Increasing evidence suggests that corporate sustainability is paradoxical in nature, as corporates and managers have to achieve economic, social, and environmental goals, simultaneously. While a paradox perspective has been broadly incorporated into sustainability research for more than a decade, it has resulted in limited improvement in our understanding of corporate sustainability paradox management. In this study, the authors conduct a systematic review of the literature of corporate sustainability paradox management by adopting the Smith–Lewis three-stage model of dynamic equilibrium. The results reveal the following: (1) Both environmental and cognitive factors manifest tensions arising from the sustainability paradox. (2) While both proactive and defensive strategies are adopted to manage the tensions embedded in the corporate sustainability, the proactive strategy is more extensively studied in the current literature. (3) Management strategies of corporate sustainability paradox are characterized as multi-level, multi-stage, and dealing with multiple paradoxes. (4) Proactive strategies enable organizations to enjoy short-term and long-term sustainability benefits. The authors call for further research explicitly addressing the following areas: (1) the paradoxical nature of corporate sustainability management; (2) corporate sustainability paradox management of for-profit organizations; (3) the micro-foundations of corporate sustainability paradox management; (4) defensive strategies and new proactive strategies; and (5) a unified standard of sustainability outcomes. The practical implications of this review are then elaborated. In practice, the results imply that organizations would best manage the corporate sustainability paradox by understanding the paradox and its equilibrium stages. This review and proposed research agenda are expected to deepen interdisciplinary knowledge and set the stage for interested scholars to undertake in their future inquiries. Although organizations have been pursuing their economic goals as primary interests for a long time, concerns about the natural environment and social welfare have become essential for organizations to deal with when considering sustainable development . Second, to depict the characteristics of these management processes, the authors analyzed the strategies and decided whether a proactive or defensive response is taken, and further coded these strategies as multi-level , multi-stage , or multi-paradox . Finally, the authors investigated the short-term impacts  and long-term outcomes  of these management strategies and their related practices. As for the initial screening of the focal articles, three of the authors independently analyzed the data. After each round of analysis, the authors discussed the coding together to resolve inconsistencies and ensure the accuracy and validity of the findings.The three stages and specific items that the authors focused on when this model was applied to investigate corporate sustainability issues are as follows: (1) the manifestation of paradoxical tensions, (2) the management of sustainability paradoxes, and (3) outcomes. First, the authors identified the factors that could render paradoxical tensions salient and categorize them into In this section, the authors answer the following questions. First, what factors make sustainability tensions salient? Second, what are the strategies  to manage sustainability paradoxes? Third, what are the outcomes (short-term and long-term)? A list of the representative articles and their coding information is found in In corporate sustainability research see , hybrid Two sets of triggering factors that could manifest organizational paradoxical tensions have emerged, consistent with Smith and Lewis'  dynamic In general, organizational paradox research has recognized the importance of environmental factors in providing the material conditions that lead to perplexing choices , and/or discussed the identified paradoxes separately. This separation tendency could constrain our understanding of the sustainability paradox's intricate nature. A few exceptional studies have appeared recently. For example, Vallaster et al.  adopt a Theoretically, proactive responses to organizational paradoxes could lead to sustainable outcomes. In the short term, individuals, teams, and organizations are likely to display excellence through enhanced learning and creativity, flexibility and resilience, and stimulated potential, which would pave the way for long-term success  requires further exploration, including theoretical clarity and empirical examination. In the mainstream organizational paradox literature, there is an ontological debate on whether paradoxical tensions are inherent in the system or socially constructed Clegg, . Smith aThis debate penetrates sustainability research and potentially shapes management strategies. With a few exceptions, such as Sharma and Bansal , who dirSecond, it is desirable to investigate the sustainability paradox management of for-profit organizations. When investigating management of the corporate sustainability paradox, the main focus of the existing literature is on social enterprises or non-profit organizations, not for-profit organizations (Smith et al., Third, the authors encourage more research on the micro-foundation of management of the corporate sustainability paradox. Organizational psychology would shed light on this line of research. There is a need to explore how the cognitions, attitudes, and emotions of managers and employees (cf., Chin et al., Furthermore, the role of employee engagement together with its underlying micro-foundation are greatly ignored when using a paradox perspective compared to those in the traditional CSR research Glavas, . Based oFourth, some researchers challenge the prevailing opinion on proactive and defensive strategies, which urges more investigations. Originally, a proactive strategy was favored as it would lead to a virtuous cycle and achieve sustainability outcomes (Smith and Lewis, Finally, a unified standard for a sustainability outcome is needed in future research. In some studies, the authors use the short-term effects to claim promising long-term achievements without substantial supporting evidence. In other cases, some researchers investigate whether the company could use its profit to cover the costs of social extension (e.g., Slawinski et al., The paradoxical nature of corporate sustainability, namely, the need to address social, ecological, and commercial concerns simultaneously, is at the core of corporate sustainability research. The literature on how corporations and firm decision-makers should address the sustainability paradox has rapidly evolved in the past decade. Hence, in this study, the authors reviewed existing sustainability research prior to 2020 when the paradox perspective received direct attention (Van der Byl and Slawinski, This review has several practical implications. Leaders of firms would best manage the corporate sustainability paradox by understanding the paradox and its equilibrium stages. Understanding how and why firms are affected by opposing tensions and the stage they are in enables firms to better position their firms and develop their own strategies, for example, by pursuing social aims associated with their core business (Kaul and Luo, For middle managers of firms, this review suggests that firms could provide development training on how middle managers attend to these tensions and how middle managers communicate this information from firm leaders and top managers to operational managers and employees. Middle managers are recognized as the major force resisting organizational change Miles, . WithoutIn conclusion, an understanding of how the existing literature has advanced knowledge of the management corporate sustainability paradox is timely for both scholars and practitioners alike. By incorporating Smith and Lewis'  dynamic The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.BL proposed the research idea and developed the initial draft with YT. EC improved the theorizing. YT, SL, and DL collected, screened, and analyzed the articles together. BL, EC, and YT revised the draft. All authors contributed to the article and approved the submitted version.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Regarded as a defining factor in resource management, it is widely accepted that visual attention and related processing will deteriorate, in a global fashion, across the lifespan and produce detrimental consequences for environmental interactions. Disproportionate detriments in such functions are also proposed to arise in the context of a range of age-related disorders. However, the nature and extent of such losses, the precise mechanisms that underlie these changes, and the origin of deficits in task performance that are used to assess this multicomponent process are still under debate. This Special Issue, comprising studies spanning healthy ageing, cognitive impairment, and dementia, represents a body of literature that fundamentally advances these points of discussion.Contrary to previous findings and prior assumptions, the research documented herein attests to a large degree of preserved task performance in ageing. First, with regard to the role of attention in working memory (WM), by simultaneously assessing participants’ ability to filter and ignore distracting stimuli, Maniglia and Souza  separateHenderson et al.  also ideFurther extending the discussion of integrity of function to those with mild cognitive impairment, Revie et al.  highlighAcross the lifespan, and between states of health and disease, the extent to which individuals are able to adequately manage attentional allocation remains contested. Extending the debate on the control of resources to the domain of divided attention, Barel and Tzischinsky  investigAdditionally, Polden et al.  comparedBuilding on the role of participant characteristics, Torrens-Burton et al.  exploredThe real-world insights advocated by the aforementioned study could produce incredibly valuable applications considering driving cessation is a major life transition, with implications for health and well-being. Huizeling et al.  enhance In recent years, non-invasive brain stimulation techniques (NIBS) have been heralded as compensation aids due to their ability to enhance neural plasticity. Use of these methods could, therefore, represent a powerful approach to sustaining cognition in ageing. However, beyond initial successes, the multitude of stimulation parameters available to researchers, especially those exploring transcranial direct current stimulation (tDCS), makes it extremely difficult to optimise protocols and maximise beneficial outcomes. Hanley et al.  address Therein, this collection of articles represents cross-modality research at the cutting-edge of advances in the multifaceted domain of visual attention-related processing. Crucially, each contribution in this Special Issue attests to the ageing process being dynamic and not simply characterised by global deficits. Similar inferences can be made in the context of various forms of cognitive impairment , where elements of preserved function have also been identified in the articles presented here. These considerations are highly important as they have direct consequences for prospective interventions, with the presence of individual differences implying that a one-size fits all approach will fail to incorporate the complexities outlined in the showcased research. By addressing the integrity of function via a unique array of approaches, encompassing novel methodologies and innovative concepts, these studies ultimately culminate in improving (1) our understanding of age-related changes, (2) the identification of potential signatures of a range of disease processes, and (3) interventions to attenuate the impact of observed decline and maintain quality of life. With a firm emphasis on the development of naturalistic applications, which test the assumptions put forward by basic research, an increase in the type of investigations advocated here will ensure the real-life relevance of future endeavours.


