Arabidopsis thaliana has allowed tremendous advances in plant genetics and physiology, in spite of only being a weed. It has also unveiled the main molecular networks governing, among others, abiotic stress responses. Through the use of the latest genomic tools, Arabidopsis research is nowadays being translated to agronomically interesting crop models such as tomato, but at a lagging pace. Knowledge transfer has been hindered by invariable differences in plant architecture and behaviour, as well as the divergent direct objectives of research in Arabidopsis vs. crops compromise transferability. In this sense, phenotype translation is still a very complex matter. Here, we point out the challenges of “translational phenotyping” in the case study of drought stress phenotyping in Arabidopsis and tomato. After briefly defining and describing drought stress and survival strategies, we compare drought stress protocols and phenotyping techniques most commonly used in the two species, and discuss their potential to gain insights, which are truly transferable between species. This review is intended to be a starting point for discussion about translational phenotyping approaches among plant scientists, and provides a useful compendium of methods and techniques used in modern phenotyping for this specific plant pair as a case study.The convenient model  The quest for drought resistant genotypes has been, for a long time now, one of the principal challenges in plant sciences: Drought stress can seriously hamper crop development leading to a decrease in yield, with serious socio-economic consequences . HistoriClimate change influence on temperature and rainfall occurrence and intensity is rapidly mutating the water balance of ecosystems, resulting, amidst other extreme climatic phenomena, in unusually extended drought periods in temperate countries . ConsequIn the beginning, new drought resilient plants were obtained by conventional breeding among promising genotypes, exploiting the genetic pools offered by natural variation . FollowiArabidopsis thaliana (Arabidopsis) has been for decades the most important model for genetics and molecular biology of angiosperms due to numerous characteristics that made it very convenient for research )Arabidopsis and Solanum [RSA phenotyping allows dynamic interactions between roots and their surrounding substrate to be understood by evaluating, e.g., fine root diameters, specific root length, root angles and root length density (reviewed by ). Unders Solanum ,134. How Solanum .Arabidopsis model is due to its compact habitus unsuitable for translation of most information in this respect. The differences in growth habitus between Arabidopsis and tomato indeed complicate a homogenization of phenotyping methods regarding vegetative growth. While the rosette-like structure of Arabidopsis allows relatively straightforward analyses, the three-dimensional structure of tomato is more difficult to parametrize. For tomato indeed, not only leaf area index (LAI), but also leaf area density (LAD) in several horizontal layers within a high-wire-system tomato canopy have been analyzed with the LiDAR-based system described above [Arabidopsis instead, 3D plant architecture analyses are not common, as its rosette-like structure is rather plain. So, the additional information on the third dimension does not seem to justify the effort needed to capture it, and stress effects can be detected as projected leaf area observed non-destructively via RGB cameras located above the plants [Leaf area densities and related source-sink relationships are known to be important for final yield in horticultural crops  and graied above . In Arabe plants .Arabidopsis [Arabidopsis [Arabidopsis flowers earlier under long-day conditions [Arabidopsis and tomato have to take day length (in-) sensitivity of the respective plant into account, either through appropriate experimental design and/or through statistical models.Early fruit set is also part of the drought escape strategy and therefore a symptom of drought stress both in bidopsis  and tomabidopsis . Many gebidopsis . Howeverbidopsis , whereasnditions . Thus, rArabidopsis [Fruit yield is a highly integrative phenotypic trait, and genetically controlled by at least 28 QTLs in tomato . Operatibidopsis  and tomaArabidopsis and tomato that has to be considered is that the short life cycle in the former ends with fruit production, whereas constant fruit production over months and theoretically over years is possible with indeterminate tomato varieties.Another important difference in reproductive physiology of After sensing drought, plants start activating a complex network of gene-expression changes affecting plant behaviour. While some of these may vary among plant species, others are pretty well-conserved, thus, representing a signature of drought stress. Transcripts of such marker genes are often quantified in physiological studies and can be used to monitor stress response intensities.Describing the specific intricacies of molecular responses during drought stress, a complex and still partially elusive network, is far from the purposes of this review; among the impressive body of literature on the topic, the reader is referred to two up to date and influential reviews ,142. HerNCED (9-Cis-Epoxycarotenoid Deoxygenase) genes, which catalyze one of the last steps of ABA biosynthesis, can be used to monitor plant sensing of drought stress in tomato and Arabidopsis. AtNCED3 is expressed quickly during drought stress [Arabidopsis leaves lose turgor [SlNCED1 and SlNCED2 seem to play similar roles [AtRD29B (Responsive to Dehydration 29 B) is typically profiled in drought stress experiments [SlRD29 [Some of the most prominent molecular responses to drought stress are governed by the stress hormone ABA (abscisic acid). Firstly, ABA biosynthesis is augmented during stress through the transcriptional induction of the genes encoding its biosynthetic enzymes. Among these, the t stress  as soon e turgor . In tomaar roles ,96. ABA-eriments ,99 and p [SlRD29 .Arabidopsis is Homeobox Protein 6 (HB6), an ABA-activated gene in drought stress that encodes a transcription factor governing several stress responses [Solyc02g084850 is a good drought marker (our unpublished data) still uncharacterized in Arabidopsis.Another commonly used drought stress marker gene in esponses ,101; howDREB2 (Dehydration-responsive Element-Binding protein 2). Both AtDREB2A and SlDREB2 expression is induced in either plant species by drought stress [DREB2 are, consequently, good putative stress markers as well.In some cases, such as the study of genotypes with disturbed ABA sensing/biosynthesis, the use of ABA-dependent stress markers may not be appropriate. In this case, ABA-independent, drought-activated genes can be used instead; one of these is t stress ,99,102, Arabidopsis and crops.Nowadays, more than 200 angiosperm species have been sequenced, and this number is predicted to increase rapidly . TogetheAs technology evolves, phenotyping facilities addressing multiple traits simultaneously are becoming the new standard in plant phenotyping ,144. TheArabidopsis model is more valuable than ever, especially if the scientific community manages to translate it to crop models from which we can obtain a real advantage, including in food, fodder or fibre. We are convinced that knowledge can be better translated between species in relation to mechanisms involved in tolerance against abiotic stresses like drought, as well as on many other plant traits, such as fruit development, light response, or resistance against pests and diseases. At present, the transferability of knowledge is still limited, as stress protocols, as well as phenotyping protocols  are often incoherent among different species. Researchers interested in translating the vast knowledge gained on Arabidopsis to crops and vice versa must carefully design their studies and ideally build interdisciplinary teams to gather knowledge on genetic background, expected and desired phenotypes and on the agricultural production systems the crops are grown in.The knowledge gathered on the Arabidopsis  may be s (e.g., , and may


The media has consistently described older adults as the population most vulnerable to COVID-19. Anti-ageism critics have taken issue with the oft-repeated statement that “only” older adults are at risk, a construction that dismisses and devalues the nuances within this population. The purpose of this study was to analyze instances of ageism in national media sources during the first month of the COVID-19 pandemic. A systematic search returned 287 articles concerning older adults and COVID-19 published in four major newspapers in the United States—USA Today, The New York Times, Los Angeles Times, and The Washington Post—between March 11 and April 10, 2020. Combining the strengths of content analysis and critical discourse analysis, we deductively and inductively reviewed the articles for patterns related to implicit and explicit forms of ageism. While ageism was rarely discussed explicitly, ageist bias was evident in implicit reporting patterns, such as frequent use of the phrase the elderly, which was often paired with statements describing older adults as vulnerable. Infection and death rates among older adults, as well as institutionalized care practices, were among the most commonly reported topics, providing a limited portrait of aging during the pandemic. While some authors utilized a survivor narrative by portraying older adults as having survived hardships, this construction implicitly places blame on those unable to do so. Older adults, when quoted directly, produced more complex and nuanced narratives of aging during the COVID-19 pandemic. Such narratives can combat societal ageism and promote self-determination and -definition.


