Positron Emission Tomography with Computed Tomography (PET/CT) is widely used in the assessment of many diseases, particularly including cancer. However, many factors can affect image quality and diagnostic performance of PET scans using FDG or other PET probes.The aim of this pictorial essay is to review PET/CT protocols that can be useful to overcome these confounding factors in routine clinical situations, with a particular focus on pharmacological interventions and problem-oriented CT acquisition protocols.Imaging protocols and representative cases will be discussed, in addition to potential contraindications and precautions to be taken. There are many factors that can affect image quality and diagnostic performance of PET/CT examinations using FDG or other PET probes. We will review pharmacological interventions (including dosage and contraindications) and use of problem-oriented radiological imaging protocols designed to optimize diagnostic information and improve the overall accuracy of PET/CT in routine clinical practice. Herein, we present representative cases with and without drug intervention to illustrate how useful these simple protocols are.68Ga-DOTA-octrotate an upper SUV threshold of 30 is typically used while for PSMA ligands the corresponding upper threshold at PMCC is 15. Lower thresholds may increase sensitivity, but this is generally at the expense of specificity, whereas higher thresholds can provide greater appreciation of heterogeneity between tumour sites but lead to faint lesions becoming inapparent.Illustrative cases are presented with a thresholding proposed by the Peter Mac Callum Cancer Centre (PMCC) team for FDG studies with the use the “rainbow” colour scale that has low activity regions displayed in the blue-green range and higher intensity regions in the orange-red spectrum. With this colour scale, the SUV threshold should be adjusted so that the liver appears blue with flecks of green. This corresponds to an upper SUV window threshold of approximately 9, except in bariatric patients where the upper SUV threshold is typically 12 [While there has been a strong professional and industry focus on harmonisation of CT windowing for various tissues, thresholding of PET images remains largely idiosyncratic and non-standardised even sometimes within departments according to individual reader preferences. At the very least, the authors recommend that each department has a consistent method of displaying images across readers and through time in order to facilitate comparative studies.The cases presented represent examples from three PET facilities primarily staffed by nuclear medicine physicians but including dual-trained radiologists. The PMCC facility operates within a combined Cancer Imaging Department with CT protocols developed collaboratively with the CT modality leads. Where variations in protocols exist, these are discussed.Hyperglycemia directly competes with FDG for uptake in normal tissues, particularly the brain, and also in tumour. This alters the tumour-to-background ratio, potentially decreasing the sensitivity of the PET scan for detection of malignancy , 3, espeSchedule FDG PET examination in the early morning in patients having received intermediate-acting or long-lasting insulin in the eveningOr schedule FDG PET examination in the early morning in patients treated with continuous insulin infusion, the insulin pump being turned-off 1–2 h prior to FDG injection and administering FDG when blood sugar levels begin to rise, indicating loss of exogenous insulin effect. Avoid corrective bolus insulin 4–6 h of FDG injection.It should be noted that even under fasting conditions, normal individuals have a basal insulin level, but this is lacking in type 1 diabetes mellitus (DM) and insufficient to overcome insulin resistance in insulin-dependent type II DM. Therefore, completely withholding insulin is such patients is inappropriate. Nevertheless, careful management to avoid acute alterations of glucose homeostasis are required to avoid impacting the biodistribution of FDG, particularly under the influence of the insulin-dependent glucose transporter GLUT-4, which is preferentially expressed in cardiac and skeletal muscle. Accordingly, management of type 1 DM or insulin-dependent type II DM should avoid FDG being injected within 4 h of subcutaneous administration of rapid-acting insulin. It is, however, preferable for diabetic treatment to be maintained to avoid hyperglycaemia. This can be achieved by;However, for patients who present with hyperglycaemia despite these recommendations, the threshold of blood glucose level (BGL) that irrevocably impairs diagnostic performance is currently unclear and strategies for cancelling imaging or intervening to correct hyperglycemia vary between organisations.The consensus clinical guidelines of major societies recommend rescheduling patients with hyperglycaemia above specified thresholds  achieved BGL < 10.0 mmol/L, there was minimal risk of hypoglycaemic events (0.7%) and high-quality PET images were obtained. It should be stressed again that giving insulin contemporaneously with FDG is contraindicated due to its ability to drive FDG into cardiac and skeletal muscle under the influence of the insulin-dependent glucose transporter, GLUT-4. Conversely, if symptomatic hypoglycemia is induced, whether requiring glucose supplementation or not, scanning should be cancelled as this generally invokes a catecholamine response that may increase brown fat activation.To lower blood glucose levels under these pre-defined thresholds, different PET scan preparation protocols have proposed the intravenous administration of rapid acting insulin. In a recent study of Pattison et al. , a persoPhysiologic uptake of FDG in brown adipose tissue (BAT) of cancer patients may confound interpretation of PET scans. It occurs in half of paediatric oncology patients and is especially common in adolescents . BAT uptBAT regulates body temperature by inducing thermogenesis. Although previously thought to be primarily an issue in cold climates, it can occur even in temperate or tropical climates when patients come into an artificially-cooled environment. Therefore, the temperature of uptake rooms should be regulated to avoid cold stress. Warm blankets may assist if air-conditioning cannot be adequately regulated.Since BAT is innervated by the sympathetic nervous system, stress or elevated catecholamines levels , may be associated with elevated BAT uptake. Accordingly, beta blockers such as propranolol have shown efficacy in reducing BAT uptake . Low bloPropranolol has been used in paediatrics to treat a variety of conditions. Although generally safe, it can cause bradycardia, hypotension and hypoglycemia. There are only few contraindications, including chronic obstructive pulmonary disease (COPD) and asthma, and various cardiovascular conditions including uncontrolled heart failure, bradycardia, hypotension, high grade AV block, sinus disease, Prinzmetal angina and Raynaud disease. It should also be avoided in patients with known or suspected pheochromocytoma / paraganglioma  unless adequately alpha-blocked due to a risk of hypertensive crisis from unopposed alpha-adrenergic activation . For aduIt is noteworthy that some groups use benzodiazepines to reduce BAT uptake, though efficacy of benzodiazepines in this setting remains questionable. In a preclinical study Tatsumi et al.  found noAdequate suppression of cardiac glucose metabolism increases the interpretability and diagnostic reliability of 18F-FDG PET/CT studies performed to detect cardiac inflammation and infection or, in rare cases, of primary or secondary malignant involvement of the heart or pericardium.There are no standardized guidelines, though prolonged fasting (beyond 24 h), carbohydrate-restricted diets, fatty meals, and heparin loading all have been proposed.At the Caen University Hospital, the protocol involves high-fat, low-carbohydrate diet on the day prior to scanning followed by prolonged fasting over 12 h. This is followed by unfractionated heparin (50 UI/kg) being injected 15 min before the FDG injection. The aim of this protocol is to decrease basal insulin and blood glucose levels and to increase blood free fatty acid (FFA) levels, which shift myocardial energy consumption away from glucose toward FFA. At the Royal Brisbane & Women’s Hospital, an even more aggressive dietary regimen is instituted to invoke ketogenesis. This involves a high-fat diet for 48 h , then heparin in patients without contraindications. Ketone monitoring (capillary and urine) has also been performed to assess the adequacy of dietary preparation for cardiac sarcoid imaging, and if not elevated, prompt more detailed assessment of dietary adherence [p < 0.0001). The main side-effect is bleeding risk and the contraindications include children under 3 years of age, severe thrombocytopenia, heparin-induced thrombocytopenia (HIT) hypersensitivity or any situation which may cause bleeding. The bleeding risk is, however, very low if used properly as a single dose. For example, there was no bleeding in the study from Scholtens et al. [Unfractionated heparin significantly improves image contrast, as demonstrated by Scholtens et al.  by usings et al.  in whichHeparin should be avoided in patients who are already receiving anticoagulant therapy or have a history of bleeding disorders. When using this protocol for PET tumour imaging, for instance for detection of myocardial metastases, attention should be paid to tumours at risk of bleeding and brain metastases. Heparin-induced thrombocytopenia from unfractionated heparin was calculated at 2.6% in a meta-analysis on thromboprophylaxis by Martel et al.  but the Physiologic smooth muscle uptake in the empty stomach of fasting patients often hampers the visualization of focal or diffuse gastric lesions by increasing apparent background activity.Based on the study from Le Roux et al. , the proOf note, because of their effect on attenuation correction CT maps, oral contrast agent can cause an increased SUV in the gastro-intestinal tract and thus causes artifacts. Therefore, we recommend keeping the same imaging protocol for follow-up imaging, for instance in the neo-adjuvant setting, so that variation in SUV between baseline and post-chemotherapy scans is not hampered by contrast media.Scopolamine (Scoburen/Buscopan) is easy to use and there are relatively few contraindications. These include known hypersensitivity, risk of acute angle-closure glaucoma, a past history of urinary retention or significant prostatism in men and this treatment is not recommended during the third trimester of pregnancy or when women are breast feeding.FDG has been used with limited success in the past in primary diagnosis and