Glucosinolate-containing foods, such as vegetables from the plant order Brassicales and its derivative products, are valued for their health-beneficial properties. The latter are linked to glucosinolate hydrolysis products, such as isothiocyanates .The Special Issue “Advanced Research on Glucosinolates in Food Products” aimed to collect the latest research on the impact of the whole food supply chain, including production, as well as domestic food preparation, on glucosinolates and the formation and chemistry of their breakdown products in vegetables and further foods. In this context, the consequences for human health are important, too. This Special Issue includes eleven research articles that cover research on the effect of pre-harvest factors on glucosinolates, their hydrolyzing enzymes, and the formation of volatile hydrolysis products ,3,4,5. F2 levels [Diplotaxis tenuifolia and Eruca sativa), demonstrating this with multiple leaf cuts, showing how the process affected the perception of pepperiness, bitterness, and hotness, which in turn reduced consumer acceptance [Given that they are evolutionary plant defense compounds, glucosinolates and their hydrolysis in Brassicaceae vegetables are strongly affected by plant genotype and ecophysiological parameters. One of the studies described by Oloyede et al. revealed that cabbage grown in the field at lower temperatures exhibited higher myrosinase activity compared to greenhouse-grown plants, but the effect on glucosinolates and their hydrolysis products differed between cabbage morphotypes . In anot2 levels . In contceptance .Tropaeolum majus L.). Next to the thermal degradation of glucosinolates to nitriles, Krell et al. showed that isothiocyanates react with the lysine residues of the wheat proteins in the bread crumb [The processing of these vegetables during food preparation can considerably affect glucosinolates, their product formation, and also the flavor of the products. Thermal treatments inactivate myrosinase, thereby stopping enzymatic glucosinolate hydrolysis, resulting in diminished isothiocyanate formation . Stir-frad crumb . Anotherad crumb .Nasturtium officinale) isothiocyanates on exercise-induced inflammation are presented in a human intervention study described by Schulze et al. [When eating Brassicales plants or products thereof, glucosinolates, and especially their isothiocyanates, are valued for their positive effects on human health. However, conversion to isothiocyanates and bioavailability determines the effects of these compounds tremendously. Shekarri and Dekker described a physiologically based model to simulate the bioavailability and absorption kinetics of sulforaphane (4-(methylsulfinyl)butyl isothiocyanate, which is found in broccoli and also in red cabbage). This model represents a preliminary step in enabling the prediction of the biological effect of isothiocyanates and can be used in the growing field of personalized nutrition . Novel ie et al.  After the et al. .Due to the inclusion of well-designed studies and well-written articles, this Special Issue offers a valuable contribution to the ambition of producing vegetables rich in valuable glucosinolates as a source of health-promoting isothiocyanates. However, the research also reveals that this ambition might negatively affect consumer liking and acceptance, and therefore this needs to be considered in the future as well. New insights into the effect of food processing on glucosinolates and the formation and fate of their breakdown in follow-up products are presented. These processes are still far away from being fully understood. Evaluating the role of glucosinolates in supporting human health still remains challenging.


Congenital diaphragmatic hernia (CDH) is a severe defect with an estimated incidence of 1:3000 live births . This anAfter several studies in animal models, a new theory was presented to explain pulmonary hypoplasia in CDH. According to this theory, the initial lesion occurs in the early stages of fetal development and organogenesis, which promotes hypoplasia of both lungs and ipsilateral lung compression by herniation of the abdominal contents. This prevents the branching of bronchi, bronchioles, and pulmonary arteries and veins, resulting in hypoplasia of the pulmonary acini. The terminal bronchioles are reduced and the alveolar septa are thickened, which results in relatively immature lungs and vascular hypoplasia. This theory explains the variability of pulmonary hypoplasia in patients with CDH .In addition to the underdeveloped lungs, CDH also involves an abnormal development of pulmonary structures, mainly vessels and bronchi. This abnormal development increases vascular resistance and culminates in significant pulmonary hypertension, which is the main cause of morbimortality in children with CDH ,2.In patients with CDH, the total number of lung vascular beds, arteries, and veins per lung unit are reduced, and vascular reshaping and hyperplasia in the tunica media of small arteries may occur. The scarcity of arteries and veins and reshaping of the lung vessels contribute to the unresponsive pulmonary hypertension, which in turn leads to refractory shock and death. The explanation for these events is the reactivity of the altered vessels due to an imbalance of autonomous innervation, vasoconstrictor mediators, and vasodilators. After birth, a combination of right ventricular hypertrophy, left ventricular hypoplasia, and an extremely altered lung architecture results in severe pulmonary hypertension, which is often unresponsive to treatment -5.Electrical impedance tomography (EIT) is a new, noninvasive, radiation-free method that can be used to assess pulmonary physiology during mechanical ventilation. EIT is a bedside method that uses high-frequency and low-amplitude electrical currents to obtain cross section images of both lungs ,7.The development of EIT for use in newborns has emerged as a noninvasive method of continuous ventilation monitoring, because it generates functional images that allow continuous evaluation of pulmonary ventilation in different lung regions and lung perfusion in real time ,8. This The electrical potentials obtained on the chest are gauged and used to determine the distribution of intrathoracic electrical impedance, and images are created to represent the air distribution area in the lungs (corresponding to areas of greater resistance to the passage of the electric current). Thus, EIT explores the differences in electrical impedance generated by the replacement of tissue conductivity obtained from the range in the tidal volume of gases during breathing .versus 2%). The distribution pattern showed compensatory ventilation in the right lung. A slight difference was observed between gravity- and non-gravity-dependent areas (56% versus 44%). Plethysmography showed an inferior tidal volume distribution on the compromised side (A 3-hour old infant with a non-corrected left CDH was referred for EIT for the observation of pulmonary hypoplasia. As shown in sed side .


Transplantation of allogeneic hematopoietic cells faces two barriers: failure of engraftment due to a host versus graft reaction, and the attack of donor cells against the patient, the graft versus host (GVH) reaction. This reaction may lead to GVH disease (GVHD), but in patients transplanted due to leukemia or other malignant disorders, this may also convey the benefit of a graft versus leukemia (GVL) effect. The interplay of transplant conditioning with donor and host cells and the environment in the patient is complex. The microbiome, particularly in the intestinal tract, profoundly affects these interactions, directly and via soluble mediators, which also reach other host organs. The microenvironment is further altered by the modifying effect of malignant cells on marrow niches, favoring the propagation of the malignant cells. The development of stable mixed donor/host chimerism has the potential of GVHD prevention without necessarily increasing the risk of relapse. There has been remarkable progress with novel conditioning regimens and selective T-cell manipulation aimed at securing engraftment while preventing GVHD without ablating the GVL effect. Interventions to alter the microenvironment and change the composition of the microbiome and its metabolic products may modify graft/host interactions, thereby further reducing GVHD, while enhancing the GVL effect. The result should be improved transplant outcome. The Iliad)“… he first commanded Bellerophon to kill that savage monster, the Chimaera, who was not a human being, but a goddess, …”  at the time of transplantation, the transplant conditioning regimen, the source of donor cells, HLA mismatch between donor and patient, and the development of (chronic) GVHD. MRD, in particular, is currently an area of extensive research. The level of detection of MRD depends upon the methodology used  –14. WhilEarly data on global T-cell depletion of the donor cell inoculum before infusion into the patient showed substantial reduction of the incidence of GVHD but also resulted in a high rate of graft failure and disease relapse . More reOne strategy is the administration of post-transplant cyclophosphamide (CY), originally for HLA haplo-identical transplants but then extended to other donor/host combinations . The redin vitro depleted of CD45RA+ T cells achieved sustained engraftment, had a very low incidence of severe acute and chronic GVHD, and were not at a higher risk of relapse than patients transplanted with T cell-replete grafts , antation ? Severalantation –35. In oantation . These pantation . Howeverhe tumor . Ongoinghe tumor .GVL effect, these patients also experienced a recipient vs. leukemia effect, suggesting activation of the patient's immune system by the infused HLA non-identical donor cells -mobilized peripheral blood progenitor cells  at various doses, from HLA-mismatched donors in patients with AML in first remission who were not given any GVHD prophylaxis . Donor cor cells . The samnovo AML , but conin vitro data from patients with MDS, which show a two-way signaling path between the clonal disease cells and non-clonal mesenchymal/stroma cells . Several reports have postulated a “leukemogenic effect” of the marrow microenvironment , 47. Is Blautia, appear to have a beneficial effect, others, for example, Veillonella or enterococcal species such as Enterococcus faecium or Enterococcus