High-performance nanomaterials have been seen as a new generation of electrochemiluminescence (ECL) probes or emitters for their finely tunable structure and concomitant remarkable properties, guaranteeing the prospective applications in the analysis and diagnosis devices with superior performances. The structure–activity relationships of ECL nanoprobes in nanoscale are presenting milestone in understanding of the ECL microscopic behaviors and mechanisms, and guide the exploitation of novel ECL probes. In this mini-review, we summarized the recent development of novel ECL probes based on the nanomaterials. The mechanism and relationships between their structure as well as the active sites and functionality were revealed. In addition, the design and regulation of the ECL nanoprobes were emphasized for the biosensing and imaging application. Finally, the potential prospect of the ECL nanoprobes, design, and their applications were discussed. For example, since 32+  with TPrA (tri-n-propylamine) as a co-reactant, it has attached much importance by the fields like clinic diagnosis, and has been one of the most advanced immunoassay systems currently. Evolving the ECL based on Ru(bpy)32+ and TPrA, typical ECL immunodiagnostic products like Elecsys® (Roche Diagnostic) are widely used worldwide, taking an irreplaceable part in the analytical chemistry and diagnosis.Electrochemiluminescence , first reported by 32+ which can generate luminescence signals during electrochemistry process are defined as ECL probes, playing the most significant role in ECL procedure. Traditionally, ECL probes can be classified as inorganic probes and organic probes in accordance with their functional component. Inorganic ECL probes, represented by coordination complexes (symbolic example like Ru(bpy)32+ and derivatives, and other probes with semblable coordination mode can see Ir(Ⅲ) complexes investigated by Uniformly, the emitters like Ru(bpy)Fortunately, ECL nanoprobes are firmly believed capable to improve such dilemmas. Recent years, increasing importance are emphasized to the development of high-performance nanomaterials. Extend to the domain of ECL, nanoprobes with smaller size, larger specific surface area, as well as higher surface activity are competent to provide more subtle optical information. Moreover, benefited from advanced synthesis methods as well as rational design and post-modify strategies, ECL nanoprobes possessing controllable morphology, tunable energy level, and component are attached more valuable features like extremer luminescence efficiency, remarkable luminescence intensity, and more flexible adjustability of photophysical properties.Thanks to the unremitting investigation and evolution of ECL nanoprobes , the numExploration, design, and application of ECL nanoprobes are of the essence for chemistry. What follows in this review would sort the novel ECL systems and application cases which got development lately, and related potential prospect would also be summarized.Along with the progress of high-performance nanomaterials’ preparation and investigations, multiple kinds of nanomaterials were found to be able to be promising candidates for high-efficiency ECL nanoprobes. At the meantime, rational design and post-modify strategies of them can usually demonstrate convincing relationships between their structure and ECL activity.32+ (or their derivatives) on nanoparticles to react with co-reactant TPrA has been developed as a mature strategy to achieve stable and enhanced ECL for better analytical performance . Meanwhile, novel illustration about luminescence pathway can also be found. Like 32+ triethoxysilane derivative (denoted by Rubpy-TES) doped core–shell silica–PEG nanoparticles will swap when the doping level of Ru(bpy)32+ changed. Such findings induced well-known systems into nanoprobes, offering persuasive evidence and results to understand ECL behaviors at nanoscale.Conventional ECL probes could be an appropriate starting point of this section. Combining with advanced nanotechnologies, traditional emitters are still of crucial value to study. For instance, doping Ru(bpy)ance see ; etc.,. 2O82-. The additional shell, ZnS, played an essential rule in this QDs ECL system for its much wider bandgap, providing an important protective barrier, thereby guarantying ECL get rid of the influence from surface traps  can usually present inspiring potential for excellent ECL properties since they possess confinement effect of electron/hole structure, different from bulk semiconductors within which electrons can transfer freely . Therefoce traps . The so-via wavelength. Besides, precise doping of Mn2+ to vacancy point, which can be treated as an efficient method of post-modify, successfully made ECL emission peak red-shift and blocked the VD-induced ECL. This work showed a valuable and reliable design strategy since all the energy-transfer paths followed energy band principles strictly. Additionally, the clear description of nanocrystals, defects, doping cases, and their corresponding ECL behaviors revealed the structure–function relationships distinctly. Following this general idea, AD-assisted enhancement of ECL  of nanoprobes are not always unfavorable to ECL. t of ECL  and monot of ECL  were alsvia host–guest recognition) showed efficient ECL with standout monochromaticity  shell–core nanohybrid ECL sensors by working with co-reactant S2O82-.Generally, recent years witnessed the revolution of ECL biological analysis, from conventional detection of biomarkers statistically to the observation of single cells or intracellular matters. Literally stating, nanoprobes played a vital role in this process. For instance, 32+@SiO2/Au nanoparticles—TPrA ECL system, Although plenty of high-efficiency ECL emitters and strategies were discovered and employed, the urgent pursuing of space-resolved ECL approaches and related probes has never been slowed down. Since As demonstrated above, such strategies showed meaningful picture of ECL in the domain of biological analysis, which can be regarded as a profit from recent improvement of nanoprobes on the one hand. On the hand, developing ECL nanoprobes also benefits the visualization of various unpredictable electrochemical mechanisms, for instance, using ECL imaging to track and evaluate electrolytic performance.32+ by enhancing the oxidation of co-reactant TPrA, in situ catalytic behaviors of Au nanoparticles, which extraordinarily expanded the application value of nanoprobes’ ECL imaging, leading to the faith in more distinct understanding of electrochemistry at nanoscale. Based on this property, the electrocatalyst screening can be evaluated by ECL techniques.Considering that Au nanoparticles would improve the generation of ECL from Ru(bpy)32+ to Ru(bpy)33+ than ordinary Au or Pt nanoparticles, which can be certified by the subsequent digital simulation. As described above, the electrochemical redox process of Ru(bpy)32+/TPrA system could generate ECL signals. Meanwhile, since nanoparticles were capable to catalyze their transformation, the total ECL intensities calculated from imaging results could straightforwardly mirror their catalytic capacity. However, in order to obtain lower ECL background at this work, the authors coated a thin Al film on the ITO electrodes, which sharply reduced ECL sensitivity, no good for the imaging quality. What is more, only one kind bimetallic probes were investigated by this work, which cannot satisfy current inquiries about increasingly more composite and doping nanostructures.Following such fundamental logic, 2O2 ECL system to generate high-intensity signals free from interference. Therefore, electrocatalytic performance of four nanostructures could be ranked based on ECL intensities, from which structure dependent electrocatalytic/electrochemical reactivity of those nanoprobes was visualized and demonstrated.Hence, based on the pursuing of greater signal to background ratio and better understanding for the catalytic activities of composite nanoparticles, A common ground between the works by 2O82- as co-reactant). Since ECL and HER both happen at the surface of electrodes, the H2 bubbles generated from HCNSs during HER affect the ECL signals of HCNSs, and the “ECL blinking” phenomenon was observed and defined. It was found that the ECL blinking behaviors were altering simultaneously with the evolutionary process  of H2 bubbles during HER. Meanwhile, both the ECL ON-time and OFF-time figured up from two HCNS nanoprobes were of proportional relation with the inverse power–law distributions at probability density dimension. Therefore, it made sense to establish correlation between such ECL imaging behavior and HER activity. Even though some HER catalysts are not able to be ECL emitters, their HER performance can also be evaluated by loading on HCNSs. Hence, this work could enlighten increasingly more electrocatalysts and electrolytic reactions be investigated.To explore the applications of ECL beyond the ECL intrinsic electrochemical system, most recently, Via the optical data originating from the ECL signals of Ru(bpy)32+-doping silica nanoparticles (RuDSNs) and TPrA, collision trajectory and collision mode  showed the tendency that more dimensions for ECL analysis are essential currently. Considering the features of wave optics, another good attempt to “increase dimensions” is polarized ECL, which was first put forward by 2O82- triggered isotropic ECL emission to polarized emission. Such polarized ECL systems might not only provide some additional information on the structural symmetry or polarization state of rather than that merely in spatial structure of probes, showing the great prospect of nanoparticles in photophysics of ECL, matching the desire of multidimensional analysis.Like introduced above, the impetus trying to build the electrochemical behaviors of nanoprobes at In this mini-review, we summarized recent progress about novel ECL nanoprobes based on traditional ECL systems, semiconductor structure probes , and some others, and emphatically highlighted the idea of “rational design” and “post-modify” to illustrate their structure–activity relationships, trying to provide some guidelines in the future preparation of ECL nanoprobes. Meanwhile, the novel analytical applications of current ECL nanoprobes were revealed by several typical instances, exhibiting the promising application prospect of ECL and nanoprobes.The future of ECL nanoprobes should still focus on the design of highly efficient and stable luminescence emitters, to satisfy the urgent need of ECL domain like high-resolution space–time imaging and dynamic analysis in complex systems. To meet the requirement, the deep understanding of the ECL mechanism in the novel system should also be emphasized. Moreover, how to develop multidimensional ECL better and make the best use of their photophysical features is worthy of our continuous considering, which will further extend the applications of ECL techniques.


