To address the complex challenges faced by our planet such as rapidly changing climate patterns, food and nutritional insecurities, and the escalating world population, the development of hybrid vegetable crops is imperative. Vegetable hybrids could effectively mitigate the above-mentioned fundamental challenges in numerous countries. Utilizing genetic mechanisms to create hybrids not only reduces costs but also holds significant practical implications, particularly in streamlining hybrid seed production. These mechanisms encompass self-incompatibility (SI), male sterility, and gynoecism. The present comprehensive review is primarily focused on the elucidation of fundamental processes associated with floral characteristics, the genetic regulation of floral traits, pollen biology, and development. Specific attention is given to the mechanisms for masculinizing and feminizing cucurbits to facilitate hybrid seed production as well as the hybridization approaches used in the biofortification of vegetable crops. Furthermore, this review provides valuable insights into recent biotechnological advancements and their future utilization for developing the genetic systems of major vegetable crops. Hybrids offer significant advantages for enhancing the economic and technological aspects of vegetable cultivation, primarily due to the phenomenon of heterosis, which confers superiority over diverse parent varieties. Hybrids’ exploitation is of the utmost importance in addressing the emerging challenges induced by climate change, including the mitigation of food and nutritional insecurities. Moreover, hybrids provide a powerful tool for breeders to maximize the yield potential of vegetable crops. Heterosis exerts a profound impact on both productivity and quality across a range of vegetable crops, enabling improvements in livelihoods by enhancing productivity and delivering high-quality products and nutritionally superior food options. Leveraging genetic mechanisms to exploit heterosis facilitates enhancing productivity, improving quality (depending on the objective), and reducing seed production costs. The growing interest in heterosis exploitation and the utilization of genetic mechanisms is evident in the intensified investigation of self-incompatibility (SI), male sterility, and gynoecism and their applications in hybrid seed production for several vegetable crops. However, it is important to note that these mechanisms are limited to specific classes of vegetable crops.Heterosis  plays a pivotal role in vegetable breeding, leading to significant improvements in yield, quality, and earliness, which are considered the desirable outcomes when genetically distant parents are used to produce hybrid off-springs in vegetable crops. However, the selection of suitable parents is crucial for achieving these results . Genetic1 hybrids over two mated inbreds. During the second decade of the twentieth century, Hayes and Jones [Cucumis sativus). Notably, the development of the commercial eggplant F1 hybrid performed in Japan in 1925 marked a significant milestone for hybridization attempts [The development of a commercially viable system for producing hybrid seeds has had a profound impact on the modern scientific understanding of crops and the agricultural industry.In 1914, George Harrison Shull, an American botanist and geneticist, while working with corn at Cold Harbor, New York, coined the term “Heterosis” ,8, whichattempts ,11. Fourattempts . InitialTo establish a successful hybrid breeding program, comprehensive knowledge of floral characteristics, reproductive features, the time of anthesis, and pollination is required. It is crucial for vegetable growers to possess in-depth knowledge of the different flower types , the time of anthesis, dehiscence timing, and the reproductive attributes of vegetable crops ,13,14 inRaphanus sativus and other Brassicaceae crops. Using the BLAST technique, 142 potential bolting- and flowering-related genes were identified in various flowering pathways, and, furthermore, out of these 142 genes, the isolation and profiling of 7 critical genes was also reported using TA cloning and RT-qPCR analysis. SQUAMOSA, GLOBOSA, DEFICIENS, AGAMOUS, and SEPALLATA1 were the previously defined groups in carrot in relation to MADS-box genes, and five genes, namely, DcMADS1, DcMADS2, DcMADS3, DcMADS4, and DcMADS5, were assigned to them [A specific class or a combination of classes of genes determines organ identities. Their role in floral development can be explained using the ABCDE model ,16, whic to them ,26. The  to them . Further to them ,28,29. T to them , while tSimilarly, in the case of cucurbitaceous vegetables, particularly in the case of melon and cucumber, male and female flower production is reported to be under the control of various genes. ACS11, the limiting enzyme of ethylene biosynthesis, which is reported to control female flower development, is encoded by the androecy gene. Male and female plants are under the control of ACS11 expression. ACS11 expression leads to female plants while its loss-of-function mutants produce male plants. CmACS11 and CmW1P1  play major roles, and their combination leads to artificial dioecy, while on an individual level, the former represses the expression of the latter to control the development and coexistence of male and female flowers in monoecious species . The mutInformation regarding pollen behavior has gained popularity among breeders as it assists their breeding plans accordingly. Complicated flower biology, sex forms such as dioecy, unsynchronized male and female flowering, and poor and irregular flower production altogether limit wide hybridization. Therefore, the improvement of vegetables through conventional breeding approaches is constrained especially in the case of yam, etc. ,36. PollVarious strategies have been proposed for hybrid seed production based on emasculation and pollination procedures. One of the most promising strategies for producing cost-effective seeds with minimal impact on seed purity is the utilization of genetic mechanisms. These genetic mechanisms, namely, self-incompatibility (SI), gynoecism, and male sterility, have proved to be reliable and economically viable options and, therefore, have gained high popularity among breeders. By leveraging these genetic mechanisms, improved yields and desirable traits can be achieved, seed purity can be concomitantly insured, and the need for labor-intensive emasculation and manual pollination can be reduced.Brassica oleracea [Self-incompatibility (SI) refers to a genetically controlled mechanism of certain plant species that prevents self-pollination . It invooleracea , which eoleracea ,79,80,81Crepis foetida, while Gerstel [Parthenium argentatum. Bateman [Brassica oleracea.Earlier studies have investigated the SI mechanism in cabbage ,82 and i Gerstel  document Bateman ,84,85 fu Bateman ,87,88 stBrassica rapa [Brassica oleracea, 30 to 40 S-alleles in Brassicaceae [Brassica oleracea and Brassica rapa have been documented by different authors. Classical genetic analysis categorized the Brassica S-alleles into 2 groups based on their phenotypic effect, i.e., with high allele activity placed high on the dominance scale (0 to 10 pollen tubes develop per self-fertilized stigma) and low allele activity considered to be recessive (10 to 30 pollen tubes develop per self-fertilized stigma). Furthermore, genetic analysis revealed that the S-locus cysteine-rich (SCR) protein and S-locus receptor kinase (SRK) were responsible for SI reactions in Brassicas [Brassica pistil [The discovery of SSI resulted in the production of a huge number of S-locus alleles. In particular, 30 S-locus alleles in ica rapa  and moresicaceae , and morrassicas ,92,93. Trassicas . Other ma pistil , the inva pistil , and inta pistil . The stuBrassica campestris. However, the co-dominance of alleles (varying in number) in both the stigmas and pollen was observed by Sampson [B. oleracea, and Hadj-Arab et al. [Brassica spp. Sampson [Brassica campestris. In relation to S-haplotypes, the S-locus revealed a dichotomy in sequence, and all class-I haplotypes were reported to show dominance over all class-II haplotypes in the determination of pollen specificity [Brassica rapa and Brassica oleracea demonstrated that dominant/recessive relationships are regulated at the mRNA level. Understanding the gene action related to SI is highly valuable, as S-alleles with higher dominance are likely to exhibit minimum selfs and sibs in hybrid seeds. Such knowledge can greatly assist breeders in identifying and isolating S-allele