The Organization for Economic Cooperation and Development defines biotechnology as the “application of science and technology to living organisms as well as parts, products, and models thereof, to alter living or non-living materials for the production of knowledge, goods, and services.” While the widespread potential of nanomaterials offered a key driving force for their research and development in the biotechnology field, the role played by many nanomaterials is still unfortunately limited today due to their hazardous properties. Gold nanomaterials often exhibit fascinating material characterizations and biofriendly properties that truly stand out from many other nanostructures; as a result, they have been extensively investigated for different bioapplications, including detection, imaging, diagnosis, drug delivery, and therapy. For their applications in various bioenvironments and aims, gold can be fine-tuned into many different nanostructures ranging from clusters and nanoparticles to layer membranes and complex microstructures. With the diverse biofunctions achieved from these structures, components, and surface chemistry, gold nanomaterials are attracting great attention from researchers to further explore their bioapplications.in vivo fluorescent temperature mapping. For example, Wu et al. reported the effect of BSA as a stabilizer of AuNPs and obtained AuNP/BSA conjugates with high colloidal stability against salt, acid, and base. They also discovered the non-thiolated DNA cannot replace proteins on the gold nanoparticles, but thiolated DNA can be controllably attached to AuNP/BSA conjugates under certain conditions. This work not only reveals interesting biointerfaces on AuNPs for DNA and proteins absorption, but also provides a controllable method to obtain stable AuNP/BSA/DNA conjugates using a protein corona as a stabilizer. Chen et al. employed ligand exchange and interfacial assembly methods to fabricate plasmonic superlattice membranes, which possessed high structural homogeneity and exhibited near-field confined hotspots uniformly fashioned over the whole substrate, enabling ultrasensitive SERS-based detection of trace chemicals and label-free direct detection of BSA. The work represents a significant step toward the exploitation of plasmonic membranes for a plethora of technical applications such as the next-generation of nanophotonics devices and point-of-care biosensors. Hua et al. synthesized renal clearable gold nanoparticles and embedded them into silk to obtain a gold nanoparticle-functionalized silk film for in vivo fluorescent temperature mapping. The renal clearance property of gold nanoparticles together with biodegradable silk significantly minimized the potential toxicity and side effects of the hybrid material, which is an important step for the future development of implantable and wearable sensor devices. Tang et al. provided an overview of research progress on the development of versatile gold nanoclusters (AuNCs) for bacterial detection and infection treatment, and highlighted the mechanisms of action of designed diagnostic and therapeutic agents.In this Research Topic, we have collected three original research articles and one review written by 27 authors. These papers cover the important bioapplications of gold nanomaterials, including the stability of gold nanoparticles in a simulated tissue environment, protein and bacterial detection, infection therapy, the metabolic process of gold nanoparticles, and In summary, the goal of our Research Topic is to emphasize the structure design, surface ligand tailoring, composite assembly, and bioapplications of gold nanomaterials. The functions of gold nanomaterials generally depend on their size, nanostructures, and surface chemistry, which can be plentifully regulated to extend their bioapplications and stride forward with their clinic translation in the future. Admittedly, the limited number of studies presented in this Research Topic cannot cover the full scope of gold nanomaterials and their bioapplications; however, we sincerely hope the papers we collected in this special edition can be beneficial to your research endeavor.All authors listed have made a substantial, direct and intellectual contribution to the work, and approved it for publication.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Internet- and mobile-based approaches have become increasingly significant to psychological research in the field of bipolar disorders. While research suggests that emotional aspects of bipolar disorders are substantially related to the social and global functioning or the suicidality of patients, these aspects have so far not sufficiently been considered within the context of mobile-based disease management approaches. As a multiprofessional research team, we have developed a new and emotion-sensitive assistance system, which we have adapted to the needs of patients with bipolar disorder. Next to the analysis of self-assessments, third-party assessments, and sensor data, the new assistance system analyzes audio and video data of these patients regarding their emotional content or the presence of emotional cues. In this viewpoint, we describe the theoretical and technological basis of our emotion-sensitive approach and do not present empirical data or a proof of concept. To our knowledge, the new assistance system incorporates the first mobile-based approach to analyze emotional expressions of patients with bipolar disorder. As a next step, the validity and feasibility of our emotion-sensitive approach must be evaluated. In the future, it might benefit diagnostic, prognostic, or even therapeutic purposes and complement existing systems with the help of new and intuitive interaction models. With a prevalence of more than 1%, bipolar disorder is one of the most common mental disorders worldwide ,3. The dBecause of the frequently severe and chronic course and the individual and social consequences, additional strategies and support options within patient care are necessary ,7. With Over the past years, the use of smartphone apps or mobile programs has been investigated with samples of patients with bipolar disorder with an often good feasibility -16. BecaHowever, the existing approaches neglect the emotional aspects of bipolar disorders. For example, during mood episodes there are typical patterns of experienced emotions: whereas manic states are often characterized by increased happiness or anger and fear, depressive states often show patterns of elevated sadness and disgust . BipolarSo far, mobile-based approaches have analyzed the keyboard activity of patients with bipolar disorder  or even EmAsIn project  we developed the first emotion-sensitive, technical assistance system for patients with bipolar disorder. Because self-assessments of symptoms are the well-established basis of mood monitoring in bipolar disorder [Story of the Day module. It analyzes audio and video data to explore the emotional experiences and expressions of patients. While many apps in this field are poorly investigated [Within the disorder ,12,17-19disorder -27, and disorder . In addidisorder . As a codisorder ,57. The disorder -49 motivstigated , we emphstigated -61. To cstigated .Our assistance system includes an Android smartphone app and a connected wearable device, which can be both code protected and password protected. It uses multichannel data acquisition to realize an early recognition of mood-state changes in bipolar disorder. It further intends to complement the rather technical exchange of information between systems and patients with new and intuitive interaction models. Therefore, it aims to recognize socioemotional cues in human communication behavior and hereby infer conclusions about emotional and mental states. To this end, the emotion-sensitive Story of the Day module analyzes the verbal and facial expressions of patients in short and actively user-triggered recordings with respect to their emotional content or the presence of emotional cues. Consequently, this module collects active and passive emotion-related data of patients with bipolar disorder and relies on its regular use (see “Story of the Day” section). If all the features of the assistance system are activated, it can gather information about mood states and the course of bipolar disorders with the aid of the following resources:daily self-assessments of patients regarding their mood, activity level, and other relevant symptoms;regular third-party assessments by relatives or other related parties regarding the most important symptoms;automatic assessments of (psycho-) physiological parameters such as heart rate or resting heart rate;automatic assessments of sleep duration and quality;automatic assessments of several behavioral parameters such as recognized activities, movement/acceleration, steps per day, range of motion, or smartphone usage behavior ;assessments of auditive information  as emotional cues and indicators of mood states;assessments of visual information  as emotional cues and indicators of mood states.mixed emotions or increased caffeine intake) or can create new items. These items are then incorporated into the daily self-assessments, where they are evaluated with yes or no. The third-party assessments are very similar to the self-assessments, but they are realized by using a separate and individually secured web application.All data resources are presented in The assistance system uses smartphone sensors to assess several of the behavioral aspects, for instance, with regard to movement or social interaction . Information about sleeping behavior and (psycho-) physiological data is continuously collected with the help of the connected wearable device, which users wear on their wrists . The system then includes the individual assignments when integrating and analyzing the assessed data. Beyond that, patients may also assign these values to the warning signs, which are then analyzed as separate factors. The system is supposed to recognize mood-state changes in patients with bipolar and to react by sending warning signals or, like other approaches ,14,28,30As opposed to earlier approaches, which analyzed ambient sound samples or voice features without processing emotional information ,32,50,51Linguistic Inquiry and Word Count (LIWC) program [EmoVoice approach [Facial Action Coding System (FACS) [The app analyzes the assessed auditive and visual information separately. The verbal information is analyzed regarding the use of emotional words, the color of the voice, its energy level , the verbal fluency, and the speech rate as well as the extent to which the story is narrated. The count of emotional words in automatic transcriptions of the used language of each recording is based on the  program  and inclapproach , a framem (FACS) . In shorInternet- and mobile-based approaches have become increasingly important to psychological research in the field of bipolar disorders. In particular, the aspiring mHealth approach benefits a consistent self-monitoring of patients with bipolar disorder ,12,17-19While the self-perception of patients with bipolar disorder is certainly the most important factor in mood monitoring, self-assessments can be less reliable in specific cases or during severe episodes ,57. BeyoOur Story of the Day module, as far as we know, is the first mobile-based approach to analyze the emotional expressions of patients with bipolar disorder. As opposed to the analysis of ambient sound samples or voice features during phone calls ,32,50,51Of course, our new assistance system comes with its limitations. Most importantly, the predictive value of our approach concerning its mood-state recognition and its efficacy and effectiveness with respect to relapse prevention has to be addressed in empirical studies with patients with bipolar disorder. In addition, not all patients approve of the involvement of relatives or related parties in their mood-monitoring approach . Our StoWhereas the EmoVoice approach  and, in With this in mind, there are still some issues to be dealt with in the further development of our assistance system and more research is needed to examine the clinical value of our system. However, our assistance system and its new and innovative features might improve the understanding of the patients’ mood state and could provide important information about the patients’ expressed emotions as well as their  interaction behavior. Considering the strong association between emotional aspects and the social and global functioning of patients with bipolar disorder, in the future, emotion-sensitive systems might be even useful during emotion-based treatment approaches in bipolar disorder -68.The mHealth approach offers many opportunities to support patients with bipolar disorder in their everyday struggle with their disease. However, the existing mobile-based approaches do not consider the importance of emotional aspects in bipolar disorder and their implications regarding the social and global functioning of patients. With our assistance system, we aim to address this issue and have therefore implemented the emotion-sensitive Story of the Day module. With the help of this module, our system analyzes the emotional experiences and expressions of patients besides regular self-assessments and third-party assessments as well as the analysis of further sensor data. In the future, emotion-sensitive approaches might not only benefit a better understanding of the patients’ emotional states, but they might also be used to complement the technical exchange of information between systems and patients with more intuitive interaction models. Moreover, they might even support emotion-based interventions in bipolar disorder.