Multipotent mesenchymal stem cells (MSCs) possess regenerative properties and have been shown to improve outcomes and survival in acute and chronic lung diseases, but there have been some safety concerns raised related to MSC-based therapy. Subsequent studies have demonstrated that many of the regenerative effects of MSCs can be attributed to the MSC-derived secretome, which contains soluble factors and extracellular vesicles (EVs). MSC-derived extracellular vesicles (MSC-derived EVs) replicate many of the beneficial effects of MSCs and contain a variety of bioactive factors that are transferred to recipient cells, mediating downstream signaling. MSC-derived EV therapy holds promise as a safe and effective treatment for pulmonary disease, but there remain many scientific and clinical questions that will need to be addressed before EVs are widely applied as a therapy. To date, the use of MSC-derived EVs as a treatment for lung disease has been conducted primarily in in vitro or pre-clinical animal models. In this review, we will discuss the current published research investigating the use of EVs as a potential therapeutic for acute lung injury/acute respiratory distress syndrome (ALI/ARDS), bronchopulmonary dysplasia (BPD), idiopathic pulmonary fibrosis (IPF), pulmonary arterial hypertension (PAH), asthma, and silicosis. Stem cell research has garnered significant attention over the last few decades, especially regarding potential applications for the regeneration of damaged or diseased tissue . Stem ceEVs are naturally occurring, cell-derived membrane-bound spherical structures that are shed or secreted from most cell types under various physiologic and pathologic conditions into the circulation or surrounding body fluids. EVs have been found in a variety of biological fluids, including blood, bile, bronchoalveolar lavage fluid (BALF), and urine . EVs areEVs range between 30–5000 nm in diameter, and until recently have been classified based on size, which included exosomes (30–100 nm), microvesicles/ectosomes (100–1000 nm), and apoptotic bodies (100–5000 nm). These categories of EVs also have distinct mechanisms of release from cells, as described below. The International Society for Extracellular Vesicles (ISEV) now recommends using “extracellular vesicle” as an umbrella term to describe vesicles of all sizes, due to the size heterogeneity and lack of specific markers for each EV size classification . The mosThe formation of EVs include three distinct biogenetic pathways. First, exosome biogenesis is dependent on the endosomal sorting complexes required for transport (ESCRT) protein for the formation of endosomal intraluminal vesicles resulting in multivesicular bodies (MVBs), from which exosomes are released via exocytosis when MVBs fuse with the plasma membrane ,15. ThisEVs express cell surface proteins similar to the cells from which they are derived and contain various cytoplasmic components, including proteins, RNA (including mRNA and noncoding RNA such as microRNA (miRNA)), lipids, mitochondria, and DNA . The conThe cellular targeting, uptake, and plasma membrane fusion of EVs is another area of ongoing investigation that is proving to be complex and multifaceted ,15,20,21The use of MSC-derived EVs as a potential therapy for lung disease is a fairly young but rapidly growing field, with current research covering a wide variety of lung diseases  22]. Th. Th22]. Acute lung injury and acute respiratory distress syndrome (ALI/ARDS) is a disease that carries a high burden of morbidity and mortality ,24,25. IEscherichia coli pneumonia murine model, Monsel and colleagues demonstrated that the intravenous (IV) administration of human bone marrow MSC-derived EVs improved survival and reduced inflammation [MSC-derived EVs have demonstrated beneficial effects in both bacterial- and viral-induced ALI. In an influenza-induced ALI porcine model, intratracheal (IT) treatment with swine bone marrow MSC-derived EVs 12 h after influenza virus infection resulted in reduced viral replication, viral shedding, and reduced production of pro-inflammatory cytokines . In an Eammation . This stammation .E. coli endotoxin-induced ALI model resulted in improvements in pulmonary edema and lung protein permeability due to reduced extravascular lung water and total protein in BALF [A variety of studies have used an endotoxin-mediated in vivo model to evaluate the effects of MSC-derived EVs for ALI/ARDS. For instance, the IT instillation of human bone marrow MSC-derived EVs in a murine  in BALF . Additio in BALF . Further in BALF . Tang an in BALF . In addi in BALF . This is in BALF . Gennai  in BALF .Bronchopulmonary dysplasia (BPD), a leading cause of morbidity and mortality in premature infants, is a chronic lung disease that is multifactorial in origin . The lunIdiopathic Pulmonary Fibrosis (IPF) is a chronic and progressive fibrosing interstitial lung disease with pathophysiology characterized by fibroblast proliferation and extracellular matrix remodeling . IPF hasA study by Mansouri et al. demonstrated that intravenous human bone marrow MSC-derived EVs could prevent and reverse intratracheal bleomycin-induced pulmonary fibrosis in mice with resulting improvements in both pulmonary morphology and lung architecture and decreased collagen deposition . They alPulmonary Arterial Hypertension (PAH) refers to hemodynamic alterations of the pulmonary circulation in which the pulmonary artery pressures are >25 mm Hg. PAH is a progressive disease characterized by the remodeling of the pulmonary arteries, increased pulmonary infiltrates, loss of vascular cross-sectional area, and elevated pulmonary vascular resistance leading to right heart failure and death ,65. A keTo evaluate the effect of MSC-derived EVs on pulmonary vasculature, Lee and colleagues used a murine hypoxia-induced pulmonary hypertension murine model . They deSilicosis is an occupational disease caused by the inhalation of dust containing crystalline silica particles, resulting in lung inflammation and leading to a progressive and irreversible lung fibrosis ,68. ChoiAspergillus hyphal extract in a murine model of severe asthma [+CD4+ T cells in BALF and in static lung elastance. This data suggests that MSCs and MSC-derived EVs have similarities and differences in their downstream immunomodulatory effects. In a different study, MSC-derived EVs were able to promote regulatory T cell proliferation by increasing the expression of IL-10 and TGF-β1 in peripheral blood mononuclear cells from asthmatic patients [Asthma is a chronic inflammatory lung disease characterized by intermittent airway obstruction, inflammation, increased mucus production, and bronchial hyperresponsiveness. There are multiple therapies on the market, but many patients continue to have poor disease control despite maximal treatment with current therapies. As a result, there is ongoing research to identify novel asthma therapies to both control and treat asthma exacerbations. For example, Cruz and colleagues successfully used MSC-derived EVs to alleviated allergic airway inflammation after the repeat mucosal exposures of e asthma . In anote asthma . While tpatients .MSC-derived EVs are now being considered as an effective and safer alternative to live MSC transplantation and have been shown to have therapeutic benefit in numerous pre-clinical models of pulmonary injury and disease. Nonetheless, the research is still in its infancy and there are still many unanswered questions that need to be addressed before MSC-derived EVs can become serious candidates as a clinical therapeutic.There will need to be standardized protocols for the production, purification, characterization, and storage of EVs, as currently there are no universally accepted protocols. The techniques used for EV quantification include protein concentration, nanoparticle tracking analysis, tunable resistive pulse sensing, vesicle flow cytometry, surface plasma resonance, and electron microscopy. There are many recent reviews describing the predominant methods currently being used in the field ,70,71,72MSCs are harvested from a variety of donors, hence a better understanding of how donor characteristics may alter EV function will also need to be explored. There is much interest in personalizing medicine, and MSC-derived therapies are ideally suited for this approach, as autologous MSCs could be used to generate personalized EVs as a therapy. Thus, fully understanding the donor effects on EVs would become very important when considering using autologous EVs as therapy.The treatment of lung diseases will require EV administration via IT instillation or IV injection. The optimal route of administering EVs and the bio-distribution of EVs based on the cellular origin of the EV and the route of administration will need to be clarified. Currently, there are no published reports of the bio-distribution of EVs after IT instillation. However, the intravenous injection of EVs into mice has been shown to initially localize primarily to the spleen and liver and then rapidly redistribute to the gastrointestinal tract and lungs, followed by renal and hepatic clearance ,75. TherIn conclusion, we have summarized the current literature on the use of MSC-derived EVs as a treatment for acute and chronic lung disease. MSCs have shown promise for their regenerative and immune modulating effects but have been questioned as a therapeutic due to possible deleterious side effects. MSC-derived EVs are shown to be able to mediate many of the beneficial effects of MSCs, and it is postulated that they have fewer negative side-effects. Pre-clinical animal models have demonstrated that MSC-derived EV-based therapy may be a viable therapeutic option for the reversal or prevention of various lung diseases. This beneficial effect appears to be the result of the activation of the signaling pathways via the transfer of EV contents containing miRNA, RNA, and proteins; however, the molecular mechanisms of the different EV components is poorly understood. The current literature using MSC-derived EVs as a treatment for lung diseases in pre-clinical animal models use different methods of quantifying the dose of EVs, making direct comparison between the studies impossible. A consensus on the best method to define the EV dose needs to be determined so the therapeutic effectiveness of EVs can be compared across studies. In addition, there remains the challenge of scaling-up the production of EVs to a level that could be produced with clinical-grade quality and quantity. Thus, there are many questions and technical issues that need to be addressed before MSC-derived EVs can make the transition from animal models to humans. If these different challenges can be overcome, then MSC-derived EVs hold great promise as a potential therapeutic for numerous lung diseases.