homozygotes/heterozygotes.Gene action studies related to the SSI system have proved helpful to breeders in many ways. Dominance, reverse dominance, co-dominance, and competitive interactions were among the main gene actions reported by different authors in their respective studies. Thompson  suggeste Sampson  in Brocc Sampson  in radis Sampson  in Chine Sampson  in cauli Sampson  in cabba Sampson  in kale, Sampson  in B. olb et al.  in caulib et al.  observedb et al.  discusse Sampson  summariz Sampson . Kakizak Sampson  reportedcificity , while Hcificity  observedcificity  working Various genetic and environmental factors can contribute to the production of self-seeds in an SI population, which is a phenomenon known as pseudo-compatibility or breakdown of incompatibility. Genetic processes involved in this phenomenon include the presence of weakly active S-alleles in the style , competiVarious methods for the detection of SI have been proposed by different researchers. These include conventional breeding approaches ,99,127, Brassica. Subsequent studies on SI feasibility in various crops of the Brassica group, such as cabbage [Brassica, single, double, three-way, and triple crosses were preferred for hybrid seed production. Double, three-way, and triple crosses proved to be more cost-effective than a single-cross hybrid seed production method. However, intensive preparatory breeding work to isolate, maintain, and increase the SI lines as well as test their combining ability was required. Moreover, the lack of uniformity in hybrids developed with SI is also a major problem. To overcome these problems, strategies such as the use of isogenic lines as parents [2) gas [Pearson  first pr cabbage ,88, broc cabbage , and kal cabbage , were co cabbage , cole cr cabbage , Brussel cabbage , and cau cabbage ,145,146. parents ,147 for  [2) gas ,149, and [2) gas ,151 to p [2) gas . Moreove [2) gas  as theirIn vegetable crops, SI and male sterility systems are mainly confined to the Brassicaceae and Solanaceae families. There are reports of such systems in other families, particularly Cucurbitaceae. The Cucurbitaceae family has been endowed with highly variable sex types and is one of the plant families characterized by predominantly unisexual flower production. Through evolutionary processes, the sex forms in cucurbits emerged as monoecious, gynoecious, andromonoecious, androgynoecious, and gynomonoecious types ,155,156.a and g, governs stable gynoecism [gy-1) controls gynoecism in bitter gourd [Well-documented studies have been conducted on the inheritance of gynoecism  in cucumber ,80,159. ynoecism . Furtherynoecism  in cucumer gourd ,161,162.er gourd . AttemptClosely linked SSR markers to the gynoecious (F) locus, namely, SSR13251 and UW020605 at 1.0 and 4.5 cM, respectively, were identified in cucumber by Jat et al. . A total3) (50–100 ppm) or silver thiosulfate (25–50 ppm) in gynoecious plants at the 2–3-leaf stage has been suggested to stimulate the production of staminate flowers. Furthermore, the use of 6 mM of silver nitrate has been suggested in bitter gourd to induce hermaphrodite flowers [The induction of male flowers in cucumber gynoecious lines for commercial-scale hybrid breeding with the application of growth regulators has been reported by Robinson . Before  flowers . HoweverWith respect to male sterility, genic male sterility (GMS), cytoplasmic male sterility (CMS), and cytoplasmic genetic male sterility (CGMS) systems have gained popularity among breeders for hybrid seed production . GMS hasSingh et al.  developePr locus that is related to the partial restoration of fertility in CMS was also reported in chili pepper (Capsicum annuum L.) [Two CMS-specific SCAR markers were developed to distinguish N-cytoplasm from S-cytoplasm and AFLP markers linked to the fertility restorer gene Rf . The CAPnuum L.) . The SCAMs gene [Ms locus identification, marker-assisted selection made it possible with RFLP [Ms locus [Ms locus [Ms locus, new PCR markers linked to the Ms locus are still being reported. For the identification of the cytoplasm, two PCR markers specific to onion cytoplasm, viz., accD, an indel marker [Ms locus determination, two PCR markers reported for the restorer-of-male-fertility (Ms) locus, namely, AcSKP1 [Ms locus. Furthermore, they observed that the limitations of PCR-based markers for the identification of the Ms locus still exist and need to be addressed by utilizing new PCR markers.Globally, S-type cytoplasm in onion is always preferred due to its stability in diverse environments and being genetically governed by a single gene, the Ms gene . PCR-basith RFLP , CAPS [2ith RFLP , SCAR [2ith RFLP , SNPs [2ith RFLP , and othith RFLP ,265,266.ith RFLP ,267 and Ms locus ,265 are Ms locus ,269,270.l marker , were ut, AcSKP1 , were em, AcSKP1  concludeThe fortification of key vitamins, antioxidants, and micronutrients has been highlighted in recent advances in conventional plant breeding. Traditional breeding methods such as variety evaluation in available germplasm, pre-breeding, selection, hybridization, heterosis breeding, mutation breeding, and polyploidy breeding are the most widely used biofortification approaches that offer long-term, cost-effective alternatives to transgenic and agronomic approaches . HoweverTraditional breeding methods, excluding mutation and polyploidy breeding, require sufficient genetic variation for traits such as carotene and other useful carotenoids, iron, zinc, and other minerals . LimitedIn cases where nutrient-rich donor parents are not cross-compatible with the recipient parents, we can alternatively transfer the genes/QTLs related to the target nutrient into a compatible variety to produce inbreds or pure lines that can be further used for hybridization to develop desirable biofortified hybrids. Information regarding the genes/QTLs for nutritional quality is given in The International Food Policy Research Institute, in collaboration with the International Center for Tropical Agriculture and the CGIAR, has started the Harvest Plus program to breed biofortified staple food crops . The majAlthough significant efforts have been made by researchers and institutions to develop hybrids through the utilization of genetic mechanisms, the real economic potential of these mechanisms remains untapped. Moreover, new and innovative traits should be introgressed into the backgrounds of strong and stable SI, CMS, and CGMS lines that can be further utilized for the development of hybrids in vegetable crops. This becomes particularly important in the face of climate change, wherein the development of hybrids, particularly for off-season production, with better adaptability, a significant number of nutraceuticals, and the ability to impart multiple resistance to biotic and abiotic stresses is of utmost importance. Molecular approaches can play a vital role in strengthening genetic mechanisms. This includes the development of novel fertility–sterility restoration systems and the exploitation of genetic tools to down-regulate specific genes  for imprSignificant progress has been achieved in vegetable hybrid breeding in the last decades through the exploitation of genetic mechanisms such as self-incompatibility and male sterility. Gynoecism has also recently gained popularity, and the application of this system in cucurbitaceous vegetable crops holds great potential. The integration of genomic tools, particularly the molecular markers that are practically feasible for the identification of GMS at the seedling stage, will help to enhance its scope. It will open new avenues to exploit monogenic recessive male sterile lines in several vegetable crops. Marker-assisted breeding, particularly the mapping of nutritional-, abiotic-, and biotic-stress-resistant genes, could be helpful to introgress such genes into newly developed genetic-mechanism-based hybrids.


Dear Editor,it is necessary to have available tests that could diagnose, in the first trimester of pregnancy, that the fetus has been affected by the virus, and that it may have important limitations, in order to subsidize the qualified discussion about abortion in these cases.” In fact, there is still no evidence that the Zika virus infection in pregnant women is the cause of abortion, and there is still no recommendation for therapeutic abortion in infected pregnant women. The important issue is the relationship between infection during pregnancy and the induction of an abnormal infant. In many regions, especially in tropical Asian countries,We read the publication on “Abortion in Cases of Zika Virus Congenital Infection.”