faecalis, favor the development or propagation of GVHD, leading to inferior transplant survival  occurred without clinical evidence of GVHD, suggesting the possibility that the presence of mixed chimerism, albeit temporary, facilitated the establishment of tolerance.Viral organisms such as picobirna viruses have also been shown to participate in these donor/host interactions , and theHomo sapiens trying to break down barriers that have evolved over millions of years. Doing so upsets the balance that we observe in healthy individuals. Of course, these therapeutic interventions are directed at the eradication of a malignant disease, which has already changed the internal milieu. A better understanding of signals that trigger the development of malignant disorders such as leukemia would allow for earlier interventions and might permit their exploitation to restrict the reactions of donor cells to the GVL effect, while preventing GVHD. Can we direct the divine ability of the chimera against the malignancy and sever the ugly head of GVHD? Current research using state of the art tools, including systems biology and machine learning, may be able to pave the way.The interactions between donor and recipient cells following allogeneic HCT are complex, and the cast of characters of this drama is not limited to donor and recipient immune cells. Additional actors include cellular and non-cellular components of the microenvironment and, importantly, the microbiome. Nature had not envisioned The original contributions presented in the study are included in the article/supplementary material, further inquiries can be directed to the corresponding author/s.The author confirms being the sole contributor of this work and has approved it for publication.The author declares that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Career sustainability is a well-researched issue in academics and other sectors. Technology advancements and COVID-19 have jeopardized career sustainability. Numerous studies have explored the influence of individual characteristics on career sustainability, but few have focused on leadership. In addition, cultural factors must be considered because leadership is rooted in culture. In particular, inclusive leadership reflects traditional Chinese culture. Therefore, based on self-determination social exchange theories, we analyzed the effects of inclusive leadership on career sustainability as well as the roles of thriving at work and supervisor developmental feedback (SDF) in career sustainability. In total, 363 samples were collected from China. The results revealed that inclusive leadership improves career sustainability through SDF and thriving at work. Theoretically, our study fills the research gap and establishes a mechanism and theoretical framework for inclusive leadership and career sustainability. Practically, we offer guidance for enterprises to cultivate inclusive leadership and improve career sustainability. Sustainability has become a popular research topic , a popular and professional online survey company in China  scale developed by Fang et al.  containsThe Supervisor Developmental Feedback (SDF) scale developed by Zhou  was alsoIn addition, the thriving at work (TW) scale designed by Porath et al.  is a devThe career sustainability (CS) scale developed by Chin et al.  is relatResearch suggests that career sustainability is affected by personal factors such as gender, education, length of employment, and nature of work . The average variance extracted (AVE) values were higher than 0.5, ranging from 0.595 to 0.727 . Therefore, the convergent validity of these measures was satisfactory. The discriminant validity of the constructs was evaluated using the approach described by Fornell and Larcker  and the Henseler et al.  introducR2 value was 0.418, and the results support hypotheses H1, H3, and H4. The results suggest that inclusive leadership had a positive effect on career sustainability (H1). In addition, the mediating role of thriving at work in the relationship between inclusive leadership and career sustainability was verified (H2), but the mediating role of SDF was not verified. Furthermore, the relationship between inclusive leadership style and career sustainability was serial-mediated by SDF and thriving at work (H4). To test the hypotheses, the bootstrap resampling method in SmartPLS was used to evaluate the PLS results, and the responses were resampled 5,000 times (Hair et al., Most relevant studies on the antecedents of career sustainability are qualitative studies conducted from the perspective of individuals (Herman and Lewis, In addition, the current study expands applications for the theory of inclusiveness and sustainability in terms of organizational behavior. The relevant literature has revealed the crucial role of inclusiveness in sustainable economic growth, ecological sustainability, and sustainable urban development, but few studies have been examined organizational behavior. The current study innovatively introduces the concepts of inclusiveness and sustainability into organizational behavior, further confirming the inseparable relationship between inclusiveness and sustainability. The application of inclusiveness in organizational behavior is mainly reflected in leadership. The impact of inclusive leadership on employee creativity, performance, and turnover rate Fang,  has beenFurthermore, in terms of the cross-disciplinary aspects of management and psychology, we revealed a mediating role of SDF and thriving at work. Specifically, we found that the relationship between inclusive leadership and career sustainability was not separately mediated by SDF but rather serially mediated through thriving at work. This may be because in the long term, career sustainability is easily affected by external environments, and employees generally have a subjective plan for their careers; therefore, developmental feedback does not considerably influence career sustainability through cognition ways. However, SDF can construct an image of organizational support for employees to reference in numerous challenging future work scenarios (Blustein, First, enterprises should cultivate inclusive leadership. The impact of inclusion on sustainability has been widely recognized, and enterprises should recognize the positive role of the inclusive leadership style in developing career sustainability. Specifically, leaders should be receptive to employees and tolerate their mistakes with encouragement and support. In addition, leaders should respect and recognize their employees, focus on employee development, and praise employees for their achievements, rather than envy them. Moreover, leaders should consider the needs of employees, treat them fairly and justly, and share benefits with them.Second, developmental feedback should be effectively utilized. Feedback has been widely used in enterprises as an effective management tool. Constructing effective feedback has always been the focus of academic and practical discussions (Li et al., Third, enterprises should actively emphasize the psychological state of their employees. Human sustainability is the basis of career sustainability, and healthy psychological and physiological conditions are the basis of human sustainability. Numerous studies have identified a close relationship between the mental state and physical health (Spreitzer et al., First, cross-sectional data were adopted in the current study, and each questionnaire was completed by employees themselves; therefore, accurately and rigorously measuring the causal relationship between variables is difficult. Future studies should consider further reducing measurement errors by issuing questionnaires at different time points and from different sources or by enhancing the control variables during experimental research.Second, the sample population mainly consisted of employees from Chinese enterprises. In the future, the sample population can include individuals from various countries and industries. A cross-layer research method can also be adopted to improve the accuracy and external validity of the data.Third, the current study only explored one of the ways that inclusive leadership can have a positive influence on career sustainability. Other influences paths may have not been discovered.Finally, the differences in individual personality characteristics of employees may also affect the significance of the study. In the future, moderating variables can be added to explore the establishment boundary of the serial mediating effect.Research has revealed that inclusive leadership plays a crucial role in promoting the career sustainability of employees. We further investigated the serial mediating effect of SDF and thriving at work in terms of cross-level interactions between individual characteristics and organizational contexts. Therefore, the serial mediating effect helped us identify the key factors of career sustainability and improved our understanding of how enterprises can fully apply these factors to improve career sustainability. More importantly, the results may help researchers outside China conduct similar studies and explore the adaptability of the research in different cultural contexts. In general, sustainability has become the focus of global practice and academia. Individual sustainable development is the original source of sustainable development in society and among enterprises; therefore, leaders must understand how to achieve corporate sustainability by helping employees improve their career sustainability. Based on valuable empirical evidence, a feasible approach was identified, which can be applied by enterprises. Furthermore, this paper indicates how enterprises can more effectively fulfill their social responsibilities and echoes the common pursuit of humans for the sustainable development of society.The original contributions presented in the study are included in the article/supplementary material, further inquiries can be directed to the corresponding author/s.Y-CF conceived and designed the research and provided guidance throughout the entire research process. Y-HR wrote the main part of the manuscript. J-YC collected the data and written the methods section. TC and C-LL helped translating and offered modification suggestions. QY participated in the literatures collecting and organizing. 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.


