Mating is essential for the reproduction of animal species. As mating behaviors are high-risk and energy-consuming processes, it is critical for animals to make adaptive mating decisions. This includes not only finding a suitable mate, but also adapting mating behaviors to the animal’s needs and environmental conditions. Internal needs include physical states  and emotional states , while external conditions include both social cues  and non-social factors . With recent advances in behavioral neuroscience, we are now beginning to understand the neural basis of mating behaviors, particularly in genetic model organisms such as mice and flies. However, how internal and external factors are integrated by the nervous system to enable adaptive mating-related decision-making in a state- and context-dependent manner is less well understood. In this article, we review recent knowledge regarding the neural basis of flexible mating behaviors from studies of flies and mice. By contrasting the knowledge derived from these two evolutionarily distant model organisms, we discuss potential conserved and divergent neural mechanisms involved in the control of flexible mating behaviors in invertebrate and vertebrate brains. Reproductive behaviors are sometimes considered to be pre-programmed, “instinctive,” or “innate,” meaning that they are controlled by genetically hardwired circuits and not dependent on previous experience . While tIntrinsic and extrinsic factors often interact with one another. For example, evidence of an available food source may drive a hungry individual to favor foraging and feeding over mating, whereas food availability may enhance courtship or receptivity in a non-hungry individual. Intrinsic factors can also be periodic  or reflect certain prior experiences, including both recent  and more distant events .Drosophila melanogaster, females extend their ovipositor to reject a courting male  have been extensively described in the literature  results in decreased sexual behaviors in both sexes  alters sex-specific social behaviors  and is critical in both males and females for the establishment of appropriate social interactions, as MOE ablation and the genetic ablation of olfactory signaling th sexes . Anotherth sexes , and theehaviors . Thus, behaviors . ThroughThe neural circuits that control mating behavior in mice have been investigated for over decades. Studies utilizing brain lesioning, electric stimulation, and immediate early gene labeling in rodents have suggested that the limbic system, extended amygdala, and hypothalamus are the crucial brain structures governing sexual behaviors in males and females , many ofDrosophila melanogaster) have been studied extensively since they were first described in detail over a century ago  and doublesex (dsx) were found to play a pivotal role in controlling fly mating behaviors , and other physiological changes that reflect changing needs . External factors include environmental cues, both social and non-social. It is important to keep in mind that internal and external factors often interact. For example, the effect of smelling male bedding  depends on the female’s estrous state  , and botSexual maturation depends mostly on internal  factors; social hierarchy, social isolation, social experience, and recent mating experience depend on interactions with the external social environment, but can also be affected by internal factors. For example, how a previous fight affects mating behaviors in the present depends on both the past interactions with the environment  and on the current state of the animal . Environmental context refers to the modulation of mating behaviors by social and non-social factors in the immediate surroundings, thus depending primarily on external factors. Food-sex interactions include both the animals’ nutritional state  and food availability . Nutritional state and food availability interact such that a hungry fly or mouse is more likely to prioritize food intake over mating when food is available.In the following sections we discuss these factors individually in mice and flies, keeping the following questions in mind:1.Which nodes in the circuitry that controls mating behaviors are modulated, and by which factors?2.Which mechanisms allow for flexibility in the control of mating behaviors over short and long timescales?3.Which mechanisms for flexible mating behaviors are shared between mice and flies, and which are not?Mating is a costly process in terms of energy consumption, competition over other needs (such as feeding or sleeping), external risks , and physiological costs  . TherefoAs mammals enter into puberty, factors including somatic growth, energy balance, and the season begin to activate the Hypothalamus-Pituitary-Gonadal (HPG) axis, the central pathway that regulates the reproductive system. In both males and females, pubertal onset is triggered by the activation of hypothalamic circuitry, which ultimately controls the reawakening of gonadotropin-releasing hormone (GnRH) neurons over the course of pubertal development. Among these circuits, kisspeptin neurons are important players that activate GnRH neurons to promote the further maturation of the HPG axis . High-frSexual maturation can be measured by the physical properties of reproductive organs. In female mice, puberty can be determined by vaginal opening, which occurs around postnatal day (PND) 26–30, or by the first estrus as assessed by cytological analysis of vaginal smears, which occurs around PND 35–40 . In maleIn addition to the maturation of the animal displaying sexual behavior, the maturation of the sexual