As life expectancy continues to increase in most EU Member States, smart technologies can help enable older people to continue living at home, despite the challenges accompanying the ageing process. The Innovation Action (IA) SHAPES ‘Smart and Healthy Ageing through People Engaging in Supportive Systems’ funded by the EU under the Horizon 2020 Research and Innovation Programme (grant agreement number 857159) attends to these topics to support active and healthy ageing and the wellbeing of older adults. This protocol article outlines the SHAPES project’s objectives and aims, methods, structure, and expected outcomes. SHAPES seeks to build, pilot, and deploy a large-scale, EU-standardised interoperable, and scalable open platform. The platform will facilitate the integration of a broad range of technological, organisational, clinical, educational, and social solutions. SHAPES emphasises that the home is much more than a house-space; it entails a sense of belonging, a place and a purpose in the community. SHAPES creates an ecosystem – a network of relevant users and stakeholders – who will work together to scale-up smart solutions. Furthermore, SHAPES will create a marketplace seeking to connect demand and supply across the home, health and care services. Finally, SHAPES will produce a set of recommendations to support key stakeholders seeking to integrate smart technologies in their care systems to mediate care delivery. Throughout, SHAPES adopts a multidisciplinary research approach to establish an empirical basis to guide the development of the platform. This includes long-term ethnographic research and a large-scale pan-European campaign to pilot the platform and its digital solutions within the context of seven distinct pilot themes. The project will thereby address the challenges of ageing societies in Europe and facilitate the integration of community-based health and social care. SHAPES will thus be a key driver for the transformation of healthcare and social care services across Europe. European citizens are living longer, with life expectancy exceeding 80 years in most European states. This increase in life expectancy is creating new challenges for health and care systems as older adults are at greater risk of cognitive impairment, frailty, and multiple other chronic health conditions. Such impairments can have negative consequences for older adults’ quality of life and independence, also placing burdens on those who care for them. Digital care technologies have the potential to improve integrated care for older adults, allowing them to live independently with multiple chronic health conditions, by monitoring their health and wellbeing, assisting with care, and creating smart home environments. SHAPES aims to empower older adults by developing and testing a range of digital technologies to facilitate long-term healthy and active ageing and the maintenance of a high-quality standard of life. Life expectancy across Europe has increased. According toThe diversity of Health and Care (H&C) systems in European countries and regions adds to the challenge. Health literacy, technology and individual involvement in care are key to making H&C more user-friendly and empowering. The redesign of care delivery suggests a benefit in shifting care from hospitals to home and the community  and citiDT-TDS-01-2019 “Smart and healthy living at home”, the Horizon 2020Smart and Healthy Ageing through People Engaging in Supportive Systems (SHAPES) project will seek to support older adults in retaining their independence and autonomy alongside the occurrence of any age-related limitations they may face. SHAPES is a four-year project (November 2019 – October 2023), which will create a unique and open pan-European Ecosystem including a standardised, interoperable, and scalable Platform for the integration of smart technologies. The SHAPES Platform has the following three core functions: to collect and analyse data relating to individuals’ health, environments, and lifestyles; to identify needs on an ongoing basis; and to provide reliable, trustworthy, and affordable recommendations and solutions in return. SHAPES Digital Solutions specifically address users' requirements and expectations on the use of innovative technologies to support and extend older adults’ independent living and active and healthy ageing at home.Responding to theThe SHAPES Consortium  consistsThe SHAPES Platform will be co-developed based on the following empirical research areas: the lifeworlds of older adults; the organisational, structural, and sociotechnical factors associated with the delivery of integrated care services; a set of technological requirements to accommodate an open and interoperable ecosystem of digital solutions specifically tailored to support and extend independent living of older individuals. The SHAPES Platform will then be validated through a multidisciplinary, large-scale piloting campaign encompassing the following seven key themes:1)Smart Living Environment for Healthy Ageing at Home,2)Improving In-Home and Community-based Care,3)Medicine Control and Optimisation,4)Psycho-social and Cognitive Stimulation Promoting Wellbeing,5)Caring for Older Individuals with Neurodegenerative Diseases,6)Physical Rehabilitation at Home, and7)Cross-border Health Data Exchange Supporting Mobility and Accessibility for Older Individuals.These pilots will involve over 2000 older adults across fifteen pilot sites in 10 states (nine EU-28 Member States (MS) and Northern Ireland), including six reference sites of the European Innovation Partnership on Active and Healthy Ageing (EIP on AHA). In addition, SHAPES will draw on expertise of professionals from a wide range of disciplinary and professional backgrounds within and outside of the consortium. Crucially, SHAPES will build and operate within clearly defined ethical and legal frameworks to protect both researchers and participants’ fundamental rights, privacy and personal data, while retaining the latter’s trust. Alongside user requirements, ethical requirements are particularly important when developing and employing solutions linked to fundamental rights and when the target group is older adults.The insights gathered throughout the lifetime of SHAPES will make a valuable contribution towards improving the lives of older adults in Europe and towards enhancing the sustainability of health and care systems in Europe. The Action will contribute to the work of the World Health Organisation’s Digital and Assistive Technology for Ageing (DATA) initiative  effects that the ageing population will have on health and care systems. This will involve new models of long-term support and care for older individuals, strong cooperation among all stakeholders of health and care, services that assist professional and informal caregivers, and integrated and economically sustainable tools.The SHAPES platform seeks to provide inclusive access to a broad range of technological, organisational, clinical, educational, and social solutions that have the potential to foster wellbeing and quality of life. It will mediate digital services and care models that put greater emphasis on inclusivity and accessibility. Furthermore, SHAPES will produce recommendations to promote the interaction between in-home and local community environments, and health and care networks. Concentrating on the needs of care receivers, integrated care involves the coordination of the processes between diagnosis and treatment, between primary care and secondary care, and between different therapeutic fields of expertise  to embed intelligence that helps with everyday practices. However, European measures remain modest, with experiments involving advanced ICT services – from smart home technologies to eHealth solutions – supporting H&C through small-scale, localised initiatives. SHAPES takes up these propositions and provides a pan-European Platform, comprising digital and technological solutions, a governance model framework, and real-life insights from relevant stakeholders. In relation to these key objectives, the project aims to construct an interoperable Platform integrating smart digital technologies which will collect and analyse data regarding older people’s health, environments, and lifestyle. This information will help to identify their needs, and to provide personalised solutions. SHAPES, which relies on the collection and analysis of personal and sensitive data, such as a person’s health information, takes a rigorous approach to ensuring that personal data are protected at all times.SHAPES digital technologies aim to support older people as they embrace independent living and active and healthy ageing at home. They will cover a wide spectrum of assistive technologies such as assisted living platforms, online communication and accessibility tools, cognitive stimulation and rehabilitation programmes, conversational assistants and chatbots, robots, telehealth and remote monitoring platforms, security solutions and COVID-19 response tools, as well as data analysis solutions in the domains of predictive systems, anomaly detection and wellbeing assessment. All of these will be brought together in a European open ecosystem that enables the large-scale deployment of a broad range of digital technologies. In order to achieve these ambitious objectives, the qualitative research and piloting activities of the project engage with over 2,000 older individuals and involve hundreds of key stakeholders.Lifeworld is conceptually understood as the immediate experiences, activities, and contacts that make up the experiential world of an individual’s life. It is the “given”, common-sense world of subjects in and beyond the physical environments, including a manifestation of collective meanings and ways of being in the world , WP3 will examine the organisational and socio-technical aspects that govern health and care systems. Seeking to develop a future-orientated concept of operations (CONOPS) , underpiWP4 and WP5, in continuous exchange with WP2, WP3, WP6 and WP8, will develop the technology for the SHAPES Platform that includes a suite of digital solutions. To be able to successfully mediate the delivery of health and social care, the Platform needs to be open, interoperable, expandable, and modular. Whether the platform can adequately fulfil these requirements will be evaluated during the piloting activities (WP6). The development of the Platform will take into consideration the functional, ethical and legal, security, health system, business and technical requirements identified in WP3, and the personas and use cases developed in WP2. Mapping these requirements to a reference architecture and to the implementation of the SHAPES technological platform (TP) and its core components will allow interconnection among the various SHAPES digital components and solutions. Integrating findings from WP2 and WP3, emphasis is given to core functional components, authentication, security elements, and interoperability services, in compliance with standards or widely adopted and emerging protocols, ensuring SHAPES’ openness, interoperability and security. Furthermore, WP4 will deal with and verify the integration of digital technologies (developed in WP5) in the SHAPES TP.WP5 will tackle the definition of the SHAPES User Experience (UX) and provide guidelines for the implementation of digital technologies specifically, those designed for older users. Moreover, SHAPES digital technologies will be adapted to comply with user requirements and follow the SHAPES architecture, benefitting from software development kits (SDKs) and application programming interfaces (APIs) developed in WP4 and WP5. The digital technologies also benefit from SHAPES’ advanced features, including speech processing and natural language processing (evolving text-based chatbots capable of engaging in human-like conversation) and hardened security and privacy .Piloting and evaluation of the SHAPES Platform and its  technologies, both under controlled conditions and in real-world contexts, is essential. The SHAPES pan-European pilot campaign (WP6) will involve both small-scale and large-scale pilots in eleven European countries . The pilot campaign is structured around