Due to the coronavirus disease (COVID-19) pandemic, medical schools have paused traditional clerkships, eliminating direct patient encounters from medical students’ education for the immediate future. Telemedicine offers opportunities in a variety of specialties that can augment student education during this time. The projected growth of telemedicine necessitates that students learn new skills to be effective providers. In this viewpoint, we delineate specific telehealth opportunities that teach core competencies for patient care, while also teaching telemedicine-specific skills. Schools can further augment student education through a variety of telemedicine initiatives across multiple medical fields. The explosion of telemedicine programs due to the pandemic can be a catalyst for schools to integrate telemedicine into their current curricula. The depth and variety of telemedicine opportunities allow schools to continue providing high-quality medical education while maintaining social distancing policies. On March 23, 2020, the Association of American Medical Colleges (AAMC) issued guidance on medical student involvement during the coronavirus disease COVID-19) pandemic, recommending that medical students not participate in direct patient care, unless there is a critical workforce need, and only on a voluntary basis [9 pandemiThe cornerstone of medical education is patient care. While direct patient care has been appropriately limited at most schools, the lessons learned from these critical patient interactions cannot be fully replaced by readings, lectures, case studies, or online modules. Certainly, direct in-person care has important educational value to the medical student. Although not an exact replacement for in-person care, institutions have a unique opportunity to bridge this current gap in clinical education with telemedicine. This viewpoint contributes to the ongoing discussion by offering specific guidance on how schools can incorporate medical students into the massive expansion of telehealth initiatives during and beyond the COVID-19 pandemic.In recent years, telemedicine has grown exponentially. Telemedicine applications have expanded from phone triage and ambulatory electronic visits (e-visits) to include mental health, postsurgical operation follow-ups, and specialty consults . In suppWith the imminent growth of telemedicine, teaching providers this unique skillset is essential for the success of future telemedicine programs. Practitioners who have not used telemedicine may find it difficult to translate their traditional interview and physical exam skills to the virtual sphere . FurtherEducation is critical to the future success of telemedicine implementation. Integration of telemedicine into undergraduate medical education allows for all future physicians to have access to this type of training. The American Medical Association (AMA) has similarly articulated the value of telemedicine curricula in medical schools and residency programs . In addiThere are a number of suggested skills associated with optimal telemedicine care. These include but are not limited to the following: communication, physical examination, professionalism, and technological literacy. Communication skills include clear enunciation and the minimization of body motions, gestures, and colloquial speech . These mSome programs have already started to institute courses to teach these telemedicine communication and evaluation skills. Among the most common curricular elements includes standardized patients ,9. HowevTelemedicine curricula could mimic traditional curricula that medical educators already employ with minimal adjustments, allowing for the continuation of clinical education during the pandemic. Students can easily access and get involved with telemedicine initiatives through technological platforms. For example, some electronic health record (EHR) platforms already integrate third-party platforms that would allow three-person calls . Other oEducating students through telehealth initiatives could parallel a majority of the 13 Core Entrustable Professional Activities for entering residency (EPAs) . These EBy integrating multiple EPAs, a single e-visit could provide students exposure to skills that they will employ throughout their careers. A telemedicine student visit could be similar to most other ambulatory student clinic visits. To ease patient burden and reduce confusion, ideally the patient, the medical student, and the physician would all be present concurrently at the e-visit. The attending could observe the student lead as much of the visit as possible, with a seamless transition when the attending takes lead of the conversation. Using telehealth technology, students could gather a history and perform a virtual physical exam (EPA 1), focusing on communication skills, rapport building, and functional physical exam maneuvers. Students could summarize their findings (EPA 6) to both the patient and attending in patient-friendly language. Depending on the comfort level of the patient, the physician could lead the student through a discussion of the possible diagnoses (EPA 2), potential diagnostic tests (EPA 3), or orders (EPA 4). This style would mimic “family-centered rounds,” providing the student the opportunity to think through an assessment and plan with direct patient input. Alternatively, the physician and student could discuss the clinical reasoning and key academic concepts after the patient’s call. Following the visit, the medical student would then document the encounter (EPA 5). To augment these visits, medical students could research topics inspired by patient cases (EPA 7) and discuss the impacts of telemedicine on health care (EPA 13). These examples show how the e-visit can provide in-depth, high-quality education. A single e-visit can provide the opportunity to teach medical students invaluable skills that they would otherwise not be able to obtain from reading case studies.With the expansion of novel telemedicine applications into more fields, including procedural fields, students can continue to learn foundational knowledge through engagement in telemedicine even beyond the stereotypical telemedicine visit. In fact, involvement in virtual opportunities exist for all EPAs , thus maEven if students are to return to the clinical setting soon, traditional clerkships may look different for them. For example, some students may have continued needs for self-quarantining while others may return to the hospital. For students in hospitals, there will likely still be patients admitted for COVID-19. In this context, telemedicine may provide a way to resume medical student education while maintaining a safe environment. For example, students can participate in consults or rounds without stepping into patient rooms through the use of tablets ,5. In faAs telemedicine is further implemented, clinical skills unique to remote care have become necessary. The rapid adoption of telemedicine due to the COVID-19 outbreak can serve as an opportunity to augment medical education curricula and to continue to provide medical students with critical educational opportunities when in-person encounters are limited. If hospital systems are already building and expanding videoconferencing and e-visit tools for providers, why not include students as well?


Cardiac magnetic resonance (CMR) has emerged as new mainstream technique for the evaluation of patients with cardiac diseases, providing unique information to support clinical decision-making. This document has been developed by a joined group of experts of the Italian Society of Cardiology and Italian society of Radiology and aims to produce an updated consensus statement about the current state of technology and clinical applications of CMR. The writing committee consisted of members and experts of both societies who worked jointly to develop a more integrated approach in the field of cardiac radiology. Part 1 of the document will cover ischemic heart disease, congenital heart disease, cardio-oncology, cardiac masses and heart transplant. Since its initial utilization during early 1980s, cardiovascular magnetic resonance imaging (CMR) has evolved from a niche modality into a new mainstream tool, changing diagnostic paradigms in various cardiovascular settings. It provides unique information to support clinical decision-making, allows accurate prognostic stratification and has proven to be highly cost-effective in different scenarios –5. Many Articles are structured to define CMR appropriateness for the first diagnosis and follow-up in various clinical scenarios.Strong recommendation: there is evidence, general agreement, or both, that the test is useful (benefit ≫ risk).Moderate recommendation: there is conflicting evidence or opinion about the usefulness of the test; the weight of evidence/opinion however, is strongly in favor of the test’s usefulness. (benefit > risk).Weak recommendation: the test’s usefulness is less well established; there is a small net benefit (Benefit ≥ risk).No recommendation: there is evidence or general agreement that the risk/harm outweighs benefits (Benefit = or < risk).Expert opinion: there is insufficient evidence or evidence is unclear or conflicting, but this is what the working group recommends. Further research is recommended in this area.As the third step, assigned scores were agreed in consensus by all authors and unanimously approved.First, the writing committee discussed the table of content and assigned referrals for each chapter. Second, each referral conducted literature searches and drafted the assigned section, highlighting indications and rating them according to the following scores:Two scenarios including patients symptomatic for stable chest pain and patients presenting as acute coronary syndrome (ACS) such as ST elevation myocardial infarction (STEMI) will be considered. The main clinical indications for ischemic heart disease (IHD) are summarized in Table CMR could be used to provide coronary artery imaging and reversible ischemia. The coronary artery imaging by CMR is achievable with non-contrast whole-heart coronary magnetic resonance angiography (MRA) that can provide visualization of the coronary tree within a single three-dimensional acquisition with an average sensitivity, specificity and negative predictive value of 88%, 72% and 88%, in a patient-based analysis, respectively . HoweverIn this setting, CMR can be helpful in both the diagnostic pathway and in prognostic stratification . CMR is CMR can be appropriately used in the assessment of cardiac anatomy and function, blood flow, and extra-cardiac vascular structures in patients with simple and complex congenital heart disease (CHD). CMR is recommended for studying pediatric or juvenile populations that often require repeated follow-up examinations over time and is preferred in this context rather than computed tomography (CT) or cardiac catheterization, to avoid the use of iodinated contrast medium and ionizing radiation. Unfortunately, routine clinical use of CMR in new-borns and small children is hampered by the need of anesthesia. Despite echocardiography is the first line test in this setting, its image quality could be limited in several cases. Consequently, also considering its elevated reproducibility in the measurement of cardiac functional parameters, CMR has become a first-line investigation for several indications, especially in follow-up of surgical patients and in the evaluation of complex anomalies . The maiCMR performs better than echocardiography in the analysis of visceral situs  and anatomy. The multi-planar nature and wide field of view of CMR enables a good appreciation of the whole thoracic and abdominal structures in a few images. CMR is