findings demonstrate that it is possible to design and implement effective interventions to motivate individuals from ethnic/racial minority groups to donate blood. One-off interventions may be as effective as multifaceted, community-based interventions. There was insufficient evidence to recommend particular interventions, and future research should empirically assess alternative interventions using robust study designs. There is an acknowledged under-representation of ethnic/racial minorities in blood donor populations in many if not most developed countries, despite otherwise well-established voluntary blood collection systems in these countries . This isIndividuals from some minority groups may differ from a country's majority population in terms of extended blood or tissue typing. This under-representation of rare blood types is of particular concern in ensuring appropriate blood supplies to avoid alloimmunisation and effectively manage conditions, such as sickle-cell disease, which require frequent transfusions and are more common among certain ethnic/racial minority populations  , interventions through minority and/or religious organisations, increased minority staff in blood donation organisations, health practitioner-led interventions, donor-recruits-donor interventions, public event recruitment, door-to-door recruitment, and public health initiatives such as cardiovascular screening , 17, 18.Research has suggested that there have been some small improvements in donor diversity; however, it is unclear whether these are the result of targeted interventions or due to demographic changes in the various communities. For example, in the USA, while the proportion of nonwhite donors increased in the decade from 2006–2015, this essentially paralleled their relative increase in the population, making it difficult to draw conclusions regarding the effectiveness of targeted recruitment campaigns implemented over the same period . A recenIn response to this gap in the literature, as well as to inform future interventions and research, this systematic review aimed to answer the following question: how effective are interventions conducted to increase blood donation among ethnic/racial minority populations in developed countries? The literature search aimed to identify intervention studies designed to increase blood donation recruitment and/or retention, regardless of study design that included adult participants with an ethnic/racial minority background living in high income countries and that reported on any outcome measure of blood donation behaviour or intentions.The review was registered on Prospero, the international prospective register of systematic reviews, prior to commencement in March 2017 (no. CRD42017058919).MEDLINE, EMBASE, CINAHL, and ProQuest databases were searched on 20 March 2017. The search combined terms related to (1) blood donation, and (2) ethnic/racial minorities. The search was kept deliberately wide by not including terms attempting to limit the results to intervention studies or to adult participants from particular countries. Instead, noneligible studies were removed from the resulting broader pool of articles during screening. A full list of MeSH terms and text words used can be found listed in The first 100 pages  from a Google Scholar search were screened for additional potentially eligible articles not identified in the database search. Additionally, the websites of the main blood collection agencies in Australia , New ZeaTo be included, studies needed to meet all four eligibility criteria listed in Studies reported only in conference abstracts were excluded as these lacked sufficient detail on interventions and outcomes. However, the authors of seven conference abstracts reporting on potentially eligible studies were contacted and copies of any subsequent publications were requested, but none were provided.All references were downloaded to EndNote. Duplicates were removed, and then titles and abstracts were screened against the eligibility criteria. Full text articles for potentially relevant references were reviewed independently by two authors.Among the articles identified as eligible through this process were four review articles –31. ThesQuality was assessed for descriptive purposes rather than to inform study inclusion or exclusion. Two authors critically appraised each of the included studies using a standardised appraisal form, which included NHMRC  level of evidence and magnitude of effect rating , and risTwo authors independently extracted relevant data as predefined in the registered protocol from intervention studies meeting the selection criteria (data extraction form available on request). This included study type and characteristics, participant characteristics, details of interventions, and details of comparison groups if