seven themes that focus on improving the health, wellbeing, independence, and autonomy of older individuals, while concurrently enhancing the long-term sustainability of European health and social care systems. It will involve older adult end-users, caregiver end-users, and care professional end-users in 25 different use cases, which will conduct their pilot activities in 15 pilot sites, including five EIP on AHA Reference Sites. The spectrum of pilot themes enables the validation of innovations that capably support, promote, and extend active and healthy ageing, and independent living of older adults who have permanently or temporarily reduced functionality and capabilities within and outside their homes, and does so in a cost-efficient, interoperable and reliable manner. To understand whether and how SHAPES might meet the needs of its end-users, the pilot campaign is founded upon the knowledge base developed in other SHAPES work packages, including the identification of SHAPES personas and use cases (WP2), understanding existing care pathways and care delivery (WP3), and the development of the SHAPES Technological Platform and solutions .. For Phase 2, SHAPES technical partners will have provided mock-ups for each use case. At this stage, participants in the pilot activities will be invited to give feed-back to be able to design the SHAPES digital solutions tailor-made to the needs of the end-users. In Phase 3, the digital solutions will have reached prototype stage. Participants will be asked to provide a second round of feed-back. Additionally, this phase will include hands-on experimentation to train the participants on the individual digital solutions. Phase 4 will involve small-scale live demonstrations, where a small number of participants will be asked to replicate the scenarios described in Phase 1. Phase 4 will act as a ‘play rehearsal’ seeking to ensure that specific combinations of different SHAPES digital solutions integrated into the SHAPES Platform work as planned in participants’ homes environments. In Phase 5, the piloting activities will be carried out in real life with approximately 450 participants. This phase will involve the testing of the platform and its digital devices in participants’ home environments, as described in Phase 1. Each pilot theme will be replicated in several pilot sites. Different pilot activities are associated with particular themes, and they are conducted by one leading pilot site (L) and replicated, in part or whole, by other pilot sites (R). Each pilot site may conduct different pilot activities addressing different themes depending on their relevance to the end-users.The SHAPES pilot campaign will be implemented in a series of phases, which altogether will include 2,000 participants. Phase 1 will involve planning of pilot activities, pilot design in consideration of end-user needs , and the specification of key performance indicators (KPIs). This phase will draw upon the detailed understanding of older adults and their lifeworlds developed within WP2 of SHAPESSHAPES will encourage the trialling of third-party technologies provided by organisations from outside the consortium, and their inclusion of external companies will be facilitated through open calls. This will contribute to both the expansion of the platform across Europe, and to evidence-based decision-making processes pertaining to the adoption and market uptake of SHAPES policies and digital technologies for active and healthy ageing (AHA).The long-term sustainability of the SHAPES is a key concern that is reflected in the high-level adoption and expansion of the SHAPES Platform and its digital solutions in the post-project phase. WP7 will define strategies to manage – i.e. shape - the market, which will inform the business models developed in SHAPES to maximise returns within the parameters of a healthy market. WP7 will develop strategies to elicit the best value for money for both users and customers, and for the internationalisation and global ambitions of SHAPES. A SHAPES ‘marketplace’ will be developed to explore the Platform’s modular architecture, enabling the establishment of a rich ecosystem of suppliers and functionalities, and obtaining a significant market advantage.The silver economy encompasses economic activity centred on the needs of people aged 50 and over, and includes an innovation and technology as a central component. It is expected that by 2025, it will generate €6.4 trillion (31.5%) of the EU’s GDP and over 88 million (37.8%) jobs . It is also expected that healthcare spending will rise from 53% to 60%, which will account for €465 billion (11.4%) of all government spending . In addressing the needs of older adults with disabilities, SHAPES activities are also carried out in compliance with the principles and the rights provided for in theSHAPES hosts an Ethical Advisory Board to provide relevant guidance to the project. The SHAPES Platform considers the relevant H&C delivery landscape to address the integrated care models and policies in Europe and the ageing in place process, as well as EU approaches to harmonise eHealth, assistive technologies, and the cross-border exchange of healthcare information. Since EU legislation takes the form of minimum standard directives, national implementation plays an important role. Within the Consortium, 13 EU Member States and Norway are represented, thus providing a wide regulatory coverage for the implementation of the SHAPES Platform. SHAPES identifies the most relevant regulatory specifications applicable to H&C information exchange cooperation, including in the context of prevailing H&C models and systems, considering personal data exchange . Further, SHAPES dedicates specific attention to EU standardisation efforts on eHealth and to the security risks and challenges related to the design and development of the SHAPES Platform and digital technologies.The knowledge gathered through the multitude of tasks and work packages of the SHAPES project will yield practical results, including i) a handbook detailing the methodology for organisational implementation of the SHAPES Platform and ii) a report capturing the key recommendations for the implementation, adoption, and scale-up of the SHAPES Platform and digital technologies across Europe.WP9 focuses on building an interactive ecosystem that comprises a heterogeneous set of stakeholders on both multidisciplinary and international level spreading over different European health and social care systems. In parallel to forming its platform, pilots and digital solutions, which are all interconnected with understanding the lifeworld of older people and substantial ethical requirements, SHAPES aims to ensure sustainability for its actions beyond the project’s runtime. The basis for this sustainability is for SHAPES to understand the ecosystem in which it operates and its key stakeholders to provide assistive services and recommendations to help facilitate active and healthy ageing. The acquired knowledge will enable SHAPES to actively form and build its ecosystem of reference to ensure uptake and long-term integration of the project outputs.In particular, WP9 aims to achieve this goal through three core activities: Running a Co-creation Think-tank for European Integrated Care that comprises internal and external members to discuss governance and data management emerging from WP3. Performing Innovation Watch, Cross-Fertilisation and Foresight Exercises to identify emerging technologies, future impacts and new societal demands and challenges that help to evaluate priorities and potential new directions in decision making. And building strong networks and liaisons to analyse, adapt and recommend standards and interoperability and to manage Open Calls for Innovation and Collaboration that will give external organisations the chance to participate in SHAPES and to enrich the portfolio of SHAPES digital solutions. The gathered information is summarised in “Influencing Factor Cards” and “Technology Cards”, that provide starting points for interested actors for more in-depth analysis into specific topics.To support the set-up of the SHAPES ecosystem, WP10 conceives seven dialogue workshops that engage external stakeholders in the project activities and are dedicated public frames where SHAPES meets a wide audience of researchers and businesses, health and social care professionals, older people and civil society. Each workshop is anticipated by an awareness campaign, circulating some key research questions of each event to its social media audience. Through the awareness campaigns, SHAPES reaches out to an informed public and confronts with it, exchanging on concepts, methodologies, outputs and results, to further deepen the project’s knowledge and outreach.SHAPES implements a co-creation methodology between the SHAPES partners and the SHAPES stakeholder ecosystem and between the Action’s social science and technological development. This occurs in six-month cycles, each delivering results and outcomes that take the Action a step closer to achieving its strategic goals.The technology developed and adapted in WPs 4 and 5, and validated in WP6, will be informed by user needs, priorities, and requirements  and is in full compliance with EU and national legal, ethical, and fundamental rights framings (WP8), in which the General Data Protection Regulation and the Directive 2011/24/EU on patients' rights in cross-border healthcare are key pillars. SHAPES adopts an ethics-based approach taking the protection of the rights of older individuals as central. The social science activities in SHAPES combine engagement in knowledge exchange with existing projects and mini ethnographic studies, lending itself to both comparability (between case-countries) and customisation (tailor-made approaches per country). 100 ethnographic case studies to understand the interconnected, contextualised, sociocultural and gendered lifeworlds of older individuals and persona-based use cases to investigate the meso and micro levels enable an exploration of differences and similarities to build local-context-specific knowledge. In-depth open-ended ethnographic interviews and participant observation, expert interviews and focus groups, and intervention-based methods are applied. Thereby, the research considers obligations emanating from relevant law, ethics and policy and provides the relevant requirements to assist the development of the technological solutions.SHAPES will develop an activity-centred concept of operations (CONOPS), which is based on the IEEE’s standard CONOPS template  and enriThe activity-centred approach to CONOPS emphasises the interconnectedness of different actors with a focus on both their shared and diverging goals and interests with a broader sociocultural and material context. This framework complements the CONOPS framework as it views the developing technological ecosystem of SHAPES as a key mediator between people and their goals and allows the SHAPES partners to view the development of the Platform with a constant eye of the intended future state. Rather than trying to ignore or avoid potential conflicts in the system, which may occur due to divergence between the goals and interests of actors, the availability and suitability of resources, and the challenges surrounding organisational change, it seeks to proactively identify such potential contradictions and position them as key design challenges. The purpose is not to eliminate all potential conflict but to develop a sociotechnical system with the flexibility and inclusiveness required to accommodate the range of diverse actors and goals as is reasonably possible. To understand and analyse end-user needs, and ultimately evaluate whether SHAPES meets those needs, the project employs a human-centred, stepwise, co-design process by developing personas, user stories, use cases and technologies. Firstly, in WP2, social scientists and partners within the SHAPES consortium produce personas from ethnographic case studies and focus groups with older adults and caregivers as well as in-depth interviews with professionals. Personas will contain the attributes, attitudes, behaviours, and characteristics of the various end-user groups