The socioeconomic impact of osteochondral (OC) damage has been increasing steadily over time in the global population, and the promise of tissue engineering in generating biomimetic tissues replicating the physiological OC environment and architecture has been falling short of its projected potential. The most recent advances in OC tissue engineering are summarised in this work, with a focus on electrospun and 3D printed biomaterials combined with stem cells and biochemical stimuli, to identify what is causing this pitfall between the bench and the patients’ bedside. Even though significant progress has been achieved in electrospinning, 3D-(bio)printing, and induced pluripotent stem cell (iPSC) technologies, it is still challenging to artificially emulate the OC interface and achieve complete regeneration of bone and cartilage tissues. Their intricate architecture and the need for tight spatiotemporal control of cellular and biochemical cues hinder the attainment of long-term functional integration of tissue-engineered constructs. Moreover, this complexity and the high variability in experimental conditions used in different studies undermine the scalability and reproducibility of prospective regenerative medicine solutions. It is clear that further development of standardised, integrative, and economically viable methods regarding scaffold production, cell selection, and additional biochemical and biomechanical stimulation is likely to be the key to accelerate the clinical translation and fill the gap in OC treatment. Synovial or diarthrodial joints comprise most of the body’s articulations and are characterised by wide ranges of near-frictionless motion, as well as load transferring capabilities during weight-bearing . In thesIn normal conditions, there is healthy homeostatic crosstalk between cartilage and bone that leads to regulated tissue remodelling and joint integrity maintenance . This reLocalised OC lesions can be created acutely or developed as a result of repetitive overloading or long-term mechanical wear, with the most common OC injuries happening in the weight-bearing joints of the lower extremity  ,11,12. SDegenerative disorders, such as osteoarthritis (OA), osteoporosis, rheumatic arthritis, and other musculoskeletal disorders, also play an important role in the development of OC lesions and subsequent joint failure. OA, the most common form of arthritis, is a chronic disease of the articular joint characterised by the degeneration of AC and malfunction of the affected joint ,22. ApprNon-operative treatment of OC lesions is usually the primary course of action for minor OC defects and includes the use of chondroprotective pharmacotherapy  injections), the administration of non-steroidal anti-inflammatory drugs (NSAIDs) to fight pain and inflammation, or physiotherapy ,27,28. IReparative methods include microfracture and drilling, as well as autologous or allogeneic OC transplantation (mosaicplasty). Microfracture and drilling are bone marrow stimulating techniques used to initiate OC regeneration . These t2) -2-chlorobenzamide (BNTA) was identified as a potential disease-modifying OA drug  copolymer, either unloaded (AC phase) or loaded with thermally stable β-TCP (SB phase). To ensure the preservation of protein integrity, given the high processing temperatures of this technique, TGF-β1 was added to each AC layer after the respective 3D printing process. These hybrid scaffolds had suitable biomechanical properties for OC applications, displaying an elastic behaviour that supported several types of deformation with no visible damage and easy recovery of their initial shape. The preservation of TGF-β1 and β-TCP bioactivity after scaffold manufacturing was first confirmed in vitro, where they respectively enhanced the expression of chondrogenic and osteogenic markers by hBMSCs. Moreover, TGF-β1/β-TCP-loaded scaffolds demonstrated remarkable bioactivity in vivo  which does not enable the consistent and reproducible manufacturing of micro- and nanoscaled devices . High-reThe extensive list of studies herein presented is proof that the treatment of OC pathological conditions remains challenging, relying still too greatly on palliative treatments rather than disease-modifying drugs and restorative therapies , and typIn a typical solution electrospinning process , charge Recently, AM methodologies also developed to enable scaffold manufacturing in four dimensions, where the fourth dimension is that of time, giving rise to 4D printing. 4D printing makes use of common 3D printing techniques to generate shape-shifting materials, where the original geometry can dynamically change upon application of an external stimulus, such as temperature , osmoticBecause in vivo testing is mandatory in the development of medical products, a suitable choice of animal models is imperative to guarantee some level of parallelism to human conditions. Several criteria must be considered when making this decision, including the type and location of the OC defect, the dimensions of the joint and cartilage thickness, and the stage of skeletal development of the animals . The firFinally, the clinical trials and regulatory obstacles that need to be overcome before a new product is applied clinically and commercialised, along with the inevitable high costs associated, are another critical aspect that contributes to an extended time-to-market . AlthougFurther technological advancements not only in the field of biofabrication but also in medical imaging and 3D modelling, will allow for the development of therapeutic scaffolds with structural and chemical characteristics closer to those observed within an OC unit. Of note, apart from achieving clinical safety and efficacy, it is also pivotal to ensure that the production of such tissue-engineered scaffolds is scalable for industrial manufacturing under cGMP and economically viable, which is still a major challenge when using cell-based therapy or constructs with intricate architecture and chemical composition. Unfortunately, it is still difficult, even using current laboratory scale techniques, to achieve artificial scaffolds with physiologically relevant sizes and appropriate micro-/nanostructures, not only to mimic the natural architecture of the ECM but also other elements such as the blood vessels present in the joint. This is especially relevant in OCTE constructs, given the lack of vascularisation of AC and its dependence on surrounding tissues for nutritional support.Thus, it is possible to conclude that the development of OCTE products is inevitably subjected to close scrutiny to ensure safety and efficacy as well as logistic and economic viability. The ideal OCTE scaffold will, hence, sustain long-term OC regeneration, promoting progenitor cell homing, differentiation, and replacement of the scaffold matrix with spatially organised bone and cartilage neotissue, while exerting no toxic effects. Moreover, it is important to ensure that the production costs will not be prohibitive for universal access to the therapy, and that large-scale manufacturing and supply will be able to meet the overall demand for OC treatment solutions.OC diseases and traumatic lesions affect a progressively greater portion of the world’s population, with considerable impacts on quality of life and significant economic burdens. Current treatment strategies are mainly palliative and have short-term efficacy, highlighting the need for therapeutic options capable of halting disease progression, ameliorating joint pain and mobility, and stimulating tissue regenerative responses. TE offers a means of generating biomimicking platforms able to reproduce the natural geometry and microarchitecture of the OC unit, while delivering progenitor cells and/or cell-guiding cues  to the lesion site. Electrospinning and AM are two types of manufacturing technology widely used for biofabrication of scaffolds with varying composition and structure, whose potential for OCTE has been demonstrated in several in vitro and in vivo studies. However, to date, no therapeutic strategy has been developed that can sustain long-term tissue regeneration and satisfactory functional recovery. On one hand, this is due to the structural, chemical, and biological complexity of the OC unit, which cannot be easily reproduced artificially; on the other hand, the absence of standardised protocols for scaffold production, biomolecule administration, and therapeutic outcome evaluation obstructs reproducibility and comparison among different products. The development of an OCTE solution capable of responding to these issues is therefore dependent not only on technological progress, but also on strengthened communication between researchers and protocol uniformisation to achieve robust, reproducible, and scalable tissue-engineered solutions.


Eukaryotes and their environments serve as petri dishes, hosting an abundant and a rich prokaryotic microbiome. The assemblage of a eukaryotic host and its microbiome is referred to as a holobiont. The holobiont’s microbiome interacts within itself, with the environment, and with the host at the chemical level through production of specialized metabolites resulting in homeostasis or dysbiosis. These interactions are triggered by a multitude of factors, such as community composition, age, presence of nutrients, xenobiotics, and change in physical conditions, such as temperature and oxygen. Understanding how holobionts respond and adapt to diverse triggers is necessary to uncover mechanisms of resilience or susceptibility to dysbiosis and to modulate the collective functioning of microbiome in health and disease. This article highlights the challenges associated with uncovering chemical contributions of individual holobiont members and the applicability of metabolomics-based approaches to uncover chemical signatures of microbial processes in the natural environment. Additional complexity in these interactions is introduced by the dynamic nature of the members of the holobiont. While some partners are essential and form core symbiotic relationships with the host, the compositions of others vary in response to various environmental and genetic triggers. Thus, the contributions of individual members of the holobiont to host health and the response to environmental perturbations are challenging to decipher within the context of the whole organism. Advancement in imaging and multiomic approaches are beginning to address these challenges. The explosion in the development of informatic tools is beginning to allow understanding of functional cross talk between members of complex ecosystems. Advancement in fluorescence-enabled imaging technologies such as combinatorial labeling and spectral imaging-fluorescent al roles . Metagenal roles . The traal roles . Moreoveal roles . MetabolThe coral holobiont represents a multipartite relationship between the coral host, the endosymbiotic dinoflagellate algae termed Symbiodiniaceae, the coral microbiome, and the dynamic coral reef environment , 6. Corain silico annotation strategies and metabolite source tracking (Disentangling the roles of the microbiome, the endosymbiont, and the coral host itself in resilience and tolerance to disease requires a multifaceted approach. Inventory of microbial community composition can reveal shifts in microbiome architectures that serve as signatures of resilience or susceptibility. Similarly, we can identify microbial community signatures that are representative of disease . The mectracking . We can Propionibacterium acnes was identified as the source of fatty acids via hydrolysis of triglycerides on human skin (in silico tools in the last 5 years, it is now possible to gain biochemical insights into the detected metabolome through substructure discovery (Spatial metabolomics, by correlating patterns of microbial community structure with metabolites, allows one to infer the biological source of metabolite and their phenotypic roles. Using this approach, man skin , microbiman skin , and locman skin . Direct man skin . Next, wiscovery , 18 and iscovery , enablinin situ community structures (in situ imaging of metabolite gradients allow visualization of microbial chemotypes and source tracking, but remain to be developed for coral holobionts (A second bottom-up approach via comparative metabolomics of isolated holobiont members and the holobiont itself presents a traditional, but a direct approach. Herein, the individual holobiont members can be fractionated, enriched, or cultured. The endosymbionts have been fractionated from the coral holobiont mechanically with the use of a high-pressure jet stream and centrifugation. This process results in a relatively pure endosymbiont fraction with minor contamination from the microbiome. The metabolome of this fraction would allow source tracking of endosymbiont-produced metabolites in the coral holobiont and probe changes in these metabolites upon perturbations. Indeed, lipid biomarkers of thermotolerant endosymbionts have been proposed using this methodology . Finallyructures  and masslobionts . We are