partner is also important, as sexual behavior toward juvenile animals is inhibited in adult males and females through sensory stimuli. A juvenile pheromone, exocrine-gland secreting peptide 22 (ESP22), is produced by young mice. ESP22 is secreted from the lacrimal gland and released into tears of 2–3 week-old mice. ESP22 is sensed by the VNO and strongly inhibits adult male sexual behavior toward juvenile mice  and suppDrosophila development: embryo, larva (three instar stages), pupa, and adult. Progression through all of these stages is dictated by pulses of the steroid hormone 20-hydroxyecdysone (20E)  .Drosophila melanogaster . DA controls JH levels by either stimulating or inhibiting its synthesis and degradation depending on the developmental stage , which promote courtship by detecting the aphrodisiac pheromone palmitoleic acid , also inMany questions remain open on this topic: Which nodes along the pathways from sensory inputs to motor outputs are controlled by JH? Does JH control all the aspects of sexual maturation, or are there parallel JH-independent pathways? Which aspects of sexual maturation in flies are sex-specific, and which are shared between males and females? Is sexual maturation a gradual or step-like process ?Social hierarchy  has been studied for decades  in humanDominant male mice exhibit higher levels of sexual behavior than subordinate males  and domiIn contrast to males, the relationship between social hierarchy and mating behavior is not clear in females. Although there is no difference in the plasma estradiol level between dominant and subordinate females, subordinate females express higher levels of estrogen receptors (ERα and ERβ) in the ventromedial hypothalamus (VMH) compared to dominant females . ERα neuDrosophila melanogaster, while both males and females compete with same-sex conspecifics for resources  (er flies . Interesd “pC2”) . These tDrosophila males and females are innately tuned to conspecific courtship song parameters (Drosophila (Drosophila courtship song) both behaviorally through a change in walking speed in response to pulse-song playback and neuronally at the activity level of the pulse-detecting pC2 cells. As sharper tuning in group-housed flies has been observed in males but not in females, this led researchers to posit that the sex-specific sharpening arises from the fact that group-housed males  are exposed to courtship song from other flies , which was previously demonstrated to be involved in modulating male sexual behavior  and consummatory (motor behaviors) components of mating behaviors. Mating behavior is governed by a complex interaction between different systems in the brain which process sensory inputs, regulate reward and motivation, integrate hormonal signals, and control copulation movement itself. All of these components are impacted by sexual experience, and sexual experience generally enhances the performance of sexual behaviors in both males and females.In male rodents, the detection of female odor is a critical trigger for the initiation of male sexual behavior. While disruption of either the olfactory or vomeronasal system has severe effects on male sexual behavior in many rodents , these eIn female rodents, the performance of sexual behavior is often assessed based on the interest shown in response to male cues (appetitive) and the lordosis reflex (consummatory). Together with exogenous hormone treatment, sexually experienced females show an enhanced preference for male urine over female urine compared to naïve females . ExpressDrosophila are largely innate. For example, male flies do not require tutoring in order to sing a courtship song  and the mating status of the female , to motor control (song production). However, many open questions related to this topic remain: How is male courtship ritual modulated by previous social experience? Does premature mating modulate female adult mating behavior ? Is therRemating is valuable for the propagation of a species if the probability of having more or better offspring is higher following remating. The value of repeated mating depends on the specific reproductive system of each organism and sex. The degree to which the amount of male ejaculation or the fertilization success of females changes as a function of the recent mating history differs between species. There is an adaptive value in having a lower sexual motivation  for repeated mating if the value of repeated mating is low, or if the cost of repeated mating is high, and this has spurred research interest in the circuits and molecular mechanisms that control sexual satiety in males and females.Male sexual behavior is strongly inhibited by sexual satiety. Male sexual satiety or exhaust occurs as a result of repeated copulation with the same receptive female, during which several ejaculatory series are achieved. This state of sexual satiety and a concomitant lack of interest in females lasts for an extended period, requiring between 3 and 15 days for the full recovery of sexual drive in rats  and seveThe neuroendocrine system is also involved in the control of male sexual satiety. Although plasma testosterone and estradiol levels are similar between non-mated and sexually satiated male rats at 24 h post-mating, AR density was reduced in the MPOA, VMH, and NAc , and theRelative to males, the mechanisms governing female rodent sexual satiety have been less well studied. Paced mating paradigms are often used to measure a female rodent’s sexual motivation and desire. This is accomplished using pacing chambers composed of two chambers separated by a divider with one or more small holes