targeted by SHAPES. Secondly, in WP2, general use cases (UCs) are developed on the basis of the needs of personas. Use cases aim to illustrate the breadth and variability of the technology use for improvements of the quality of life of older adults. Lastly, in WP6, on the basis of the more general use cases developed in WP2, specific use cases are developed for each of the pilot themes (UC-PTs). In these UC-PTs, the needs of end-users and the user stories are merged with specific digital tools and solutions of the SHAPES Platform.For the technological development and deployment activities, SHAPES applies a three-stage methodology: 1) Prototyping – to consider the end-users requirements and deliver hands-on experience of mock-ups for early validation by end-users; 2) Development – design, implementation, adaptation and integration of the Platform, improving existing solutions; and 3) Deployment – installation of the Platform and digital technologies in users’ real-life environment.In the background of the SHAPES architecture  lies theThrough the SHAPES concept of a Marketplace, such extended new types of solutions can be advertised, discovered and consumed by stakeholders as well as combined together to address new and emerging e-Heath needs, an example being the current COVID-19 pandemic that has identified a need for new types of e-Health solutions targeting large–scale infectious diseases. To offer technological capabilities that address such generic user needs, a practical implementation of the Technological Platform differentiates device, middleware and service layers at the bottom of the diagram that are all associated with symbIoTe platform as the core IoT interoperability platform. Using its multiple levels of interoperability compliance, devices, platforms, and services are seamlessly and flexibly registered to symbIoTe, match their individual data information models to produce conversion rules, allowing them to directly and bilaterally exchange their information. This way privacy of data can be achieved compliant with GDPR regulations. Such a mediation framework offered by symbIoTe further permits flexible addition of new types of solutions thus supporting expandability of the SHAPES system to new solutions in the future.Using 3rd-party expandability capabilities using concepts of “enablers”, representing approaches ranging from software development kits (SDKs) and application programming interfaces (APIs) to explicit reusable code offered as Docker components, new solutions, services and applications can be grown on top of the SHAPES technological platform to address new and emerging e-Health domains. The core architecture defines the underlying system organisation and its components whose purpose is to support seamless connectivity and data exchange/sharing among interconnected devices, services, platforms and applications (as described in the concept architecture earlier), all of which being referred to as Digital Solutions. Such an architecture has been defined to offer the following basic functionalities:1. Single sign-on to all Digital Solutions compatible and registered to SHAPES2. Data authorisation managed by users and on Digital Solutions where data is stored3. Seamless integration of devices, services, platforms and applications4. Standard-independent interoperability w.r.t IoT data and Medical Information5. Advanced data analytics of data shared among diverse Digital Solutions6. Management of medical data over diverse Digital Solutions for same individual7. GDPR-grade protection of private identifiable private data8. Future extendibility to additional medical standards and data models9. Optional sharing of captured anonymised data sets for research purposesSHAPES will deploy these digital solutions in the homes and communities of end-users of the Platform. Table-top exercises, implemented at the early stages of the Action, will validate initial concepts and approaches, by recreating realistic scenarios. Mock-up or prototype validation, performed at the early stages of the Action, will assess user acceptance and feedback on user experience and other non-functional elements, such as accessibility and inclusion features. Hands-on experiments, conducted at the mid and late stages of the Action will validate functional elements of SHAPES components and selected digital technologies and gather user feedback on performance, shortcomings and new additions. Based on the feedback, the digital technologies will be adapted and improved and be tested again by the older adults.The SHAPES Platform must be validated at the European scale, across different regions, cultures and health and care organisational models and on the basis of its overall aims. Evaluation will be facilitated by a range of both quantitative and qualitative methods and analyses and, rather than focusing merely on individual technologies or themes, SHAPES will be evaluated as a platform and ecosystem, and as a comprehensive set of potential solutions to a set of complex issues. Care technologies must meet the needs of their target beneficiaries, and do so effectively.For each use case, key indicators relating to health, quality of life, and wellbeing, will be developed and the constituent technologies, will be evaluated in relation to their impact in such indicators. As the ecosystem aims to be greater than the sum of its constituent parts, in addition to data collection for assessment of outcomes of individual pilots, a core set of harmonised psychosocial data will be collected across the pilots. This core set of data is also used together with additional use case specific data to evaluate the outcome of each use case with MAST. MAST is an established method to assess the effectiveness and contribution of telemedicine applications to quality of care , incorporating gender sensitivity , and gender specificity . The promotion of health and quality of life is bound by politico-economic and fiscal imperatives, and socio-economic sustainability and cost-benefit analyses will be performed within SHAPES.Health technologies, although they may be promising, are often not adopted by end-users to a substantial degree, are not accepted by end-users, are not scaled-up within organisations or contexts, do not spread to other organisations and contexts, and are not sustainable  optimisation of a growing SHAPES ecosystem, b) building of critical mass, c) cost reduction through the expansion of integrated technology-mediated care delivery, which may lead to greater market competitiveness, d) accelerated transfer of knowledge and technologies, and e) facilitating access to non-EU markets. Key to achieving this is the delivery of new and innovative digital solutions and services.SHAPES will take a system-of-systems ecological approach, which facilitates the analysis of systems as a network of interrelated relationships rather than siloed units. This approach recognises the role of decentralised services and local structures in accessing innovative products and services. It is expected to result in greater autonomy at local level emphasising the role of community-based services, and to strengthen the agency of service users.Systems Market for Assistive and Related Technologies (SMART) Thinking Matrix tool  for the SHAPES Platform and its digital solutions to ensure their competitive advantage. The market analysis will investigate European and global markets of digital technologies that support active and healthy ageing and facilitate independent living. This analysis will draw upon the business expertise within SHAPES, supported by the wider ecosystem of SHAPES stakeholders, and will take into consideration tools and models such as SWOT and PESTEL, the KSFs and prevailing market barriers.In addition, SHAPES will conduct an objective cost-benefit analysis of the daily living activities of its users and day-to-day integrated care delivery activities. Based on the outcomes of the SHAPES pilot campaign, a set of indicators for the socio-economic sustainability of the SHAPES results will be defined. Highlighting the importance of balancing social, economic and environmental objectives, these indicators are intended to be used to assess the impact of the SHAPES adoption and scaling-up initiatives. Including forecasts for the next five to ten years, the SHAPES economic sustainability analysis will vary the SHAPES approaches  to identify assumptions, bottlenecks, intellectual property aspects, economic outcomes and alternative development and funding strategies. Platform) can be commercialised successfully to ensure that access to SHAPES is cost-effective, will maximise returns for SHAPES partners and will serve the needs of older adults and health and care systems. The SHAPES Uptake Roadmap, which will be informed by the SMART Thinking Matrix , as well as defining ethical requirements as features of the proposed SHAPES solution during that development and implementation work . SHAPES pilots are multifaceted from the viewpoint of ethics, since they concern ethics of development work, validation of the ethical features of SHAPES and the use of SHAPES in real-time settings. In the pilots, validation activities will be conducted in an environment that expects the development version to be piloted itself, fulfilling the minimum legal requirements. Efficient and compliant management of the SHAPES data is required to increase the sustainability of project results and enhance the integrity and validity of the project methods. As the project progresses, any newly identified ethical issues will be addressed by mitigating options to the Consortium.One of the key challenges for the SHAPES project is the need to balance users’ rights to privacy and data protection, while simultaneously allowing for the use of big data analytics to create user profiles and deliver personalised solutions feeding into the SHAPES Platform. The debate about balancing accessing and using personal data, in particular, health data and protecting users’ privacy rights has gained fresh momentum since the introduction of the GDPR and adoption of the European Commission’s Digital Strategy . HoweverThroughout the lifetime of the project, all SHAPES partners commit to upholding ethical research standards, including the European Code of Conduct for research integrity. They are committed to delivering high-quality scientific outputs and to be transparent, ensuring deliverables’ reliability and impact. These features of deliverables are validated as part of the quality management procedures. The principles of maximising benefit and minimising harm, of social responsibility, of dignity of persons, of fundamental human rights and other issues mentioned in the Horizon 2020 ethical self-assessment are supported during the R&D work by taking into use ethical self-assessment procedure as part of the SHAPES governance structure. This ethical self-assessment is based on the Horizon 2020 template, but it is further modified for the specific purposes of the SHAPES ethics governance.At the time of submission of this protocol article, the project is halfway through its lifespan. Much of the groundwork – such as the ethnographic investigation of older adults’ lifeworlds, persona development and use cases and the analysis of the trends in health and social care provision across Europe for the evolving CONOPS – has been undertaken successfully. Results, which form the basis for the development of the platform, have been shared with all SHAPES partners. Ethical, technological, social and legal requirements for the SHAPES Platform have been developed as the groundwork for the implementation of the Platform. Furthermore, methodologies for testing pilot use cases have been developed, and digital solutions and necessary technological prerequisites have been created and cross-aligned among all partners. A suitable ethical framework has been established and is guiding the piloting activities. Moreover, we are in the process of developing the SHAPES governance model, which will address questions related to business, enterprise and data governance, and will be aligned with the newly launched European Health Data Space. Midway through the project, the focus is now on piloting activities. Other activities