locoregional staging of urinary bladder cancer or other urothelial tumours, mainly because of the pharmacokinetics of renal excretion.Indeed, pooling of urinary radiotracer in the ureters, bladder or urachus can obscure urinary tract lesions even when they have high metabolic activity, which is typically the case for transitional cell carcinoma (TCC). While FDG PET/CT remains useful for detecting nodal and distant spread of TCC, without alteration of scanning protocols the detection of primary disease is compromised.This is optimally achieved by diluting activity in the bladder by using a diuretic drug such as furosemide (Lasilix/Lasix) in addition to water oral intake (> 500 mL). Diuretics can be used even in patients with kidney failure but at increased doses –25. DelaThe contraindications are kidney obstruction, acute kidney injury, hepatic encephalopathy, dehydration, severe hypokalaemia, severe hyponatraemia, severe kidney failure (eGFR < 30 ml/min), hepatitis and liver failure.Focal urinary collection of activity in the ureters of radiotracers that are renally-excreted can be mistaken for nodal disease. This is particularly common at the pelvic brim and in the lateral pelvis where the ureters cross major vessels. Primary genito-urinary tumours including prostate, endometrial and cervical cancers often involve pelvic, iliac and retroperitoneal nodes in close proximity to the ureters as well as directly involving these structures. With the increasing use of imaging of prostate cancer using prostate-specific membrane antigen (PSMA) agents, use of a delayed CT urogram protocol can assist in increasing the confidence with which nodal disease can be diagnosed or excluded . This inThe non-medication intervention of use of a vaginal tampon can be proposed to women prior to the image acquisition. The tampon stretches the vagina and allows better delineation of the inferior margin of the uterine cervix. There is no specific contraindication but this intervention should be used with caution in symptomatic patients after radiotherapy or in old women with vaginal dryness, since this may cause pain and bleeding.Of note, the vaginal tampon should be inserted properly, as it has been shown by Burger et al. that tampons inserted below the pubococcygeal line can lead to contamination by urinary radiotracer . As for In patients receiving metformin, high bowel activity may hamper interpretation of FDG PET images, either by mimicking pathological uptake or by masking lesions located within or near the gut. This may compromise evaluation of primary colorectal cancer or detection of incidental colonic neoplasia, which is not an uncommon benefit of FDG PET/CT . In the Metformin decreases glucose transport from food to plasma and enhances glucose consumption by enterocytes. It is known from preclinical studies that metformin-induced increased uptake selectively involves the colon wall without any significant contamination of the bowel content.Several studies have evaluated the discontinuation of metformin for periods of 1 to 3 days, with conflicting results. While Oh and colleagues  found a In a prospective randomized single centre trial comparing the effects of a 24H- and 48H metformin discontinuation, Hamidizadeh and colleagues  providedHigh FDG uptake in the normal cerebral cortex and basal ganglia decreases the ability to detect and assess the extent of primary and secondary brain tumours. Although F-18 fluoroethyl-tyrosine (FET) PET/CT overcomes this limitation, when FDG is also required for systemic staging, suppressing cortical uptake of FDG can be advantageous. In the paediatric population, general anaesthesia can markedly reduce uptake in the brain, which may also be desirable from the perspective of minimizing radiation exposure to the developing neurological system but must be carefully balanced against the risks of anaesthesia itself. In adult, use of conscious sedation with midazolam or benzodiazepines may be an alternative. These should be avoided in patients with risk of airway obstruction. The degree of suppression of cortical activity with these interventions is relatively limited and the combination of MRI and FET PET/CT are preferred methods for evaluation of brain lesions.Movement is probably the most common cause of degraded image quality and is exacerbated by patient discomfort. This is particularly a problem in patients suffering from cancer. Attention to patient comfort is paramount to achieve high-quality PET/CT studies but judicious use of pain modifying medication should also be considered. It must, however, be recognized that these medications can increase the risk of falls when getting on or off the scanner and careful supervision of patient transfers is required. It is noteworthy that patients in pain may also benefit from fast or even ultra-fast PET imaging, which is achievable on modern PET/CT .The methodology used for the correlative CT component of PET/CT studies remains one of the more contentious debates in cancer imaging. The incremental radiation dose and risk of contrast reactions associated with diagnostic protocols may be offset by the possibility of improving diagnostic certainty in certain circumstances but, in others, these potential adverse effects serve to add nothing beyond the localising information that can be provided by a low-dose, non-contrast CT. Unfortunately, sometimes the debate has been driven by professional rivalries and not by evidence or even by rational argument. Some PET/CT services are provided entirely within radiology departments and routinely perform full diagnostic CT whereas in other jurisdictions, nuclear medicine physicians never use other than low-dose CT, primarily for attenuation correction purposes . We takeA further area of controversy is whether or not to include the brain when acquiring oncological PET/CT studies. An argument against doing so is that it potentially provides a false sense of security that brain metastases have been excluded and that by not including the brain, that oncologists are compelled to do the more appropriate investigations of either MRI or contrast-enhanced CT. However, a counterargument is that incidental metastases can be detected, either as hypermetabolic lesions or due to hypometabolism related to vasogenic oedema. Incidental detection of features indicating neurodegenerative or cerebrovascular disease may also impact treatment choices. With tracers that have low or no significant uptake in the brain, inclusion of the head can detect incidental abnormalities that might also provide complementary information to formal neuroimaging .While slightly increasing the complexity or time investment in performing PET/CT scans, the pharmacological and radiological interventions described above can help imaging specialists to optimize diagnostic information with limited impact on throughput in busy PET units and generally much less impact than having to repeat a non-diagnostic scan or perform another investigation to clarify equivocal findings. Amongst these interventions, management of the diabetic patients and avoidance of BAT uptake are the most frequently encountered issues in PET units and their management is now well documented. When using PET for therapy monitoring purposes, forced diuresis and gastric distension protocols should ideally be kept identical  between


Cellular senescence has been recognized since the 1960s as a cell biological program of aging. It also serves physiological functions in organismal development, regeneration and tissue repair. Senescence is triggered by replicative telomere attrition but also stress such as DNA damage, hypoxia, nutrient deprivation, mitochondrial impairment and oncogene activation . In respThe diverse biological functions of senescence in aging, developmental processes, tissue repair, cancer growth and chronic diseases suggest that the senescent state reflects a dynamic cellular stress program depending on the cell type, nature of inducer and extent of senescence with highly variable SASP composition , 3, 5. DThe timely and spatially controlled induction of senescence is part of a developmental program where the secretory function of senescent cells serves to fine-tune cell fate determination and tissue patterning . In highIn tissue remodeling and repair, senescence is induced locally in a well-controlled manner , 11. TheIn aging and in chronic diseases, senescent cells accumulate and persist in the tissue . SenesceSenescence is a well-recognized feature of lung fibrosis. Patients with mutations in telomere genes, which causes replicative senescence, develop familial forms of idiopathic pulmonary fibrosis (IPF) indicating that senescence causally contributes to the development of lung fibrosis in this subgroup of familial IPF . CellulaThe recent scRNA seq data from mouse models of lung repair fundamentally challenge the traditional view that senescent cells are irreversibly growth arrested and act mainly in a paracrine fashion to promote proliferation and reprogramming in neighboring cells. Strunz et al. identified a transitional stem cell state involved in alveolar repair of bleomycin-injured mouse lungs . These KRemarkably, a very similar signature of de-differentiation, stemness, and senescence was detected in aberrant basaloid cells of irreversibly remodeled IPF lungs , 41. The+ T cells (Given this astonishing overlap of gene expression programs between aberrant basaloid cells in IPF and the newly identified alveolar repair cells, i.e., de-differentiation, stemness and senescence, it is tempting to speculate that the IPF-specific basaloid cells are “locked in repair” . Locking T cells . Togethe T cells  this wil T cells .Our concept predicts that repair-locked senescent cells represent an Achilles heel for the development of pulmonary fibrosis. Novel therapeutic concepts should then aim at targeting these cells in IPF by either eliminating them or putting them back on the right repair track. Such approaches could have both cell-autonomous as well as non-autonomous effects due to the expected modulation of the SASP.The key issue is to understand what locks these cells in their senescent repair modus. Is this an irreversible state? Are there any means to unlock them? Comparison of these basaloid cells in IPF that seem to be permanently locked in repair with their repair competent counterpart in the bleomycin mouse model might help to shed light on possible approaches, including epigenetic signatures, to unlock these cells.3 histone signatures (As the senescence program is characterized by extensive chromatin remodeling with predominant H3K9megnatures , 3, 15 ignatures . Among tgnatures . Indeed,gnatures . Howevergnatures . This apgnatures . It's tiAll 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.