This chapter is a proactive risk exploration of hypothetical remotely operated search and rescue (SAR) ships in the Canadian Arctic. The harsh and remote environment in the region, combined with complicated coastlines and many uncharted or poorly charted traffic routes, makes it one of the most challenging SAR areas. Canada has committed itself to safety, environmental protection and sovereign presence in the area by maintaining joint SAR centres of federal government departments and mobilizing private volunteers. The characteristics of Canadian SAR response in the Arctic rest with its high dependency on heavy equipment such as aircraft, helicopters and icebreakers, entailing prolonged hours of response time. As recent climate change impacts and maritime traffic increase in the northern waters disclose safety gaps, innovation in SAR assets is anticipated. The safety gaps may be filled by state-of-the-art remote control technology. This chapter discusses remotely operated unmanned ships for SAR response, exploring their opportunities, risk dimensions and governance implications. Yara Birkeland, an 80 m-long autonomous cargo ship, is expected to be in service within a few years , which extends 1450 km, is a uniquely complex navigational area consisting of multiple routes as shown in Fig. Most importantly, the Canadian Arctic is not only fundamental to Canada’s national identity but is the homeland of Indigenous peoples across the Yukon, the Northwest Territories and Nunavut. Canada has maritime boundary disputes and incomplete boundaries with the United States in the Beaufort Sea and Denmark in the Lincoln Sea  due to the vast area and because the average speed of vessels on Arctic routes is known to be around 7 to 13 knots, compared with 21 to 25 knots in open sea  the limited hydrographic survey and nautical charting of marine routes, (2) “dead zones” of radio communications, (3) the lack of trained SAR personnel, (4) ageing equipment, (5) insufficient icebreaking services, (6) the lack of land base connectivity through fibre optics, (7) the low bandwidth of satellite communications and (8) prolonged SAR time  , information technology (IT)  and communication technology (CT)  in Canadian Arctic operations. Any failure of these technical components of RO-SARS could cause serious and unanticipated hazards to the safety of the SAR operation. In addition to hull structure, engines, machinery, electrical systems and conventional ship equipment, special attention will need to be paid to remote technical elements, including sensor technology, communication links, cyber safety and cybersecurity, and the interface between RO-SARS and remote controllers. Each of these four elements is discussed immediately below. More significantly, practical rescue functionality should be added to the conceptual design of RO-SARS. After all, human and technical elements of RO-SARS will have to prove that their reliability is as capable as the ordinary practice of conventional maritime rescue responders in the Arctic . Even if the federal government approves the operation of RO-SARS, other Arctic states might not welcome the novel technology in waters under their sovereignty or jurisdiction for safety and security reasons  in the near future. Fourth, a prototype of RO-SARS should be repeatedly tested, inspected and surveyed through sea trials to verify their human and technical seaworthiness and effective interaction with ships in distress, other SAR units and relevant technologies, including drones. Fifth, remote controllers should be qualified, trained and licensed seafarers for SAR operation under new legal standards that would need to be developed, possibly by the IMO as well as under Canadian federal law. Sixth, stakeholders should increase multitier satellite and terrestrial supports and data transmitters so that the interconnectivity of OT, IT and CT can meet the ordinary practices of SAR responders in the Canadian Arctic. Seventh, a safety management system specifically for unmanned RO-SARS should be put in place with approved training simulators and mandatory procedures Dasgupta . Eighth,Given the Canadian Arctic context, unmanned and remotely controlled ships could considerably enhance and complement Canadian SAR capabilities, particularly rescue operations, by reducing response time, infrastructural costs and life risks to responders. However, there are complex and uncertain risks that can be identified under the IRGC risk framework: the qualification and certification of remote controllers, the technical reliability of sensor technology, the stability of communication links, the hazards arising from the breach of cyber safety and cybersecurity, the probable interface errors between RO-SARS and remote controllers and the new design requirements for remote rescue functions such as the remotely operated ship-to-ship personnel transfer crane. These risks should be addressed under multilevel governance systems comprising international and national and public and private stakeholders. Most significantly, it is clear that unmanned RO-SARS are in line with the international conventions concerning SAR operations, that there are multilayered regulatory regimes applicable to these novel ships and that these vessels and craft should gain political and social license from the Arctic states and northern communities.Nonetheless, the risks of deploying unmanned RO-SARS in the region should not be treated as simple. These risks feature a combination of intrinsic heterogeneities such as remote technology, the SAR operation itself and the extreme environment, all of which are complex and uncertain in nature under the terms of the IRGC risk framework. Indeed, although the opportunities for RO-SARS look promising, the required technical reliability and actual SAR practicability are unproven in the Arctic context. Hence, it is premature to characterize overall risks as intolerable, tolerable or acceptable. However, the minimal exploration of the risk dimensions taken in the foregoing discussion might trigger feasibility studies and dedicated ship design approaches accounting for the different hazards originating from this novel technology concept, for which new design approaches for unmanned vessels could be applied.


Prevention of exposure to the COVID-19 virus in the general population is an essential strategy to slow community transmission. This paper shares the experiences and challenges of community engagement in COVID-19 prevention in the Kilimanjaro region, Northern Tanzania implemented by our team from the Institute of Public Health (IPH), Kilimanjaro Christian Medical University College (KCMUCo) in collaboration with the COVID-19 response team in the Moshi Municipality. We conducted an education session with the COVID-19 response team and together brainstormed transmission hotspots and which interventions would be most feasible in their settings. The first hotspot identified was crowded local market spaces. Suggested interventions included targeted and mass public health education through the engagement of market opinion leaders, public announcements, and radio shows. We conducted participatory rural appraisal techniques to enable market vendors and clients to visualize two-meter distances and provided a prototype hand-washing facility that was foot operated. We found mass public health educational campaigns essential to inform and update the public about COVID-19 pandemic and to address rumors and misinformation, which hampers compliance with public health interventions. Coordinated efforts among stakeholders in the country are necessary to develop context-specific prevention and case management strategies following the national and international guidelines. Local ownership of recommended interventions is necessary to ensure compliance. Tanzania had its first reported COVID-19 case on March 16, 2020, a returning national who re-entered the country through the Kilimanjaro International Airport (42kms from Moshi Town) [Community engagement activities: the IPH-KCMUCo team took a 5-steps approach to assess, engage, implement, and sustain public health prevention interventions within the Moshi municipality  held on April 22, 2020, and the other with all market leaders (at one of the big markets) held on April 24, 2020. During these meetings, the team conducted an education session, followed by brainstorming with the group to identify which of the recommended interventions  would beThe IPH-KCMUCo team, together with health officers from the municipality, and ward executive officer, visited the three big markets 2-7 days after the meetings with leaders to assess progress and provide practical advice to address implementation challenges. During these visits, IPH-KCMUCo donated a hand-washing device that uses a foot pump for dispensing both the water and liquid soap, instead of taps requiring users to touch with possibly contaminated hands . The staObservations in the largest market in town  revealed that despite the size, the arrangement of tables within the space still allowed for physical distancing strategies to be implemented. However, social distancing was a challenge in more crowded market areas. Our team faced challenges in introducing the same public health measures in the third and most crowded market in Moshi. This market has no walls or gates and, therefore, no physical reminders of what is expected of clients or traders. During the follow-up visits, we observed compliance with some interventions. For example, all traders in markets had face masks. However, they were not wearing them at all times. The same problem was witnessed amongst customers. Therefore, the leadership of the markets instigated a fine for non-compliance to wearing face masks equating to approximately US$3 and two bottles of liquid soap. This was applied to both traders selling goods and customers buying without wearing a face mask. Customers who refused to wash their hands were not allowed inside the market. The sustainability of such an intervention is yet to be established. The team, in collaboration with the Moshi municipality, developed a monitoring checklist to assist health officers in assessing the uptake of recommended interventions, progress, and bottlenecks at different hotspots, particularly bars and restaurants, salons, markets, and public transport stands in their areas. Our team also developed preventive messages in Swahili for each hotspot that can be used by any person involved with COVID-19 prevention and control. A copy of the messages was shared with the municipal COVID-19 response team, ward health officers, and market leaders.Our activities highlighted the need for public awareness campaigns to focus on creating awareness regarding the potential benefits of the preventive approaches, seriousness, susceptibility of COVID-19, and addressing potential barriers to uptake of the interventions -8. As weReligious leaders were also crucial stakeholders in COVID-19 prevention. The IPH-KCMUCo team had meetings with individual religious leaders who are known by members, to discuss potential preventive measures during religious programs. Sessions were held with leaders from different denominations . The recommended interventions, i.e., hand-washing, maintaining a physical distance, disinfection of benches, and surface after services and wearing face masks, have been implemented in the specific places of worship. The collection of these activities contributed significantly to the mass public education and awareness of COVID-19 in the region. Finally, the IPH-KCMUCo team summarized key challenges and provided feedback to the Moshi municipal COVID-19 response team for informed decisions and actions. To this point, there are deliberations about the way forward in this district. The reach of IPH-KCMUCo was further widened through requests from neighboring districts, to provide education to the ward health officers and appear on other local radio stations. The COVID-19 response team in a neighboring district also recommended the formation of a WhatsApp group to include members of the IPH-KCMUCo as well as district officials to be used for sharing experiences, challenges and asking questions. The WhatsApp group facilitated the monitoring of implementation.Lesson learned, success, challenges, and way forward: In summary, the key lesson learned included: 1) Educational interventions are essential to COVID-19 prevention. During the implementation of these activities, the team was able to educate the district through the office of the DMO, motivating uptake of interventions by stakeholders such as health officers and market leaders, seeing the support from local government. 