involve the development of the SHAPES ecosystem, its marketplace and think tank to ensure the lasting impact of the SHAPES project in the health and care sector.SHAPES commenced only four months before the onset of the coronavirus pandemic and the work of the SHAPES consortium was considerably affected by Covid-19. The ways in which the consortium worked, both internally and in gathering first hand data, had to be adjusted and had to respond to the methodological challenges relating to lockdowns and the fact that the population at the heart of this research was at high risk from Covid-19. However, while the pandemic challenged all staff to work even harder to find solutions to continue the work and research in SHAPES, all deliverables have been submitted on schedule and with the highest quality.Throughout the lifespan of the project, SHAPES will engage in dissemination activities to communicate with academics, policymakers, the health and care sectors, and public audiences. Using social media and a website as well as a blog called #shapesstories, results from and the process of research within the Innovation Action are processed in a way to be both intellectually stimulating and easily accessible. SHAPES communicates its findings, including real life experiences of older adults and key questions that need answering in innovation research, at various conferences across Europe and beyond via paper presentations, blog entries and panel discussions as well as through accessible SHAPES dialogue workshops.Publications are an integral part of the SHAPES dissemination strategy. A number of scientific articles in a range of open access and academic journals have been published to date, and many more are planned for dissemination over the next two years. Additionally, two manuscripts for books are in preparation: the edited volume “Rethinking Smart and Healthy Ageing in Europe“  and the monograph “From Legacies to Futures. Lifeworlds of Older Adults in Europe” . established by the European Commission to connect the main Horizon2020 funded research projects, SHAPES, the work group on dissemination, fosters capacity building and the continuous collaboration and exchange with other leading researchers in the field.The SHAPES platform as well as the SHAPES governance model, ecosystem, and marketplace will all provide opportunities to disseminate results and findings and to ensure access to SHAPES data and findings for future researchers and contribute to market shaping. Furthermore, through webinar activities, SHAPES engages with the European Parliament and with the WHO to foster its impact. Embedded in the Health and Care ClusterSmart and Healthy Ageing through People engaging in Supportive Systems” (SHAPES) project will create the first European open Ecosystem that will enable the large-scale deployment of a broad range of digital technologies for supporting and extending healthy and independent living for older individuals. The Ecosystem will build on an interoperable Platform integrating smart digital technologies to identify older individuals’ needs and to provide personalised solutions that uphold the individual’s data protection and trust. The SHAPES Platform will offer a combination of devices, software, and accessible modes of interacting within the living environment that can adapt to the needs and priorities of older individuals, including those facing permanent or temporary reduced functionality and capabilities. Beyond the lifespan of the project, the SHAPES Platform will sustain increased efficiency gains in health and care delivery across Europe, bringing improved quality of life to older adults, their families, caregivers and care service providers.In innovative ways, the “Using an inclusive, action learning approach allows for the inclusion of bottom-up insights from empirical data collected in the ethnographic lifeworld research as well as the pilot site validation that will make the project innovative and responsive to the needs and discoveries during its four-year lifespan. Rather than imposing fixed research structures, the project partners will work closely and collaboratively with each other to ensure effective cross linking of all work packages and their associated activities.The SHAPES Platform will be responsive to sociocultural and economic contexts and the users’ physical and mental states, therefore being able to adapt to their pressing needs and concerns and use nudges to encourage users to make smart, healthy and active decisions, as well as achieve improved levels of independence, active social participation and quality-of-life. In other words, the SHAPES Platform will drive interconnection and integration, physical activity, personal agency and in-person human and community relationships. In addition, the SHAPES Platform will bring effective and efficient mechanisms to augment social interactions between individuals and, where applicable and consented, caregivers and care and health service providers, with rich communications accessible to all  and ancillary data  improving diagnosis, risk assessment and complications prediction to enable early intervention and avoid corrective actions.As such, the SHAPES Platform will be intelligent, processing user data in real-time and adapting itself as a suite of resources and capabilities to flexibly provide the human, digital and mechanical wherewithal to improve a wide variety of real-life environments. Powered by a big data and artificial intelligence (AI) engine, the Platform will incorporate an understanding of the evolving environment in which older individuals actually live and iteratively follows-up and evolves  based on how the resources are being used. Building an ecosystem attractive to European industry and policy-makers, SHAPES develops value-based business models to open and scale up the market for digital technologies focused on active and healthy ageing and provides key recommendations for the far-reaching deployment of innovative digital health and care solutions and services supporting and extending healthy and independent living of older adults in Europe.No data are associated with this article. The partners are described in detail, but the activities in which each of them will work have not been specified.It would help if, in the introduction, when talking about the topics, the objectives that will be worked on in SHAPE were included and, in turn, related to the WP. All of this, in turn, could be connected to the sustainable development goals.The titles "Understanding older adults, their lifeworld, and social contexts" and etc. they should be listed in Figure 3 as objectives . In this way, I suggest that the WP be mentioned with the full name so as not to have to constantly look at the table. The WPs could be developed in an orderly fashion, instead of mixing the description, separating paragraphs and ignoring the following WPs, that is, concentrating on the description, sample and detail of who is going to carry out said activities in each WP.In the ethics of activities and, given the possibility of developing a completely transferable system, the possibility of a medical device must also be considered. In this sense, it would be necessary to apply and comment on the ethics of medical devices, as well as all the regulatory requirements and aspects.The architecture of Figure 5. generically comments on market aspects, but feedback through the health product is not reflected. For this reason, it is also convenient to mention not only the layout of the database, but also how it can be suggested through new technological tools related to the topics discussed in the introduction. The contribution and possible technological transfer, as well as the TRL that the system could reach, once the investigation is finished. All this can be connected with the section "Market shaping to inform SHAPES' business models and socio-economic impact" and Figure 6. Instead of commenting on market aspects (since they could correspond to future phases that are not scope in this project) mention should be made of TRLs upon completion. On the other hand, if despite everything you want to continue commenting on market aspects, you should mention real indicators, for example, CARG, demand, etc... instead of replicating the theory at the system, meso, macro level, and micro, which is discussed in the social literature. In short, it is necessary to consider critical market, economic and socioeconomic indicators, as well as technological ones.The risks of the project do not appear at any time. A risk analysis would be useful as a form of evaluation to achieve the intended objectives. A risk analysis is suggested from market aspects , not only social.Thank you very much for your article. I greatly appreciate your research "Implementation of a pan-European ecosystem and an interoperable platform for Smart and Healthy Ageing in Europe: An Innovation Action research protocol" and am very grateful to be able to review it. Here are some suggestions to improve it and I hope that all this can facilitate a more visible article from an interdisciplinary approach:Is the study design appropriate for the research question?YesAre sufficient details of the methods provided to allow replication by others?PartlyIs the rationale for, and objectives of, the study clearly described?YesAre the datasets clearly presented in a useable and accessible format?PartlyReviewer Expertise:Economics and Markets, Technology, Engineering, Sustainability, Sociology.I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard. This protocol article reported the SHAPES project's objectives, methods, structure, and expected outcomes. The article provides a comprehensive SHAPSE Consortium framework and structure of the projects and described stakeholder engagement well. The evaluation plans are well-rounded in user case-level, theoretical aspects, and systematic levels. Under the evaluation methods, please elaborate on the tools/instruments that would be used to evaluate the key indicators.  Please discuss possible influences of internal and external events that may happen in designated field sites.Please discuss plans to maintain the sustainability of the project.   I only have a few comments for the authors to consider:Is the study design appropriate for the research question?YesAre sufficient details of the methods provided to allow replication by others?YesIs the rationale for, and objectives of, the study clearly described?YesAre the datasets clearly presented in a useable and accessible format?YesReviewer Expertise:Public health sciencesI confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard. WP2 aims to explore the lifeworlds of older individuals, and it could be useful to specify whether specific attention will be paid to the use of technology by these adults, since this is the topic of the current project.The difference between WP4 and WP5 could be made clearer.When describing WP5, the authors might want to mention whether older adults themselves will be involved in the UX development process, at which stages and to what extent.The authors might want to add to the "Dissemination of results" section the address of the project website and some examples of published papers from the project.The paper presents an impressive project spanning a large range of experts from different backgrounds and countries, aiming to create an open pan-European ecosystem including a standardised, interoperable, and scalable platform for the integration of smart technologies. This platform can be used to promote the health and well-being of older adults. The project seems to be making good progress, despite a rocky start due to the COVID-19 pandemic. The project is adequately described in detail, and I only have a few small comments.Is the study design appropriate for the research question?YesAre sufficient details of the methods provided to allow replication by others?YesIs the rationale for, and objectives of, the study clearly described?YesAre the datasets clearly presented in a useable and accessible format?YesReviewer Expertise:Aging adults (gerontology), social relationships at old age, cognitive aging, ageism.I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard.