Early in the development of the COVID-19 pandemic, it was evident that health care workers, first responders, and other essential workers would face significant stress and workplace demands related to equipment shortages and rapidly growing infections in the general population. Although the effects of other sources of stress on health have been documented, the effects of these unique conditions of the COVID-19 pandemic on the long-term health and well-being of the health care workforce are not known.The COVID-19 Study of Healthcare and Support Personnel (CHAMPS) was designed to document early and longitudinal effects of the pandemic on the mental and physical health of essential workers engaged in health care. We will investigate mediators and moderators of these effects and evaluate the influence of exposure to stress, including morbidity and mortality, over time. We will also examine the effect of protective factors and resilience on health outcomes.The study cohort is a convenience sample recruited nationally through communities, professional organizations, networks, social media, and snowball sampling. Recruitment took place for 13 months to obtain an estimated sample of 2762 adults who provided self-reported information administered on the web through structured questionnaires about their work environment, mental and physical health, and psychosocial factors. Follow-up questionnaires will be administered after 6 months and annually thereafter to ascertain changes in health, well-being, and lifestyle. Participants who consented to be recontacted form the longitudinal cohort and the CHAMPS Registry may be contacted to ascertain their interest in ancillary studies for which they may be eligible.The study was approved by the Institutional Review Board and launched in May 2020, with grants from Travere Therapeutics Inc, McKesson Corporation, anonymous donors, and internal funding from the M. Louise Fitzpatrick College of Nursing at Villanova University. Recruitment ended in June 2021 after enrolling 2762 participants, 1534 of whom agreed to participate in the longitudinal study and the registry as well as to be contacted about eligibility for future studies.The CHAMPS Study and Registry will enable the acquisition of detailed data on the effects of extended psychosocial and workplace stress on morbidity and mortality and serve as a platform for ancillary studies related to the COVID-19 pandemic.ClinicalTrials.gov NCT04370821; https://clinicaltrials.gov/ct2/show/NCT04370821DERR1-10.2196/30757 Globally, SARS-CoV-2 has infected more than 224 million individuals, with more than 41 million cases in the United States . Experie, for example, have had to reconfigure patient-care spaces and restructure clinical teams rapidly to address the increase in the number of patients with COVID-19 [Health care systems in numerous countries were overwhelmed by the COVID-19 pandemic, which has caused increased pressure on frontline health workers . HospitaCOVID-19 .Concerns of health care workers that they might not only become infected with COVID-19 but also transmit it to family members and friends add to their psychological burden ,12.In the context of the COVID-19 pandemic, we anticipated that health care workers would face anxiety and depressive symptoms due to traumatic patient-care experiences and the risk of SARS-CoV-2 infection to family, friends, and colleagues . FrontliEven under normal circumstances, workplace stress has effects on physical health , and theEmotional stress and stressful life events have been shown to contribute to the six leading causes of death in the United States—cancer, coronary artery disease, accidents , respiratory disorders, cirrhosis of the liver, and suicide—as well as to type 2 diabetes, sleep disorders, and other health conditions . WorkplaThe effects of stress on long-term physical and mental health can take many years to become manifest. Military combat deployment ,24 and eResearch on the effects of stress on the COVID-19 workforce  is under(CHAMPS) during later phases of the pandemic. The CHAMPS Study and Registry is designed to ascertain the time course and magnitude of these changes on the long-term mental and physical health of essential workers in the health care environment.Based on the extensive body of literature on the psychosocial influences on health, it is likely that the COVID-19 pandemic will have deleterious effects on the health of health care workers and support staff in a broad range of professions . Over tiThe objective of the CHAMPS Study and Registry is to assess the short- and long-term physical, social, and behavioral health of personnel  involved in supporting or delivering care for patients with COVID-19. Included are first responders, maintenance and support staff working in clinical settings, and health care professionals of all specialties and services. At enrollment, baseline data were obtained on the study participants’ working environment, lifestyle, and mental and physical health. Subsequent waves of data collection will obtain follow-up information after 6 months and then annually through 2024. The study was approved by the Villanova University Institutional Review Board (IRB) in May 2020.The primary objectives of the CHAMPS study are to (1) determine the extent to which working in health care affects future physical and mental health; (2) evaluate trends over the time course of the evolving pandemic; (3) identify exposure variables that are associated with incident changes in health over time; and (4) identify factors that influence short- and long-term health. Specific hypotheses include the following:Measures of mental health symptoms of anxiety, traumatic stress, depression, insomnia, disordered eating, resilience, burnout, and poor self-reported health will be more prevalent in essential workers than in the general population and greater in magnitude than reported in similar populations before the COVID-19 pandemic.The severity of mental health symptoms will be associated with prevalence of SARS-CoV-2 infection rates in the geographic region in which the study participants work.Mental health symptoms will persist longer among study participants with more severe symptoms.Essential workers who report elevated symptoms of anxiety, traumatic stress, or depression will experience exacerbation of existing physical health conditions over time and higher incidence of new physical health conditions.Severity and progression of mental and physical health conditions will be exacerbated by the degree of perceived exposure to adverse working environments, such as availability of personal protective equipment, extended working hours, and adequacy of staffing levels.CHAMPS consists of three components: (1) a cross-sectional study of respondents who consented to participate in a single assessment of health; (2) a longitudinal study of participants who consented to repeated waves of data collection; and (3) a registry comprised of participants who enrolled in the longitudinal study. The registry serves as a source of study participants who may be eligible for future ancillary observational studies as well as randomized clinical trials.Measures section. Demographic, occupation, work environment, and geographic questions were written by professionals who work in health care.To address the specific hypotheses, we selected validated instruments available for mental and physical health variables of interest. They are described in detail in the Eligible participants were adult essential workers involved in health care. For the purposes of the CHAMPS study, essential workers were defined as adult (aged 18 years or older) health care personnel, support personnel, and first responders working in any health care facility or in the community. Included were those working in facilities that are involved in patient care, such as hospitals, clinics, private practices, or screening facilities; those directly involved in patient care ; laboratory staff; service employees ; individuals working in long-term care facilities; and first responders . Collectively, we refer to these as “essential workers.” Enrollment took place over 13 months, from May 2020 through June 2021, throughout the United States.Recruitment included advertising through local and national professional and trade organizations, news, social and professional media, alumni organizations, and word of mouth to describe the study and disseminate the internet link that provided access to the informed consent form and study questionnaires. Professional and trade organizations were identified and contacted. Website posts, e-newsletters, and social media were used to reach members of eligible occupations, with news or social media posts either by their group administrators or by the study team. The wider social media universe was reached through posts on university channels on various platforms, university-affiliated influencer posts, and a limited paid media campaign. Social media posts and outreach to organization leaders continued periodically throughout the recruitment period.An internet link provided access to the informed consent document that described the purpose of the study, time required to complete the questionnaire, duration of the study, and options for cross-sectional or longitudinal study participation. An affirmative consent response opened the baseline questionnaire. At the end of the initial survey, participants were asked to provide their email contact information for the longitudinal study and registry, which would be used to provide them with future questionnaires and information about studies for which they might qualify. The consent form also stated that information would be used only for medical research and shared only with other medical researchers. Participants were informed that they could end participation at any time.CHAMPS has been registered at ClinicalTrials.gov (NCT04370821).The baseline questionnaire consisted of questions related to the participants’ demographic information, occupation, and work environment, followed by validated instruments on