2) Routine follow-up is necessary for hotspots such as markets and places of mass gatherings to enforce uptake of the interventions and to overcome the initial resistance to change. These visits should also include mobilization of opinion leaders and groups of people resisting changes. 3) WhatsApp groups are an effective mechanism to share experiences and challenges among health officers. 4) Support from respective local governments such as the district commissioners, the district/ municipal executive directors, and the DMO is necessary to support and strengthen the implementation of preventive interventions. 5) Question and answer sessions during radio sessions are essential not only for health education but also to allow the public to get context-specific clarifications from health experts. The media stakeholders are willing to participate in disease prevention activities, given that they are timely engaged. 6) The public prefers pictures with brief descriptions or explanations for COVID-19 prevention education over and above short messages alone. There were various successes and challenges in the implementation of community engagement activities in the Kilimanjaro Region. The success included compliance with physical distancing, hand washing, and the wearing of face masks in some settings. The health education through radio shows might have contributed to this success. The radio programs contributed to addressing misinformation and ensuring the public have accurate information and counterarguments against false information [ormation ,9. The cormation . LocallyMass public health educational campaigns are essential to inform and update the public about COVID-19 pandemic and to address rumors and misinformation, which may hamper the efforts to fight against the virus. Local-level coordination of contextually appropriate strategies is likely to be the cornerstone of the national response. Coordination of regional and district/ council stakeholders such as the regional and district commissioners, local government health management teams, religious leaders, political leaders at all levels, healthcare workers, public health practitioners, the media, and non-governmental organizations, is critical to the effective implementation and ensuring compliance to preventive strategies. Coordinated efforts among stakeholders in the country are necessary to contain COVID-19 in our communities through context-specific prevention strategies. Such interventions should also focus on ensuring compliance with the national and international recommended preventive measures. Until the point of writing this paper, IPH-KCMUCo continued to provide health education to the public through the local radios. However, due to the evolving nature of this pandemic, it is clear that messaging needs to be dynamic and continuously updated in line with growing evidence and changing recommendations .


Exosomes are nano-sized biovesicles of endocytic origin physiologically released by nearly all cell types into surrounding body fluids. They carry cell-specific cargos of protein, lipids, and genetic materials and can be selectively taken up by neighboring or distant cells. Since the intrinsic properties of exosomes are strictly influenced by the state of the parental cell and by the cellular microenvironment, the analysis of exosome origin and content, and their cell-targeting specificity, make them attractive as possible diagnostic and prognostic biomarkers. While the possible role of exosomes as messengers and a regenerative tool in cardiovascular diseases (CVDs) is actively investigated, the evidence about their usefulness as biomarkers is still limited and incomplete. Further complications are due to the lack of consensus regarding the most appropriate approach for exosome isolation and characterization, both important issues for their effective clinical translation. As a consequence, in this review, we will discuss the few information currently accessible about the diagnostic/prognostic potential of exosomes in CVDs and on the methodologies available for exosome isolation, analysis, and characterization. Since their discovery in the early 1980s , it has While the potential of exosomes as biomarkers is becoming important in several clinical settings ,14,15,16The possible role of exosomes in cardiovascular diseases (CVDs), particularly in ischemic heart disease, as messengers or as a regenerative tool is actively investigated by several groups ,21,22,23In this review, we will critically discuss the methodologies currently available for exosome isolation and analysis of their cargo since they represent the most challenging issues for the clinical transition of exosomes, including in the cardiovascular field. In addition, we will present recent evidence about the role of exosomes in CVDs, focusing on their diagnostic and prognostic potential.Exosomes are generated by invagination of late endosomal membranes that results in the formation of intraluminal vesicles (ILVs) within multivesicular bodies (MVBs) . Upon exThe endosomal sorting complex required for transport (ESCRT) is a key mediator of MVB biogenesis. The ESCRT is a complex machinery composed of four separate proteins (0 through III) that works cooperatively to facilitate MVB formation, vesicle budding, and protein cargo sorting ,26. The It is likely that the mechanisms of exosomes biogenesis and release in the context of CVDs are similar to those above described. Nevertheless, as already said, exosome generation and release are highly sensitive to several factors and presumably also to cardiovascular system alteration. Clear examples of this aspect are provided by works on cultured cardiomyocytes showing an alteration of exosome generation and release upon glucose deprivation  or treatThe isolation of pure exosomes is a critical step to understand their physio-pathological role and for their clinical application in several diseases, including CVDs. However, exosome isolation is challenging and there are no currently reliable protocols able to isolate with absolute precision exosomes from other extracellular vesicles in a given biological specimen. Especially when working with biological fluids, the enrichment/purification of exosomes from a given sample is necessary to enhance the exosome signal over non-exosomal noise; however, each methodology currently used for exosome purification likely exerts distinct bias contingent on the sample used and on possible contaminants co-purified with the exosomes. Moreover, the method used for exosomes isolation highly impacts on the quality of exosomes isolated and thus on the downstream analyses. Hence, the implementation of the purification techniques is a fundamental prerequisite to obtain reliable data in exosome research. The most commonly used techniques for exosomes isolation, as well as their advantages and disadvantages, are summarized in g. Finally, the isolated exosomes are resuspended in the appropriate medium and used for downstream analyses. Despite the wide use of ultracentrifugation for exosome isolation, it requires costly instrumentation and is time-consuming. Moreover, ultracentrifugation is also not always applicable to clinical samples due to the required volume of starting material and it is not able to efficiently separate exosomes from other co-isolated biomolecules . In addition, repeated “washing” ultracentrifugation steps can reduce the amount of co-isolated contaminants but they can damage the vesicles and reduce their yield [Differential ultracentrifugation is currently the most commonly used method for exosomes isolation. It usually consists of a series of centrifugation cycles of different centrifugal force and duration to separate exosomes from other sample components . The difir yield ,63.Variants of ultracentrifugation also exist, such as density gradient ultracentrifugation that exploits differences in vesicle size and density through the creation of a discontinuous density gradient in a centrifuge tube with progressively decreased density from the bottom to the top . The samg, 4 °C for 90 min  is an additional size-based separation technique applied to exosome isolation that uses a stationary phase consisting of resin particles with known porous size.Larger particles are excluded from entering the pores and thus are eluted from the column earlier. Molecules and small particles are longer retained into the pores of the stationary phase and elute later. Similarly to density gradient centrifugation, SEC has been shown to allow reduction of contaminant proteins in the exosome population ,71,72,73Exosome membranes are known to contain large quantities of proteins and thus immune-affinity capture may be used to isolate them, exploiting the interactions between these proteins (antigens) and specific antibodies  Figure . Immuno-Precipitation methods represent an easy and fast approach for exosome isolation which is mostly exploited by commercial kits and is becoming largely employed with clinical samples. Precipitation-based methods work by altering the solubility of exosomes, thus allowing their isolation from the solution . For thiNew techniques are essential to meet the challenge of providing high-purity exosomes for clinical practice. Recently, microfluidics-based technologies have become important techniques for the microscale isolation and detection of exosomes. This technique exploits both physical and biochemical properties of exosomes together with innovative sorting mechanisms, such as acoustic, electrophoretic, and electromagnetic manipulations ,81. The Other devices use the microfluidics-based membrane filtration approach isolating exosomes by their size. The first example of such device is a nanoporous membrane with an adjustable pore size that has been inserted in a microfluidic chip, and the authors developed two kinds of devices exploiting pressure- or electrophoresis-driven filtration to separate EVs . In addiThe