In schizophrenia and related disorders, a deeper mechanistic understanding of neocortical dysfunction will be essential to developing new diagnostic and therapeutic techniques. To this end, combined transcranial magnetic stimulation and electroencephalography (TMS/EEG) provides a non-invasive tool to simultaneously perturb and measure neurophysiological correlates of cortical function, including oscillatory activity, cortical inhibition, connectivity, and synchronization. In this review, we summarize the findings from a variety of studies that apply TMS/EEG to understand the fundamental features of cortical dysfunction in schizophrenia. These results lend to future applications of TMS/EEG in understanding the pathophysiological mechanisms underlying cognitive deficits in schizophrenia. Schizophrenia is a debilitating psychiatric disorder with a mean lifetime prevalence of 1% . PatientNeuroimaging studies in schizophrenia patients have consistently observed reduced cortical gray matter volume in temporal and prefrontal areas, an anatomical change that may reflect neuronal cell loss, lowered dendritic complexity, and/or synaptic density changes . HoweverTranscranial magnetic stimulation (TMS) has emerged as a powerful tool to non-invasively study cortical physiology in humans . While eTranscranial magnetic stimulation uses an electromagnetic coil to create a time-varying magnetic field within the cortex. This changing magnetic field then induces an electrical current, transiently affecting the firing of cortical neurons close to the coil . Thus, TWhen TMS is applied to the motor cortex, electromyography (EMG) of the associated muscle shows a motor-evoked potential (MEP) . If TMS Neural oscillations are rhythmic fluctuations in the electrical activity of populations of neurons, and these synchronized oscillations may allow more efficient communication between different brain regions . Neural Relative to the study of strictly task-based biomarkers, TMS/EEG offers a more direct and spatially localized probe of cortical oscillations. In one early TMS/EEG study, schizophrenia patients and healthy controls underwent single-pulse TMS at the premotor cortex, and high-density EEG showed that evoked gamma oscillations in frontal areas of schizophrenia patients were significantly decreased in amplitude and synchronization . This fiSimilar findings have been observed in preclinical models of schizophrenia; in two separate putative mouse models of schizophrenia , mice displayed a deficit in evoked gamma band activity together with deficits in the reliability of neuronal coactivity patterns that could not be explained by alterations in single neuron activity. Interestingly, acute ketamine administration or acute pharmacogenetic suppression of PV-INs did not have the same effect , suggestResonant frequencies offer a complimentary approach to studying neural oscillations in schizophrenia. Upon a pulse of TMS stimulation, different cortical areas tend to oscillate at specific frequencies, with more frontal areas oscillating at higher frequencies and more posterior regions oscillating at lower frequencies. Even as the intensity or location of the TMS pulse is varied, the EEG signal at each region continues to oscillate at the same frequency. Thus, region-specific resonant frequencies appear to be a deliberate physiological property reflecting variations in regional neuronal organization and thus may offer a useful tool for studying cortical region-specific disturbances . In one Gamma oscillations are produced by the firing patterns of inhibitory PV-INs, cells which play a role in maintaining an appropriate excitatory/inhibitory balance in the cortex in mice . Thus, sTo study cortical inhibition via TMS/EEG, many studies make use of a paired-pulse paradigm. Two separate TMS pulses (the conditioning stimulus and the test stimulus) are applied to the cortex; if they are applied within a 1–4 ms of each other, the response to the test pulse is typically diminished relative to the conditioning pulse and termed short interval intracortical inhibition (SICI). In contrast, with intervals set to 50–200 ms, the resulting inhibition is called long interval intracortical inhibition (LICI) . These tRecent studies have applied paired-pulse TMS/EEG to measure inhibition of TEPs in the prefrontal cortex . One stuIn addition to alterations in excitation/inhibition imbalance, schizophrenia has also been broadly conceptualized as a failure of efficient communication between neural systems required for cognition and perception. Resting state fMRI analyses have provided strong support along these lines, demonstrating alterations in functional connectivity between the prefrontal cortex and other brain regions  in patients with schizophrenia . In addiTo measure the functional connectivity between two cortical regions with TMS/EEG, studies can apply a TMS pulse to one area and then use the EEG data to determine how that pulse spread to another region. Unlike approaches that rely on temporally correlated patterns of activity , responses to experimentally induced TMS pulses can provide inferences about causality and directionality of cortical information spread . One stuBy transiently modifying oscillatory activity, TMS/EEG can offer causal insights into the role of beta/gamma desynchronization in schizophrenia. For example, one study applied TMS at varying frequencies to the prefrontal cortex and simultaneously measured memory formation in healthy subjects. Memory formation was most impaired when the cortex was stimulated at a beta band frequency, implicating desynchronization of beta activity in the process of normal memory formation . SimilarIn the ways described above, TMS-EEG offers a unique opportunity to directly test a variety of specific hypotheses pertaining to cortical dysfunction in schizophrenia. By analyzing TEPs across multiple domains see . FinallyAll authors discussed the reviewed research, contributed to manuscript revision, and approved the submitted version. AV wrote the first draft of the 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.


This rationale for athlete self-regulation in practice could improve athlete performance by enhancing problem solving, engagement with constraints of learning designs and supporting better attunement to contextual information abundant in a competitive environment. Finally, by introducing this unified and multidisciplinary DoM, specialist coaches, team coaches and sport science support staff, within the organisational structure, can collaboratively debate and co-design individualised athlete training programmes to enrich skill adaptability and performance functionality. To underline these contentions, three high-performance sport case studies from Australian Football: goalkeeping in Association Football and Rugby League are presented.With increasing resources in sports organisations being allocated to the development and preparation of individual athletes and sub-groups with specialist performance roles, the work of coaches, specialist (role) coaches and support staff needs to be functionally and coherently integrated. This integration of sport science support and coaching can be administered by staff in a Department of Methodology (DoM). Particularly, in this paper, we propose how specialist coaching can be situated in a DoM, presenting a model advocating effective functioning in high-performance team sports organisations. Using principles of ecological dynamics, we provide a rationale for a functional methodology for the design of practice tasks in a DoM that views learners as  Underpinned by principles of ecological dynamics, this opinion piece proposes a model for how specialist coaching in high-performance team sports organisations can be situated in a Department of Methodology (DoM).The presented model illustrates processes for (1) reviewing the athlete development programme and training environment; (2) examining the athlete-support structure and (3) establishing a DoM structure to cohesively guide athlete development strategies.To underline all contentions, three high-performance sport case studies from Australian Football; goalkeeping in Association Football and Rugby League are presented.Recently, Otte et al.  proposedAccess to sport science support and coaching can be administered by staff in a Department of Methodology (DoM), where service unification is guided by principles of ecological dynamics , 4. Yet,The introduction of specialist coaches purported to enhance athlete development in high-performance sport has contributed another layer of complexity to the multidisciplinary nature of performance preparation. Addition of more sub-discipline specialists to an often already large multidisciplinary team of practitioners  might actually hinder, rather than enhance, athlete performance. Although this level of expertise may provide an illusion of integration in a sports organisation, sports practitioners have traditionally operated in a non-integrative way, with sub-discipline specialists working in silos, leading to disjointed athlete preparation . Major cIt has been argued that the DoM concept can alleviate issues associated with disciplinary integration . A DoM iNext, we discuss how the integration of sub-discipline specialists within a complex system, such as a DoM, can be achieved through the conceptualisation of ecological dynamics. This, we contend, will enable the coordination of common approaches, co-designed principles and language used by specialist coaches to integrate and enhance performance preparation.complex adaptive systems with inherent self-organising tendencies, which support the emergence of synergies  guided by contextual information detected from performance environments. The detection of information is important for regulating actions, and the skilled coupling of perception and action throughout the performance is a hallmark of expertise in sport ).GK-specific training and team-based training  outline the invaluable need for collaboration between head coaches, support staff and specialist coaches. In detail, this coordination between coaches appears critical in order to ensure facilitation of effective training designs . Whilstsessions ), the exIn order for professional staff members to adequately monitor, periodise and plan  athlete loading and successive training sessions, multi-directional information exchange and communication about compiled data across all staff members become paramount. Linking this idea back to an overarching DoM, this case study reiterates one critical notion: the imperative need for coaching staff, specialist coaches (e.g. GK coaches) and support staff (e.g. sport scientists and psychologists) to co-design training and constantly exchange information.In Rugby League football, ‘metres gained’ and ‘making quick ground’ with the ball are important predictors of successful match outcomes , 25. TraTo improve a team’s ability to gain metres onfield in possession of the ball, players could develop the physical, perceptual and emotional skills to collectively perceive and use relevant environmental opportunities offered to advance the ball forward. Therefore, all players must be afforded representative opportunities to perceive shared information sources in practice, which are related to physical and task constraints on action in competition . HoweverIn this opinion piece, we proposed how a DoM could functionally integrate the work of specialist (role) coaches in coordinated, collective organisational collaborations, underpinned by the theoretical framework of ecological dynamics. This type of organisational design for support staff in high-performance sports organisations could empower specialist coaches to contribute meaningfully to individual athlete development and preparation for performance, without feeling marginalised. Indeed, further empirical work is needed to understand how such an integrated collective system of support staff can best enrich athlete development and engagement in the learning process across the course of their careers.