behavioral health, described below. Instruments were selected to balance validity and brevity. An open-ended question provided an opportunity to describe personal experiences of working during the COVID-19 pandemic and participants’ interest in being interviewed. The measures and timepoints of data collection are outlined in These will be ascertained by specific questions related to age, sex, occupation, self-reported general health, and infection status at the time of enrollment.Anxiety symptoms will be assessed by the Generalized Anxiety Disorder-7 (GAD-7) , and depThe Oslo Social Support Scale-3 (OSSS-3)  will be The Insomnia Severity Index-7 (ISI-7) will be used to measure sleep quality. The ISI-7 has been validated as a useful clinical tool to quantify perceived insomnia severity for primary as well as secondary insomnia and in young and older patients . It has traumatic stress with the Impact of Events Scale - Revised (IES-R) [stress specific to COVID-19, we adapted scales from Wu et al [Stress has been associated with elevated risk of cardiovascular, metabolic, and musculoskeletal disorders . Essenti (IES-R) , which aWu et al  and ChonWu et al  to assesThe last question on the questionnaire provides the participants with an opportunity to describe their service experiences during the pandemic in narrative form for qualitative analysis, with no limits on the time or length of the narrative.The first follow-up occurred 6 months after enrollment and completion of the baseline questionnaire. Assessments included resilience, burnout, eating habits, and alcohol and substance use; physical activity; nutrition; height, weight, and weight discrimination items ,42; and Resilience will be measured by the Brief Resilience Scale (BRS), which is designed to assess the ability to bounce back from stress and to be brief . It inclProactive coping is defined as a forward-looking strategy to address anticipated stressors or hardships, and it will be measured by the Proactive Coping Subscale of the Proactive Coping Inventory (PCI). We hypothesized that proactive coping would serve as a moderator of the consequences of stress. The Proactive Coping Subscale of the PCI consists of 14 items that combine autonomous goal setting with self-regulatory goal attainment cognitions and behavior. The scale showed high internal consistency, as seen in reliability measures of .85 and .80 in two samples during its development ,45.Burnout will be assessed by the Oldenburg Burnout Inventory (OLBI). Burnout is a psychological condition that follows highly demanding or stressful physical and mental work conditions that may be accompanied by inadequate resources ,46. HistIncreased caloric intake is a common response to mental stress . We chosSelf-reported height and weight were obtained. Weight discrimination history, namely whether the participant experienced any bias based on body weight, was assessed by a single question ,42.We hypothesized that alcohol and substance use may increase in response to the stress of workplace demands, but we thought that a thorough screening would not be in the best interest of the overall study. We decided to limit our data to answers to two questions, namely, whether the participants’ alcohol and substance use increased, was unchanged, or decreased since the start of the pandemic.We chose to assess physical activity with two brief questions: one on the amount of time the participants spent in moderate-to-vigorous physical activity, and one on whether their amount of physical activity had changed since the pandemic.The relationship among stress, major life events, and other biopsychosocial factors has a long history of study in epidemiology and public health, with a sound mechanistic basis ,53,54. WWe will define and quantify exposure to COVID-19 service burden by summing participants’ scores on the Perception of Stress at Work scale developed by Wu and colleagues  in 2009 For this purpose, perceived stress was deemed to be an outcome of exposure, but it can also be included as an exposure variable in models that ascertain incidence of health conditions over time.Annual follow-up questionnaires will include incidence of conditions and diseases, as well as selected instruments from the baseline and 6-month follow-up questionnaires. Additional items may be added in response to the evolving research literature on COVID-19 and that in other fields. During enrollment participants were given the option of providing contact information for follow-up questionnaires, which also enrolls them in the CHAMPS Registry. The CHAMPS Registry enables recruitment of eligible individuals for clinical trials of interventions relevant to COVID-19 or other disorders or conditions, as well as for ancillary studies of basic behavioral research or observational studies that require new data collection. Anticipated ancillary studies include a detailed assessment of lifestyle factors, interactions with family life, and wellness interventions. Ancillary study proposals will be reviewed by the CHAMPS Steering Committee for relevance, scientific interest, and participant burden.Data are collected through the web-based Qualtrics system. SPSS (IBM Corporation) and SAS  will be used for data analysis. Initially, we will perform an exploratory analysis to establish patterns of missingness and distributions of the variables being measured. The registry data will be characterized with descriptive analyses, including frequency distributions, prevalence rates of COVID-19 testing and SARS-CoV-2 infection, and means and standard deviations of scores on the scales measuring work environment, general health status, anxiety, depressive symptoms, social support, traumatic stress, social support, sleep quality, and stress. Multiple logistic and linear regression methods will be used to assess baseline associations of health and well-being with estimates of exposure to COVID-19 infection and COVID-19–related stress at work. Models will be adjusted for sex, age, race/ethnicity, and job classification. Other preliminary explorations of associations between measured variables will be performed to aid the development of substudies from the registry. Follow-up health status questionnaires will enable analysis of associations between incident health events and baseline variables.Narrative responses to the final question on the questionnaire will be analyzed using a deductive approach and thematic content analysis techniques. Multiple investigators will read the narratives provided by participants to perform coding of the significant statements made by participants. After multiple readings, investigators will generate data categories and clusters of statements and will develop themes.We will determine whether essential workers with higher COVID-19 service exposure burden will experience higher rates of stress, depression, anxiety, and substance use than the general population at the time they enroll in CHAMPS, and higher than in similar populations before the COVID-19 pandemic. We will compare exposure by type of service, occupation, and place of service , controlling for age, self-reported sex, and race. We will conduct analyses to identify mediators, such as racial status, and moderators, such as proactive coping and resilience.We will determine whether the severity of mental health symptoms will be associated with changes in the prevalence of SARS-CoV-2 infections by geographic region.We will ascertain whether exposure to COVID-19 service burden will affect physical and mental health over time and how these are influenced by psychosocial variables . We will analyze whether longitudinal health outcomes are associated with type of service; severity and duration of mental health symptoms; and age, sex, race, and other variables. We anticipate that some analyses will depend on the adequacy of the sample sizes in various demographic subgroups.We will obtain longitudinal data to determine whether mental health symptoms will persist longer among participants with more severe symptoms and whether these symptoms will be associated with higher incidence of physical health conditions.We will ascertain the extent to which the thematic contents of the narrative responses to the final question of the questionnaires are correlated with measures of distress  and moderated by measures of resilience. We will ascertain whether thematic content is a moderator or mediator of health outcomes and is associated with retention of participants over time.A review of the published literature on relationships between stressful life events and health led to the hypothesis that stress experienced by essential workers during the COVID-19 pandemic would affect their health status over time. In response, CHAMPS was designed to ascertain work-life experience during the first year of the pandemic, exposure to stress, and self-reported mental and physical health, as well as to determine effects of service during the pandemic on health over the subsequent 5 years. The CHAMPS longitudinal cohort serves as a registry for recruiting participants in future ancillary studies. After IRB approval, 2762 participants were enrolled between May 2020 and June 2021, with 1534 providing contact information for future studies and follow-up questionnaires.The longitudinal CHAMPS Study and Registry will provide important prospective data about relationships between extended health care–related service during a global pandemic and the future health of the health care–related workforce. Of special interest are first responders and service workers, who are less often included in such studies, as well as a broad ethnic diversity of participants. The lengthy enrollment period as the pandemic progresses should result in a sample with a wide range of exposures to various working conditions, thereby providing insights into the potential attributes that contribute to changes in health over time. Assessments of an extensive range of psychosocial factors, such as lifestyle habits, resilience, and pre-existing conditions, will enable assessment of their influence on mental and physical health.The CHAMPS Registry is designed to provide opportunities for investigating additional questions and potential extension of data collection for specific hypotheses. The CHAMPS Registry anticipates ancillary studies of behavioral and biological mechanisms of action, studies of lifestyle changes and health, the effects of the pandemic on extended families, and potential randomized trials of interventions.Altogether, CHAMPS has the potential to provide advances in knowledge about the role of chronic stress in health. The value of the study for some questions, such as identification of associations with various demographic variables, depends on the availability of a sufficient sample size of participants. We anticipate that over time, the study findings will inform public health policy relevant to national health emergencies.This is a convenience sample, which limits the generalizability of the results to the entire population of essential workers. Some occupation categories will have small sample sizes; thus, we may not have sufficient representation for individual analyses. Self-selection bias may also be a factor if some individuals decline to participate due to reluctance to be perceived as not supportive of their institutions. The measurement of COVID-19 service exposure relies on an algorithm that should be evaluated against an independent data set. Finally, the planned longitudinal analyses will depend on retaining a sufficient proportion of participants over time.