accurate determination of exosome characteristics is required to determine their biological meaning but also their feasibility as a potential biomarker in the clinical routine. So far, several techniques were employed for exosome characterization but, as for isolation methodologies, their use is still under debate and each of these techniques has its own limitations that must be taken into consideration. At the present, the potential use of exosome as a biomarker of diseases focused on exploring their concentration and specific protein/miRNA cargo ,5,7,8, aNanoparticle tracking analysis (NTA), first applied to exosome characterization in 2011, is a light-scattering technique that allows sizing and enumeration of nanoparticles in liquid suspension . NTA, usDynamic light scattering (DLS) is an alternative technique for measuring the dimension of exosomes. In this technique, a monochromatic and coherent laser beam passes through a suspension of particles and, when it encounters a particle, the laser light is scattered in all directions . ParticlTunable resistive pulse sensing (TRPS) measures physical properties of exosomes, such as absolute size, concentration, and surface charge  ,98. ThesTransmission electron microscopy (TEM) is currently the most used imaging methodology to gain information about exosome morphological structure. The working principle of TEM is the creation of images generated from a beam of electrons passing through a sample, where a secondary electron is generated. These electrons are collected and magnified using special lenses. The great advantage of TEM is its high resolution power that allows to discriminate vesicles from other similar-size non vesicles. Nevertheless, sample preparation for TEM requires dehydration, chemical fixation, or staining of samples that may induce change in the morphology of the exosomes. Moreover, in some cases, the electron beam may also damage biological samples. Through TEM it is possible to label specific exosome antigens using immunogold staining, and TEM images can also be used to determine particle size.Cryo-electron microscopy (cryo-EM) is a variant of electron microscopy where the samples are analyzed at cryogenic temperature. This EM technique presents major advantages over conventional TEM and is considered the best method for visualizing nanoparticles without dehydration and fixation artifacts . MoreoveAtomic force microscopy (AFM) represents a unique alternative to electron diffraction techniques for studying exosomes . By deteOver the last two decades, flow cytometry has been regarded as one of the most commonly used techniques for EVs analysis . AlthougRaman spectroscopy is a vibrational technique based on Raman scattering that is becoming popular in many areas, including biology and medicine. In this methodology, the sample interacts with incoming monochromatic laser light and the photons change their energy by exciting vibrational modes of molecules. This vibrational technique measures the inelastically scattered photons, resulting in a Raman spectrum that corresponds with known vibrations of specific chemical groups . This meRecently, NanoView Bioscience has developed the ExoView R100 platform which is an affinity-based technology specifically dedicated to exosome cargo analysis. Briefly, exosomes are immobilized on ExoView chips by affinity capture usually against exosomal transmembrane proteins. Once captured, exosomal cargo can be studied permeabilizing the sample with the ExoView Cargo Kit to enable access of antibodies. This platform allows specific populations of exosomes to bind in a multiplexed manner to a functional chip and to measure both internal and external proteins, as well as to perform co-localization studies on single exosome. In addition, exosome subpopulations can be quantified in terms of vesicle size and number. The sample volume required is small and it must be underlined that purification biases are reduced, since the sample can be directly read without the preliminary step of vesicle isolations.Most of the current available works characterize exosome by widely used approaches. Specifically, the protein content is usually quantified by common assays  and the expression of specific proteins and/or exosomal markers is usually investigated by Western blot analysis or by enzyme-linked immunosorbent assay (ELISA). Finally, omics approaches, including proteomic analysis as recently described by Mallia and colleagues , are freDespite the progress in prevention and treatment of CVDs, this pathology still represents the most common cause of death and comorbidity in industrialized countries. Thus, further exploration of the pathophysiology of CVDs is important to provide new targets of disease, as well as novel biomarkers and/or therapeutic approaches. Increasing evidence in the field of CDVs strongly suggests a role for exosome-mediated intercellular communication in maintaining the homeostasis of the cardiovascular system, thus representing a potential therapeutic target ,116,117.In CVDs context, growing evidence suggests a role for these nanovesicles in cardioprotection and, thus, their potential use as a therapeutic tool. In myocardial infarction, reperfusion of the occluded coronary vessel leads to (ischemia-)reperfusion injury (I/R injury) and reduction of this detrimental effect is crucial to improve the survival of AMI patients. Interestingly, several studies demonstrated that the effect of remote ischemic conditioning (RIC), a strategy to protect the heart against I/R injury, is strongly related to the recruitment of circulating exosomes ,119,120.In the last years, the scientific interest toward exosomes is enormously increased for their possible implication in clinical application. The number, origin, and cargo of circulating exosomes differ under pathological conditions, suggesting their potential as a biomarker of disease. Although evidence about the importance of exosomes in cardiovascular disease is continuously expanding, their clinical application as markers for a specific disease is still strongly limited especially by the technological difficulties. Indeed, most of the current studies are performed in vitro and/or in animal models, whereas works about exosomes characteristics in cardiovascular patients are still scarce.Coronary artery disease (CAD) is the most common type of heart disease and it represents the leading cause of global mortality. Despite advances in treatment and prevention strategies, patients with established CAD still face a high residual cardiovascular risk, demonstrating the great need of novel, reliable tools for risk stratification to guide secondary preventive therapies. The principal cause of CAD is atherosclerosis, a chronic pathological condition characterized by the accumulation of lipids and fibrous elements in the large arteries. The mechanism underlying atherosclerosis is complex and involves multifactorial causes among which oxidative stress, endothelial dysfunction, and inflammation are considered the most important. Exosomes may contribute to atherosclerosis development and progression by means of their role as intercellular messengers. For instance, exosomes released by endothelial cells (ECs), upon activation of CD137 inflammatory signaling, promote plaque formation by inducing proliferative and migratory phenotype in vascular smooth muscle cells (VSMCs) as well as intimal hyperplasia after arterial injury . ExosomeAs above mentioned, only few studies explore the potential diagnostic implication of exosomes in CAD and in its acute clinical manifestation, the acute coronary syndromes (ACS). All these works are mainly focused on the identification of specific miRNA or lncRNAs in exosomes, that have been demonstrated to play a critical role in cardiovascular development and disease and that may be potential markers of CAD ,131. IndAt this regards, Wang and colleagues analyzed the exosomal expression of miRNAs previously related to the pathogenesis of CAD and they detected a higher expression of miR-30e and miR-2a in plasma exosomes of patients with coronary atherosclerosis compared to healthy controls . SimilarSeveral exosomal miRNAs have been also related to ACS. In particular, miRNA-208a expression is significantly higher in the serum exosomes of ACS patients compared to healthy controls, and ACS patients with higher expression of miRNA-208a have a reduced survival rate during one-year follow-up. Interestingly, a positive association between serum exosomal miRNA-208a expression and Killip class, troponin peak, and LDL has been found in the ACS group . This miAnother important aspect is that hypoxia has been demonstrated to alter exosome cargo. Indeed, cardiomyocytes under hypoxic conditions release exosomes carrying the cytokine TNFα  and the This in vitro evidence paves the way to the use of exosome-associated proteins (and miRNAs) as “pre-necrotic” biomarker(s), a signal actively released by the injured heart before necrosis begins.Many CADs progress up to chronic heart failure (HF), and the number of patients developing HF is increasing each year. Despite the significant advances in therapies and prevention, mortality and morbidity in patients suffering from HF are still high and quality of life is poor . HF is aRecently, Matsumoto and colleagues suggested that exosomes-bound miRNAs can be predictors of ischemic HF in post-AMI patients . They shExosomes content may be used as a valuable biomarker also in cerebrovascular diseases. For instance, in patients with acute ischemic stroke, serum concentration of exosomes, as well as exosomal levels of brain-specific miRNAs (miR-9 and miR-124), increase compared to controls. Moreover, the level of miR-9 and miR-124 positively correlate with scores of the National Institutes of Health Stroke Scale (NIHSS), the infarct volumes and the IL-6 serum levels , suggestThe role played by exosomes in other CVDs, such as cardiac arrhythmia, valve diseases, cardiomyopathies, is poorly understood, and available data are very limited. However, there are recent in vitro and animal studies suggesting a potential role of exosomes in the pathophysiology of the above-mentioned diseases. Cardiomyocytes derived/isolated from CAD patients or from the ischemic rat model release exosomes rich in miR-1 and miR-133 ,136, twoIn regard to valve diseases, the evidence of a possible role of exosomes is further limited. Mitral valve prolapse (MVP) is a common valvular heart disease that can lead to mitral regurgitation and subsequent congestive heart failure. Naturally occurring myxomatous mitral valve disease (MMVD) in dogs closely resembles MVP in humans and it has been associated with increased levels of plasma-associated exosomal cfa-miR-9, cfa-miR-181c, cfa-miR-495, and cfa-miR-599. Interestingly, in this canine model of disease, the expression levels of these miRNA change with disease progression and development of heart failure, but also exhibit changes as a function of age .In conclusion, more studies are needed to examine the role of exosomes as mediators of these CVDs and further research towards these directions is needed to broaden the application range of exosomal biomarkers.Research on the biology, pathophysiological function, and potential clinical application of exosomes has increased exponentially over the past years and they have emerged as potential biomarker reservoirs for several pathological processes. In this review, we decided to focus mainly on the potential application of exosomes as a diagnostic/prognostic tool in CVDs and, as consequence, we discuss the pros and cons of methodologies actually used in exosome research. Exosomes provide several advantages as a potential biomarker: (1) they are extremely specific and sensitive since they carry biological cargo that reflect the pathophysiological condition of the parental cells; (2) they can be isolated from easily obtainable biofluids such as blood and urine ; (3) theDespite the promising potential of exosomes both as therapeutic and diagnostic tool in CVDs, the challenge to their real clinical translation is represented by technical limitations. The isolation and characterization and they should be standardized and simplified to ensure their feasibility in daily clinical practice. Moreover, these techniques should be suitable to the small volumes that can be obtained from patients in the clinical context. Furthermore, the isolation of exosomes without contaminating plasma/serum proteins and lipoproteins is crucial for their downstream molecular analysis and reproducibility. Along with isolation/characterization methodologies, also nomenclature should be standardized. Despite the recent guidelines of the International Society for Extracellular Vesicles (ISEV) , a lot o