Gut peptide hormones are one group of secretory factors produced from gastrointestinal endocrine cells with potent functions in modulating digestive functions. In recent decades, they have been found across different brain regions, many of which are involved in autism-related social, emotional and cognitive deficits. Clinical studies have revealed possible correlation between those hormones and autism spectrum disorder pathogenesis. In animal models, gut peptide hormones modulate neurodevelopment, synaptic transmission and neural plasticity, explaining their behavioral relevance. This review article will summarize major findings from both clinical and basic research showing the role of gut peptide hormones in mediating autism-related neurological functions, and their potential implications in autism pathogenesis. The pharmaceutical value of gut hormones in alleviating autism-associated behavioral syndromes will be discussed to provide new insights for future drug development. Autism spectrum disorder (ASD) affects more than 1.5% of children worldwide . It is eGut peptide hormones refer to a group of polypeptides derived from GI tract and can exert multiple physiological functions via binding with specific receptors. Gut peptides have a long history since the identification of its first member, secretin, at almost 100 years ago. Currently dozens of molecules have been classified into this group, including cholecystokinin (CCK), vasoactive intestinal peptide (VIP), pituitary adenylate cyclase activating peptide (PACAP), peptide YY (PYY), secretin, pancreatic polypeptide, ghrelin and so on . Those sBesides the feeding and metabolic regulation, the potential role of gut peptide hormones in other behavioral paradigms is emerging in recent decades. Notably, some gut peptides are involved in social functions. Examples include secretin, whose gene knockout in mice leads to social interaction deficits , and VIPIn this mini review, for each gut peptide hormone we will start with its distribution and functions in the central nervous system, following with the evidence of its involvement in ASD. We will also discuss about the neurobiological mechanisms of those gut-brain peptides, with particular interests in mediating social and cognitive deficits as well as stereotypic behaviors, all of which are the core behavioral symptoms of ASD. Lastly, the potency of drug development targeting those gut peptides and their receptors in alleviating ASD-related symptoms will be discussed.in vivo recording, opto-/chemo-genetics have made it possible to delineate specific functions of those peptides within certain neural circuits including those related with ASD.Gut peptide hormones exert various neuromodulating effects as suggested by their wide distribution across different brain regions. Recent advances in neural science techniques including gene knockdown, As one widely studied gut-brain peptide, CCK can be found within a certain group of inhibitory neurons and has known effects in mediating appetite and food intake . PreviouSome studies have already been done to investigate the potential neurological mechanisms of CCK in social behaviors. Targeted mutation of CCK receptor alters social isolation-related behaviors in female mice . The selPACAP is one polypeptide hormone sharing high sequence similarity with secretin and is also involved in social modulation. Genetic study investigated more than 1,000 ASD children with their normal siblings and identified one specific loci (rs1557299) at the downstream of PACAP gene  which suElectrophysiological studies reveal that PACAP improves synaptic plasticity in mouse hippocampus , and incVIP is mainly expressed in the hypothalamic nuclei such as suprachiasmatic nucleus (SCN) and tuberal hypothalamus as well as hippocampus and cerebral cortex , b, and Recent evidence suggested a close relationship between VIP and ASD. Newborns with higher VIP concentrations in the cord blood were more likely to develop autism in later periods . PrelimiO-a-cyl-transferase (GOAT) from DAG. Only AG is able to bind to the growth hormone secretagogue receptor type 1 a (GHSR1a), a G protein-coupled receptor, to execute their biological functions  . The hypunctions , 2001. Gunctions . Those eRecent study revealed the close relationship between ghrelin and ASD. For example, the expression level of un-acylated ghrelin acylated ghrelin in blood is lower in children with autism . GhrelinSecretin is one classical gut-peptide hormone to modulate pancreatic secretion as well as body energy homeostasis. Several lines of evidence from rodent and human studies show that secretin and its receptors are widely distributed in the brain areas from cerebral cortex, hippocampus, amygdala, hypothalamus to cerebellum . The neuIn 1998, a case series reported secretin in amelioration of the symptoms in three ASD children . HoweverFor investigating the molecular and cellular mechanism, secretin has been found to induce the spinogenesis of hippocampal neurons , and thein vivo and low efficiency for penetrating the blood brain barrier (BBB). Large amounts of works are thus required to overcome those disadvantages and to generate drug candidates with satisfactory efficiency, persistency and safety.Based on the inter-correlation between ASD pathogenesis and potentially neurological mechanisms in regulating social, emotional and repetitive behaviors by gut peptides, it is expected that those hormones may work as drug candidate or targets for developing novel drugs in alleviating ASD symptoms. However, failures are frequently reported when directly trying the administration of those peptides to ameliorate autistic behaviors. For example, one clinical trial failed to detect significantly improvement of social phobia by using CCK-tetrapeptide (CCK-4) , and manin vivo. Such limitation can be addressed by two approaches: the development of stable analog or receptor agonist, or the application of bioengineered sustained drug delivery system. The former approach is based on molecular design and screening of small molecules, or the modification of existing receptor ligands, to generate long-lasting effects as those of natural ligands. Alternatively, one can adopt nanoparticles system to chronically release peptide drugs such as those used in immunomodulation therapy for releasing autoantigens (The short half-life is one common property of gut hormones due to their peptide composition. For example, secretin has less than 5 min half-life under normal physiological conditions , making antigens . In drugantigens . Those dGut peptide hormones are now recognized as having prominent brain distributions and various behavioral modulatory functions. Clinical studies with ASD subjects have revealed that the genetic polymorphisms of gut peptides such as CCK, PACAP, and VIP are related to ASD. Altered expression levels of the peptides including CCK and ghrelin are found in ASD children. Additionally, more animal studies suggest that these gut peptides are actively involved in the modulation of social, emotional and stereotypic behaviors. In future, one should perform more site-specific studies to illustrate the precise mechanisms of gut peptides in mediating social or emotional or cognitive behaviors for potential drug targets. Further drug development can be pursued to generate stable analogs that can modulate gut peptide pathways in the brain, aiming to relieve behavioral symptoms of ASD.X-RQ wrote the manuscript with the help from LZ. LZ revised the 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.


In recent years, non-coding RNAs (ncRNAs) have gained unprecedented attention as new and crucial players in the regulation of numerous cellular processes and disease responses. In this review, we describe how diverse ncRNAs, including both small RNAs and long ncRNAs, may be used to engineer resistance against plant viruses. We discuss how double-stranded RNAs and small RNAs, such as artificial microRNAs and trans-acting small interfering RNAs, either produced in transgenic plants or delivered exogenously to non-transgenic plants, may constitute powerful RNA interference (RNAi)-based technology that can be exploited to control plant viruses. Additionally, we describe how RNA guided CRISPR-CAS gene-editing systems have been deployed to inhibit plant virus infections, and we provide a comparative analysis of RNAi approaches and CRISPR-Cas technology. The two main strategies for engineering virus resistance are also discussed, including direct targeting of viral DNA or RNA, or inactivation of plant host susceptibility genes. We also elaborate on the challenges that need to be overcome before such technologies can be broadly exploited for crop protection against viruses. Due to climate change and alterations in cropping systems, plant viral disease outbreaks will likely have increasingly negative impacts on food production, which will bring new challenges to current agricultural practices worldwide. To advance virus disease management, rapid, efficient, and safe responses are required to minimize viral threats. The use of virus-resistant crop varieties is a traditional and efficient way to reduce losses caused by viral infections. However, conventional antiviral breeding strategies, even augmented with modern molecular techniques such as quantitative trait locus (QTL) mapping, marker-assisted selection, and whole-genome sequence-based approaches, are slow and laborious and require monitoring of large crop populations over multiple generations .Advances in genetic engineering and plant transformation techniques in the early 1980s enabled the rapid production of virus-resistant transgenic plants . Since tIt should be noted that historically, mainstream approaches to engineering plant virus resistance have been driven by a classical view of the central dogma of molecular biology, in which the genetic information contained in DNA is transcribed into mRNA which in turn directs the synthesis of particular proteins, and variations in proteins give rise to different traits. Accordingly, many transgenic plants were generated to express virus-encoded proteins , with the expectation that any resistance obtained would be protein-mediated. However, in many cases, the resistance had been surprisingly proven to be RNA-mediated. This phenomenon was named RNA silencing or RNA interference (RNAi) . This isWhile the small siRNAs and miRNAs constitute important classes of regulatory ncRNA, many other types of ncRNAs are known, and all of these form the majority of the transcriptome, given that only about 2% of transcribed RNAs encode proteins . ncRNAs Thus, while the central dogma of DNA>RNA>protein still holds true, ncRNAs can greatly influence this information flow and are, therefore, now an emerging major new source of next-generation targets which may be exploited in diverse applications, including conferring plant resistance against pathogens and viruses in particular .RNA technologies provide us not only with new sources of resistance but also with powerful tools to manage plant defense responses. With regards to antiviral resistance, RNAi-based approaches have long been used in the form of transgene-induced RNA silencing to target the degradation of diverse viral RNAs or to inactivate virus susceptibility genes in various crops ,10,11,12RdRp), and serve as secondary siRNAs that systemically spread throughout the plant; which is crucial for efficient RNAi-based antiviral defense [Of the different types of ncRNAs, siRNAs have attracted considerable attention due to their