present. Primary outcomes extracted were reported blood donation behaviour and blood donation intention. Secondary outcomes were attitudes towards blood donation. The data extraction form was piloted with two of the included articles to ensure the two authors understood and agreed on the process prior to extraction of data from the remaining studies.Outcomes were tabulated and summarised in a narrative synthesis of findings. The incomplete and inconsistent reporting of different outcome measures that could not be standardised across studies meant that the results could not be pooled in a quantitative meta-analysis.After removal of duplicates, 2,912 titles and abstracts were screened against the eligibility criteria, resulting in 188 potentially relevant references. Through review of full text articles for these references, eight intervention studies meeting the eligibility criteria were identified, reported in nine publications  –42. PRISThe most common reasons for exclusion of full text articles were (a) articles did not report on the evaluation of an intervention (95 articles) and (b) the studies were not conducted in a high income  country (41 articles). The complete list of reasons for exclusion of full text articles is included in Four studies reported on a one-off information or one-off education intervention. One trialled a face-to-face educational session and video , one wasAll interventions could be classified as using motivational (cognitions-based) techniques, according to Godin et al.'s typology of blood donation recruitment and retention interventions . Some stAll eight trials reported positive outcomes in blood donation and/or intention to donate, some with statistical comparisons with a control group or with a historical comparison period . Seven tp=0.39) and no significant difference in the donation presentation rate during the subsequent 76-day period .One study tested the effectiveness of a recruitment e-mail and video link tailored specifically to African American potential donors, compared with generic versions of the same materials . While bt (146) = −1.455, p=0.148. Robbins et al. )The Action Plan developed by the Missing Minorities group of the European Blood Alliance could act as a useful starting point for targeting activities . The ActIn conclusion, the results of this systematic review suggest that interventions aiming to increase blood donation among ethnic/racial minorities can be effective. Broadening participation in blood donation would have the advantage of increasing the overall blood supplies, as well as improving the ability to match rare blood types where required . Includi


Lipid cell membrane composed of various distinct lipids and proteins act as a platform to assemble various signaling complexes regulating innumerous cellular processes which are strongly downregulated or altered in cancer cells emphasizing the still-underestimated critical function of lipid biomolecules in cancer initiation and progression. In this review, we outline the current understanding of how membrane lipids act as signaling hot spots by generating distinct membrane microdomains called rafts to initiate various cellular processes and their modulation in cancer phenotypes. We elucidate tangible drug targets and pathways all amenable to small-molecule perturbation. Ranging from targeting membrane rafts organization/reorganization to rewiring lipid metabolism and lipid sorting in cancer, the work summarized here represents critical intervention points being attempted for lipid-based anticancer therapy and future directions. Lewis Thomas in the Lives of a Cell  undersco in vitro growth of cancer cells, there are abundant nutrients and these cells synthesize fatty acids de novo. But, under conditions of stress, the cancer cells usually adapt to the cell growth by scavenging extracellular lipids [Lipids form a considerable part of the dry weight of mammalian cells. A substantial supply of lipids is required for cell proliferation . Usuallyr lipids , This scr lipids . Cancer r lipids . Due to r lipids . But ther lipids , due to Lipids fulfill many critical requirements in the cell including composing membrane bilayer, storing energy due to their reduced state, acting as first and second messengers in signal transduction, providing functional implementations of membrane-proteins structure and function, and finally recognition processes. Advanced mass-spectroscopy and analytical techniques have allowed sensitive and highly selective analysis of lipids of diverse chemical structures within complex biological samples and testified to their intimate involvement and their aberrations in many diseases such as neurodegenerative disorders, infectious diseases, and cancer. Interestingly, lipidomics have revealed cancer type-specific alteration in the lipidome of cells