that only the female can pass through . The malIn a paced mating paradigm, both in rats and mice, the pattern of approach toward and withdrawal from the male is dependent on prior interactions. Females are more likely to leave the chamber with the male as the intensity of the preceding male mating behavior increases (mount without intromission < intromission < ejaculation). The latency to return to the male chamber also depends on the preceding stimulation, with the return latency being longer following an ejaculation than following a mount or an intromission . These dMuch as in males, monoamines and opioids have been reported to inhibit lordosis or potentially related to sexual satiety in female rats. The activation of delta opioid receptors in the MPOA inhibits lordosis , and thedsx-expressing pCd neurons and a set of neuropeptide F (NPF)-expressing cells. The NPF-pCd recurrent loop is also connected to the P1 neurons. Interestingly, this courtship circuit mechanism appears to be under homeostatic control, as copulation-reporting neurons (CRNs) in the abdominal ganglion suppress the NPF signaling to the DA SMPa neurons, reducing the mating drive. The CREB2-dependent transcription of specific K + channels (TASK7) in the NPF-pCd recurrent loop leads to persistent satiety following mating. Another aspect of male mating motivation relates to the drive of a given male to maintain copulation for multiple minutes despite competing stimuli. Males retain copulation motivation even when facing a threatening stimulus. This motivation slowly decays down to a low level around 6 min after the initiation of copulation, which corresponds to the time it takes to transfer the sperm to the female. This effect depends on a slow decay of CaMKII kinase activity in the courtship circuit  also depends on her recent mating history. A mated female shows rejecting behaviors toward a courting male . This efDsx-expressing pC1a neurons in the female central brain (Dsx (Doublesex). Taken together, this suggests that subsets of the sexually dimorphic pC1 cells control mating motivation in both sexes , and are involved in mediating reduced mating motivation following recent copulation. Recurrent connectivity between pC1 cells and other groups seems to play a role in controlling persistent mating motivation in males, and in controlling a persistent aggressive state in females  occurs in response to the male courtship song and is dependent on the mating status of the female. Therefore VPO depends on the integration of both exteroceptive  and interoceptive (mating status) inputs . The sexal brain . Through females . Whether females , it remaBeyond the role of the sex peptide, it has also been shown that the sensory experience of copulation is sufficient to induce a reduction in female receptivity. Neurons expressing the mechanosensory channel Piezo transmit the sensory detection of copulation to a pair of ascending LSAN neurons, which relay the information to Myoinhibitory peptide (MIP)-expressing neurons in the female brain . On the Both social cues  and non-social cues  in the immediate environment can modulate mating behaviors. While sexual behavior is often measured in the lab in isolation  and under controlled conditions , the environmental context has clear ethological relevance for mating decisions. Here, we focus on a few examples that exemplify the impact of such context on mating behaviors in mice and flies.The social environment influences sexual maturation, reproductive state, and sexual behaviors. In house mice, the sexual maturation of juvenile females is delayed by the presence of group-living adult female mice . In adulExposure to predator-related cues can also alter mating behaviors in males. Prolonged exposure to cat urine, for example, reduces male mating behaviors, likely through vomeronasal signaling . PredatoDrosophila melanogaster are highly dynamic with respect to their social environment  and food availability affect mating behaviors in mice and flies. The impact of food access on mating behaviors depends on a given animal’s hunger state. Food availability may signal a good substrate for egg-laying in flies or the feeding of progeny in mice, as well as a higher probability of a prospective sexual partner being well-fed, therefore enhancing mating probability. On the other hand, starved or recently mated flies may prefer food over sex, in which case food availability may reduce mating probability. Lastly, as mating and post-mating behaviors require energy, this hunger state is expected to influence mating decisions. Below we discuss how hunger and food availability shape mating decisions in mice and flies.Nutritional states have notable impacts on reproductive functions in rodents. Pre-pubertal food restriction leads to delays in puberty onset in both male and female rats . A low eNutritional state also affects reproductive functions at the behavioral level. Acute food deprivation or chronic food restriction suppresses receptivity (lordosis) in female rodents . In maleVarious metabolites that reflect nutrition/hunger state have been suggested to modulate sexual behaviors. Ghrelin is a hormone mainly produced by the stomach that is released at higher levels under conditions of food restriction and promotes food intake . The intBoth mating and feeding are critical behaviors for the survival of the species, raising the question of how behavioral decisions are made when these two conflicting needs coexist. Hunger states change odor preferences. Both male and female mice are equally attracted to food odors and pheromones (opposite sex urine) when they are