particularly accurate in identifying malformations and assessing the systemic venous return during preoperative evaluation .CMR is reliable in the diagnosis and overall assessment of atrial septal defects (ASD), although transoesophageal echocardiography (TOE) represents the gold standard in this setting. CMR also correlates with cardiac catheterization for the invasive quantification of shunts. CMR overcomes the limitations of the other imaging modalities in the presence of atypical defects, like sinus venosus or when associated with a partial anomalous pulmonary venous return (PAPVR) . AccordiCMR is accurate in the study of discordant atrio-ventricular connections, valve atresia or atrio-ventricular defects and ventricular septal defects (VSD). However, it does not add significant data to echocardiography except for shunt quantification. CMR is useful in the presence of VSD associated with complex anomalies and in the preoperative evaluation of complex CHD. CMR may provide additional information in Ebstein’s disease  and may be indicated in the follow-up of right ventricular structure and function .CMR is able to accurately quantify valvular regurgitation and stenosis by using phase contrast (PC) imaging sequences with adequate correlation to other traditional imaging modalities and it is highly reproducible. For this reason, CMR plays a central role in the serial follow-up of pulmonary regurgitation for corrected Tetralogy of Fallot (TOF) patients as well as for valvular or surgical conduit stenosis .CMR is an extremely accurate method for studying all the diseases of the aorta, allowing precise measurements of aortic dimensions and the extent and morphology of the disease. MRA is well suitable for studying aortic arch anomalies, particularly utilizing 3D electrocardiogram (ECG) -gating with respiratory navigator sequences . LocatioThis is the most well-established indication of CMR given its high reproducibility in studying biventricular function and considering that serial functional evaluation is crucial in the follow-up of these patients. Right ventricular function evaluation is of outmost importance in the follow-up of surgically managed CHD, such as TOF, pulmonary atresia with VSD, and transposition of the great arteries (TGA) treated with atrial switch. Other information exclusively provided by CMR is the depiction of myocardial fibrosis by LGE imaging, which stratifies risk of arrhythmias for these patients . CMR is Coronary MRA allows an accurate depiction of the origin and proximal course of the coronary arteries without the administration of contrast medium and ionizing radiation , 32.Figure Advances in cancer treatment led to improved survival of patients but have also increased morbidity and mortality due to treatment side effects. Cardiotoxicity can be classified in type 1  characterized by dose dependent myocardial injury, more likely to be irreversible, and type 2  with a higher likelihood of recovery after discontinuation of the offending agent. Anthracyclines are one of the most studied examples of cardiotoxicity as this class of drug has high efficacy for treatment of solid tumors and hematological malignancies but may also cause irreversible cardiac damage, which can be acute, early or late, affecting prognosis .Cardiotoxicity is defined as a decline of left ventricle ejection fraction (LVEF) ≥ 5% from baseline in symptomatic patients or ≥ 10% in asymptomatic patients to less than 55%. The 2014 Expert Consensus Statement for Multimodality Imaging Evaluation of Adult Patients during and after cancer therapy from the American Society of Echocardiography and the European Association of Cardiovascular Imaging updated the definition of cancer therapeutics-related cardiac dysfunction as a decrease in LVEF of > 10% to a value < 53%.CMR is recognized as a method to screen for chemotherapy-related cardiotoxicity in case of poor transthoracic echocardiography (TTE) image quality due to its accuracy, reproducibility and ability to detect subtle changes in right and left ventricular function . CMR alsDiffuse myocardial fibrosis induced by anthracycline therapy can occur several years after completion of treatment and it can be assessed by T1 mapping . HoweverSome small studies using CMR have also shown myocardial edema early following anthracycline therapy by using T2-weighted sequences. The presence of edema has been associated with persistent reduction in RV function in follow-up examinations .Finally, ECV is also increased in patients after anthracyclines therapy as compared to healthy controls .CMR is also appropriate in monitoring cardiac involvement in cancer-related treatment, providing distinct bio-signatures of early inflammatory involvement (raised native T1 and T2) and interstitial fibrosis and remodelling (raised native T1 but not T2), thus providing an algorithm allowing to identify susceptible myocardium to potentially guide cardio-protective treatment measures .Moreover, radiation therapy improves cancer-related outcomes in a variety of malignancies such as lymphoma, breast, lung, and head and neck cancers. Radiation-induced heart disease is a serious side effect of cancer treatment, which may manifest as pericarditis (acute and subacute), pericardial effusions or late effects (10–15 years after exposure) related to cardiovascular fibrosis which can lead to diverse clinical manifestations including heart failure, constrictive pericarditis, restrictive cardiomyopathy, valvular abnormalities, premature coronary disease and arrhythmias . The maiCMR offers the most comprehensive approach to cardiac masses as it allows to determine the location, pathological substrate, lesion mobility, dynamic perfusion and hemodynamic impact of a suspected cardiac tumor.Differentiation between non-tumoral versus tumoral lesions: anatomic pitfalls refer to the presence of normal cardiac structures or embryological remnants, which can easily be misinterpreted as cardiac masses. These include, among others, the presence of prominent or hypertrophic structures like the moderator band, Eustachian valves, Chiari network, crista terminalis or false cordae tendine. Among pseudomasses , the list of differentials is long and covers a large and heterogeneous series of congenital and acquired conditions. Most common pseudo-tumors comprise intracavitary thrombi, areas of lipomatous hypertrophy, infective or coelomic cysts and abscesses and large hiatal hernias . CMR canDifferential diagnosis between benign and malignant lesions Fig. : besidesSurgical planning: due to the high risk of distal embolization, sudden cardiac death (SCD), and hemodynamic collapse, surgical treatment is often indicated in cardiac tumors and require careful preoperative staging. Patients with primary malignant diseases or metastases may undergo surgery for symptomatic and palliative treatment  whereas radical resection can often be obtained in benign masses. CMR is fundamental to evaluate tumor characteristics, for planning of the preferred surgical approach and reconstruction of the cardiac chambers .Detection and follow-up of tumor recurrences: incidence of local recurrences largely depends on tumor histology and adequacy of surgical excision, reaching a cumulative incidence of up to 13% for cardiac myxomas . CMR supDetection and characterization of myocardial damage following oncological treatments .CMR appropriateness is limited in the evaluation of small lesions attached to fast moving structures. This can be observed in the evaluation of valvular masses, like papillary fibro-elastomas or infective vegetations, which are barely visible if smaller than 10 mm, because of the intrinsically low temporal resolution of cine sequences, as compared to an echocardiographic approach. The main clinical indications for cardiac mass evaluation are summarized in Table CMR is recommended in the diagnostic process and its contribution in the clinical workup is summarized as follows:Heart transplantation is a life-saving therapy for individuals with end-stage heart failure, congenital heart diseases, restrictive cardiomyopathy and infectious cardiac diseases. Acute rejection and accelerated coronary artery disease, considered as chronic rejection, represent the most common clinical problems. Despite stringent selection criteria and significant advances in anti-rejection therapy, the mortality rate is still very high. As result of a non-uniform pathologic process and of a difficult histological interpretation, surveillance with endomyocardial biopsies can underdiagnose the rejection. In the early stages it may occur without major cardiac dysfunction, therefore echocardiographic measurements may lack sensitivity; diastolic measures have shown correlation with acute rejection, however without uniform consistency .In this context, the tissue characterization ability of CMR has suggested its use as a non-invasive tool to diagnose acute rejection. Compared to endomyocardial biopsy, CMR has shown positive results ; howeverThe value of T2-weighted imaging has been evaluated both in animal and human studies with inconsistent results , 48. HowLate gadolinium enhancement is not sensitive and has not been widely investigated. In a small study including patients with different grades of rejection, a higher relative myocardial signal intensity in the early phase post-contrast, a marker of inflammatory, hyperemia, was observed .The new mapping techniques may have an emerging role in the diagnosis of cardiac transplant rejection . MyocardMyocardial ischemia, a component of the rejection process and transplant arteriopathy, lacks clinical symptoms. Adenosine stress perfusion shows a reduction in patients with a prior history of rejection compared with those without .The main clinical indications for CMR in post-cardiac transplantation are summarized in Table


Twenty years after landmark publications, there is a consensus that the somatic hypermutation (SHM) status of the clonotypic immunoglobulin heavy variable (IGHV) gene is an important cornerstone for accurate risk stratification and therapeutic decision-making in patients with chronic lymphocytic leukemia (CLL). The IGHV SHM status has traditionally been determined by conventional Sanger sequencing. However, NGS has heralded a new era in medical diagnostics and immunogenetic analysis is following this trend. There is indeed a growing demand for shifting practice and using NGS for IGHV gene SHM assessment, although it is debatable whether it is always justifiable, at least taking into account financial considerations for laboratories with limited resources. Nevertheless, as this analysis impacts on treatment decisions, standardization of both technical aspects, and data interpretation becomes essential. Also, the need for establishing new recommendations and providing dedicated education and training on NGS-based immunogenetics is greater than ever before. Here we address potential and challenges of NGS-based immunogenetics in CLL. We are convinced that this perspective helps the hematological community to better understand the pros and cons of this new technological development for CLL patient management. Twenty years after landmark publications by the Chiorazzi and Stevenson groups , 2, therIGHV gene SHM status is one of the most robust prognostic markers in CLL, readily identifiable at diagnosis and independent of clinical stage or other biomarkers. More importantly, it remains stable over time, thus contrasting all other well-established prognostic markers, including genomic aberrations, which are affected by or reflect disease evolution . Furtherwww.ericll.org) has been at the forefront of establishing initiatives related to IG gene sequence analysis, particularly with regards to promoting good practices, while