implying critical roles of lipids in cancer progression and initiation –15. Admi exoplasmic leaflet of the lipid bilayer. Underpinning this concept is the propensity, in vitro, of the saturated hydrocarbon chains intercalating strongly with cholesterol molecules inducing liquid ordered lo phases [d phase. Thus, the raft domains can be imagined as platforms of lo phases dispersed in the ld matrix of unsaturated glycerolipids. These raft-like assemblies are ordered and tightly packed but are still fluid due to lipid acyl chain packing differences. This, in turn, is governed by the saturation level of the hydrocarbon chains in the rafts compared with the unsaturated state of fatty acids of phospholipids in the ld phase. These rafts act as membrane scaffolds to house various proteins, organize receptors and their downstream molecules, and hence regulate a number of membrane-associated signaling pathways [High compositional complexity modulates interactions and localizations of lipids in membrane and befittingly influences the diverse phases lipid can form . The samo phases –42. The pathways , 44. In pathways . Their ipathways –49 but tOn the mechanistic level, a cellular signaling event originates from the raft domains at the membranes with transmission of signals  through membrane-bound receptors, e.g., receptor tyrosine kinases (RTKs). Stimulation by the diverse signals promotes receptor dimerization enabling their phosphorylation mostly via their intrinsic tyrosine kinase activity. The resulting phosphorylated residues exposed to the cytoplasm act as docking sites for effector proteins that harbor specialized membrane-loving domains—fostering their recruitment to the membrane. This chain of events induces allosteric changes in the activity and/or affinities on another module of the recruited molecule conveying signals deep into the cell and finally leading to regulation of processes like proliferation, migration, division, and differentiation, among others. Malfunctioning of the cellular signaling both inherent and induced at any given step in space and time results in unchecked downstream cellular functions culminating into various diseases, especially cancer. As an example, we focus on the most fundamental signaling cascade mediating proliferation and differentiation—mitogen-activated protein kinase cascade, MAPK signaling, EGFR stimulation promotes binding to adaptor proteins GRB2, which further binds Son of Sevenless (SOS) . This biPathological, pharmacological, and nutritional situations strongly regulate lipids in cell with profound biological implications. With advances in high-throughput lipidomics, precise characterization of lipid structures is revealing critical lipid alterations in composition and abundance among various cell types and cancers and surprisingly during varied cellular processes as well , 58. ForLipid membranes of cancer cells possess relatively higher negative charge due to increased abundance of PS and PE phospholipids on the exoplasmic membrane surface , 62. On  in vivo have transformed the field of eukaryotic cell biology. Small-molecule-mediated inhibition of the function of specific proteins has enabled cell biologists to query their functional roles. Most classic example in this regard is of colchicine and paclitaxel as tubulin depolymerizes and stabilizers, respectively, which have provided unprecedented insights into the function of this cytoskeletal protein [Small molecules targeting specific biomolecules and modulating their structure and activity protein , 19. Dev protein , 77.Membrane rafts regulate key signaling molecules and proteins implicated in cancer by modulating their association with and localization with lipid membranes including interactions with other membrane-bound proteins , 78, 79.Central roles in the initiation and progression of many tumor types responsible for the alteration of cell cycle, cell adhesion, cell migration, and programmed cell death are regulated by various factors. Lipid rafts and membrane microdomain or compartments play an active role in each of these cell processes by mainly regulating the downstream intracellular signaling pathways , 22, 43.