fed. However, mice investigate food odorants more than pheromones when they have been starved for 24 h. This hunger-dependent food odor attraction is mediated by agouti-related peptide (AGRP)/NPY neurons in the arcuate nucleus (ARC) through the projection to the paraventricular thalamus (PVT) . A 48-h Drosophila melanogaster are often referred to as “fruit flies” as they (and other closely related species) are associated with the presence of over-ripened fruit and vegetables, where they aggregate, feed, and mate  expressing ORNs caused a dramatic reduction in male courtship activity. IR84a-expressing neurons are activated not by fly-derived chemicals but by the aromatic odors phenylacetic acid and phenylacetaldehyde, which are widely found in fruit and other plant tissues. Projection neurons downstream of the IR84a ORNs converge in the pheromone processing region of the lateral horn, possibly promoting male courtship behavior by enhancing the response to pheromones secreted by the female, although this was not shown directly in this study. While the olfactory responses of IR84a ORNs are not significantly different between sexes, Ir84a mutant females do not show overt defects in reproductive behaviors, including in copulation latency, success, or duration  neurons project to the AVPV and this projection is necessary to display sexual receptivity in OVX plus hormone-primed (estrus) females. The presynaptic terminal density of VMHvlPR neurons in the AVPV and functional connectivity between VMHvlPR neurons and AVPV neurons are three times higher in OVX plus hormone-primed females relative to OVX females (diestrus). This is an interesting example of how the modulation of mating behavior circuits can also occur through the periodic anatomical remodeling of circuits, rather than just as a result of the modulation of excitation/inhibition balance in an anatomically stable circuit. Another example of the mouse-specific modulation of mating behavior relates to the ability to recognize an individual partner. Specifically, the motivation of the male to remate is higher when the prospective female is a novel female as compared to remating with the same female. While it is possible that flies are capable of distinguishing between individuals  reportedly has a stronger impact on males relative to females in both mice and flies, with the winners being more successful in mating in both organisms. Group-rearing, as compared to rearing in isolation, has profound effects on the circuits controlling mating behaviors in both species, from sensory processing to behavioral control. For example, social experience has been shown to modulate the responses of female mice to urine odors and the responses of female flies to male courtship song. Strikingly, there is evidence that social and sexual experience may overcome even severe circuit manipulations: While disruption of either the olfactory or vomeronasal system can have a strong adverse impact on male sexual behavior in many rodents, these effects are less severe if subjects are sexually experienced when they undergo disruptive manipulation. In Circuit nodes controlling mating behaviors can be broadly divided into three levels: sensory processing, behavioral decision making, and motor execution. As is evident from the studies of flies and mice reviewed above, internal and external factors influence neural responses at all three levels. While a simplified feed-forward model of information flow from the sensory periphery to the motor periphery is often used as in , the celHormones and neuromodulators seem to play pivotal roles in the flexibility of mating behavior, though likely to a greater extent in rodents than in flies. DA  has been reported to be a currency of sexual motivation in both species, where DA levels in specific circuits link recent mating experiences with sexual satiety/motivation. In both species (and in both sexes) low nutritional levels reduce mating motivation. Relatively small populations of neurons  integrate multisensory information and control flexible mating behaviors in these two species. As previously noted, these similarities may be superficial and coincidental, or may represent conserved or analogous modules for the high-level control of flexible mating behaviors in both model systems. Another plausible mechanism for incorporating direct mating cues regarding a potential mating prospect and other internal or external factors is a balance between inhibition and excitation that inputs to a common node. In flies, for example, inhibitory and excitatory pathways converge on the command neuron P1. P1 neurons in male flies control the initiation and persistence of courtship, and the balance between the excitatory and inhibitory inputs to this population determines the level of P1 activity. These inputs to P1 carry both pheromonal and olfactory information about the prospective mate, leading to a higher net response of P1 to an appropriate mating partner. It is tempting to speculate that a balance between inhibitory and excitatory inputs may also serve to integrate information about the environment  and about a mating prospect , thus allowing for flexible mating decisions. NPY and its fly homolog NPF are associated with both feeding and mating motivation, possibly linking the two behaviors.Mating behavior is fundamental and universal among all species, including humans. Based on the assumption that mating is a primitive behavior that is essential for the survival of the species, the control of mating behavior may be relatively preserved over the course of evolution. Efforts should be made to leverage the