direct targeting of viral RNAs for degradation as a part of the natural RNAi-based antiviral immune response ,30. For  defense . RNA sil defense , which sFor DNA-containing plant viruses  the precursors of siRNAs are presumably produced by DNA-dependent RNA polymerase II (Pol II) which mediates bidirectional transcription in both sense and antisense orientations on the viral DNA. Such transcripts can pair virus-specific mRNAs and hence form perfect dsRNAs to be processed by some DCLs into siRNAs .To successfully overcome siRNA defenses and infect plants, viruses have evolved suppressors of RNA silencing (VSRs) which may target key steps in the siRNA pathways by inhibiting siRNA production, sequestering siRNAs, or preventing the spread of RNA silencing signals . Some otThus, siRNA-triggered RNAi represents a robust host defense mechanism against plant viruses which has been proven for numerous viruses in many plant species and crops ,20. WithFundamentally, miRNAs and siRNAs share some similarities in size, structure, and functions. Both are short duplex RNA molecules that may exert gene silencing effects at the RNA level and are derived from double-stranded regions of RNA precursors. They both require DCL and AGO proteins for processing, maturation, and action, although the species of DCL and AGO may be different. However, there are many important differences between miRNAs and siRNAs. A major distinction between them is that siRNAs are processed from foreign dsRNAs  whereas miRNAs are derived from endogenous precursor transcripts containing double-stranded  regions encoded by miRNA genes (MIR genes). miRNAs are derived from longer, primary transcripts termed pri-miRNAs. The pri-miRNAs contain an RNA hairpin in which one of the two strands includes the mature miRNA fragment. This hairpin then is cut out by a microprocessor enzyme complex which includes DCL1 nuclease (a member of an RNase III endonuclease family) and HYL1, a dsRNA binding protein ,39, whicPlantae from mosses to monocots and eudicots and can be grouped in two classes: one which is conserved and ancient, and one which consists of miRNAs that are less conserved and younger [In toto, miRNAs constitute a highly diverse set of molecules which may match nearly all RNAs encoded in a plant genome, thereby controlling almost any aspect of the plant life cycle; playing key roles in growth and development, signal transduction, innate immunity, and responses to various ecological biotic and abiotic stresses [Many miRNAs are evolutionarily conserved across major taxa of the Kingdom  younger ,40. In tstresses .Arabidopsis plants, thus elevating the accumulation of its gene target transcripts MYB33 and MYB65 . This consequently led to the production of severe viral symptoms [With regards to virus infections, information on miRNAs has mainly focused on their role in the induction of viral disease symptoms and antiviral defense mechanisms. It is well documented that some viruses may control accumulation rates of miRNAs involved in plant symptom development . For exasymptoms . Similarsymptoms . It has symptoms . Interessymptoms . Some ofsymptoms . Why theAGO or RdRp genes, which are involved in siRNA biogenesis and activity, are themselves natural targets of various plant miRNAs. For example, the antiviral AGO1 is the target gene for miR168. To prevent degradation of AGO1 transcripts and thereby alleviate AGO1-mediated antiviral defense, plants have evolved a decoy protein gene AGO18 [AGO18 transcripts compete with AGO1 transcripts for miR168 to attenuate the cleavage of AGO1 mRNA. Expression of AGO18 in rice was shown to be activated upon RSV infection, conferring resistance against this virus through partial miR168 sequestration [RdRp1-mediated RNA silencing pathway in rice by targeting RdRp1-inhibiting proteins [Many components of the antiviral silencing machinery such as ne AGO18 . AGO18 tstration . miR444 proteins .N gene encoding the resistance TIR-NB-LRR protein gene that mediates hypersensitive response against tobacco mosaic virus (TMV). Overexpression of these miRNAs decreases levels of TIR-NB-LRR transcripts and attenuates N-mediated resistance to TMV, suggesting that miR6019 and miR6020 play an important role in controlling resistance to TMV [It has also been shown that some miRNAs can target genes that trigger antiviral resistance . For exae to TMV ,49.AGO18, resulting in increased AO activity and elevated ROS accumulation; rendering plants more resistant to the virus [TCP21 and JA concentrations in rice had elevated abundance after infection by rice ragged stunt virus, leading to a more virus-susceptible phenotype [General plant responses contributing to antiviral resistance are often orchestrated by the rapid production of reactive oxygen species (ROS) and several phytohormones such as salicylic acid (SA), jasmonic acid (JA), and auxin ,51. It hhe virus ,53. miR3henotype . Recentlhenotype .As exemplified by these case studies, miRNAs are currently regarded among the most important gene regulators. As discussed, these small RNAs are involved in many pivotal aspects of plant-virus interactions and are emerging as the next generation targets for engineering plant virus resistance.7 G cap, 3′ poly (A) tail, and exon-exon junctions [Arabidopsis under stress conditions [Another group of non-coding RNAs playing important roles in many biological processes is lncRNAs. LncRNAs are defined as transcripts longer than 200 nt with little or no coding potential. Similar to coding mRNAs, most lncRNAs are transcribed by Pol II. They may also have typical mRNA-like structures such as 5′munctions ,56. In anditions ,58. In gcis and that of distant genes in trans. Cis–acting regulatory lncRNAs modulate the expression of the neighboring target genes either by recruitment of Pol II or through the event of getting transcribed, which results in local chromatin conformational changes that facilitate downstream mRNA transcription [Based on genomic origin, lncRNAs can be classified as intergenic, intronic, or exonic regions in sense or antisense orientation . Besidescription .Trans-acting lncRNAs may act as scaffolds to facilitate the recruitment of protein(s) and the formation of RNA protein complexes. Such interactions are usually mediated by specific lncRNA domains and may regulate gene expression either through sequestering proteins and preventing them from reaching their target (as a decoy) or by delivering them to specific target sites (as a guide) [a guide) ,9. LncRNa guide) . For exaa guide) . The subThe repertoire of biological processes and mechanisms of action of the lncRNAs are rapidly growing. Plant lncRNAs are known to play pivotal roles in the regulation of flowering time, modulation of reproductive organ development, leaf development, auxin signaling, photomorphogenesis, and responses to biotic and abiotic stresses .It is conceivable that lncRNAs may also play important roles in plant-virus interactions. Indeed, it has been shown that both RNA-containing tobacco rattle virus  and DNA-The ELENA1 lncRNA identified in another recent study was shown to take part in controlling plant defense by modulating the SA pathway . It was Nicotiana benthamiana with a CTV isolate deficient in LMT1 production (CTV-LMT1d) led to an elevated SA accumulation  encodes its own subgenomic lncRNA, LMT1 . Infectisistance  and a cosistance . This stsistance . FurtherCircular RNAs (circRNAs) are a group of endogenous non-coding ssRNAs that have a closed-loop structure . CircRNAArabidopsis, miPEP171b from Medicago [Brassicaceae [Despite their definition as non-coding RNAs, a growing number of reports have suggested the existence of stably expressed and functional peptides (or microproteins) translated from lncRNAs . These pMedicago , and miPsicaceae . These pThere is a growing body of evidence that SEPs play an important role in animal cell immunity , suggestAs mentioned above, other classes of non-coding RNAs include rRNAs, tRNAs, snRNAs, and snoRNAs. There are no doubts that some functional crosstalk may exist between various activities of these RNAs and virus infections. A striking example of such interplay is that sophisticated functional mimics of tRNAs (transfer RNA-like structures) are found at the 3′-ends of the genomes of some plant positive-strand RNA viruses , which mPioneering work published in 1986 by Powell Abel et al.  demonstrTo address these hurdles and public concerns, another approach termed “spray induced gene silencing” (SIGS) has been developed whereby RNA silencing-based antiviral protection is induced by spraying plants with exogenous dsRNA (or hpRNA), complementary to viral RNAs . In addiPMMoV, pepper mild mottle virus; TEV, tobacco etch virus; AMV, alfalfa mosaic virus; PSTVd, potato spindle tuber viroid; CEV, citrus exocortis viroid; CChMVd, chrysanthemum chlorotic mottle viroid; SCMV, sugarcane mosaic virus; PRSV, papaya ringspot virus; CymMV, cymbidium mosaic virus; CMV, cucumber mosaic virus; ZYMV, zucchini yellow mosaic virus; BCMV, bean common mosaic virus; ToLCV, tomato leaf curl virus; PVY, potato virus Y; PPV, plum pox virus; PSbMV, pea seed-borne mosaic virus; LDH, layered double hydroxide clay nanosheets.It is also worth noting that the antiviral RNAi-based resistance mechanism induced by the exogenous dsRNA may be further enhanced via pattern triggered immunity (PTI). PTI is a general plant response that is triggered by a conserved microbe—or pathogen—ssociated molecular patterns  as the first step of plant defense . RecentlAltogether, these scientific achievements create new opportunities for practical applications of dsRNA-based approaches for crops. Recently developed breakthrough technological platforms for large-scale cell-free or bacterial production  and purification of dsRNA at as low a cost as $0.5–1/g , greatlyAnother approach to engineer virus resistance using SIGS is to silence virus susceptibility genes in host plants. Among the known susceptibility genes required for successful infection with some plant viruses, such as potyviruses, are those encoding the translation initiation factors eIF4E, eIF(iso)4E, and eIF4G . InactivAlthough this has not yet been experimentally verified, one can suggest that exogenous dsRNA designed to silence a particular host susceptibility gene target can be applied to intact plants without the need for plant transformation. In addition, modification of individual cultivars is not required due to the sequence similarity of targeted genes; however, only a few such genes have been identified. Therefore, finding novel universal and effective targets among host genes and non-coding RNAs is a challenge for further development and marketing of the SIGS technology.Until now, only exogenous dsRNA has been used in SIGS as a silencing trigger. In the future, the arsenal of RNA triggers for SIGS could be significantly expanded by adding next-generation tools such as artificial miRNAs (amiRNAs), artificial trans-active siRNAs (atasiRNAs), and target mimics of several kinds . A corolThe miRNA mimic technology (miR-Mimic) is another novel approach for gene silencing. miRNA mimics are artificially synthesized dsRNA molecules imitating miRNA duplexes. Once delivered into the cell, these RNA fragments mimic natural miRNAs which bind to the target mRNA molecules and block their expression either by degradation or by translational repression .Although the exogenous applications of ami/atasiRNAs or miRNA mimics have not been reported so far, we believe that these types of RNA molecules will be assessed and exploited in SIGS