β-cyclodextrin-MβCD [ Rheum palmatum, inhibits cancer cell migration by suppressing the PI3K-Cdc42/Rac1 signaling pathway and leads to inhibitory action on cell invasion and cell migration [κB (nuclear factor kappa-light-chain-enhancer of activated B cells) signaling [ 9 (MMP-9) [β1 clustering and its colocalization with membrane rafts as judged by cell-based microscopy assays in breast cancer model cell lines [β1, and focal adhesion complex (FAC) from cytoplasm to membrane rafts, mainly attributed to reduced cholesterol levels in the membranes, thus hampering efficient raft-signaling-platform formation [β-cyclodextrin (MβCD), a water-soluble polymer and a cyclic oligosaccharide, absorbs cholesterol from the cell membrane and has been shown to impair actin polymerization, cell migration, Akt  phosphorylation, protein kinase C translocation, and EGF-induced cell adhesion in selective cancer cell models [βCD, this molecule has been chemically modified to serve as platform for cellular lipid shuffling enabling generation of asymmetric model membrane systems [βCD is an acute process due to comparatively short term of treatment [Membrane rafts are involved in various surface receptor signaling pathways in tumor and lipid depleting agents causing disruption of rafts domains lead to changes in signal response of each , 43. Memrin-MβCD    as shownll lines . Furtherormation . Thus, tormation  as shownl models , 87, 88. systems –91. The reatment . Thus, sreatment , 93.κB signaling culminating to apoptosis [Statins, the first committed inhibitors of the mevalonate pathway, act at an early step in the synthesis of cholesterol . Statinspoptosis , 29 and Recent single molecule tracking techniques have elucidated that actin-based cytoskeleton structures on the cytoplasmic surface of the plasma membrane are also a key player in inducing membrane domain organization or partitioning into small compartments underpinning the cellular dynamics of protein and lipid lateral diffusion. Such a model implicit in the picket fence model states that the membrane-actin skeleton interactions induce temporary confinement of transmembrane proteins thus generating transient domains that function as signaling host spots similar to rafts. Many receptors like the G-protein coupled receptor (GPCR), transferrin receptors, etc. have been assigned to such transient domains with the membrane skeleton mesh and are also enriched with cholesterol and sphingolipids –100. SpeRecent studies have elucidated membrane rafts to form signaling platforms capable of activating pro- and antiapoptotic pathways susceptible to pharmacological perturbations aimed at stabilizing these special apoptotic-linked raft domains . ActivatCancer cells display a highly distinct metabolic growth profile compared with nontransformed normal cells. The metabolic reprogramming of the enzymes of various pathways of cell growth forms the underlying basis of cancer. One of the most implicated pathways that are heavily tinkered within cancer is lipid metabolism. Lipid metabolism is linked closely with the glycolytic pathway by virtue of it providing the required starting substrate—acetyl-CoA—for fatty acid (FA) synthesis. Lipids play key roles in this network, as they are crucial for the formation of cell membranes and also act as signaling messengers. Due to the enormous upregulated growth rate of cancer cells, relatively larger amounts of lipids are required to keep up with alarming rates of growth, proliferation, energy storage, and production of signaling molecules , 114. Taβ-oxidation of FAs, where it facilitates the movement of FA-CoA from the cytosol to the mitochondria across the mitochondrial membrane. Etomoxir and perhexiline are two small molecules shown to be effective against tumors and curb their proliferation via targeting FA oxidization mediated through CPT1  is an enzyme involved in the ugh CPT1  [122, 12 in vivo . Cycloti in vivo , 128. Th in vivo , 130.Lipids, apart from being intimately involved in cellular functions and cellular signaling as isolated modules, add another level of complexity by their covalent attachment to proteins—posttranslational protein lipidation—that forms the heart of membrane-associated signaling in cells, e.g., small GTPases, such as Ras, Raf, and ARFs. A classic example of addressing oncogenic signaling involving lipidated proteins is via targeting the protein lipidation leading to improper membrane-raft localization of these proteins causing nonfunctional signaling platforms and hence subdued oncogenic signaling. This aspect is greatly exemplified by the class of lapidated protein-Ras. The lipid moieties attached to the protein consist of a palmitoyl group and 1-2 farnesyl lipid anchors. Ras was the first oncogene to be discovered and is involved in many human cancers; however small-molecule targeting of Ras still remains an unmet task in cancer therapy. In 1989, one of the first drugs to be thought to target Ras was lovostatin. Farnesyl pyrophosphate farnesylates Ras and it is a key intermediate of the mevalonate-cholesterol biosynthetic pathway. Being a HMG-CoA reductase inhibitor, lovostatin was shown to block the mevalonate-cholesterol biosynthetic pathway and hence the farnesylation lipidation of Ras. However, requirement of much higher dosage for selectively blocking farnesylation leads to adverse effect on cholesterol biosynthesis and unspecific cell death, thus making the journey of lovastatin quite short-lived regarding clinical targeting