knowledge gained from animal studies to shed more light on human sexual behavior and the underlying neural circuitry with the goal of answering questions including: How do social isolation, sexual experience, and social context modulate sexual behaviors in healthy and non-healthy human individuals? How is sexual behavior modulated by internal motivational states? What goes wrong in this system in the case of pathological behaviors?A better understanding of the relationship between the mechanisms governing flexible sexual behaviors and non-adaptive or pathological behaviors is missing in both clinical and non-clinical human populations. Particularly in disorders characterized by rigid and/or repetitive behaviors, such as obsessive-compulsive disorder or autistic spectrum disorders, social challenges are common and often hamper sexual relationships . In thesWhat is the neural basis of maladaptive sexual behaviors? The dysfunction of sexual behavior can arise at different stages of the sensorimotor pathway. For example, individuals that experience compulsive sexual behavior (CSB) have been reported to exhibit enhanced attentional bias to explicit cues , possiblvia a closed loop system that computes the difference between the animal’s need and the current state at any given point in time  are more likely to occur in response to different factors or at different levels of the circuitry. More broadly, it also remains uncertain whether there are any unifying principles for the control of flexible mating behaviors across taxa. Different conceptual models have been suggested for the neural circuit mechanisms that encode motivational states. According to Tinbergen’s hierarchical model for behavioral decisions , the hig in time . UncertaIn both rodents and flies, there have been significantly fewer studies addressing behaviors and neural circuits in females relative to the numbers performed in males. There are a few reasons for this bias. First, as in many species, including mice and flies, the males actively court and the females are considered more passive, such that quantifying mating phenotypes in males  is easier. Second, in some species, including mice, the female estrous cycle is considered to add an experimental factor that must be controlled for, whereas this can be avoided by using males as experimental subjects. As the brain, body, and associated physiology are subject to sexual dimorphism, the knowledge derived from studies of males cannot simply be applied to females, and therefore more research is necessary to comprehensively understand the biology of mating behaviors and reproduction. While significant progress has been made in dissecting the neural circuitry underlying mating behaviors in female mice and flies , this gaWhile using genetic models such as mice and flies to study the neural basis of behavior has clear advantages, a comparative approach involving a wider range of animal models is critical if we aim to understand how nervous systems have developed to enable adaptive mating behaviors in different species and under different evolutionary constraints . MoreoveAs mating behaviors are critical for animal survival, they must have a pre-programmed component, but at the same time, they must be flexible enough to allow the circuits to adapt to changing needs and environments. While much progress has been made in our understanding of the neural basis of mating control in genetic model systems including mice and flies, many questions remain open. We believe that the emergence of new experimental and computational tools, the study of males and females of diverse model species, and new theoretical models that take into account the broad ethological context of mating behaviors will ultimately lead to exciting new findings in this fundamental field of neurobiology in the near future.Both authors listed have made a substantial, direct, and intellectual contribution to the work, and approved it for publication.


Phenology refers to the seasonal timing patterns commonly exhibited by life on Earth, from blooming flowers to breeding birds to human agriculture. Climate change is altering abiotic seasonality  and in turn, phenological patterns contained within. However, how phenology should evolve is still an unsolved problem. This problem lies at the crux of predicting future phenological changes that will likely have substantial ecosystem consequences, and more fundamentally, of understanding an undeniably global phenomenon. Most studies have associated proximate environmental variables with phenological responses in case-specific ways, making it difficult to contextualize observations within a general evolutionary framework. We outline the complex but universal ways in which seasonal timing maps onto evolutionary fitness. We borrow lessons from life history theory and evolutionary demography that have benefited from a first principles-based theoretical scaffold. Lastly, we identify key questions for theorists and empiricists to help advance our general understanding of phenology. Phenology—the seasonal timing of biological events on scales ranging from individual life cycles to global cycles—is a universal feature across plants and animals, from ecosystems  to human systems  –3. PhenoPhenology—regardless of scale or system—describes cyclical patterns in the dimension of time . HistoriClimate change influences cyclical timing patterns in 2 main ways. The first is via overall warming , e.g., iPhenological shift: directional change in the timing or phase duration of life cycle schedules within the context of geophysically fixed annual oscillations in the