technology in the future.As mentioned above, CRISPR-Cas technology has emerged as an expanding toolbox with which to generate antiviral resistance in various crop plants. In this regard, two of the most common approaches using CRISPR-Cas to control virus diseases is either by using it to directly target and inhibit virus machinery or by mutating “susceptibility” genes in host plants , the designed sgRNAs were shown to be able to directly target various specific regions of the viral DNA genomes and consequently reduce or abolish disease symptoms caused by such geminiviruses as WDV ), tomatoThe discovery of CRISPR-related sgRNA-guided endonucleases specifically targeting RNA molecules such as FnCas9 and Cas13 , opened These examples show that direct manipulation of viral DNA or RNA genomes by CRISPR-Cas may represent new technologies to engineer antiviral resistance. However, this approach requires persistent maintenance of the CRISPR-Cas components in the plant which thus far can only be carried through their transgenic expression. Due to GMO regulations, this may preclude or significantly limit practical applications of CRISPR antivirals.To mitigate GMO-related barriers in engineering virus resistance, several other CRISPR-Cas approaches avoiding the use of GMO have been developed, which target host plant genes involved in regulating virus susceptibility. In the first approach, CRISPR-Cas components are initially expressed transgenically to modify a host “susceptibility” gene leading to virus resistance, and these transgenic components are then segregated out following sexual reproduction . AlthoueIF4s and coilin genes were previously used for the development of resistance in HIGS experiments  which can affect virus susceptibility have been exploited for CRISPR editing to engineer virus resistance within a rather narrow range of viruses, such as potyviruses and rice tungro spherical virus ,118,119.e above; ,106).As discussed, both RNAi and CRISPR-Cas have been successfully used as powerful tools for engineering plant virus resistance. However, the question arises as to which of these seemingly competing technologies is the right option for a particular application. There are several technical and methodological aspects to be addressed before making a choice:GMO. As noted above, some CRISPR applications, in particular, direct targeting of plant viruses, require the permanent presence of CRISPR-Cas components in plants which is only possible in transgenic plants, whereas SIGS RNAi-based technology is transgene-free.Plant regeneration. To inactivate host plant virus “susceptibility” genes, the constant presence of the Cas protein and sgRNA is not needed and hence, the use of transgenic plants can be avoided either by eliminating the Cas9/sgRNA transgenes or by delivering editing reagents as mRNAs or RNPs. However, the associated steps for the regeneration of whole plants from the edited cells with consequent identification of the edited lines are usually time-consuming, technically demanding, and costly. Moreover, many crop plants and varieties are recalcitrant to regeneration. So far, CRISPR transgene-independent techniques are only feasible within a limited range of plant species and varieties. In contrast, the RNAi-based SIGS approach does not require regeneration steps at all.Individual cultivars. With CRISPR technology, each cultivar should be edited and tested independently whereas RNAi tools developed for targeting conserved gene sequences could be applicable to many cultivars.Time. Preparation of RNAi tools (weeks) is significantly faster than production and selection of edited plants (months or years).Nature of phenotype. Existing (transgene-free) CRISPR technologies induce permanent and irreversible gene edits in plants, whereas RNAi induces transient knockdown of gene expression. In many cases, transient perturbation of gene transcripts may be preferred, particularly when the complete knock-out of those genes that are essential could be lethal , but partial loss of them would not be. For example, virus “susceptibility” genes such as coilin genes (and others) possess many important cellular functions, and therefore their expression should be modulated in a temporary manner during virus attack only.This analysis suggests that RNAi engineering of antiviral resistance may currently offer a number of significant advantages over CRISPR. While CRISPR technology has been a game-changer that led to remarkable progress in the development of new methods for targeted therapy of human diseases , the appOne of the major problems in topical applications of naked dsRNA molecules is their instability in the environment, which is typically manifested in a short antiviral protection window when applied against virus infections . TherefoAnother concern is the capacity of RNAi compounds to penetrate cells, particularly where the large complexes which protect the RNA from degradation may significantly reduce permeability. Plants have evolved stringent security control systems (barriers) which govern specific signaling and transport pathways that prevent foreign molecules from penetrating and transporting through the plant tissues. The cuticle and cellulose cell wall are the first barriers typically encountered which are inhibitory to cell entry . The nexVarious factors, such as climate change, decreases in agricultural land and global population growth as well as hurdles associated with GMO and current chemical pesticide regulations, have promoted the development of new biosafe and eco-friendly technologies for crop protection against pests and pathogens. Among them are RNA-based biocontrol technologies, such as RNAi and CRISPR. Although both of them are still in the research and development pipeline, they are well anticipated to be exploited for engineering plant virus resistance in agricultural practice. In spite of the popularity and great capabilities of the CRISPR-Cas system, RNAi approaches seem to be preferable at least for direct targeting plant viral pathogens. Commercial expectations surrounding RNAi applications are rising, with great interest towards sprayable RNA-based techniques from agribiotech giants such as Bayer Crop Science, Monsanto (now acquired by Bayer) and Syngenta, and many other newly developed start-up companies . This trIn addition, we are well aware that the emergence of CRISPR-Cas editing tools will significantly strengthen the arsenal of applications for genetic crop improvement. Unprecedented advances have been achieved by using CRISPR-Cas technology in basic plant biology. CRISRP-Cas has been proved to be a vital tool for plant functional genomics, including the generation of CRISPR/Cas mutant libraries for developing genome-wide mutation screens . AnotherHowever, there are some common technical challenges and societal problems that must be addressed for both RNAi-SIGS and CRISPR-Cas technologies before they take a certain unique place in engineering plant virus resistance.Identification of novel host susceptibility targets among coding or non-coding RNAs that should be inactivated or modified by RNAi or CRISPR would be a crucial next step to further develop basic engineering strategies for virus resistance. It could also be useful to expand a potential list of gene targets by identifying gene/ncRNA targets in genomes of insect vectors to prevent transmission of plant viruses. RNAi has recently been successfully used to control insect pests .Another critical factor to be considered is the off-target effects of both RNAi and CRISPR technologies. Exogenously applied long dsRNAs can produce many different siRNA species with a higher chance of causing off-target host gene regulation . HoweverThe rapid progress in RNAi and CRISPR applications also desperately requires the improvement of protective delivery platforms that allow efficient uptake and dissemination of RNA silencing or CRISPR editing components by plant cells. Cooperative efforts between plant biologists, geneticists, and nanotechnologists may provide a real game-changer in such crop protection technology.Finally, clear and standardized regulatory guidelines are required for the development of non-transgenic externally applied RNAi and CRISPR tools to enable them to progress to agricultural markets.


Mesenchymal stromal cells (MSCs), also known as mesenchymal stem cells, have been intensely investigated for clinical applications within the last decades. However, the majority of registered clinical trials applying MSC therapy for diverse human diseases have fallen short of expectations, despite the encouraging pre-clinical outcomes in varied animal disease models. This can be attributable to inconsistent criteria for MSCs identity across studies and their inherited heterogeneity. Nowadays, with the emergence of advanced biological techniques and substantial improvements in bio-engineered materials, strategies have been developed to overcome clinical challenges in MSC application. Here in this review, we will discuss the major challenges of MSC therapies in clinical application, the factors impacting the diversity of MSCs, the potential approaches that modify MSC products with the highest therapeutic potential, and finally the usage of MSCs for COVID-19 pandemic disease. Mesenchymal stromal cells (MSCs) are pluripotent non-hematopoietic stem cells with self-renewal capability  and beinOver 300 clinical trials of MSC therapies have been completed in patients including but not limited to degenerative or autoimmune diseases , increased cell density, and/or serum deprivation can induce high expression of MHC-II in MSCs, while TGF-β suppresses MHC-II expression . The immMSCs of high quality is the first step to ensure the safety and efficacy in clinical trials. Understanding the molecular and cellular mechanisms underlying the immune incompatibility of MSCs will help to improve the manufacture of MSC products.MSCs have mesodermal lineage differentiation potential and the potential to regulate tissue regeneration by mediating tissue and organ repair, as well as replacing damaged cells . DiffereSeveral laboratories have analyzed the proteome modifications associated with MSCs differentiation , 26. TheSerial passaging in long-term culture could negatively affect the expression of stemness genes , 49. A pTo provide sufficient MSCs for clinical trials, MSCs need to be amplified in a large scale, which will inevitably face the issue of MSCs senescence and subsequent modifications of gene expressions . TherefoHeterogeneity of MSCs is determined by multiple factors including but not limited to donors and tissue sources, cell populations, culture conditions, cell isolation techniques, cryoprotective and thawing protocols –64 Fig..Fig. 3MSMSCs were defined as adherent cells with a spindle-shaped morphology in standard culture conditions according to the minimal criteria developed by the International Society of Cell Therapy in 2006 . They weTo identify specific cell subsets in heterogeneous MSCs, researchers have been continuously exploring characteristic cell surface markers and molecular signatures. Single cell-derived colony with rapidly dividing cells shows high colony-forming efficiency. STRO-1, CD146, and CD271 have been identified as cell surface markers for this subset . HoweverThe therapeutic efficacy of MSCs is highly dependent on their in vivo migration and homing capacities. The migrating direction is determined by chemokine receptors expressed on MSCs and chemokines in tissues . FreshlyHowever, the expression profile of chemokines in