of Ras Farnesylation . This faδ, in cells that sustains the spatial orientation of the Ras superfamily of proteins [δ towards K-Ras trafficking to reach plasma membrane rafts to initiate signaling was demonstrated [δ leading to K-Ras mislocalization and downregulated cancer signaling leading to reducing cell proliferation and finally cancer cell death [However, amidst such failed attempts, recently Waldmann and coworkers have demonstrated an exciting alternative to target Ras-associated cancer by mislocalizing Ras lipoprotein not channelized via blocking the lipid attachment but by an innovative chemical biology approach . Lipidatproteins . Recentlnstrated , 140, anll death , 141. Alll death . This le in vitro.Although a lot of work has to be yet done in identifying membrane specific small molecule against cancer, the effects of presently available small molecule on the membrane specific organization and signaling are proven as effective against the maladyThe quest for targeting cancer using varied chemical and genetic approaches still is faced with enormous hurdles and generates an unmet need to develop therapeutic approaches inspired by careful inspection of modulated cancer cell attributes. One of the aspects gaining considerable attention recently has been the altered lipid repertoire of cancer cells leading to modulated membrane-dependent cellular processes including membrane organization and cellular signaling, strongly contributing to tumor growth and metastasis and understanding the underlying mechanism behind the same to elucidate potentially novel targets and pathways against cancer. In this review we focused on some of the most promising lipid associated candidates and processes for anticancer targeting by small molecules. Ranging from targeting of lipid enzymes involved in the lipid metabolic pathway to the proteins and lipids that help in lipid organization, oncogenic lipoprotein sorting and signaling membrane micro-domains-rafts have proven to be highly crucial to not only contemplate their therapeutic aspect but also address and unveil specific mechanisms of lipid deregulation in cancer. The various small-molecule-based drugs and tool compounds that have been discussed so far have shown promising results in targeting lipid-dependent processes in cancer cells. Drugs like Salirasib that inhibit Ras function have shown the ability to act in multiple ways and have been used in Phase I clinical trials showing good results with the drug being well tolerated. Deltarasin which targets K-Ras downstream signaling and K-Ras localization may have potential to achieve better efficacy in the long run. Although Statins look the most prospective in cancer treatment, there are side effects to their use. In addition, they all act predominantly by remodeling membrane rafts composition and organization but lead to distinct downstream effects on the oncogenic signaling in various cancer models. This brings to the forefront membrane rafts as “selective cancer therapeutic targets,” with the structure, function, and associated raft-signaling pathways being subject of extensive studies. As such, a better understanding of the structural, conformational, and functions aspects of raft biology would foster exploitation of membrane rafts for developing personalized cancer therapy for targeting distinct raft-associated oncogenic signaling in various cancers. In this regard, recent surge in technological advancements in super resolution microscopy (SRM) is already providing invaluable information on the distribution and organization and dynamics of plasma membrane components to bendings  that occFurther chemical biology investigations on the regulation of membrane-lipid-dependent signaling pathways in cancer cell may provide novel targets for therapy and elucidating the role of distinct lipid signaling molecules will offer innovative therapeutic opportunities for development of anticancer drugs. However, discovery of lipid-dependent novel targets and novel signaling pathways in cancer biology strongly lies at the hand of discovering innovative small molecules. Generally, small molecules with well-defined targets permit obtaining novel insights into biological processes and extensive analysis of their structure-activity relationships permits chemical modification for improving efficacy of such candidates. Though nature is a comprehensive source of molecules with a variety of bioactivities, in recent years, a burst in organic synthesis strategies and synthesis of organic molecules generated via innovative hypothesis generating platforms, such as diversity oriented synthesis (DOS) biology oriented synthesis (BIOS), and activity oriented synthesis, have enriched the pool of small molecules in an informed manner, thus promoting their use as research tools to explore previously uncharacterized biology and elucidate novel targets for drug discovery. Given the limited number of drug targets addressed till today in cancer and immunology, including protein kinases, G-protein coupled receptors, and ion channels, development of new small organic molecules rightly fits the criteria for meeting the ever-increasing need for new therapeutic targets. Finally, identification of the molecular targets of such new compounds still remains a major bottleneck, underscoring the demand for appropriate methodologies to elucidate the targets of small molecules in a relatively unbiased and timely fashion.