Neurological diseases severely affect the quality of life of patients. Although existing treatments including rehabilitative therapy aim to facilitate the recovery of motor function, achieving complete recovery remains a challenge. In recent years, regenerative therapy has been considered as a potential candidate that could yield complete functional recovery. However, to achieve desirable results, integration of transplanted cells into neural networks and generation of appropriate microenvironments are essential. Furthermore, considering the nascent state of research in this area, we must understand certain aspects about regenerative therapy, including specific effects, nature of interaction when administered in combination with rehabilitative therapy (regenerative rehabilitation), and optimal conditions. Herein, we review the current status of research in the field of regenerative therapy, discuss the findings that could hold the key to resolving the challenges associated with regenerative rehabilitation, and outline the challenges to be addressed with future studies. The current state of research emphasizes the importance of determining the independent effect of regenerative and rehabilitative therapies before exploring their combined effects. Furthermore, the current review highlights the progression in the treatment perspective from a state of compensation of lost function to that of a possibility of complete functional recovery. Neurological diseases are major sources of burden to both patients and caregivers. Furthermore, the associated financial burden is immense, with major countries such as the U.S. spending over $800 billion each year on related expenses . Among nIn recent years, steady progress in the clinical applications of regenerative medicine has occurred for a variety of diseases. Cell transplantation has emerged as a promising treatment for stroke recovery. Integration of these cell into functional neural networks and generation of microenvironments in which they can work effectively are necessary to obtain desirable results. Furthermore, it is necessary to retrain the neural systems that were lost. Rehabilitative interventions are expected to play an important role in this process, which was been discussed in the Stem Cell Therapies as an Emerging Paradigm in Stroke (STEPS) guidelines ,5. HowevIn the present review, we (i) discuss the current status of research on cell therapy for stroke recovery and challenges therein; (ii) outline the findings from the field of regenerative rehabilitation that could hold the key to unraveling these challenges; (iii) present the findings of past animal studies, evaluating the effects of approaches combining cell therapy and rehabilitation interventions; (iv) summarize the findings of pre-clinical research using brain stimulation, a method expected to yield drastic results when combined with cell therapy in the future; and (v) discuss future expectations and challenges faced in treating stroke with a combination of cell therapy and rehabilitation.Cell transplantation therapy is a promising and appealing treatment method for stroke recovery . IntrapaA study of the optimal time window for therapeutic intervention after brain injury by Peron et al. showed that transplantation of cells into the brain one week after injury is effective . MeanwhiIn addition to the transplantation site, the source of the transplanted cells is also important for the reconstruction of normal neural networks. When E14 embryonic motor cortical tissue was transplanted into the injury sites of mice with motor cortex injury, the tissue integrated successfully into the host motor cortex and was able to develop fibers projecting towards the sensorimotor cortex, the same as those seen in a healthy mouse . ImportaStudies on cell transplantation methods utilizing tissue engineering strategies are also making great strides. Because age is strongly involved in predicting the functional prognosis of patients with stroke, the ability to recreate microenvironments similar to those of young individuals holds the key to supporting the functional recovery . Tissue The above-mentioned findings demonstrate the continuous advances in current research and support the application of cell therapy as a promising treatment for stroke recovery. Establishing cells to be transplanted and the transplantation method, as well as further clarification of the mechanism of action of cell therapy, remain to be clarified for clinical application of cell therapy. Additionally, rehabilitation intervention post-stroke, discussed in further detail in subsequent sections, is one factor leading to uncontrollable outcome variation in clinical trials ,5. It isAlthough expectations are high for cell therapy to be a completely new treatment method for many diseases, such methods unfortunately cannot be called universal at present. Cell therapy generally requires cell engraftment, survival, and differentiation into matured cells. Additionally, the transplanted cells need to reconstruct tissue form and must further be functionalized as a tissue or organ. However, there is limited research on post-transplantation therapy , and the lack of information on how to support engrafting, survival, differentiation, and functionalization after transplantation remains a challenge.The concept of regenerative rehabilitation was developed to solve this problem. Regenerative rehabilitation can be briefly defined as “the application of rehabilitation protocols and principles together with regenerative medicine therapeutics toward the goal of optimizing functional recovery through tissue regeneration, remodeling, or repair” . The conResearch on regenerative rehabilitation for stroke recovery is still in its infancy, and there is limited evidence of its effects. The therapeutic interaction of cell therapy and rehabilitation, not specifically for stroke, can be generally grouped into five categories: synergistic, additive, subadditive, antagonistic, and overlapping . A synerSeveral studies examined the effects of combining intravenous cell transplantation with rehabilitation intervention. Zhang et al. studied the therapeutic effects, particularly the neuroprotective effects, of a combination of MSC transplantation and treadmill exercise in MCAO model rats . The resIntravenous transplantation of MSCs or ADSCs has the potential to synergistically improve motor function when combined with rehabilitation intervention. Research suggests the involvement of various mechanisms such as cytoprotection, increase in synaptic plasticity, and angiogenesis in this improvement of motor function, although there remains some discrepancies. Multiple studies have reported the effects of combining local cell transplantation into the cerebral parenchyma with treadmill exercise or EE interventions.A study by Seo et al. investigated the effects of combining treatment with transplantation of human adipose stem cells (hASCs) to the striatum and EE in hypoxic-ischemic brain injury model mice . Their sHicks et al. published three papers examining the effects of combination therapy with EE using a variety of animal models of cerebral infarction and cells intended for transplantation ,36,37. IVerifying the effects of combining two therapies is only possible if the therapeutic effects of each of them are independently assured. We must continue to optimize the cell transplantation therapy method and the rehabilitation intervention method.The above experiments produced different results, perhaps because of the fact that intraparenchymal cell transplantation is technically more difficult than intravenous transplantation. Other reasons could be the higher number of examined variables, as well as the use of different cells, which could be one of the cause of variability in outcomes. Before exploration of the effects of combination with rehabilitation can advance, it is essential to further standardize cell transplantation protocol.Treatment with intraparenchymal cell transplantation therapy alone had limited effects on motor function recovery and required the addition of rehabilitation interventions ,40. On tBrain stimulation has been studied as a method of neurorehabilitation for regulating and enhancing brain function . Many stMany animal experiments revealed that CS enhances the neural reorganization effects induced by rehabilitation training. In rat models of cerebral infarction, CS increased motor cortical evoked potentials in the stimulated region  and densRepetitive transcranial magnetic stimulation (rTMS) is a stimulation method in which a sequence of magnetic stimulation is repeatedly administered to a region of the cortex. It has been demonstrated that this method is able to temporarily regulate the excitability of the stimulated cortex, and can thereby modulate neuroplasticity of the cerebral cortex . ConventRecent in vitro and in vivo research has begun to clarify details of synaptic and cellular changes induced by rTMS ,66,67,68Unlike clinical studies, most of the studies using animal models of stroke begin with the evaluation of the effects of rTMS in the acute phase. Below, we outline the effects of rTMS in animal models of stroke and the related mechanisms of action.