damaged tissues is often not compatible with that of receptors on MSCs. For instance, CXCL1, CXCL2 and CCL7 increased in infarcted myocardium, while expression of corresponding receptors (CCR1 and CXCR2) on MSCs was very low, resulting in low efficiency in the migration of MSCs to infarct sites . To imprThe delivery of MSCs emerges as a prerequisite to the unfoldment of their full therapeutic potential. Different delivery routes could affect cell homing, survival, and paracrine function. Systemic delivery is considered a reasonable approach. However, the reported effect in terms of homing rate, survival rate, and maintenance of cellular function was modest and transient  for reas2 compared with 20 cells or 2000 cells per cm2 [2) was suggested to support better proliferation [Theoretically, MSCs can be expanded in vitro in traditional culture plates and flasks to any amount that meets experimental purpose. However, with prolonged culture duration and increased passage numbers, MSCs reach the Hayflick limit, exhibiting a marked decrease in proliferation with a transformation in morphology from a thin spindle shape to a flattened square shape. The cell density seeded in the culture containers also plays a role in the senescence of MSCs. Neuhuber et al. found the optimal cell growth of rat MSCs at 200 cells per cm per cm2 . In otheferation . AlteratLarge-scale expansion in 2D plates over long term also impacts stem cell characteristics of MSCs. According to Zhao et al., hUC-MSCs at various passages have multiple mutation spectra on signatures and functions, and cells at high passage showed declined therapeutic effect in aGVHD mouse model . It has Moreover, MSCs have shown the capacity to differentiate into numerous cell types such as neural cells, hepatocyte-like cells, and pancreatic islet-like cells , 90. TheTaken together, developing the technique that can produce a huge number of cells rapidly and cost-effectively with guaranteed cell quality is paramount for the clinical progress of MSCs.The secretion of cytoprotective factors by MSCs was first reported by Gnecchi and colleagues. They observed that Akt-MSCs (MSCs overexpressing Akt) prevented ventricular remodeling and improved the heart function following surgical myocardial infarction (MI). Since cell transplantation and myogenic pathways would be ineffective over such a brief interval, a new mechanism was proposed that the injected MSCs might act through releasing trophic factors that contribute to myocardial protection following an ischemic insult. This hypothesis was then confirmed by evident improvements in cardiac performance following injection of conditioned medium (CM) collected from hypoxic Akt-MSCs into an induced MI model, which protected ventricular cardiomyocytes with less apoptosis when subjected to a hypoxic condition .In 2007, Dai et al. observed that MSCs-CM had a similar, albeit less intense, effect of MSCs in myocardial infarction, indicating that at least part of the effect observed following MSCs injection could be attributed to soluble factors . In the The paracrine effects of MSCs as an initial mechanism of action inspired further biological analysis of MSCs secretome . SubsequAlthough there were no attributable serious adverse events after MSC therapy, fever within 24 h and temporary pain at the injection sites are commonly occurred. Here we summarize four strategies to limit adverse events related to MSC treatments and improve the therapeutic outcomes, including genetic modifications or priming strategies to change the inherent characteristics of MSCs, and biomaterial strategies to modify the outside circumstances, and the usage of MSCs secretome  in the host immune system, which can potentially result in fibrosis and failure of the implantation. Therefore, biomaterials suitable for MSCs were constructed to ameliorate the FBR and subsequent fibrotic encapsulation . For exaMSCs are typically delivered to a graft site using a decellularized extracellular matrix (ECM) scaffold. The advent of synthetic polymers has revolutionized tissue engineering. These polymers are highly tunable, homogenous, and cell-free materials and have a high batch-to-batch consistency taking the form of porous hydrogels, sponges, plates, or membranes , 107. HoTo further optimize the therapeutic efficacy of MSCs, MSCs have been genetically engineered to produce trophic cytokines or other beneficial gene products in numerous preclinical models by transfecting MSCs with viral or non-viral vectors. Over the last few decades, these MSCs have successfully been engineered to express therapeutic peptides and proteins in animal models . For insHowever, both viral and non-viral vectors have some limitations. Non-viral vectors present transient gene expression and low-transfection efficiency, while viral transduction is associated with a higher risk of chromosomal instability, insertional mutagenesis, and proto-oncogene activation despite the inherent high transfection efficiency . The advSome studies made attempts on human-induced pluripotent stem cell (iPSC)-derived MSCs to obtain improved expandability. Actually, therapeutic transgenes could be inserted into iPSC-derived MSCs before MSCs derivation. This strategy could eliminate insertional mutation as well as guarantee stable expression of transgenes during prolonged expansion . So iPSCWith CRISPR/Cas9 technology, genetic modification of MSCs can be done with higher efficiency and specificity . CompareCRISPR/Cas9-mediated gene knockdown in MSCs has been proved effective in treating diseases such as myocardial infarction . TargeteDespite the specificity of CRISPR/Cas technology in gene delivery , only onGiven current manufacture of MSCs cannot meet the requirement for clinical trials in terms of production scale, the alternative is to boost the function of limited cells through priming MSCs. Priming has also been referred to as licensing or preconditioning, which is a concept commonly used in the field of immunology, and it has been adapted to the scope of stem cells , 145. On“Priming” MSCs resulted in exogenously boosted therapeutic function in comparison with original state. Several “primed” MSC products have been applied clinically, with the most notable being NurOwn from Brainstorm Cell Therapeutics Company. NurOwn boosted the expression of multiple neurotrophic factors (NTFs) including GDNF, BDNF, VEGF, and HGF . When adHowever, priming approaches of MSCs still have many limitations in clinical translation, such as induction of immunogenicity, high costs, variable effects, and lack of good manufacturing practices (GMP) suitable for clinical application . MoreoveThe “secretome” of MSCs, including secretory proteins such as growth factors, cytokines, and chemokines and EVs such as microvesicles  and exosomes (40–150 nm diameter), has been shown to exhibit many of the therapeutic properties of MSCs. For example, MSC-derived EVs have demonstrated similar or even superior therapeutic capacity for autoimmune diseases and neurodegenerative disorders compared with their parental MSCs , 177. ThSeveral studies have demonstrated the clinical effectiveness of MSC-EVs. For example, hBMMSC-EVs revealed significant improvements in patients suffering from refractory graft-versus-host disease . In anotDigital technology and AI are driving the revolution of healthcare industry . The druAI can find new molecular compounds and emerging drug targets much faster than traditional methods, thus speeding up the progress of drug development , 185. AtTo understand the essential elements in MSCs treatment, AI may recognize the dynamic molecular characteristics of essential elements, which include different protein sequences, molecular structures, as well as the binding forces and stabilities between targeted molecules and cell receptors. These data could be used to train a predictive model to the utmost accuracy . PredictAt present, the pharmaceutical world is increasingly engaged in technologies to shorten the time required to identify new drugs and repurpose current drugs. Since MSC therapies showed beneficial effects with complex undetermined components, AI may be well-suited to analyzing and revealing essential elements. Companies such as Merck, GSK, and Roche have developed partnerships with AI companies to construct suitable platforms , 190. HoParacrine effect was discovered to mediate MSCs therapeutic efficacy in previous studies –193. EVsFirst of all, the manufacture of large scales of MSC-EVs with high purity is difficult. MSC-EVs are isolated from MSC culture media, of which conditions including the seeding cell number, media volume, and isolation method and time of EVs can influence both the quantity and quality of EVs . TherefoSafety and efficacy of MSC-EVs in various disease conditions need to be ensured in further preclinical and clinical evaluation. In vivo distribution analysis of fluorescence-labeled EVs has shown that MSC-EVs might have homing capacity for injured or tumor-bearing sites comparable as MSCs . Long-teAfter the disclosure of precise mechanisms of action or key therapeutic factors in MSC-EVs therapy, targeted-EVs could be expanded in uniform proliferative cells such as fibroblasts via gene modification technology. Therefore, with big data-based analysis of transcriptome and proteome, engineered EVs may be manufactured with desired elements. For instance, Thomas C. Roberts et al. engineered EVs to express IL6 signal transducer (IL6ST) decoy receptors to selectively inhibit the IL6 trans-signaling pathway. Treatment in the Duchenne muscular dystrophy mouse model with these IL6ST decoy receptor EVs resulted in a reduced phosphorylation of STAT3 in muscles; further functional studies verified the in vivo activity of the decoy receptor EVs as a potential therapy . SimilarPandemic diseases like 2019 novel coronavirus disease (COVID-19) have dramatically increased the number of sickness and death worldwide. Though vaccines have been developed recently, the viruses are still rapidly mutating and expanding, and the available specific and effective treatment options are currently very limited . For sevMSC therapeutics may be the ideal candidates for handling the broad spectrum of COVID-19 symptoms due to their multifactorial mode-of-action . They caCOVID-19 has been the top priority of global healthcare systems since its emergence. There have been more than 160 vaccines in development and more than 60 clinical trials are ongoing, and now, only a few vaccines have been approved . The repAlthough MSCs therapies have achieved tremendous advancements over the past decades, substantial challenges remain to be overcome. The main challenges include the immunocompatibility, stability, heterogeneity, differentiation, and migratory capacity. More and more studies are focusing on the attempts to overcome these shortcomings. Although the detailed mechanism of MSCs immunomodulatory effects is still elusive and any attempts to improve MSCs efficacy are still lack of evidence, the preclinical studies are developing rapidly and more standardized clinical trials are wildly carried out. It might be expected that the conversion to canonically registered MSC therapies will flourish with time. The lessons from the current MSCs investigations may provide critical guidance for investigators pursuing further translational processes. With the clarification of MSCs effectors and the emergences of new technologies assisting in-depth studies, MSCs are promising to be proved as effective treatment options for a variety of devastating conditions.