Nanomedicine has demonstrated substantial potential to improve the quality and efficacy of healthcare systems. Although the promise of nanomedicine to transform conventional medicine is evident, significant numbers of therapeutic nanomedicine products have failed in clinical trials. Most studies in nanomedicine have overlooked several important factors, including the significance of sex differences at various physiological levels. This report attempts to highlight the importance of sex in nanomedicine at cellular and molecular level. A more thorough consideration of sex physiology, among other critical variations , would enable researchers to design and develop safer and more-efficient sex-specific diagnostic and therapeutic nanomedicine products. Given the early failing of therapeutic nanomedicine in the clinic the importance of understanding bio-nano interactions and biosystem parameters has gained much attention. Here, the authors review the work to date that looks at the impact of sex on the applications and outcomes of nanomedicine. Nanomedicine employs nanotechnology for medical applications including diagnosis and treatment of diseases2. As an example, for drug delivery, nanomedicine aims to design and develop biocompatible nanoparticles (NPs) that can protect payloads from degradation and unwanted interactions with biosystems  and deliver them to the desired biosystems 3. The purpose of this review is to explore the relevance of sex at cellular and molecular level for the laboratory and clinical research in the general field of nanomedicine.According to the British Standards Institute, nanotechnology refers to “intentional design, characterization, production, and applications of materials, structures, devices, and systems by controlling their size and shape in the nanoscale range (1–100 nm)”4. Remarkably, the trials of some nanomedicines have failed to show any improvement in pharmacokinetics or drug accumulation within tumor tissues, as compared to the parent drugs4.Although the emergence and development of nanomedicine have demonstrated promising (and sometimes even tremendous) positive results in in vitro environmental and animal studies, therapeutic nanomedicines such as those for cancer faced significant translational challenges. More specifically, a 2019 clinical trial analysis of 75 cancer nanomedicines, that were registered through clinicaltrials.gov database, revealed that cancer nanomedicine products have had a success rate of 14% in phase 3 trials, which largely reflects the efficacy of nanomedicine products6. Therefore, identifying and gaining a much deeper understanding of these factors is an essential step in improving successful clinical translation of nanomedicine products.These disheartening clinical translational results are driven at least in part by neglect and/or a failure to fully understand several crucial/influential factors  at play in both nano- and bio-systems and in nano-bio interactions8, nanomedicine may be following a similar trajectory, but perhaps for different reasons. It is essential, therefore, that all factors critical to therapeutic efficacy–including differences in biological cellular responses between males and females—be robustly considered and incorporated into both laboratory and clinical research. This will enable development of more effective drug delivery systems and more reliable diagnostics and therapeutics.Monoclonal antibodies failed their first decade of clinical trials owing to their murine-derived origin; only when humanized/human monoclonals were used did clinical trials succeed. Based on current evidence9. However, most researchers are still using only one sex of biosystems  and applying the results to both males and females. With the emergence of nanomedicine, the importance of sex in biomedical research has become even more obvious. In a recent review article, Liyod-Parry et al.10 provide a comprehensive overview of the role of nanotechnology and nanomedicine in women’s health research. The authors present both the benefits and shortcomings of the application of nanomedicine to both laboratory-based research and clinical trials. Most importantly, in recent years, the importance of sex at the cellular level has also been recognized in biomedical research. Nevertheless, it is still the case that only a small number of published articles report the sex of biosystems used in their study11.The importance of sex as a biological variable in laboratory and clinical experiments has been recognized by the biomedical research community since the 1990sThis review focuses on how sex-dependent physiological differences Fig.  can affe12. In another study, amorphous silica NPs triggered more-severe inflammation and lung tissue damage in female rats compared to male rats13.Although only a handful of nanomedicine studies have focused on the sex-dependent effects of NPs, results indicate that the same NPs have different therapeutic and toxic impacts in male and female animals. For example, PEG-coated gold NPs show different toxic effects in male and female mice, producing more severe kidney damage in females, but higher liver toxicity in males15. However, despite the extensive deployment of therapeutic nanomedicines in both the laboratory and clinical trials, they have had very limited success in reaching clinical practice. The safe and effective use of therapeutic nanomedicine remains a growing concern among biomedical and clinical researchers16. Among the many reasons for the frequent failure of clinical trials involving nanomedicine is our lack of a deep understanding of the mechanisms of interaction between NPs and body fluids, extracellular matrix, and cellular components. Another important stumbling block is the fact that most laboratory and clinical studies involving cellular interactions with nanomaterials do not take the significance of sex  into account. This is more obvious in studies involving diseased cells and tissues.With the emergence of nanomedicine, a wide range of NPs has been developed for diagnostics/therapeutics application17. For example, the level and pathway of NP uptake and intracellular trafficking in human amniotic mesenchymal stem cells and cancer cells are strongly sex dependent; in addition, the composition of the biomolecular/protein corona  is affected by sex-specific paracrine factors17. Furthermore, it has been found that toxic silver NPs significantly alter the bacterial species harbored in male zebrafish, but not in females19. It is clear that challenges to the progress of NP-based nanomedicines will not be overcome until biological variable aspects are taken into consideration, including alteration of the biomolecular/protein corona on the surface of NPs and different structural and molecular responses to functionalized NPs by male and female cells.Recent studies provide convincing evidence that sex differences can alter NP efficacy at the cellular level26. For example, in cancers, sex significantly influences tumor incidence, growth, cellular and molecular phenotypes, and therapeutic efficacy29. This is at least partially due to sex-related genetic differences: abnormal reactivation of X-linked genes30, differences in duplication/deletion of segments on X/Y chromosomes28, differences in mutated genes in tumors31, transcription factor-binding patterns29, and sex-specific roles of hormones32. Drugs also have different pharmacodynamic and pharmacokinetic patterns in males and females and, therefore, exert sex-dependent therapeutic and toxic effects35. Male and female cells of different origin may respond differently to the same NPs. For example, quantum dots have higher uptake in female than in male human amniotic stem cells; however, the same particles have shown lower uptake in female salivary gland primary cells compared to their male counterparts17.It is noteworthy that the extent of sex-related differences in biosystem responses to NPs/drugs depends strongly on the type of disease and tissue involved. Sex difference of the whole organism at the physiological level is one of the most important biological variables determining the incidence, prevalence, and severity of many diseases, including cancer, neurodegenerative, cardiovascular, autoimmune, and psychological diseases18. Most studies in nanomedicine have not properly reported/considered biological variable factors in the in vitro and in vivo microenvironment that determine the fate, safety, and efficacy of NPs in both animal and human clinical trials8. Corrective efforts have mainly involved: (1) identifying important biological variable factors in the NP microenvironment; (2) improving NP toxicity assays; (3) optimizing in vitro protocols mimicking in vivo conditions; and (4) more-accurate computational modeling approaches to identify NP mechanisms of action37.Once exposed to biological fluids, the surface of NPs becomes rapidly covered with lipids, metabolomes, proteins, and other biomolecules; this biomolecular/protein corona alters the original surface of the NPs and gives them a new biological identity6. Exposure of NPs to proteins in the plasma of healthy individuals vs. patients with cancerous conditions significantly affects the composition of biomolecular/protein coronas, pointing to NPs’ therapeutic and diagnostic properties40. It has been shown that changes in plasma composition during the disease development process substantially alter corona composition and thus NP biological function43. Other recent findings have revealed the role of sex-dependent secretion of paracrine factors in NPs’ interactions with cells17.Human plasma is the most suitable biological fluid to study biomolecular/protein corona. One of the major sources of misprediction of the biological safety and efficacy of NPs is conflicting biomolecular/protein corona data, originating from the use of various biological fluids, including (1) aqueous non-biological fluids, (2) semi-biological fluids such as cell culture media, and (3) animal serum/plasma. Even when using human plasma, characteristics of the donor  can significantly affect the composition of the biomolecular/protein corona. Unfortunately, much of the current literature on the corona does not report the details of the plasma used; in fact, in most cases, pooled plasmas from multiple individuals are employed44. The outcomes of this meta-analysis have revealed robust and reproducible sex differences in 77 of the biomolecules. Among these, 40 have higher concentrations in females , sialic acid-binding immunoglobulin-type lectin-6 (decreases from 10 to 22 weeks and increases from 23 to 40 weeks), placental growth factor (increases from 10 to 31 weeks), prolactin (increases from 10 to 35 weeks), and interleukin-1 receptor 4 (increases from 25 to 40 weeks)47.Based on both proteomics and metabolomics data, the composition of female and male plasma differs significantly, with significant variations in the abundance of 231 proteins. Another study analyzed 174 serum biomolecules in 196 male and 196 female human samples, from nine independent cohorts, and probed sex-specific differences50 significantly affects NP’s biomolecular/protein corona profiles. Therefore, one can expect that sex-based biomolecular differences in any given plasma sample to be reflected in NP biomolecular/protein corona composition. Although corona profiles have been widely investigated under a variety of conditions6, to the best of our knowledge, few reports have considered the effect of sex on the composition and decoration of biomolecular/protein corona. For example, it has been demonstrated that incubation of identical 70-nm SiO2 NPs with plasmas of male and female zebrafish leads to the formation of coronas with different compositions;51 the vitellogenin family  was found to represent one major difference in male and female corona compositions51. Blood cells therefore interact differently with NPs exposed to