+-calmodulin-dependent protein kinase II (CaMKII)–cAMP-response element binding protein (CREB) signaling pathway, thereby promoting BDNF expression and synaptic plasticity [Most studies report the effects of rTMS in the acute phase, which begins within 24 hours after stroke. Ipsilesional high-frequency (20 Hz) rTMS that was started one hour after cerebral infarction reduced apoptosis and infarct volume and improved the neurological outcome . Similarasticity .The rTMS interventions resulted in beneficial effects, even when started 1 to 7 days after the onset of stroke. Administering high-frequency (10 Hz) rTMS 4 to 18 days after MCAO reduced apoptosis and improved motor function . IpsilesInterestingly, administering rTMS before transient ischemic attack has been found to induce ischemic tolerance, thus preventing neuronal cell death in the hippocampus . This stThese studies demonstrate the extremely wide range of effects of rTMS in animal models of stroke, including motor function recovery, reduction of infarct volume, mitigation of cerebral edema, anti-apoptosis, elevation of neurotrophic factors, facilitation of neurogenesis, promotion of proliferation and migration of neural precursor cells, and furtherance of neuronal differentiation and inhibition of glial differentiation.Transcranial direct current stimulation (tDCS) is a noninvasive method used to stimulate the brain activity and modify the brain behavior by applying a weak direct current from outside the skull. Because the necessary equipment is highly safe and portable, tDCS has been actively studied in numerous conditions, including stroke, epilepsy, Alzheimer’s disease, and Parkinson’s disease . While tSimilar to studies on rTMS, several studies verified the effects of tDCS intervention using animal models of stroke at the beginning of the acute phase, unlike clinical studies. In the following section, we will outline the effects of tDCS in animal models of stroke and related mechanisms of action.Cathodal tDCS administered immediately after MCAO reduced infarct volume . InfarctStudies have also found that tDCS initiated 1 to 7 days after the onset of stroke is useful. One study examining the effects of ipsilesional anodal tDCS initiated 1 or 7 days after infarction in MCAO model rats found motor function improvement in both starting conditions . ExpressSimilarly to rTMS, these results confirm the wide range of effects of tDCS in animal models of stroke, including improvement of motor function, reduction of infarct volume, mitigation of cerebral edema, anti-apoptotic and anti-inflammatory effects, increase in dendritic spine density, neurogenesis, and facilitation of stem and progenitor cell migration.As seen in the above examples, most studies found that the rTMS and tDCS groups showed a better post-stroke tissue preservation and functional improvement than did the control or sham stimulation groups. However, research methods used for animal experiments and clinical trials differ in several important ways. In most animal experiments, only stimulation of the lesioned hemisphere is studied, and most studies on rTMS used ipsilesional high-frequency stimulation. However, clinical studies have demonstrated the effectiveness of low-frequency stimulation of the contralesional hemisphere as well . It willAs noted above, brain stimulation not only reduces infarct volume, prevents apoptosis, increases neurotrophic factor levels, and improves synaptic plasticity, but also facilitates intrinsic neurogenesis and cell migration to the site of injury. Crossing these characteristics with those of cell transplantation therapy is expected to yield a synergistic effect . Althoug5 rat femur-derived MSCs to the left corpus callosum  one day after infarction in right-MCAO model rats [5 human dental pulp stem cells into the striatum and cortex of normal rats and performed low-frequency TMS on the hemisphere ipsilateral to the transplantation site every second day for two weeks, beginning two days after transplantation [5 hNSCs into the striatum four days after infarction in MCAO model rats [A study by Morimoto et al. investigated the effects of combining CS and transplantation of 2.5 × 10del rats . A cathodel rats . Kremer antation . TMS wasdel rats . The rTMThe above-mentioned studies demonstrate that combining brain stimulation with cell transplantation can facilitate synergistic motor function recovery through the promotion of neuronal differentiation and migration of transplanted cells to the target area. As improper stimulation conditions can lead to negative effects, it will be essential to optimize stimulation parameters and consider factors such as injury site, transplanted cell type, and the timing and conditions of stimulation. Lastly, it may be possible to enhance the effects of cell therapy further by combining this technique with brain stimulation and intensive exercise therapy, which have already shown clinical results . This wiThis paper discussed the effects of combining cell therapy and rehabilitation for stroke recovery. Many studies report findings that suggest synergistic effects and promising clinical applications of these therapeutic strategies, but it is necessary to accumulate further evidence in this regard. Furthermore, the methods of brain stimulation introduced here have an extremely wide range of effects for stroke treatment, and evidence suggesting the beneficial effects of their combination with cell therapy is gaining momentum. The impact of such evidence could drastically change the very concept of therapy for stroke recovery. In the recent past, it was thought that the central nervous system could not regenerate, and the focus of rehabilitation was on the ability to somehow compensate for the loss function using the function that remained. In addition to advances in neuroscience, new findings have been integrated into rehabilitation to give rise to the concept of neurorehabilitation. Methods such as constraint-induced movement therapy and task-oriented training, which attempt to facilitate changes in synaptic plasticity based on Hebb’s rule and activate reorganization of brain mapping, have been developed. With the recent advent of cell transplantation therapy, facilitating a marked suppression of inflammation, protection of cells and promotion of functional compensation have been achieved. The next goal is to achieve true regeneration of lost function based on these methods. To do so, discovering a method to induce the reconstruction of neural circuits from transplanted cells and endogenous neural stem cells will be the key. One particular challenge will be the distance required to be travelled by transplanted cells for the reconstruction of the corticospinal tract. In studies of natural stroke recovery, there is an association between better outcome and less corticospinal tract injury ,94. In aCell therapy and rehabilitation share much of the same extremely wide range of underlying mechanisms of action . HoweverThe findings discussed in this paper demonstrate that interdisciplinary collaboration centering on regenerative medicine, rehabilitation medicine, and engineering is essential for regenerative rehabilitation research. Consequently, connecting these fields is a topic that must be addressed in the future .Lastly, we should mention the translational issues of combining cell therapy and rehabilitation for stroke recovery. Pre-clinical research should take into account many potential factors that influence experimental stroke therapies, such as age, sex, and comorbidities ,5. Rehab


The hydrogen production reaction of the proton exchange membrane (PEM) water electrolysis cell stack is the reverse reaction of the fuel cell, but the water electrolysis operation requires high pressure, and the high pressure decomposes hydrogen molecules, thus aging or causing failure in the water electrolysis cell stack. In addition, there are five important physical parameters  inside the water electrolysis cell stack, which can change the performance and shorten the life of the cell stack. However, the present techniques obtain data only by external simulation or single measurement; they cannot collect the internal real data in operation instantly and accurately. This study discusses the causes for aging or failure, and develops an internal real-time microscopic diagnosis tool for accelerated aging of the PEM water electrolysis cell stack. A flexible integrated  microsensor applicable to the inside  of the PEM water electrolysis cell stack is developed by using micro-electro-mechanical systems (MEMS) technology; it is embedded in the PEM water electrolysis cell stack for microscopic diagnosis of accelerated aging, and 100-h durability and reliability tests are performed. The distribution of important physical parameters inside the PEM water electrolysis cell stack can be measured instantly and accurately, so as to adjust it to the optimal operating conditions, and the local aging and failure problems are discussed. Therefore, renewable energy has become the key point of current research, including hydrogen energy, solar energy, hydraulic power, wind power, geothermal, tide and biomass energy. Hydrogen is one of the optimal sustainable energy carriers, and hydrogen is a zero carbon energy carrier; it is applicable to low-carbon transportation and industrial decarbonization. Generally, burning fossil fuels to produce hydrogen will generate a lot of CO2, so carbon-free hydrogen production is the present research objective. Electrolytic technology can transform low-carbon electricity, which is unlikely to be stored and transmitted into hydrogen. It is friendlier to the environment than traditional hydrogen production methods . In addition, the intermittent power problems in wind energy and solar energy can be solved, and hydrogen is generated continuously, so as to relieve the outage problem. Renewable energy and electrolytic technology have major significance for promoting the high quality development of the hydrogen energy industry, promoting mass renewable energy consumption and speeding up energy structure transformation [With the progress of human civilization, as energy becomes more and more important, the demand increases greatly, and people use fossil fuels, such as petroleum, natural gas and coal, to meet the huge demand, so that a lot of COormation . At presormation ,4,5,6,7.ormation . The chaormation ,16,17,18ormation . The efformation . Hydrogeormation . The proormation . Most peormation . Howeverormation ,16,17,18ormation . HoweverThe micro current, voltage, flow, pressure and temperature sensors are integrated into a 5-in-1 microsensor, as shown in The sensing principle of micro temperature sensor is that when the ambient temperature rises, as Au has a Positive Temperature Coefficient (PTC), the resistivity increases with temperature; this characteristic results from the “temperature coefficient of resistance” (TCR) of the conductor.The electrode form of the capacitive micro humidity sensor used in this study is the interdigitated electrode structure. There is a humidity sensitive thin film above the electrode. When the water vapor absorbed by the humidity sensitive thin film increases, the dielectric constant increases with ambient humidity; the humidity can be obtained by calculating the changed capacitance value.The current, voltage, flow, pressure and temperature sensing structures are integrated by using MEMS technology in this study. The process of the flexible integrated microsensor equipment is shown in (a)PI thin film cleaningThe PI (Polyimide) thin film substrate is cleaned with organic solvent ethanol, and then it is put in the hot organic solvent acetone, and the acetone is volatilized.