Filoviridae and cause severe haemorrhagic fever in humans. Due to high mortality rates and potential for spread from rural to urban regions, they are listed on the WHO R&D blueprint of high-priority pathogens. Recent ebolavirus outbreaks in Western and Central Africa have highlighted the importance of diagnostic testing in epidemic preparedness for these pathogens and led to the rapid development of a number of commercially available benchtop and point-of-care nucleic acid amplification tests as well as serological assays and rapid diagnostic tests. Despite these advancements, challenges still remain. While products approved under emergency use licenses during outbreak periods may continue to be used post-outbreak, a lack of clarity and incentive surrounding the regulatory approval pathway during non-outbreak periods has deterred many manufacturers from seeking full approvals. Waning of funding and poor access to samples after the 2014–2016 outbreak also contributed to cessation of development once the outbreak was declared over. There is a need for tests with improved sensitivity and specificity, and assays that can use alternative sample types could reduce the need for invasive procedures and expensive equipment, making testing in field conditions more feasible. For MARV, availability of diagnostic tests is still limited, restricted to a single ELISA test and assay panels designed to differentiate between multiple pathogens. It may be helpful to extend the target product profile for ebolavirus diagnostics to include MARV, as the viruses have many overlapping characteristics.Ebolaviruses and Marburg virus (MARV) both belong to the family  Filoviridae, cause severe haemorrhagic fever in humans and are considered high-priority pathogens by WHO; recent ebolavirus outbreaks in Western and Central Africa have led to rapid development of a number of diagnostic tests for ebolaviruses.Ebolaviruses and Marburg virus (MARV), both of the family Despite these advancements, gaps in diagnostic preparedness for ebolaviruses remain, including difficulties obtaining full regulatory approval for tests approved under emergency licenses, lack of incentive to continue development after an outbreak is over, poor specificity and sensitivity of some existing tests, and a need for tests able to use different sample types.For MARV, availability of diagnostic tests is still limited; target product profiles for ebolavirus diagnostics should be extended to include MARV and should consider persisting challenges to diagnostic preparedness for ebolaviruses.Filoviridae and can cause severe haemorrhagic fever in humans and non-human primates. Filoviruses are filamentous, negative-sense RNA viruses, with a genome that encodes seven structural proteins: nucleoprotein (NP), polymerase cofactor (VP35), matrix protein (VP40), glycoprotein (GP), replication-transcription protein (VP30), minor matrix protein (VP24) and the non-structural protein RNA-dependent RNA polymerase (L) , Sudan ebolavirus (SUDV), Bundibugyo ebolavirus, Tai Forest ebolavirus (TAFV) and Reston ebolavirus. The marburgvirus genus consists of Marburg marburgvirus (MARV), including the MARV POPP, MARV Angola, MARV Durba, MARV Ozolin and MARV Musoke variants, and Ravn virus.Ebolaviruses and Marburg virus (MARV) belong to the viral family rase (L) .1 The viSince the first report in Sudan and Zaire in 1976, ebolavirus disease (EVD) has caused more than 30 outbreaks in the subsequent 40 years . EVD is Sporadic outbreaks of EVD and MVD typically occur and are limited to countries in sub-Saharan Africa.The fruit bat is thought to be the native host for filoviruses ,Filoviruses are primarily transmitted to humans through close contact with blood, secretions, organs, or other bodily fluids of infected humans or animals.The incubation period for EVD and MVD is between 2 and 21 days, with an average of 3–10 days.Treatment options for EVD and MVD are limited and rely on supportive therapy.Two of the most promising EBOV vaccine candidates, rVSV-EBOV and ChAd3-EBO-Z, underwent phase II/III efficacy trials in Liberia, Sierra Leone and Guinea during the peak of the 2014–2016 epidemic, showing high efficacy and long-term antibody responses.www.finddx.org) has