also ensuring the widest possible dissemination globally. ERIC accomplishments in this field include: (1) establishment of the ERIC IG Network (http://www.ericll.org/ignetwork/), which aims to promote awareness throughout the hematological community about the need to apply standardized and consistent analytical methods, based on the state-of-the-art in immunogenetics and the most innovative bioinformatics tools: the ERIC IG Network currently comprises 7 European reference labs and more than 100 labs across five continents, certified by ERIC for performing accurate immunogenetic analysis in CLL; (2) organization of educational events that combine lectures, computer-based practical sessions, interactive discussions, and expert panel-led debates on topics pertinent to immunogenetics in CLL; (3) providing the community with an online expert forum (http://www.ericll.org/pages/submission_form) to discuss general queries on IG gene sequence interpretation in CLL or to analyze and provide advice about complex IG gene rearrangement sequences that can be difficult to interpret in everyday practice; (4) creation and maintenance of the ERIC-IMGT/CLL-DB (http://www.imgt.org/CLLDBInterface/query), the largest database of IGHV–IGHD–IGHJ gene rearrangement sequences from patients with CLL, currently holding sequence data from over 32,000 patients collected from 33 institutions spanning Europe, the USA, and further afield; (5) frequent publication of recommendations for the determination of IGHV gene SHM status in CLL, complemented by instructions detailing how to handle analytically challenging cases or cases difficult to categorize , thus hSecond, low-throughput approaches are inherently limited with regards to accurately characterizing: (1) the clonal composition  of a given case and the intraclonal temporal dynamics  ; and,  into their existing workflows. With the introduction of NGS for immunogenetics analysis (collectively termed Repertoire Sequencing (RepSeq)), deeper investigations of IG (and similarly T-cell receptor (TR)) gene rearrangements are now within reach, which could have a profound impact on all applications of immunogenetic analysis, including IGHV gene SHM analysis in CLL .In essence, NGS could offer solutions to the analytical problems mentioned above and, moreover, assist in addressing open issues in immunogenetic analysis in CLL Table . TherefoTP53, but potentially also including NOTCH1, ATM, SF3B1 [when and if these biomarkers will be shown to carry a value for decision-making in CLL], could be obtained simultaneously in a single NGS sequencing run. Indeed, libraries specific for IGHV-IGHD-IGHJ gene rearrangements and a panel of genes meaningful for CLL prognostication or treatment decision can be prepared independently and then pooled for combined sequencing on the same run. An appropriate ratio of these libraries will however need to be determined in order to ensure a sequencing depth appropriate for each gene. On the one hand, the sequencing depth and coverage should be sufficient to facilitate the detection of minor oncogenic mutations, such as those occurring within the TP53 gene, which may influence disease evolution and treatment decisions, while on the other hand attaining deep sequencing is less relevant for the determination of IGHV gene SHM status due to the size of the dominant CLL clone. Generally speaking, such combined workflows would be very attractive, both from a laboratory organization as well as a cost-efficiency perspective.Besides the abovementioned considerations there is another frequently used argument for switching from classical Sanger-based analysis to NGS. Now that an increasing number of molecular diagnostic assays are transformed into NGS-based protocols, it could be very advantageous to combine multiple assays into a general NGS workflow. In the case of CLL, the need to have reliable assays for IGHV gene SHM analysis and IGH marker identification for minimal residual disease (MRD) purposes could in principle be combined via an IGH leader-based NGS strategy. This combination of assays could be extended such that the IGHV gene SHM status as well as the mutational status of several (onco)genes, such as No matter how attractive and promising NGS-based immunogenetic analysis in CLL could be, there are several technical pitfalls and biological questions, which deserve attention before NGS-based immunogenetics can be safely implemented within routine diagnostic laboratory testing.An interesting feature of NGS is that it provides quantitative results, e.g., one can determine clonotype size based on read numbers . HoweverA major concern for using NGS-based methods for clinical applications, such as the determination of IGHV gene SHM status in CLL, pertains to the robustness of the methodology. This is essential, considering that this biomarker is not only prognostically relevant but also predictive , 12, 13,Robust IG/TR gene analysis with NGS critically relies on the availability of dedicated bioinformatics tools. In contrast to other genes, one cannot use tools based on simple comparison with a reference genome. This is due to the fact that: (1) antigen receptor variable domains are created by the assembly of 2 (V and J) or 3  types of genes, and, (2) random nucleotides are deleted and/or inserted at the junctions between these genes, thus resulting in a huge variability of sequences. With the growing interest for immune RepSeq and multiple applications in various scientific and/or medical fields, there has been an intense development of bioinformatics tools designed to address all different issues related to this topic. However, most of these developments stem from research laboratories and require extensive expertise in bioinformatics, which is often limited in clinical laboratories. Moreover, to fully enter the clinical arena, bioinformatics tools need to be compliant with all the requirements of quality assessment schemes, including, to name a few, security issues related to patient data transfer and storage, software maintenance and upgrades.Most Sanger-based protocols rely on direct sequencing, where only the most prevalent sequence appears on the pherograms, provided that the cell population is monoclonal and bears only a monoallelic rearrangement as demonstrated by GeneScan analysis. NGS-based assays offer a far better resolution and may depict a much more complex reality.First, sequence variations of the tumor clonotype will become obvious, even if these differences account for only a minor proportion of sequences, whereas they were, mostly not detected by the low-throughput direct Sanger sequencing technology Fig. . How mucSecond, the higher resolution afforded by NGS methodology will offer the possibility to detect independent, unrelated clonotypes that emerge above what can be considered as “polyclonal background.” Such clonotypes may go undetected by Sanger direct sequencing due to either their small size or inefficient amplification Fig. . Relevanhttps://euroclonalityngs.org/usr/pub/pub.php), which was launched in 2012 and consists of several EuroClonality laboratories experienced in designing assays for detecting IG/TR rearrangements, supplemented by laboratories with expertise in IG/TR gene-based MRD studies, IG/TR  repertoire studies, immunoinformatics, and bioinformatics. In recent years, ERIC and the EuroClonality-NGS Working group have been collaborating systematically on the development of a robust pipeline for NGS-based determination of SHM status in CLL for application with in a routine diagnostic setting, covering both in vitro and in silico aspects. The former pertains to amplification and sequencing while the latter concerns novel bioinformatics solutions that would be “user-friendly” , offer intuitive graphic visualization of the results and also operate with reasonable speed compatible so as to meet the time-sensitive requirements of clinical reporting. This joint initiative has reached a mature stage, whereby the developed end-to-end pipeline is tested and refined by expert laboratories from both organizations.NGS immunogenetics is the focus of the EuroClonality-NGS Working Group (http://www.ericll.org/ongoing-projects/), we are seeking to generate sufficient data that will shed light on open issues regarding both the biological and clinical implications of the findings, more specifically: the true meaning of minor expanded clonotypes unrelated to the dominant one; the extent and clinical impact of intraclonal diversification; reviewing the cutoffs for discriminating U-CLL from M-CLL and determining whether there is a need for setting novel clinically relevant germline identity cutoffs.ERIC and the EuroClonality-NGS Working Group are also placing great emphasis on the correct interpretation of the obtained results. Capitalizing on the availability of well-annotated primary patient samples, facilitated by the ERIC database, a standardized registration system holding clinicobiological information from patients with CLL (NGS has heralded a new era in medical diagnostics and immunogenetic analysis is following this trend. There is indeed a growing demand for shifting practice and using NGS for IGHV gene SHM assessment, although it is debatable whether it is always justifiable, at least taking into account financial considerations for laboratories with limited resources. While NGS will probably become the method of choice, the traditional Sanger sequencing is still the standard method for IGHV gene SHM assessment. Nevertheless, as this analysis impacts on treatment decisions, standardization of both technical aspects and data interpretation becomes even more essential. Therefore, the need for establishing new recommendations and providing dedicated education and training on NGS-based immunogenetics is greater than ever before.


Cancer represents one of the most pernicious public health problems with a high mortality rate among patients worldwide. Chemotherapy is one of the major therapeutic approaches for the treatment of various malignancies. Platinum-based drugs  are highly effective chemotherapeutic drugs used for the treatment of several types of malignancies, but their application and dosage are limited by their toxic effects on various systems, including neurotoxicity. Simultaneously, researchers have tried to improve the survival rate and quality of life of cancer patients and decrease the toxicity of platinum-containing drugs by combining them with non-chemotherapy-based drugs, dietary supplements and/or antioxidants. Additionally, recent studies have shown that the root cause for the many side effects of platinum chemotherapeutics involves the production of reactive oxygen species (ROS) in naive cells. Therefore, suppression of ROS generation and their inactivation with antioxidants represents an appropriate approach for platinum drug-induced toxicities. The aim of this paper is to present an updated review of the protective effects of different antioxidant agents  against the neurotoxicity induced by platinum-based chemotherapeutics. This review highlights the high potential of plant antioxidants as adjuvant strategies in chemotherapy with platinum drugs. A “honeymoon” with platinum-based compounds in chemotherapy that officially started by pronouncing cisplatin as “the drug of the century” more than 50 years ago has gradually been overshadowed by very serious therapeutic limitations of those drugs. Besides the inherited resistance to treatment with any of the currently approved platinum agents, each of them has a number of clinically confirmed side-effects, ranging from minor to dose-limiting. The fact that almost half of all the patients who receive anticancer chemotherapy are treated with a platinum drug  gives a