Respiratory viruses can be transmitted by multiple modes, including contaminated surfaces, commonly referred to as fomites. Efficient fomite transmission requires that a virus remain infectious on a given surface material over a wide range of environmental conditions, including different relative humidities. Prior work examining the stability of influenza viruses on surfaces has relied upon virus grown in media or eggs, which does not mimic the composition of virus-containing droplets expelled from the human respiratory tract. In this study, we examined the stability of the 2009 pandemic H1N1 (H1N1pdm09) virus on a variety of nonporous surface materials at four different humidities. Importantly, we used virus grown in primary human bronchial epithelial cell (HBE) cultures from different donors to recapitulate the physiological microenvironment of expelled viruses. We observed rapid inactivation of H1N1pdm09 on copper under all experimental conditions. In contrast to copper, viruses were stable on polystyrene plastic, stainless steel, aluminum, and glass, at multiple relative humidities, but greater decay on acrylonitrile butadiene styrene (ABS) plastic was observed at short time points. However, the half-lives of viruses at 23% relative humidity were similar among noncopper surfaces and ranged from 4.5 to 5.9 h. Assessment of H1N1pdm09 longevity on nonporous surfaces revealed that virus persistence was governed more by differences among HBE culture donors than by surface material. Our findings highlight the potential role of an individual’s respiratory fluid on viral persistence and could help explain heterogeneity in transmission dynamics.IMPORTANCE Seasonal epidemics and sporadic pandemics of influenza cause a large public health burden. Although influenza viruses disseminate through the environment in respiratory secretions expelled from infected individuals, they can also be transmitted by contaminated surfaces where virus-laden expulsions can be deposited. Understanding virus stability on surfaces within the indoor environment is critical to assessing influenza transmission risk. We found that influenza virus stability is affected by the host respiratory secretion in which the virus is expelled, the surface material on which the droplet lands, and the ambient relative humidity of the environment. Influenza viruses can remain infectious on many common surfaces for prolonged periods, with half-lives of 4.5 to 5.9 h. These data imply that influenza viruses are persistent in indoor environments in biologically relevant matrices. Decontamination and engineering controls should be used to mitigate influenza virus transmission. This seasonal burden and the threat of future influenza virus pandemics make it critical to develop strategies to impede influenza virus transmission.In 2009, an H1N1 influenza virus emerged from swine and caused a pandemic, with 60.8 million cases in the United States in the first year  on surfaces, including fomite material. Influenza viruses deposited on porous surfaces, such as fabrics and wood, exhibit reduced persistence of infectivity compared to those on nonporous surfaces, including steel and plastic , 14, 15.Environmental conditions such as humidity and temperature also affect the stability of expelled influenza viruses , 11, 19.20–The matrix composition of droplets is known to influence viral stability on surfaces. Many prior studies examining influenza virus stability on surfaces used viral stocks propagated in egg allantoic fluid or tissue culture monolayer cell lines, such as Madin-Darby canine kidney cells. The resultant viral suspensions do not mimic the biochemical composition of virus-laden respiratory secretions expelled from an infected individual. Adding exogenous mucin to cell-grown viral stocks has been reported to have no effect on influenza virus stability , 11. HowWhile many studies have looked at the individual effects of humidity, surface material, and respiratory droplet composition on influenza virus infectivity, the interplay between these factors remains understudied. A better understanding of this relationship could improve transmission risk assessment and evidence-based mitigation. To address this, here we examined the stability of viruses grown in different HBE cultures on a variety of nonporous materials at RHs ranging from 23% to 98%. The use of HBE cultures better mimics the composition, complexity, and heterogeneity of secretions expelled by infected individuals. We found that virus viability depends not only on the surface tested but also on the RH used. Importantly, we also found that the HBE patient culture influenced viral stability, suggesting that heterogeneity in human respiratory secretions may contribute to persistence of infectious influenza viruses in the environment.Environmental factors such as temperature, humidity, and the material of the contaminated surface can impact the survival of influenza viruses , 30. We 6.8 to 107 50% tissue culture infective doses (TCID50)/mL were deposited onto the surface materials of polystyrene (PS) plastic, stainless steel, aluminum, glass, acrylonitrile butadiene styrene (ABS) plastic, and copper. Droplets were then incubated for 2 h inside a desiccator chamber with a RH of 23% to model a dry indoor environment, such as that produced by heating and insulation during a temperate region winter   relative to the sealed control, as previously observed (10 TCID50/mL. Decay was significantly higher on ABS plastic (1.5 ± 0.39 log10 TCID50/mL) and copper, where the deposited virus decayed to undetectable levels  compared to low (23%) or high (>80%) RH . However10 TCID50/mL, even on ABS plastic , a surface effect (assumed to apply across cultures), and a single intercept . This simple model assumes that the culture effect and surface effect act independently to modify the half-life of the virus.Using this approach on our experimental data, we report the predicted half-life for the virus in each given culture on a hypothetical “neutral” (average) surface—that is, with zero surface effect. Similarly, we report estimated surface effects as predicted half-lives on each surface given a hypothetical neutral (average) culture—that is, with zero culture effect .The median viral half-life of a neutral culture on different surface materials (excluding copper) ranged between 4.5 and 5.9 h . In contIn this study, we investigated influenza virus stability on different types of surfaces using H1N1pdm09 propagated from three-dimensional patient-derived lung cultures. Stability of H1N1pdm09 in the presence of airway surface liquid appears to be surface dependent under various common indoor RHs tested . The notable exception was deposition on copper, which completely inactivated virus at all RH levels and as quickly as 30 min at room RH. On the other nonporous surfaces tested (excluding copper), the half-life of HBE-propagated H1N1pdm09 at 23% RH was similar on PS plastic, stainless steel, aluminum, and glass. Interestingly, a subsequent analysis revealed that the half-life of infectious H1N1pdm09 at 23% RH varied substantially among the different HBE patient cultures that were used to propagate the virus. This final observation suggests that the respiratory secretion composition may have a profound impact on the persistence of viruses in the environment and that individual variation may matter a great deal.The rapid inactivation of HBE-propagated H1N1pdm09 observed on copper at all RHs tested indicates that this effect is mediated by the surface itself and is robust against other environmental conditions. Noyce et al. also described inactivation of 20-μL droplets of A/PR/8/34 (H1N1) after 6 h on a copper surface , whereas34–41–Environmental conditions such as RH could affect the transmissibility of influenza virus fomites by influencing virus stability. Relevant RH conditions include those similar to indoor RH during temperate-zone winters (approximately 20%) and those similar to indoor RH during temperate-zone summers (40 to 60%). Previous work has suggested a U-shaped relationship between RH and virus stability for enveloped viruses, including SARS-CoV-2 and influenza virus, where virus stability is lowest at midrange RHs , 33. HowMost surprisingly, the half-lives of HBE-propagated influenza viruses at 23% RH appeared similar across surfaces, but droplet composition from HBE cultures impacted H1N1pdm09 stability more . This suThe emergence of multiple respiratory pandemic viruses in the last few decades has highlighted the importance of understanding virus transmission pathways  and of determining the impact of virus traits, environmental conditions, and host factors on each mode . Persistl-glutamine, and 1% penicillin/streptomycin. Four primary human bronchial epithelial cell (HBE) cultures were differentiated from lung tissues of four different patients and were maintained at an air-liquid interface in Transwells (Madin-Darby canine kidney (MDCK) cells (ATCC) were maintained in Eagle’s minimum essential medium supplemented with 10% fetal bovine serum, 2 mM answells . Each HB10 TCID50 of virus was added to each Transwell from the apical side after the cells had been washed once with phosphate-buffered saline (PBS). The inoculum was removed after 1 h, and the HBE-propagated virus was collected with 150 mL of PBS every 24 h up to five times. Viral washes collected at the same time from different Transwells of the same HBE culture were pooled and titrated on MDCK cells. Up to four viral washes with the highest titers from the same HBE culture were combined. To make an abundant HBE-propagated virus stock without diluting physiological components in the suspension, the combined virus wash was diluted 1:1 with the cell wash from the same HBE culture, which had been collected prior to infection. To account for patient-specific variations, four stocks of HBE-propagated virus from different cultures were made. The stocks were aliquoted and stored at −80°C before use.The 2009 pandemic H1N1 influenza A virus, A/California/07/2009 (H1N1pdm09), was acquired as previously described . To prepSix surface materials were selected because of common use: PS plastic, stainless steel, aluminum, glass, ABS plastic, and copper. Disposable PS plastic was readily available as the flat bottom of a 6-well tissue culture plate . Disposable glass cover slides  were used as glass surfaces without additional preparation. Circular, smooth-surface coupons of 304 stainless steel (20 gauge), 6061 aluminum (1.6 mm thick), ABS plastic , and 110 copper (0.6 mm thick) with a uniform diameter of 20 mm were purchased through Alumagraphics. The coupons were cleaned with an interfering-residual-free detergent (Alconox) prior to sterilization  or by disinfection with isopropanol (ABS plastic). The coupons were reused after cleaning and sterilization (or disinfection) following the assay.3COO for 23%, K2CO3 for 43%, Mg(NO3)2 for 55%, and K2SO4 for 98%  to desired levels: KCH for 98% . The tem50 assay .The decay of H1N1pdm09 estimated in 50 assay . SpecifiThe results of the virus decay were analyzed with GraphPad Prism 8. Statistical analysis excluded results from copper, because the virus decay was mostly beyond the detection limit. At 23% RH, one-way analysis of variance (ANOVA) using surface material as the variable was performed with Tukey’s multiple-comparison test to define surface-based variations in virus decay within 2 h. Two-way ANOVA using RH (or incubation time) as the second variable was performed with Tukey’s multiple-comparison test to define the effect of RH (or incubation time) on virus decay. Due to the interactive effects of RH and surface material on virus decay, the analysis of surface-based variations at different RH was adjusted with Tukey’s multiple-comparison tests following two-way ANOVA and summarized in TC 1-step kit (Thermo Fisher). Primers specific to the matrix gene (M) were used at a concentration of 0.1 μM (M25 F [5′-AGA TGA GTC TTC TAA CCG AGG TCG-3′] and M123 R [5′-GC AAA GAC ATC TTC AAG TCT CTG-3′]) and an M64 probe (6-carboxyfluorescein [FAM]-TCA GGC CCC CTC AAA GCC GA-NFQ) was used at 0.25 μM. All reactions were performed in duplicate. The thermal cycling step was conducted on a StepOnePlus real-time PCR system (Applied Biosystems) with StepOne software (version 2.3) following the manufacturer’s recommendations. A standard curve was generated with five 10-fold serial dilutions of viral RNA extracted from a stock virus with a known TCID50 titer.The RNA of H1N1pdm09 was extracted from virus samples using a PureLink viral RNA/DNA minikit (Invitrogen). For quantitative reverse transcription-PCR (qRT-PCR), 5 μL of the viral RNA was analyzed using a TaqMan RNA-to-To estimate the effects of surface and culture on virus half-lives  and 4, w50 from the distribution of positive and negative wells observed at various dilutions of the sample. We assume that the number of virions inoculated into any given well is Poisson distributed, with a mean given by the virus concentration in the diluted sample used for that well. This is the same Poisson single-hit assumption that underlies traditional endpoint titration statistical approaches, such as the Spearman-Karber and Reed-Muench methods. The difference is that by implementing this in a Bayesian framework, we are able to connect our model directly to a regression for estimating half-lives, and we are able to obtain principled estimates of uncertainty  for each estimated quantity, rather than just a single number.The models quantify virus from positive or negative readouts of inoculated wells by treating well inoculation as a “single-hit” process: if at least one virion successfully infects a cell within the well, the well will show evidence of infection. We then estimate the virus concentration in TCIDTo estimate virus half-lives, we coupled our titer estimation model to a simple regression model in which virus is assumed to decay exponentially over time. The slope of a regression line predicting log titer as a function of time therefore gives an estimate of the exponential decay rate, which can be converted directly into a half-life. Our Bayesian approach allows us to account in a principled way for various sources of noise, such as variation in the initial quantities of virus deposited onto individual surface coupons.Finally, to estimate the effects of individual surfaces and cultures on virus persistence, we used a regression model in which we treated log half-life of the virus in a given culture on a given surface as depending on three quantities: a culture effect (assumed to apply across surfaces), a surface effect (assumed to apply across cultures), and a single intercept . The model assumes that the culture effect and surface effect act independently to modify the half-life of the virus. We estimated these culture and surface effects, and the intercept, from our data.To aid interpretability, we report estimated culture effects in units of half-life. Specifically, we report the predicted half-life for the virus in the given culture on a neutral (average) surface—that is, with zero surface effect. Similarly, we report estimated surface effects as predicted half-lives on the given surface given a neutral (average) culture—that is, with zero culture effect.(i) Notation. In the model notation that follows, the symbol “~” indicates that a random variable is distributed according to the given distribution. Normal distributions are parameterized as ~normal . Positive-constrained normal distributions  are parameterized as ~Pos-normal . Beta distributions are parameterized as ~beta , with canonical shape parameters α and β > 0 .(ii) Titer inference and model description. We inferred individual titers directly from titration well data according to a Poisson single-hit model . Therefore, f = −ln(1 − fpp).The one variation from the model described in reference (iii) Prior distributions. We assigned a weakly informative normal prior to the log10 titers iv : iv ~ normal. We placed a beta prior on the false-positive probability fpp, assuming it to be small but allowing it to be nontrivial: fpp ~ beta .(iv) Predictive checks. We assessed the appropriateness of prior distribution choices using prior predictive checks. The prior checks suggested that prior distributions were agnostic over the titers of interest.(i) Model description. To infer half-lives of viable virus in the various experiments, we used a regression model.i, we have two sets of measurements: those for treatment samples deposited on surfaces and incubated at a given temperature and humidity, and those for control samples kept in bulk solution at room temperature. We assume that each sample j for experimental condition i, whether treatment or control, had some unknown initial log10 titer ijv0 at time zero.For each experimental condition 10 titer vi: ijv0 ~ normal (vi).We assume that all of these initial values are normally distributed about a mean initial logi for treatment samples in condition i and λi for the corresponding control samples. For the treatment samples, we assumed there was an experiment-specific mean amount of virus lost during deposition (ℓi).We modeled loss of viable virus as exponential decay at a rate δijv of virus sampled at time ijt is given byIt follows that the quantity i, λi, and ℓi under the assumption that our observed well data reflected the corresponding predicted titers ijv.We then used the direct-from-well data likelihood function described above, except that instead of estimating individual titers independently, we estimated the values of δ(ii) Prior distributions. We placed a weakly informative normal prior on the mean initial log10 titers vi: σvi ~ Pos-normal . This allows either for large variation (more than ±0.5 log10) about the experiment mean or for substantially less variation, depending on the data.i and Λi, where Δi = ln{[log10(2)]/δi} and Λi = ln{[log10(2)]/λi}. We made the priors weakly informative ; we verified this with prior predictive checks: Δi ~ normal  and Λi ~ normal .We placed normal priors on the log treatment and control half-lives Δi): ℓi ~ Pos-normal .We placed a Pos-normal prior on the experiment mean deposition losses (ℓfpp was as in the titer estimation model (see “Titer inference and model description”).The prior for the false-positive probability (iii) Predictive checks. We assessed the appropriateness of prior distribution choices using prior predictive checks and assessed goodness of fit for the estimated model using posterior predictive checks.i and Λi as in the half-life inference model (detailed above), but instead of placing priors directly on them, we assumed that the log half-lives Δi could be predicted according to a linear equation with intercept n, and surface-specific effects μm: Δi = C(i) + μS(i) + ϵi, where C(i) is the culture for experiment i, S(i) is the surface for experiment i, and ϵi is a normally distributed error term with an estimated standard deviation σΔ: ϵi ~ normal .To estimate and compare the effects of surface and culture on virus persistence, we used a simple regression model. We estimated treatment and control log half-lives Δi were normally distributed about an unknown mean Λ: Λi ~ normal (Λ).We assumed that the control log half-lives Λi and Λi in the half-life inference model: Δi ~ normal  and Λi ~ normal .We placed normal priors on the intercept of the log half-life for a neutral surface, n and the surface effects μn with standard deviations designed to rule out implausibly large effects: χn ~ normal  and μn ~ normal(0. 0.5).We used normal priors centered on zero for the culture effects χΔ and σΛ: σΔ ~ Pos-normal and σΛ ~ Pos-normal .We placed Pos-normal priors on the standard deviations σThese allow either for substantial (1-log) variation compared to the regression prediction or for minimal variation.All other priors were as in the half-life estimation model.We assessed the appropriateness of prior distribution choices using prior predictive checks and assessed goodness of fit for the estimated model using posterior predictive checks.