male and female plasma. The NPs with coronas containing proteins present in female plasma are preferentially recognized by leukocytes compared to their male counterparts. This finding suggests that blood circulation time and pharmacokinetics of identical NPs, together with the host immune system response to them, may differ significantly between males and females.It has been increasingly recognized that plasma composition 54. Low concentrations  of silver NPs delay the differentiation of female mESCs by interfering only with the X chromosome inactivation process. The same NPs, however, show no interference with the self-renewal process and exert no significant cytotoxic effect. It is well understood that even slight interference in X chromosome inactivation considerably impairs programmed ESC differentiation by regulating the expression of relevant genes55. The random inactivation of one X chromosome of the female cell is known to equilibrate gene content between the sexes57. Xist and Tsix are noncoding RNAs playing critical roles in silencing and activating the X chromosome, respectively. A low concentration of silver NPs interferes with the expression of Xist and Tsix and disrupts the optimal Xist/Tsix ratio for X chromosome inactivation. Silver NPs also prevent the expression of genes responsible for triggering programmed ESC differentiation by enhancing the trimethylation of histone . Therefore, even a low concentration of silver NPs can cause a series of transcriptome and epigenome changes interrupting X chromosome inactivation and subsequent ESC differentiation in females. The most striking observation has been that such epigenome and transcriptome changes are absent in male mESCs treated with low concentrations of silver NPs, revealing programmed differentiation even after long exposure to silver NPs. As X chromosome inactivation is a female cell-specific phenomenon, agents interfering with this process impair the differentiation of female cells only54.One of the main challenges to the vigorous study of NP interactions with biosystems is the selection of appropriate cells that originate at a very early stage of development, prior to the initiation of hormonal changes and genetically and structurally driven sexual dimorphisms. The use of embryonic cells is one strategy to address this issue. For example, male and female mouse embryonic stem cells (mESCs) have been used to determine the sex-specific effects of silver NPs on stem cell differentiation17. Exposure to quantum dots (QDs) reflects significant sex-dependent differences in cellular uptake efficacy in human amniotic stem cells (hAMSCs): female cells take up more QDs than male cells. Aside from sex-dependent biomolecular/protein coronas, sex-based differences in the structure and function of cells also play key roles in their response to NPs. It was shown that sex-dependent uptake of QDs is caused by differences in arrangement, shape, and distribution of actin filaments (cytoskeleton), which regulate the endocytosis and cellular trafficking of QDs. Remarkably, male and female cells show different capabilities in reprogramming hAMSCs into pluripotent stem cells when treated with nanosized Sendai virus commonly used for the preparation of pluripotent stem cells. The Sendai virus shows more effective transfection—and hence, better reprogramming efficacy—into female hAMSCs compared to male hAMSCs17.As another example, human amniotic stem cells (hAMSCs), one of the earliest sources of somatic stem cells, have been employed to probe the effect of cell sex on NP uptake59. Therefore, small variations in hormone production/balance may produce significant changes in susceptibility/resistance to diseases and therapeutic drugs/NPs. For example, it is well understood that cell sex characteristics and sex hormones have significant impacts on both normal lung physiology and lung diseases60. New findings from the NELSON Trial provide evidence of sex-specific differences in lung cancer screening and survival61. Notably, the results of related research suggest that lung cancer progression may be more rapid and lethal in females. These important factors need to be considered in interpretation of the toxicity and therapeutic outcomes of NPs being used for tissues  that are highly affected by sex-specific phenomena. In addition, sex-specific disease-dependent biomolecular changes in plasma can significantly affect the biomolecular/protein corona profiles of NPs and thus their interactions with biosystems. For example, different concentrations of vitellogenin in male and female zebrafish plasma significantly affected the composition of the corona formed on the surface of SiO2 NPs51. In addition, since the protein metabolome profiles of males and females reflect considerable differences in amino acids, fatty acids, and lipids62, their blood plasmas also have different metabolome and lipidome profiles. Depending on the sex and age of a person, different metabolic pathways are activated and different plasma metabolite association networks form63. As the metabolome can affect protein-NP interactions64, one could expect metabolomics variations to affect the biomolecular/protein corona composition of NPs in a sex-specific manner.For accurate evaluation of the interactions of NPs with non-embryonic cells, the possible roles of sex-specific hormonal effects on both cell behavior/characteristics and biomolecular/protein corona of NPs should be considered in detail. Hormones are sex-specific chemical messengers regulating the duration and level of crucial processes  in tissues and organs65. Indeed, female mice and rats have more CD45+ leukocytes and macrophages in both naive peritoneal and pleural cavities66. In the respiratory tract, sex hormones including estrogens, androgens, and progesterone regulate the number and function of innate immune cells in a sex-specific manner67. Sex can also affect patient responses to cancer immunotherapy23. Sex-specific differences have also been observed in the functions of other immune cells such as T-lymphocytes, B-lymphocytes, astrocytes (in the brain), and natural killer cells68. Therefore, males and females show different innate/adaptive immune responses to antigens/infections. The immune response to self-antigens, leading to autoimmune diseases, is more prevalent in females; ~80% of patients diagnosed with autoimmune disease are females69. As stated above, it is legitimate to hypothesize that immune system responses to identical NPs are sex-dependent. For example, female mice have stronger immune responses  to PEG-coated gold NPs compared to male mice12. In addition, male mice treated with gold NPs show more significant infection and inflammation compared to female mice12.Immune system response to NPs is an important factor in their toxicity and therapeutic efficacy; and sex-related physiologic differences significantly affect host immune system responses to foreign materials including NP/biomolecular therapeutics. In rodents, for example, the phagocytic activity of macrophages is stronger in females than in males70. In addition, in some cancers such as glioblastoma, current treatments show better outcomes in females than in male patients71. For example, the effectiveness of immune checkpoint inhibitors , among the most efficient therapeutics, is sex-dependent74. One of the recent promising applications of NPs in cancer is immunotherapy75; here again, sex may have significant effects on the therapeutic efficacy of nanomedicine products, in an organ-dependent manner, for several reasons, including sex-related differences in tumor microenvironments76; sex-associated molecular differences in response to cancer immunotherapy;77 and the role of NPs in the efficacy of the immune system in tumor microenvironments78. For example, it was shown that ferumoxytol, a Food and Drug Administration (FDA)-approved iron supplement, could increase the number of pro-inflammatory M1 tumor-associated macrophages in female mice, demonstrating intrinsic inhibitory effects on the growth of early mammary cancers together with lung cancer metastases in liver and lungs78. Although the study was limited to female mice, one may expect to observe different therapeutic outcomes with ferumoxytol in male mice, as the production of cytokines and chemokines by macrophages differs between males and females79.The immune system plays a critical role in many diseases, including cancers, in a sex-specific manner. Due to their genetic and epigenetic mosaicism due to random inactivation of one of the X chromosomes in XX female cells, females are more resistant and show stronger immune response to cancer compared to males80. The placentas of male and female fetuses use different mechanisms to suppress oxidative stress and show different responses to identical stimuli. NPs encapsulating MtiQ, an antioxidant suppressing mitochondrial oxidative stress, have been used to treat fetal hypoxia and prevent subsequent placental dysfunction in animal models, showing sex-specific therapeutic effects81 in favor of female fetuses. Therefore, sex differences should be considered throughout the development of nano-based drugs for treatment of placenta- or embryo-related disorders.NPs not only exert different toxic effects in males and females, but also show sex-dependent variations in therapeutic efficacy. The fetal hypoxia that often occurs during pregnancy may induce oxidative/nitrosative stress, which increases the risk of catastrophic diseases in mature offspring84, differences in therapeutic efficacies of drug/NPs between males and females should be considered in an organ-specific manner. This is because male and female cells have different sex chromosomes, display dissimilar gene expression patterns, and activate distinct sex-specific signaling pathways in response to the same treatment86. For example, the body’s natural healing process in response to cardiac diseases  is sex specific89. Although nanotechnologies have been widely used to address cardiovascular diseases91, the role of sex in therapeutic efficacy has not been critically considered. It is noteworthy that even circulating biomarkers and/or biomolecules associated with cardiovascular diseases are sex dependent92, which may affect the biomolecular/protein corona of therapeutic NPs. Such variation in circulating biomolecules may also be related to the gut microbiota93, whose composition depends strongly on host sex94. Although NP-based oral treatments may affect the composition of the gut microbiome in a sex-dependent manner 19, we do not consider them in this review.Since sex differences affect the prevalence and progression of many diseases96. This will significantly affect the delivery of identical NPs to the brain tissue in a sex-dependent manner. It has been demonstrated in transgenic mice models of TBI that PEGylated polystyrene NPs were more effectively delivered to female than to male brain tissue97. Although not evaluated in previous studies, the therapeutic efficacy of the NPs that accumulate in brain tissue is assumed to be sex-specific, at least in part due to the sex-specific response by brain immune cells. Recent reports have revealed significant differences in function, morphology, abundance, and gene expression profiles between male and female microglia, which are the main immune cells involved in central nervous system maturation, signal transduction, and clearance of protein/cell aggregates/debris in the brain101. As sex differences in microglia are involved in the cause of differing incidence and prevalence of neurodegenerative diseases such as Alzheimer’s, Parkinson’s disease, Amyotrophic lateral sclerosis, and frontotemporal dementia between male and female patients, the successful use of therapeutic nanomedicines for these diseases must also be sex-specific105. Microglia are the resident immune cells in the brain and maintain their homeostasis and