cis-diamminedichloridoplatinum II), carboplatin , and oxaliplatin , while some other platinum-based therapeutics are approved only in individual countries, such as heptaplatin (Korea), lobaplatin (China), miriplatin (Japan), and nedaplatin (Japan) [For decades various platinum-based compounds were employed as an important part of the combination chemotherapy regimens used to treat different types of solid tumors. However, there are only three platinum-based medications that are used throughout the world for the cancer treatment: cisplatin  in the treatment of various malignancies: lung , ovarianLike for the other chemotherapeutic drugs, the basic cytotoxic effect of platinum-based compounds (DNA damage) is not restricted only to target tissue (tumor cells), but is also affecting numerous organ systems in patients receiving chemotherapy, resulting in a variety of side effects. Based on a similar pathophysiological background, the adverse effects of platinum-based chemotherapeutics may be classified in certain categories of toxicities according to their clinical manifestations. The most commonly described types of side effects associated with platinum-based treatment are usually classified as nephrotoxicity, hepatotoxicity, neuro- and ototoxicity, cardiotoxicity, hematological toxicity, and gastrointestinal toxicity. However, patients’ responses to chemotherapy toxicity, including adverse effects of platinum-based compounds, are significantly determined by several factors, such as age, gender, drug administration schedule and performance status .Nausea and vomiting are considered as the most common clinical manifestations of side effects following cisplatin administration. Strongly depending on the applied dose, this effect of cisplatin, which can be successfully abolished by antiemetic action of 5-HT3 antagonists , is founNephrotoxicity represents the main limitation for chemotherapeutic protocols that involved platinum-based compounds which is not surprising due to the fact that kidneys are the main route for platinum compounds excretion. Although the platinum-based drugs affect all three key kidney functions , the two most common nephrotoxic side effects of cisplatin are acute kidney injury  and hypomagnesemia, which is reported to affect up to 90% of cisplatin-treated patients . HoweverHepatotoxicity is also considered as one of the most frequent adverse effects of platinum-based compounds administration in clinical practice. Morphological alterations accompanying platinum-based compounds application are manifested as necrosis, degeneration of hepatocytes, and increased inflammatory response , as wellNeurotoxicity in response to platinum-based therapy is the leading clinical entity, aside from nephro- and hepatotoxicity that usually hampers platinum-based chemotherapy . The mosWhen comparing to the individual extension of neurotoxic manifestations for platinum-based drugs, it has been shown that carboplatin neurotoxicity is negligible compared to cisplatin and oxaliplatin. However, carboplatin, particularly when applied in high doses, can lead to the development of neurotoxic manifestations that may become irreversible in 30-50% of patients.The evaluation of mechanisms underlying platinum-induced peripheral neurotoxicity (PIPN), in both in vivo and in vitro studies, confirmed that those compounds after passing the neuronal membrane initiate several proapoptotic phenomena including the activation of p53, Bax translocation, cytochrome c release, as well as the activation of caspase-3 and 9 . SimilarInterestingly, although it is still very questionable how cisplatin passes through an intact blood-brain barrier, there are certain literature data considering the central manifestations of neurotoxicity induced by platinum compounds. It has been shown that cisplatin administration strongly affects CNS progenitor cells and oligodendrocytes inducing the apoptotic events in hippocampal dentate gyrus and corpus callosum . Only a Since there is a plethora of evidence that oxidative damage is crucially involved in the mechanisms of platinum-based compounds toxicities, it is not surprising that an enormous effort has been put in order to increase the safety of platinum-based therapy regimens by promoting the antioxidant supplementation as potentially protective interventions during the chemotherapy protocols-based platinum-containing antitumor agents.The root cause of many side effects and organ injury of platinum-based drugs is the generation of reactive oxygen species (ROS) . The incOxidation reactions are very common in our body and they have a role to change a chemical substance, but also could produce highly reactive free radicals . Xenobiode novo antioxidants. There is also an adaptation mechanism of antioxidant defense that is referred to as ‘the fourth line of defense’ [There are different definitions of antioxidants; the most comprehensive definition describes antioxidants as substances that directly scavenge free radicals, inhibit their production, or indirectly act to up-regulate antioxidant defenses. Furthermore, antioxidant compounds should have the ability to form a new stable radical that is inactive for further oxidation, after scavenging the free radical ,39. Antidefense’ ,41.The first line of defense includes preventing mechanisms for biological molecules damage by suppressing or preventing the formation of free radicals and their derivatives. This defense line involves enzymatic antioxidants  that catThe human antioxidant defense mechanisms include both non-enzymatic and enzymatic antioxidants groups classified according to the molecular structure which forms endogenous antioxidant systems. But the human endogenous antioxidant system does not suffice, and the production of non-enzymatic antioxidants can be hardly producing in the human body. Therefore, there are exogenous non-enzymatic antioxidants  such as According to the World Health Organization recommendations nutrition is one of the major modifiable determinants of chronic disease . Some esVitamin E  is a potRetinoic acid (Thiamine pyrophosphate and water-soluble formulation of provitamin coenzyme Q10 (WS-CoQ10) has also been studied for neurotoxicity prevention. The neuroprotective effect of WS-CoQ10 was tested on PC12 cells exposed to cisplatin. These cells possess neuronal cell features and respond positively to nerve growth factor. WS-CoQ10 showed neuroprotective effects on the PC12 cells, protection from cisplatin-induced DNA damage, and an increase in the total intracellular GSH . The appSelenium is a trace element that has an important role in cellular redox regulation and the protection of cellular components from oxidative damage. Selenium was shown to possess moderate neuroprotective effects on the cisplatin neurotoxic effect in rats. The study showed that selenium administration partly reverses nerve conduction velocity, the amplitude of compound action potential, and the number of axons in experiments with cisplatin application in rats .Calcium and magnesium (Ca/Mg) infusions were suggested as supplementation in platinum-based drugs-induced peripheral neuropathy. Oxaliplatin is the most researched among them as chemotherapeutic agents because its metabolite oxalate has the potential to chelates both Ca and Mg ions disrupting the function of ion channels in nerve membranes . The hypAlpha lipoic acid (ALA) is one of the most studied dietary antioxidant supplements in the literature. This molecule  is essenMelatonin is a pineal hormone, also known as a free radical scavenger, which manifested protection activity against oxaliplatin-induced alterations in motor activity and muscular strength in rats. Besides, this supplementation inactivated Bcl-2 and caspase 3 apoptotic protein, as well as decreased Cytochrome c release and modulated oxidative stress in rats’ brain caused by oxaliplatin application . Tuncer Yehia et al.  reportedD-Methionine is also one of the effective antioxidant supplements against cisplatin-induced neurotoxicity in experimental animals. It showed a positive influence on the cortical neurons damage in an in vitro model of cortical networks , preventThere are some studies about the use of different antioxidants which are usually not present or which are present in small amounts in the human diet, against cisplatin-induced neurotoxicity. Song et al.  reportedThe injurious effects of platinum-based drugs, especially a sensory peripheral neuropathy, are primarily dose-limiting, but they also can affect the quality of life of the patients. There are many possible therapies for the treatment of PIPN although none of them is completely efficient. That is the main reason for numerous investigations, presented in Amifostine is an organic thiophosphate that has cytoprotective and detoxicant activities. It is generally an inactive prodrug which activates after dephosphorilation in plasma membrane with alkaline phosphatase. When active metabolite enters the cell it act like free radical scavenger protecting DNA and cellular membranes . There aCisplatin-induced neurotoxicity in rats was treated with oxytocin, a neurohypophyseal nonapeptide synthesized in the hypothalamus . The resN-Acetylcysteine (NAC) is a precursor of glutathione which can increase the concentration of glutathione in blood and thus increase the antioxidative defense in the organism . The eff2+ as a soft acid  has a high affinity for chelation with compounds possessing soft bases (soft ligands) [2+ ions, increasing renal excretion of platinum thereafter influencing on neurophysiological aspects of patients treated with cisplatin. But in the randomized placebo-controlled multicenter evaluation of DDTC in cisplatin treatment, it was shown that DDTC was not able to alleviate cisplatin-induced toxicities [2+ ions in the dorsal root ganglion and where they can bind to proteins, that lead to the development of oxidative and nitrosative stress [2+ has an affinity for DPDP, and the newly formed complex of metal with DPDP has fast excretion and low toxicity which are positive aspects for use of DPDP as chelation therapy of PIPN. Another chelation complex of manganese and the ligand fodipir, mangafodipir, displayed significant results in the study conducted by Coriat et al. [2+/Mn2+) was patented in 2015 by Karlsson et al. [Many studies have dealt with the neuroprotective activity of different medications against platinum drugs-induced neuropathy. Several compounds were considered to have a number of favorable properties. For example, an chemoprotectant BNP7787 showed good neuroprotective effects in phase II randomized study . A theraligands) . For exae stress . In the e stress  was showt et al.  on oxalin et al.  as a newn et al. . The autn et al.  criticizn et al. .Xaliproden, a 5HT1A agonist that possesses neuroprotective activity, showed a reduction in grade 3–4 neurotoxicity by activation of MAPK pathways in oxaliplatin-induced neuropathy without affecting FOLFOX4 antitumor activity in first-line treatment of patients with metastatic colorectal cancer (MCRC) . Some neCurcuma longa (Zingiberaceae) rhizomes. It showed protective activity in vitro, in terms of neurite outgrowth inhibition in PC12 cells treated with cisplatin, but on the other hand, curcumin had not affected the anticancer activity of cisplatin in HepG2 tumor cells [Compounds of the natural origin are in use in traditional and modern medicine regarding their numerous beneficial effects. They can be used in the prevention and/or therapy of various pathological conditions such as cardiovascular diseases, metabolic disorders, carcinogenesis, and neural impairments . The mosor cells . The in or cells  in rats or cells ,113.O-rutinoside)   were tesPhenolic acids are known for their significant antioxidant, antitumor, and