Mobile and smartphones are owned and accessed by many, making them a potentially optimal delivery mechanism to reach pediatric patients with socially complex needs .To address the specialized needs of youth from such groups, this review synthesized the literature exploring the use of phone-based delivery to access pediatric populations with socially complex needs, targeting mental and behavioral health outcomes. The purpose of this synthesis was to provide recommendations for future research developing phone-based interventions for youth with socially complex needs.A trained medical librarian conducted the search strategy in the following databases: PubMed, Scopus, CINAHL, PsycINFO, Cochrane CENTRAL Register of Controlled Trials, Cochrane Database of Systematic Reviews, and Google Scholar. Studies targeting youth with socially complex needs were defined by recruiting samples that were primarily from traditionally underserved populations . A systematic narrative framework was utilized and the Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines were followed (registration number CRD42020141212).A total of 14 studies met the inclusion criteria, with 3 depicting the use of phones to complete assessment and tracking goals and 11 to intervene on mental and behavioral health targets.The literature indicates important directions for future research, including (1) involving diverse and representative teens , stakeholders, and clinical/research staff; (2) integrating evidence-based therapies with minority-focused theories; (3) harnessing mobile device capabilities; and (4) considering and assessing for potential costs in phones as delivery mechanisms.PROSPERO CRD42020141212; https://www.crd.york.ac.uk/prospero/display_record.php?RecordID=141212 Pediatric populations with socially complex needs are likely to have their behavioral health negatively impacted—from having a lower health status than peers to being less likely to receive appropriate physical and mental health diagnoses . The terThe ubiquity of mobile and smartphone access has promoted refrains about the promise of digital mental health tools to overcome access barriers to pediatric behavioral health interventions . While sWhile more research is critically needed, some work to harness phones as a delivery mechanism specifically for socially complex populations has begun. As the field shifts to adapt such tools to be more inclusive, synthesis of the small but existing literature may be beneficial. Indeed, this synthesis may promote increasing adaptations of such tools for pediatric populations with socially complex needs and avoid potentially superfluous evaluations that would delay deployment to youth in need of support and care. Therefore, to provide recommendations for future research developing inclusive interventions, this study systematically reviewed the literature for multiple criteria. First, a focus on phone-based interventions  was made. While the field has moved toward a focus on digital health technologies , telehealth interventions that involve calling participants—even on landlines—were included. These criteria were used because interventions using telephone calls to reach youth may still be applicable as smartphones have phone call capabilities. Second, this review focused on mental  and behavioral health targets  for pediatric patients with socially complex needs. As any pediatric patient is likely to have socially complex needs , we operationalized studies that targeted pediatric patients with socially complex needs as those that recruited samples primarily comprising  youth from traditionally underserved populations . The syThe review was conducted and reported following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) statement and checklist  and was For inclusion in the review, studies were required to (1) specifically target a pediatric population with socially complex needs ; (2) utLiterature search results were uploaded into Covidence, a not-for-profit, online systematic review service partnered with Cochrane . From thReviewer teams  extracted data  independently and in duplicate from each eligible study using an online extraction form designed by the lead author (CS-S) using Google Forms. Discrepancies were, again, resolved through consensus.The Cochrane Collaboration’s tool for assessing risk of bias in randomized trials was used to assess the risk of bias for selection, performance, detection, attrition, and reporting . Specifipopulation-specific tailoring, evidence-based skills and theories (interventions only), use of phones for delivery, and additional support. These categories were selected prior to data collection as they would provide key insights from the literature into development decisions made for specific user needs relating to pediatric populations with socially complex needs. Finally, to contextualize the findings, study characteristics, primary outcomes, and usage and acceptability were also included.Because of the variability in outcome measures and methodologies, a meta-analytic approach was deemed inappropriate for the current review. Alternatively, a systematic narrative framework was utilized, with results classified under “Tracking and Assessment” or “Intervention.” To best inform the design of future interventions for pediatric populations with socially complex needs delivered through phones, the systematic narrative synthesis included Tracking and Assessment studies will be discussed, followed by findings from the 11 Intervention studies, and finally, the outcomes of the quality of all included studies will be presented.Following the removal of duplicates identified by Covidence, 4626 titles and abstracts were independently reviewed in duplicate by 2 reviewers. A total of 69 full-text articles were reviewed in duplicate for inclusion, with 14 articles selected for data extraction. See Three studies targeted mood, anxiety, or wellness outcomes for socially complex pediatric populations through tracking and assessment. Specifically, 2 studies focused on pain tracking for African American youth with sickle cell disease (SCD) ,25. The Of the 3 studies, only Bakshi et al  directlyThe 3 studies utilized mobile phones or smartphones in some way to deliver their assessment. Bakshi et al  employedSupport beyond the described use of phones was included in the methodology of 2 studies. First, Bakshi et al  describeThe primary outcome for the studies tracking SCD symptoms was pain. However, the 2 studies reported these findings differently. Bakshi et al  reportedUsage and acceptability reports were also variable across studies. EMA was completed the majority of time for Bakshi et al , with moEleven studies reported interventions targeting mood, anxiety, or wellness outcomes for pediatric populations with socially complex needs. Specifically, interventions targeted (1) sexual risk behaviors in (i) African American adolescent females , (ii) atFour studies explicitly described tailoring of the intervention to the targeted population. DiClemente and colleagues  utilizedThree studies cited previous work depicting iterative design input from potential end users. While Leonard et al  reportedFour studies provided vague or no descriptions of tailoring the intervention to a specific population. While Perry et al  providedThe included studies reported the use of evidence-based skills and grounding in multiple theories. The most common evidence-based treatments were cognitive behavioral therapy (CBT) ,33, skilConsistent with the inclusion criteria, all studies used phones for some means of intervention delivery. Included in 5 studies, smartphone apps were the most commonly reported phone-based delivery mechanism -31,33,35With one exception , all stuP=.04) . Fu. Fu23]. This study synthesized the literature on the use of phones  as a mental and behavioral health delivery mechanism for pediatric populations with socially complex needs. There was high variability in methodological approaches and reporting of data, negating the possibility of a meta-analytic approach to this systematic review. The studies that met the inclusion criteria were primarily intervention based and occurred mainly in 2005-2007  . SamplesThe purpose of this synthesis of the literature was to provide recommendations for future research developing phone-based interventions for youth with socially complex needs. The following sections will therefore be used to discuss implications of the current findings for the development of future interventions targeting such pediatric groups. Specifically, we discuss (1) uses of phone-based delivery practices, (2) culturally specific tailoring practices, (3) applications of evidence-based skills and grounded theories to inform design, and (4) additional supports that promote comfort, use, or intended intervention outcomes for pediatric populations with socially complex needs.Given the ubiquity of mobile phones and smartphones , the useApps and SMS text messaging were the most commonly employed method to access youth. This is consistent with current usage trends, such that youth are more likely to use SMS text messaging or social media to communicate than phone calls . While aRelevant to the use of phones, potential costs associated with the use of phones as mental and behavioral health delivery mechanisms are also worth noting. Indeed, this consideration is particularly crucial in trying to access certain socially complex populations who are more likely to be impacted by lapses in service or burdened by the cost of data packages/SMS text messaging plans ,18. SeveInforming design with the feedback and preferences of likely end users is an integral aspect of user-centered design practices . CulturaInvolving representative end users  in design decisions may be achieved through multiple means. For example, the current literature detailed the use of semistructured individual interviews, focus groups, membership in a CAB, and stakeholder involvement. Recruiting youth to participate in such activities likely requires multimethod strategies, including flyers or targeted electronic chart messaging from pediatric primary care, school-based health centers, or specialty clinics. Engaging community organizations in partnership to engage youth will also increase the likelihood of receiving input from populations who have been historically less involved in research. Examples from the current literature depicted recruitment through specialty clinics  or community settings to aid in tailoring messages or determining workflow of the interventions. Message tailoring was achieved through engaging representative groups, using messages authored by individuals for themselves , or by aAlso integral to the methodologies of interventions targeting pediatric populations with socially complex needs is grounding the design in evidence-based treatments. Skills grounded in CBT were most commonly employed to achieve the intended behavioral change in the studies meeting inclusion criteria. However, theories relating to minority populations  were also used to guide the interventions. As members of underserved populations tend to have intersectional identities , grounding in theory likely also requires an intersectional approach. For example, in targeting sexual risk behaviors in African American adolescent females, the HORIZONS intervention was grounded in multiple theories, incorporating both an evidence-based treatment ,44 and aAdditional supports, whether automated, human, or both, also appear to be important for interventions targeting pediatric populations with socially complex needs. The potential for automated responses promotes the scalability of interventions for specific population targets. However, relating to the specialty needs of varying pediatric populations who may also be from underrepresented groups, the ability to have support as needed or ongoing human support may also be important. Fortunately, the incorporation of human support further opens up the possibility of increased diversity in the workforce that designs and deploys such interventions. Indeed, the US Department of Health and Human Services’ Office of Minority Health argues that one way to improve health disparities is by increasing clinical workforce diversity . IdeallyThis systematic review should be interpreted in light of specific limitations. First, the studies meeting inclusion criteria were incredibly variable in their methodologies and reporting strategies. This variability precluded a meta-analytic data approach to data synthesis and we were hesitant to overinterpret outcomes and usage patterns. Related to the variability in the studies, the search for research about “pediatric populations with socially complex needs” comprises a broad group. The current findings should be interpreted in terms of broad application to these pediatric populations. Second, the inclusion criteria for this systematic review led to the exclusion of more broad applications of mHealth and telehealth interventions for pediatric populations . It is unclear how larger reviews of the literature for pediatric populations may generalize to the populations targeted in this review, and vice versa . Third, Repeated calls have been made to better target the behavioral health needs of pediatric patients with socially complex needs. Mobile devices are often owned and utilized, and therefore may be an optimal delivery mechanism to access youth from such groups. Given the need to focus such interventions to the specialized needs of socially complex youth, this study systematically reviewed the literature of phone-based interventions  aimed at mental and behavioral health targets for pediatric populations. The synthesis highlighted the importance and potential opportunities of (1) the involvement of representative end users, stakeholders, and clinical/research staff; (2) integration of evidence-based therapies with minority-focused theories; (3) harnessing the capabilities of mobile devices, including SMS text messaging; and (4) considering and assessing for potential costs related to phones as delivery mechanisms. Future research should promote such practices and explicitly detail population-specific tailoring, usage, and acceptability of interventions delivered via mobile devices.