launched initiatives for needs assessment and partnerships across a broad range of diseases in endemic-prone countries. This landscape analysis describes the current state of filovirus diagnostics, and identifies remaining needs for further research and development.Due to the sporadic nature of outbreaks, high mortality rates and potential spread from rural to urban regions, filoviruses are some of the high-priority pathogens identified in the WHO R&D Blueprint,The diagnosis of ebolaviruses and MARV is based on direct identification of the viral particles, proteins or specific RNA in a suspected case from whole blood (WB), serum or plasma. Since ebolaviruses and MARV are classified as biosafety level 4 agents due to their propensity for human-to-human transmission, high risk for laboratory-acquired infections, and lack of a specific and safe vaccine, patient samples present an extreme biohazard risk and special laboratory procedures must be in place to safely confirm infection.Confirmation can only be obtained by virus isolation. However, other assays such as electron microscopy, histological techniques and specific detection of nucleic acid, immunofluorescence and immunoassays of both antigen and antibodies provide putative positive identification; an orthogonal approach using multiple assay types can be used to increase confidence in results.et al, there are a number of laboratory designed tests (LDTs) using reverse transcriptase (RT)-PCR, quantitative RT-PCR (qRT-PCR) and RT-loop mediated isothermal amplification to detect ebolaviruses and MARV,Common targets for both ebolaviruses and MARV NAAT testing include the NP, L and GP genes.The 2014–2016 West Africa ebolavirus outbreak resulted in the rapid development of new commercially available benchtop and point-of-care  NAATs to allow for rapid detection of ebolavirus infection in lower resource laboratories.9 copies/mL, while survivors’ viral titre ranges around 107 copies/mL.Quantification of the viral load is useful for patient management as it can potentially predict the probability of recovery since fatal cases often exhibit rapid increases in viraemia with viral load in blood for ebolaviruses reaching approximately 10A major limitation of the majority of the available NAATs is their limited strain coverage. Only a few NAATs are able to test for multiple ebolavirus or MARV subtypes, which can potentially result in false negatives during outbreak responses or surveillance activities. Further development is needed for easily deployable, POC NAATs that can distinguish multiple ebolavirus and MARV subtypes for future outbreaks and improved surveillance.Serology is useful in epidemiological studies of infected hosts by detection of ebolavirus-specific or MARV-specific antigen or IgM and IgG antibodies. Detection of anti-ebolavirus-IgM or anti-MARV-IgM indicates recent infection and can be detected as early as 2–4 days after symptom onset, while anti- ebolavirus-IgG or anti-MARV-IgG can be detected around 8–10 days after symptom onset and persist for up to 2 years after infection . IgG detThere are currently no SRA-approved benchtop ELISAs for ebolavirus or MARV detection. Two commercial ELISAs are available for research use only to detect EBOV VP40  and IgG/10.1136/bmjgh-2018-001112.supp1Supplementary dataRapid diagnostic tests  can leverage the same antibody/antigen capture agents as an ELISA but in a lateral flow strip format, allowing faster results with minimal specimen processing .The 2014–2016 ebolavirus outbreak resulted in the rapid development of new RDTs. As of May 2018, five EBOV RDTs were approved for the detection of EBOV by SRAs ,Implementation requirements of the different filovirus diagnostic types are shown in Viral haemorrhagic fever (VHF) refers to a diverse group of animal and human illnesses in which fever and haemorrhage are caused by viral infection. In regions where VHF viruses may be endemic and maintained through natural reservoirs, a panel for distinguishing the viral family causing infection and, potentially, co-infection will be useful for surveillance as well as appropriate isolation and infection prevention and control measures. While the majority of ebolavirus outbreaks are caused by EBOV and SUDV, current tests do not have full strain coverage. A multiplexed, syndromic approach is a more useful and efficient strategy for surveillance, differential diagnosis and outbreak determination than relying on single, pathogen-specific assays. Currently, there are few commercially available molecular diagnostics that can detect multiple fever-causing agents.During the 2014–2016 West Africa ebolavirus outbreak, WHO established an EUAL procedure for in vitro diagnostics (IVDs) in order to expedite the introduction of critically needed technologies during an outbreak setting.However, on 29 March 