environmentPhenophase: the duration of a categorically distinct phase of a life cycle, such as adolescence or reproductive periodProximate phenological causality: system-specific triggers that induce the expression of phenological traitsUltimate phenological causality: broad evolutionary forces that influence the seasonal timing of universal life history traits such as birth, growth, reproduction, and death, considering that the timing of each trait contributes to fitness and all are constrained by trade-offs with one another Life history evolution: the evolution of the holistic suite of life cycle traits in the context of the ecology of populations as well as predictable fluctuations in the environmentEvolutionary demography: selection dynamics that produce, as well as directly result from dynamics in the size and structure  of populationsEco-evolutionary dynamics: the concurrent and reciprocal dynamics of ecological and evolutionary processes where one shapes the context of the other, usually described as a feedbackPerhaps the most unresolved conundrum is that the same change in climatic growing seasons often induces very different phenological shifts between organisms occupying the same habitat in both direction (when) and magnitude (by how much). Discrepancies in shifts are observed among individuals , betweenIn summary: The rapidly growing body of top-down (observations first) studies of phenological change is brimming with contrasting effects and specific explanations, which makes it difficult to generalize the eco-evolutionary links between seasonality change and phenological change across systems ,46,47. WIn our first section, we argue that a first principles view of phenological evolution starts with the recognition of a simple truth in any system: the fitness consequences of some key phenological traits vary over time within a year. In other words, if any phenological manifestation confers an equivalent consequence for fitness, there would be no discernible phenological pattern around the planet. Drawing analogies from spatially oriented theory, we highlight that discretizing the dimension of time in bounded units enables quantitative conceptualization of how phenological variation maps to variation in fitness. Then, we outline how this variation in fitness produces selection pressures on phenological variation, first at the scale of individuals and populations, and then at the scale of multispecies communities. Our overarching goal is to introduce theorists to the unsolved puzzle of general selective forces acting on phenology around the world and invite empiricists to test those emerging hypotheses to advance cross-system understanding.Patterns, in any dimension, can only be quantified and systematically compared in the context of defined bounds and scales. Drawing analogies from the more familiar spatial dimension helps crystallize this point . EcologiFocusing on the temporal bounds that encompass annual patterns might be an important step for first principles theory development because bounds are one of few parameters that are universal. In other words, any system that exhibits cyclical phenological patterns has a beginning and an end to the seasons that constrain the sequence. The relationship between the size of the bounded domains in natural systems—whether in space (area) or time (duration)—and the biological patterns they contain is often complex, and thus inevitably requires mathematical modeling. For example, in space, the size of islands or habitat patches nonlinearly determines biodiversity, distribution, and coexistence patterns contained within that space ,10,49–52Arabidopsis show that phenological traits like flowering time are expressed in the laboratory even in the absence of climatic cues characteristic of the natural populations’ localities and can even be mapped to specific genes under selection [In theorizing the causality behind any change in temporal patterns, it is also important to keep in mind that cyclical phenological patterns are distinct from emergent “phenomenological cycles” that arise from internal systems dynamics  or Markovian transition processes . In contrast, phenological patterns are evolutionarily adaptable strategies that are repeatedly expressed within periods of geophysical environmental cycles  in time, from past to future. The anisotropic nature of temporal patterns makes causal influences stronger in time than in space since effects from multiple directions can be counteracted or obfuscated in space . In otheA collection of species that exhibits a repeatable phenological pattern year to year in the same space may consist of strikingly different generation times or activity schedules; hence, those species’ population dynamics actually operate on very different scales of time . AnnualTowards that goal, we ask a general guiding question: How do organisms that live in environments with periodic time windows evolve to utilize nonrandom portions of the windows? We break the question down to 2 key hierarchies: single-species phenological evolution and community interactions that influence multiple coexisting phenologies.Phenological timing is typically studied as a variable responding to seasonal transitions in the abiotic environment . Responses to seasonal environmental variables, often involving plastic expressions of traits ,72, consA sense of what constitutes “correct” timing, or the baseline null expectation of how phenological timing should change given some change in the environment, is currently not theoretically generalized. Expectations are often set by intuitions that can arise from