(b)Evaporate metalThe metal is evaporated by electronic beam evaporator as shown in (c)PhotolithographyThe positive photoresist is coated on the sample uniformly by spin coater, as shown in (d)Metal etching® P4620, Merck Co., Darmstadt, Germany) can form a wet etching mask after the photolithography process; the unwanted Au/Cr is corroded by Type-TFA Au etching solution which is manufactured by Transene Electronic Chemicals , and by Cr-7T Cr etching solutions, manufactured by OM Group Incorporated , and then the photoresist is removed by acetone.The photoresistor (AZ(e)~(i)Dielectric layer definition, evaporate metal, metal etching and protection layer definitionThe pattern of the dielectric layer is defined through the aforesaid photolithography process, the Cr/Au metal is evaporated again, and the unwanted metal is removed by wet etching process. Finally, the LTC9320  with high mechanical strength and fit for high chemical environment is used as a protection layer, so as to avoid the flexible integrated microsensor being destroyed by the closing pressure of end plate inside the PEM water electrolysis cell stack. Afterwards, the sensing area of micro voltage and current sensors is defined by using the photolithography process. The stereogram of the flexible integrated microsensor is shown in After the sensing structure is successfully integrated into the PI film, the temperature and flow of flexible integrated microsensor are corrected, and the reliability is validated. The flexible integrated microsensor is connected to NI PXI 2575 data capture equipment of National Instruments  to capture the microsensor data instantly, as well as the resistance and current of micro temperature and flow sensors. The system is measured and controlled by LabVIEW system design software, and the signals are processed and analyzed and exported to the computer. The correction curves of temperature and flow are drawn, respectively, as shown in In order to observe the local physical change inside the PEM water electrolysis cell stack, the flexible integrated microsensor is embedded above the runner plate of PEM water electrolysis cell stack without influencing the performance of the PEM water electrolysis cell stack . FinallyThe performance of the PEM water electrolysis cell stack is tested at different temperatures , as shown in 2 after about 40 min, and then the reaction is stabilized. The performance begins to decline at about 55 h, meaning the MEA (Membrane Electrode Assembly) and runner plate of PEM water electrolysis cell stack have aged gradually, the external glass tube with Deionized water (DI) water has been blackened, and the dissolved graphite has been discharged from the outlet, as shown in After the 100-h accelerated aging test for the PEM water electrolysis cell stack, the PEM water electrolysis cell stack is disassembled to ascertain the internal condition, as shown in This study has successfully developed a flexible integrated microsensor by using MEMS technology. This flexible integrated microsensor has five sensing functions, it is resistant to high-temperature electrochemical environment, it can perform real-time measurement, and it can be placed in any position inside the PEM water electrolysis cell stack.The flexible integrated microsensor was successfully embedded in the PEM water electrolysis cell stack, and the local important information inside the PEM water electrolysis cell stack was extracted successfully without influencing the operation. A 100-h accelerated aging test was performed. The test result showed that the key factors influencing the PEM water electrolysis cell stack were temperature and voltage. As the PEM water electrolysis cell stack aged internally, the nonuniform distribution of temperature and flow resulted in large temperature difference, lower current density, graphitization of the internal runner plate and carbon paper pyrolysis. The most severe condition occurred between the downstream inlet and the midstream.


In recent years, knowledge on the biology and pathobiology of extracellular vesicles (EVs) has exploded. EVs are submicron membrane-bound structures secreted from different cell types containing a wide variety of bioactive molecules  and mitochondrial DNA). EVs have important functions in cell-to-cell communication and are found in a wide variety of tissues and body fluids. Better delineation of EV structures and advances in the isolation and characterization of their cargo have allowed the diagnostic and therapeutic implications of these particles to be explored. In the field of liver diseases, EVs are emerging as key players in the pathogenesis of both nonalcoholic liver disease (NAFLD) and alcoholic liver disease (ALD), the most prevalent liver diseases worldwide, and their complications, including development of hepatocellular carcinoma. In these diseases, stressed/damaged hepatocytes release large quantities of EVs that contribute to the occurrence of inflammation, fibrogenesis, and angiogenesis, which are key pathobiological processes in liver disease progression. Moreover, the specific molecular signatures of released EVs in biofluids have allowed EVs to be considered as promising candidates to serve as disease biomarkers. Additionally, different experimental studies have shown that EVs may have potential for therapeutic use as a liver-specific delivery method of different agents, taking advantage of their hepatocellular uptake through interactions with specific receptors. In this review, we focused on the most recent findings concerning the role of EVs as new structures mediating autocrine and paracrine intercellular communication in both ALD and NAFLD, as well as their potential use as biomarkers of disease severity and progression. Emerging therapeutic applications of EVs in these liver diseases were also examined, along with the potential for successful transition from bench to clinic. Knowledge of the pathobiology of extracellular vesicles (EVs) has expanded significantly in the last decade ,2. IndeeNovel insights into the biology of EVs show that these particles regulate critical biological functions and may act as contributors to disease pathogenesis, and may also serve as disease biomarkers, with the virtue of the relative simplicity of EV isolation from different biofluids . In addiIn the field of hepatology, EVs have recently emerged as novel players in the pathogenesis and progression of several conditions ,12,13, iDetails on the formation and secretion pathways of EVs have been recently reviewed elsewhere ,20,21, aIn general, EVs are classified according to size and biogenetic pathway, such as exosomes, microvesicles, and apoptotic bodies . Exosomehttps://www.isev.org/) that is constantly updating existent information on technical issues in order to homogenize the data being generated in different laboratories. To that end, the ISEV released a document entitled Minimal Information for Studies of Extracellular Vesicles (“MISEV”) Guidelines, to help researchers to better isolate and characterize their EV preparations, as well as suggesting protocols to document specific EV-associated functional activities [The impact of EV research is highlighted by the existence of a dedicated scientific society , microRNA (miRNA), ribosomal RNA (rRNA), transfer RNA (tRNA), noncoding RNAs (ncRNAs), DNA, mitochondrial DNA (mtDNA), and metabolites ,30. LipiIn the liver, both parenchymal (hepatocytes) and non-parenchymal cells  have been found to release EVs in both physiological and pathological states . HoweverHepatocellular damage determines the release of a number of signals into the extracellular environment that can contribute to tissue inflammation . Some ofCollectively, these findings suggest that EVs released from injured hepatocytes have an important role in modulating the inflammatory response during liver damage through intercellular communication between different cell types, with potential contributions from other cell-derived EVs . SpecifiPersistent liver fibrogenesis and the development of cirrhosis are responsible for the liver-related morbidity and mortality associated with chronic liver diseases . ActivatDynamic changes of EV generation in pathological conditions and the accessibility to measure and analyze them in biological samples  make EVs good candidates to be used as disease biomarkers. The relative accessibility of testing and the ability to perform repeated measurements over time could facilitate early diagnosis, disease monitoring, and the development of personalized medicine. Furthermore, refinement of techniques allowing isolation and in-depth characterization of EV cargoes could lead to identification of disease-specific molecular signatures or profiles, providing ample opportunities for EVs to be used as suitable, non-invasive biomarkers .EVs have been studied as potential biomarkers of liver injury in the settings of ALD, NAFLD, drug-induced liver disease, and cholangiopathies ,27,55,56From a therapeutic perspective, EVs, either unmodified or engineered, can be utilized for therapeutic purposes . With reRegarding the use of EVs as therapeutic agents, most of the available evidence has been generated using mesenchymal stem cell (MSC)-derived EVs obtained from human umbilical cords or human embryos that have been tested in numerous preclinical liver disease models  with promising results ,53. DataA third, and still nascent, approach related to EV-based therapy is based on the concept that interfering with EV secretion or uptake may attenuate harmful effects on target cells ,64. In tA number of studies have demonstrated a role of EVs in both the pathogenesis and progression of NAFLD ,15. TrigStudies carried out in mouse models of NASH have shown that total circulating EVs and particularly hepatocyte-derived EVs are elevated early in the disease process, while other cell-derived EVs  appear in the circulation later, likely reflecting the ongoing inflammatory process ,76. ProtTherapeutic efforts involving EVs in the field of NAFLD/NASH are nascent. Attempts to halt inflammation and fibrosis in rodent models of NAFLD/NASH using EVs as therapeutic agents have been published recently. EVs obtained from amnion-derived MSCs (AMSCs) were used to treat rats with either NASH or liver fibrosis induced by the hepatic toxicant CCL4. AMSC-EVs were given intravenously in one or two doses, and amelioration of inflammation and fibrogenesis was observed . More reRecent studies have focused on the role of EVs in ALD ,14,84,85From a clinical point of view, there is currently no biomarker able to assess the early stages of ALD. EVs have been shown to correlate with the diagnosis and prognosis of alcoholic hepatitis . AdditioThe field of EVs in fatty liver diseases is rapidly evolving as the important functions of these particles in cell-to-cell communication and in the pathogenesis of both ALD and NAFLD, the most prevalent liver diseases worldwide, are unveiled. The release of large quantities of EVs by stressed/damaged hepatocytes contributes to inflammation, fibrogenesis, and angiogenesis, fueling liver disease progression and provides opportunities for intervention. The identification of specific molecular signatures of released EVs is promising in the search for disease-specific biomarkers, although more data are needed to validate these markers in larger cohorts and in a rigorous manner. EVs may have potential for therapeutic use, but this field is still nascent. More research is needed for a successful transition of current EV knowledge from the bench to the clinic.