antimicrobial activity and thanks to that they display many benefits on human health ,142. ManNigela sativa (black cumin) seed oil is thymoquinone  of the hippocampus. Moreover, this compound exhibited the ability to rescue the cholinergic neuron function in rat brain and also lowered oxidative stress and neuroinflammation.In the study of Li et al.  cyanidinoquinone . Due to 2S [2S and modulating Kv7 channels [Silybum marianum that possesses antioxidant and antineoplastic activities. In this study, silibinin reduced oxidative damage of oxaliplatin in rats at a concentration of 100 mg/kg. It also recovered motor coordination and showed a reduction in oxaliplatin-dependent pain induced by mechanical and thermal stimuli. Silibinin is one of the components in silymarin complex mixture which also contains three more flavonolignans  and its use is mainly focused on liver disorders treatment [Isothiocyanates, a group of natural bioactive compounds mostly present in cruciferous vegetables, are characterized by their antioxidant properties . That is2S . Similarchannels . The samchannels  conductereatment . Silymarreatment .Astragalus membranaceus radix and Huangqi Guizhi Wuwu Decoction that contained a mixture of Astragalus membranaceus radix, Cinnamomum cassia, Paeonia lactiflora, Ziziphus jujuba, and Zingiberis recens rhizoma. The common feature for both mixtures was Astragali radix (Huang Qi). The extracts of Astragalus roots, aqueous and two hydroalcoholic extracts, were tested in vitro against oxaliplatin-induced neurotoxicity in the neuronal-derived cell line SH-SY5Y and in primary cultures of rat cortical astrocytes [Astragali radix extracts exhibited strong antioxidant activity, ameliorating the lipid peroxidation, proteins, and DNA oxidation. The 50% hydroalcoholic extract was dominant in the prevention of caspase-3 activation and it stimulates astrocyte viability.Many herbal mixtures are in use for relieving chemotherapy-induced neuropathic pain. One that showed significant improvement regarding the negative effects of platinum-based drugs is goshajinkigan, a traditional Japanese Kampo medicine consisted of ten medicinal plants that is used clinically to treat pain in Japan. Ushio et al.  showed ttrocytes . The AstSalvia miltiorrhiza, mainly used in the treatment of cardiovascular diseases and neurasthenic insomnia [Matricaria chamomilla). The ethanol extract of chamomile flowers showed anti-inflammatory effects and reduction of pain responses in the formalin test in mice with intensive analgesic effect [Hypericum perforatum L. (St. John’s wort) is well-known for its antioxidant, anti-inflammatory, analgesic, and neuroprotective effects [Satureja hortensis aerial part extract, applied in rats at three different concentrations  along with cisplatin injection, exhibited strong anxiolytic activity and lowered apoptotic parameters in rat hippocampal tissues [Another immensely important medicinal plant in traditional Chinese medicine is Danshen or insomnia . Neverthc effect . Hyperic effects ,152,153. effects . Saturej tissues . MoreoveVitis vinifera L., the common grapevine, is known for many benefits on human health, particularly for the antioxidant potential of its main bioactive compounds proanthocyanidins but also many phenolic acids and flavonoids [V. vinifera hydroalcoholic extract in the model of oxaliplatin-induced neurotoxicity showed extensive activity towards the reduction of superoxide anion concentration and lipid peroxidation in rat astrocytes [Camellia sinensis) can attenuate toxicities linked to treatment with platinum-based drugs  [Lithospermum erythrorhizon is a plant used in traditional Chinese medicine for eczema and other skin diseases as well as for wound healing. However, it was shown that Lithospermi radix extract can be an excellent neuroprotective agent against oxaliplatin-induced peripheral neuropathy in both in vitro and in vivo models [avonoids ,141. Thetrocytes . It alsotrocytes . The maiiplatin) . Its extiplatin) . Lithospo models .In summary, we intentionally presented the variety of previously reported beneficial effects of antioxidant supplementation in the treatment of neurotoxicity, as one of the most frequent dose- and time-limiting factors in chemotherapeutic protocols that include platinum-based compounds. It seems reasonable that future investigations that should analyze the protective role of specific antioxidants more precisely may result in achieving even more subtle coverage of chemotherapeutic protocols with adverse effects preventing strategy.


Circular RNAs (circRNAs) are an evolutionarily conserved novel class of non-coding endogenous RNAs (ncRNAs) found in the eukaryotic transcriptome, originally believed to be aberrant RNA splicing by-products with decreased functionality. However, recent advances in high-throughput genomic technology have allowed circRNAs to be characterized in detail and revealed their role in controlling various biological and molecular processes, the most essential being gene regulation. Because of the structural stability, high expression, availability of microRNA (miRNA) binding sites and tissue-specific expression, circRNAs have become hot topic of research in RNA biology. Compared to the linear RNA, circRNAs are produced differentially by backsplicing exons or lariat introns from a pre-messenger RNA (mRNA) forming a covalently closed loop structure missing 3′ poly-(A) tail or 5′ cap, rendering them immune to exonuclease-mediated degradation. Emerging research has identified multifaceted roles of circRNAs as miRNA and RNA binding protein (RBP) sponges and transcription, translation, and splicing event regulators. CircRNAs have been involved in many human illnesses, including cancer and neurodegenerative disorders such as Alzheimer’s and Parkinson’s disease, due to their aberrant expression in different pathological conditions. The functional versatility exhibited by circRNAs enables them to serve as potential diagnostic or predictive biomarkers for various diseases. This review discusses the properties, characterization, profiling, and the diverse molecular mechanisms of circRNAs and their use as potential therapeutic targets in different human malignancies. Circular RNAs (circRNAs) are single-stranded non-coding RNAs that are covalently linked to form a continuous closed-loop and participate in the regulation of transcriptional and post-transcriptional gene expression . In receCircular RNAs were initially thought to be unable to translate through cap-dependent mechanisms due to their lack of 5′ cap structure and poly-A tail. But recent studies have shown the ability of circRNAs to translate in prokaryotes by mimicking DNA rolling circle amplification and association of circRNAs with translating ribosomes and the ability of circRNAs to generate proteins from circRNA minigenes . The traDrosophila showed inhibition of spliceosome components by U2snRNP depletion or inhibition that caused increased circRNA generation compared to its linear counterparts  and spliceosome RNA helicase UAP56 (for longer circles) . Exonic in situ hybridization and is a useful technique for understanding its biology by co-localizing potential miRNA sponges , cell proliferation and evasion of apoptosis by activating the Ras/ERK signaling pathway  and carbohydrate antigen 19-9 (CA19-9) might improve CRC diagnosis . A studyMET gene expression by sponging miR-198 and promoted the migration, invasion and metastasis of ESCC cells through the activation of RAS/MAPK and PI3K/Akt signaling pathways, thus showing its potential as a diagnostic target in ESCC  . circ-00via the MACC/MET/ERK or AKT pathway ( pathway . Further pathway . Yang F.in vivo (Many circRNAs are associated with resistance in acute myeloid leukemia (AML) . Shang ein vivo . Moreovein vivo .BCR-ABL1 is a fusion protein kinase derived from a reciprocal translocation between chromosomes nine and 22 and is a crucial protein for Chronic Myeloid Leukemia (CML) pathogenesis . TyrosinNumerous studies have demonstrated that circRNAs has emerging potential as a clinically relevant and disease-specific molecular biomarker in cancer and other complex diseases. Growing evidence suggests that circRNAs can be considered as promising biomarkers for the early diagnosis, metastasis, prognosis, and drug resistance of tumors due to their stable structure, the long half-life, tissue specificity, and abundance, and their presence in body fluids . CircRNAMoreover, circRNAs play a key role in tumor metastasis, circHECTD1 can directly bind to MIR142, resulting in astrocyte activation and contributing to cerebral infarction. Silencing of circHECTD1 expression can ameliorate cerebral infarction by inhibiting astrocyte activation , which can activate the autoimmune pathway   . Many sttis (OA) .Many researchers confirmed that circRNAs play roles in bone marrow failure and hematologic malignancy. in vitro into mammalian cells can induce the expression of innate immunity genes and thus protect against viral infection. This suggests that cells can distinguish endogenous and foreign circRNAs, dependent on the intron encoding the circRNAs , a protein that decreases rapidly in Alzheimer’s disease (AD) and other neurological diseases . MoreoveOver the last few decades, the significance of circRNAs in numerous pathological processes has gradually been realized; while initially known as splicing redundant by-products, clinical studies have managed to uncover the hidden characteristics and ability of circRNAs to serve as ideal biomarkers in different pathologies. Unlike other RNA species, circRNAs have prognostic or diagnostic value due to high stability and specific cancer expression. Understanding the molecular mechanisms/regulatory roles of circRNAs in various transcriptional and translational processes in the eukaryotic machinery is essential to elucidate their human diseases’ roles, most importantly, cancer. Apart from the notorious role of circRNAs as miRNA sponges, further studies are required to reveal other mechanisms manifested by circRNAs. Although the expression of circRNAs has been extensively studied, their function remains inconclusive in normal physiological processes and human diseases. Many mechanistic aspects of how circRNAs potentially work in various cancers by affecting oncogenic pathways need to be better clarified, and we are still a long way from their development as a therapeutic or prognostic target that warrants thorough analysis of their mechanism of action. Research in this area must step away from simple quantification and therapeutic inference into mechanistic studies with a translational benefit. Developing technologies that enable sufficient single-cell level identification and modulation of circRNAs without disrupting their linear RNA partners may help gain more insight into the regulatory functions of circRNAs and thus help enhance the production of strategies that target the circRNA network in various human diseases. As circRNAs are generated from pre-mRNA back splicing, different bioinformatics algorithms have been designed to characterize circRNAs using different annotations to read back splicing sites. It will always make sense to develop the algorithm based on specific circRNA to avoid false reads and correlate the results using independent analysis by different platforms.SN, AB, MS, IA, and TK: collected the literature, wrote the manuscript, and generated the figures. AR, SH, PB, RR, FJ, SU, GC, DB, WE-R, and MF: critical revision and editing of the scientific contents. AB, IA, TK, MM, and MH: conceived and designed the review contents and contributed to the manuscript writing and editing. All authors read and approved the final manuscript.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.