system-specific knowledge, e.g., food availability for birds . HoweverLife history theory and evolutionary demography  have conSimilar to the phenotype-to-fitness mapping considered by life history and demography theorists, timing of occurrence or trait expression within seasonal windows is an axis that covaries with fitness ,93,94. TAnalyzing phenological traits as independent response variables oversimplifies the manner in which life cycles are structured by trade-offs and contingencies between life history traits ,102 Fig. The expLife history optimization studies historically assumed constant environments, but theorists recognize that stochasticity in the environment can produce very different outcomes ,106–109.2 cycle [However, the manner in which life history timings are shaped by nonrandom cyclical temporal structures in the environment—such as those governed by Earth’s rotation around the sun which is geophysically locked—remains much less well understood theoretically . Specifi2 cycle ) and phe2 cycle .Above examples of studies that incorporated life history interdependencies and demographic structure into phenological analysis demonstrate that phenology is a highly eco-evolutionary process  and woulIn testing phenological evolution theory using the common types of phenological data, a nuanced conceptual gap that needs to be bridged is one between how “rate”  evolves and how “timing”  evolves. Rates are the parameters typically manipulated in demographic and life history models due to the time differential nature of dynamical systems modeling. Such models ask what happens over a fixed time step, whether that be a large step  or an infinitesimally small one . As a sPhenological evolution occurs in the context of ecological communities. The challenge is to understand how periodic interactions between coexisting species influence each species’ adaptive occupation of different portions of seasonal windows. Empirical evidence shows that different types of ecological interactions such as competition, invasion, or consumer-resource dynamics can alter the occurrence or trait expression timing of species in a community. Broadly, periodic interactions can favor overlap  or segresensu [The key ecological consequence of the differential expansions, contractions, and shifts among species’ phenologies is that the interaction potential between combinations of species can change within seasonal time windows ,123,124.sensu ) can forsensu  increasesensu . Among ssensu . Similarsensu , anuranssensu ,129, vassensu ,130,131,Within our framework of conceiving phenological phenomena as fitness-related distributions in time windows , we propTo what extent are parameters of abiotic environmental cycles  themselves agents of phenological selection? In other words, how do seasonal cycle parameter changes map to fitness landscape change? Does this perspective help explain phenological shifts in the context of global change, wherein parameters of environmental cycles are being altered?How do life history traits—which are intricately interdependent due to covariances and trade-offs—evolve upon a template of environmental cycles, which are themselves structured by temporal autocorrelation and lag effects? Recognizing connections among phenological traits, borrowing from life history theory, will advance our understanding of phenological shifts beyond correlative approaches that focus on single traits. We have highlighted that marrying concepts of timing and rate in models will be important.Temporal autocorrelation can occur at various resonances that may constructively or destructively interfere with seasonal cycles . How do multi-resonance regimes influence the evolution of seasonal phenology?Do phenologies of organisms with different numbers of generations per seasonal window evolve in fundamentally different ways, as the windows expand, contract, or become otherwise distorted ?How can we better integrate empirical approaches to enhance our general understanding of phenological evolution? As an example, can models that make predictions about optimal phenology  as a function of environmental cycle parameters  be tested by manipulating those parameters and measuring genetic and phenotypic frequency change simultaneously in experimental populations?How is the timing of a trait that is important from the perspective of the community  controlled simultaneously by life history optimization at the species level and periodic interactions with other species that favor co-occurrence or temporal segregation? Are there general rules regarding if and when single-species evolution or multispecies interaction is a stronger driver of phenological selection in nature?How do changes in seasonal time window parameters alter interaction potentials in ecological communities, create novel no-analog communities in different portions of the season, and affect phenological diversity?Phenology is ubiquitous. Species around the planet have evolved a panoply of physiological, genetic, behavioral, plastic, and neuronal mechanisms to strategically utilize seasonally available time windows ,132–135.While the sheer ubiquity of phenology itself warrants theoretical synthesis, global change makes it urgent. Perturbations to the cyclically occurring seasonal windows—analogous to spatial perturbations to habitats such as deforestation—are altering the familiar phenological patterns contained within those windows in perplexing ways ,126. CorUSA National Phenology Network and the Pan-European Phenology Database, and will make such interdisciplinary and comparative investigations possible.The microevolutionary process