2016, WHO announced that the ebolavirus situation in West Africa no longer constituted a Public Health Emergency of International Concern. While previously submitted and approved products under WHO EUAL may still be purchased and used, both WHO and FDA recommend test manufacturers to submit for full regulatory approval.In addition, as the ebolavirus outbreak waned, so has access to confirmed positive samples, which are required in order to fully validate an assay for regulatory approval. These circumstances make the regulatory approval process after an outbreak a significant challenge that must be addressed in order to ensure access to reliable and robust diagnostics in the future.The need for rapid, sensitive, safe and simple ebolavirus diagnostic tests was highlighted in a November 2014 Call for Diagnostics by WHO since efforts to contain the ebolavirus outbreak were hampered by cumbersome, slow, complex and costly diagnostic tests. In order to stimulate product development, WHO issued a detailed product profile of the ‘ideal’ rapid, sensitive, safe and simple diagnostic test considered most likely to accelerate interruption of virus transmission in severely resource-constrained settings.Of the commercially available tests with publicly available performance data, two ebolavirus tests listed meet the ‘acceptable’ TPP criteria  and one meets the ‘desired’ TPP criteria .In terms of MARV, although a TPP for diagnostic tests to detect MARV does not exist, it may be helpful to extend the ebolavirus TPP criteria to evaluate the MARV tests, considering the viruses are within the same family and have many overlapping characteristics. Unfortunately, none of the MARV tests would be classified as ‘acceptable’ according to the ebolavirus TPP  due to the lack of deployable tests in areas with limited infrastructure and resources.Specifically for MARV, there are limited recommendations from WHO regarding the diagnosis of MVD. The Centers for Disease Control and Prevention recommends that ELISA, PCR or virus isolation be used to diagnose MVD within a few days of symptom onset. The IgG-capture ELISA is recommended when testing patients later in the course of the disease or post-recovery.The usual clinical samples for testing for EVD and MVD are WB, plasma and serum. However, venepuncture is an invasive procedure with high risk for both the patient and the health worker. Moreover, blood separation into plasma or serum often requires additional equipment  or expensive collection tubes. With the difficulties of blood collection and POC analysis under field conditions, less invasive, simpler to obtain and more stable clinical specimens must be considered for new diagnostic development.There is evidence that ebolaviruses can also be detected in saliva, breast milk and semen.In an effort to avoid manufacturer fatigue and promote development of novel diagnostics for outbreak response, new funding mechanisms and partnership models should also be considered. Many developers who commenced development of a diagnostic test for the detection of EVD ceased development as the 2014–2016 ebolavirus outbreak waned, as access to samples became challenging and funding slowed. Incentives such as advanced market commitments or volume guarantees for manufacturing scale-up could be considered in order to stimulate innovation in disease areas with small and volatile markets, such as EVD and MVD,Leveraging existing supply chains, operator training on a specific platform technology, and product registration and regulatory experience of manufacturers with multiple tests on the market may also optimise programme efficiency for ebolavirus and MARV disease surveillance and outbreak management. In particular, platform technologies commercialised by manufacturers with products on the market for common infections and chronic diseases that require routine purchasing and training  could be highly cost-effective and improve outbreak response times.The magnitude of the 2014–2016 West Africa ebolavirus outbreak brought to light the need for new diagnostic tests to rapidly assess and respond to VHF outbreaks to interrupt transmission. This review has identified several commercially available diagnostics and laboratory designed tests for filovirus detection.However, significant gaps still remain . While t10.1136/bmjgh-2018-001112.supp3Supplementary data10.1136/bmjgh-2018-001112.supp4Supplementary dataIn regions of the world where infrastructure and resources are limited or in situations where rapid response to decrease transmission is a priority, the use of simple, deployable tests is needed. Concerted, collaborative efforts in research, development and systems strengthening must occur to rapidly halt transmission and to prevent future transmission of filovirus diseases.10.1136/bmjgh-2018-001112.supp2Supplementary data