functionality. The role of sex in microglial cell functions and its effect on physiological and pathological conditions in the brain has been discussed in a recent review article by Yanguas-Casás106.Some injuries may also have sex-specific physiological effects that may significantly affect NP delivery to the injured tissue. For example, it was shown that within the first couple of hours following traumatic brain injury (TBI), the blood–brain barrier (BBB) often undergoes less damage in females than in malesTaking sex into account will have a significant impact on the success of both laboratory and clinical research in nanomedicine, encompassing increased safety and efficacy, identifying sex differences in cellular characteristics, plasma composition, and secreted factors that affect NPs’ biomolecular/protein coronas and functions 108; premenopausal and postmenopausal females have different hormone profiles108; and blood concentrations of progesterone, estriol, and prolactin change considerably during pregnancy108. All these variables may have significant effects on the biomolecular/protein corona profile of NPs and their interactions with biological systems.Compared to male biosystems, female biosystems  have more physiological complexity, due to conditions such as menstruation, pregnancy, lactation stage, and menopause, among others. These complexities may make the outcomes of studies involving females less reliable and/or reproducible than the findings of studies on males. Age also needs to be properly considered in study designs, as many sex-specific hormones are strongly dependent on age and individual health status. For example, during puberty, hormones differently induce the development of sex organs in males and femalesTo obtain useful biomolecular/protein corona data, researchers need to have full information on the age, sex, health, and physiological condition of plasma donors. Researchers should avoid using pooled plasma and/or plasma with no comprehensive information on the donor , as the outcomes may not be conclusive. When seeking to determine the role of sex in the biomolecular/protein corona profile of NPs, the characteristics/specification of plasma donors  need to be the same for both sexes. In addition, other influential factors  need to be carefully considered in the experimental setup. The best approach to considering the effect of female-specific physiological conditions  on the biological efficacy of NPs is to use plasma samples from the same individuals in a prospective manner.6. Even in the limited number of publications that do report cell sex, other crucial information 36 are often missing. The number of cell passages is critically important, as some male cell lines widely used in in vitro studies gradually lose their Y chromosome with increasing numbers of passages109.Results published from both in vitro and in vivo studies should include full details on the sex of the cells and/or animals. Unfortunately, in most studies, the sex of cell lines used in in vivo and in vitro  are not appropriately considered and reported in the current literatureResearchers need to have full information about the cells they intend to use for in vitro and/or in vivo studies, including their sex, origin, passage numbers, and storage conditions. In addition, to completely investigate the effect of sex on cellular responses to NPs. The selected male and female cells should (i) have the same origin , (ii) have been extracted from similar donors , and (iii) have had minimal exposure to sex-specific hormones. For in vitro evaluations of the effects of sex on NPs, researchers must ensure that their culture media are free from sex-specific hormones that may differentially affect the behavior of male and female cells. Another major problem is the limited number of comparable male and female cells offered by vendors/distributors of cells . Therefore, efforts by bioresource companies, as important stakeholders, should be made to provide resources for studying sex as an important biological variation. In the absence of such resources, researchers may have to access suitable cells through their clinical collaborators.110. Although the mechanism formation of these clusters and their function is unknown, the composition and concentration of these impurities associated with the corona formed by exposure of NPs to male and female plasma could be different.As discussed above, achieving rigorous and concise information on the composition of biomolecular/protein corona is essential to predict NPs’ interactions with biosystems and their biological impact. Recent findings obtained by combining cryo-electron microscopy, cryo-electron tomography, and image simulation have revealed that the biomolecular/protein corona formed on the surface of NPs contain some impurities in the form of non-specific clusters of biomolecules. This will influence the accuracy of the results of proteomics analysis of the biomolecular/protein corona111.More in-depth attention needs to be paid for investigation of the purity of protein corona prior proteomics analysis. Another crucial aspect is experimental obstacles affecting the characterization of NPs and their corona profiles. Using diluted NPs to study biomolecular/protein corona is one practical approach that can minimize such impurity issuesAccess to materials and data are easily available through extensive studies of differences between the sexes on the reproductive systems. Lack of availability of resources on sex differences in nonreproductive biology, where differences are much less recognized, creates critical challenges in the field. Although most of the differences in sex in nonreproductive areas of biology are difficult to detect, some of these differences can have significant implications for clinical and laboratory research within the discipline of nanomedicine. An additional problem is the fact that many researchers may not have access to comparable male and female biosystems  for a variety of reasons, including lack of a clinical collaborator/center.Researchers should thoroughly report all available information, including sex, on the biosystems they study and be specific about their claims/conclusions. For example, if studies are performed on male plasma and/or cells, that should be clearly conveyed in an article’s conclusion to avoid apparently conflicting results arising from sex-specificity; that is, the results might not be reproducible if female plasma and/or cells are used by other researchers.6.Beyond conventional medicine and commercially available parent drugs, therapeutic and diagnostic applications of nanomedicine have yet to be developed to the point of clinical use against devastating diseases such as cancer, heart disease, and neurological diseases. Among the reasons for slow progress in many areas of therapeutic nanomedicine are the lack of comprehensive, basic scientific research and an insufficient understanding of the mechanisms involved in cellular interactions with nanomaterials and functionalized NPs together with other complex biological variables, including sex, age, race, health status, and comorbidities112, interlaboratory procedures and instrumentations should be standardized for assessment of the role of sex on diagnostic and therapeutic nanomedicine.To accelerate successful clinical translation of diagnostic and therapeutic nanomedicine, future studies should consider the importance of overlooked factors described in this review and report critical information including the alteration of physicochemical and biological properties of NPs upon exposure to body fluid and most importantly, the significance of biological variables including sex. More specifically, to robustly consider and understand the role sex plays in the safety and therapeutic efficacy of nanomedicines. There is a need for advancement of our understanding of (i) the effects of ethnicity, disease stage, age, sex, and physiological conditions on the biomolecular/protein corona profiles of NPs, (ii) the interaction of NPs with sex-specific elements of the immune system , and (iii) the interactions of NPs with sex-specific disease environments . As different laboratories may have significant variations in the characteristics readouts of identical NPs114, the NP-based vaccines developed by Pfizer/BioNTech and Moderna performed slightly better in males [95.4% (Moderna) and 96.4% (Pfizer/BioNTech)] than in females [93.1% (Moderna) and 93.7% (Pfizer/BioNTech)]114. While monitoring of the sex-specific efficacy outcomes of these vaccines should be continuously monitored and reported, more fundamental studies are required to be conducted on the sex-specific efficacy of the NP-based vaccines at cellular and molecular level. The outcomes of such efforts will help the scientific community to better define potential sex-dependent therapeutic/toxic effect of COVID-19 NP-based vaccines which in turn pave a way for development of sex-specific vaccine design, development, and administration.The importance of the topics outlined in this article is evident by the application of nanomedicine-based approach to develop and produce an effective vaccine against COVID-19. According to the initial reports115)19. When only statistically significant results are published, it is obvious that the outcomes misrepresent the real findings116. According to Joober and co-workers117, “withholding negative results from publication—publication bias—could have major consequences for the health of millions”. The negative impact of publication bias has been studied and validated in most of the scientific fields including chemistry, medicine, and biomedical sciences123. Due to the multidisciplinary nature of nanomedicine, it is obvious that nanomedicine literature has the same issue which has been poorly considered so far124; despite extensive number of publication since its emergence, addressing the literature bias in nanomedicine has been ignord124.The other important factor that should be considered in the sex-specific meta-analysis of nanomedicine is to minimize publication bias  should be considered, as an essential parameter, in nanomedicine studies. Such consideration will be critical in addressing the reasons behind the alarming signals of failure in therapeutic cancer nanomedicine125. Unfortunately, most of the recommendations outlined in this book have been ignored so far. Such a framework of integrated responding of all involved stakeholders may increase responsibility and response-ability in understanding and solving problems in timely and efficient manner126. Considering the current challenges facing nanomedicine, it would be helpful to establish a similar undertaking to overcome some of the shortfalls in nanomedicine. Inclusion of sex-specific biological factors at cellular and molecular is essential for advancements of nanobiotechnology and nanomedicine for both sexes. Undoubtedly, nanomedicine holds the promise to improve the health care system in near future; therefore, it is important for the scientific community to pay more attention to the importance of sex differences in their studies. Last, but not least, if we are to move closer to the lofty goal of personalized nanomedicine, the role of gender must also become a central consideration in future studies.A collective national and international effort among all involved stakeholders  is required to introduce policies, guidelines and regulation to promote inclusion of sex-specific factors in nanomedicine research. A good example is the initiation of the Committee on Understanding the Biology of Sex and Gender Differences that was constructed by the National Academy of Science to address biology at the cellular, developmental, organ, organismal, and behavioral levels with emphasis on Sex and Gender Differences. The results of this comprehensive study have been published in a book by T.M. Wizemann and M-L. Pardue