of phenological evolution will be another frontier of investigation. Population genetic processes such as recombination and drift are constrained by the seasonally fluctuating probabilities of encounter among individuals, which limit periodic opportunities for mating and gene exchange, and thus the inheritance of phenological traits. Studying how all of the above processes per species are simultaneously influenced by interactions between species in ecological communities will be the next challenge. We have strived to distill key questions for future investigators, particularly those interested in developing general theory . EmpiricFinally, the general concepts about time windows we discussed extend to a fundamental theme in ecology and evolution, beyond just the scale of seasons. While phenological research has focused on the seasonal scale, cycles in the physical environment in fact exist on many other temporal scales such as daily, tidal, and multiannual , with “phenological” scales to match . Geophy


Leber’s Hereditary Optic Neuropathy (LHON) is the most common primary mitochondrial DNA disorder. It is characterized by bilateral severe central subacute vision loss due to specific loss of Retinal Ganglion Cells and their axons. Historically, treatment options have been quite limited, but ongoing clinical trials show promise, with significant advances being made in the testing of free radical scavengers and gene therapy. In this review, we summarize management strategies and rational of treatment based on current insights from molecular research. This includes preventative recommendations for unaffected genetic carriers, current medical and supportive treatments for those affected, and emerging evidence for future potential therapeutics. Inherited optic neuropathies have been estimated to affect 1 in 10,000 individuals and are an important cause of visual impairment . ClinicaTo date, treatments have been limited and there is no curative therapy. The European union has authorized the use of Idebenone in patients with LHON and current consensus guidelines recommend its use in affected individuals . AlthougLHON is uniquely placed for scientific study as most cases occur from identifiable point mutations, it affects a unique and measurable cellular line, and is one of the more commonly inherited mitochondrial disorders. Clinicians and scientists have been working on further defining the clinical disease course of LHON in combination with the molecular impact of genetic and environmental factors. This review will discuss the natural disease course and molecular mechanisms in the development of LHON to allow for an in-depth review of the current and future treatment strategies in LHON. It is this multifaceted understanding of specific pathways of damage and their influence on LHON and visual outcomes which have provided insights to allow for the creation of novel therapeutic strategies. Additionally, as LHON has become a model for mitochondrial disorders, the advancements in this field serve to inform research of other related disorders.LHON has been subcategorized into disease states to help further define and approach clinical interventions . It is iCurrently, the asymptomatic phase is defined as any individual who is a carrier of one of the known causative mutations but is not experiencing visual loss. Due to highly variable, incomplete penetrance, LHON individuals may never develop vision loss. Carriers and pre-symptomatic patients may demonstrate clinical findings in the absence of subjective vision changes. Inferotemporal retinal nerve fiber layer (RNFL) swelling can be seen on fundoscopic exam or Optical Coherence Tomography (OCT) and is not indicative of progression to vision loss ,7. Pre-sThose who convert from asymptomatic to symptomatic states typically do so in their second or third decade of life . Diseasem.11778G>A mutation have been found to have the worst visual outcomes [After about 1 year stability occurs, and it is at this time patients are considered to have transitioned to the chronic phase of the disease ,19. Someoutcomes ,21. MostIt should be noted that the disease state categories above are applied to help define disease process for clinical and scientific trials; however, it is becoming further elucidated that the LHON phenotype is fairly heterogenous. There is variability between each mutation in regard to timing, severity and outcomes. Additionally, within specific subgroups of genetic mutations there is variability given environmental and genetic heterogeneity within the population, for example mtDNA haplogroup polymorphisms .There appears to be two important subcategories of clinical manifestation that has been termed LHON Type I and Type II. Type I is likely to be a mtDNA mutation in the context of nDNA which is susceptible. This gene-gene process may be inevitable. Type II may be Type I cases in which environmental factors are key and lead to conversion. This might explain several distinctions in presentation. Type I is abrupt (sub-acute), and Type II is more insidious. Type I occurs about age 20 and Type II often after age 40. Type II is almost always associated with a major exposure to smoke or smoking. Type II is more likely to recover if the patients risk factor is removed. Most importantly, in Type I there is profound loss of both structure (OCT-RNFL) and function (VFs), but in Type II there is often a structure-function mismatch (RNFL is partly preserved) . m.3460G>A), ND4 (m.11778G>A), and ND6 (m.14484T>C) subunits. Complex I then uses two co-factors: coenzyme Q  and Cytochrome c (cyt c) to transfer electrons to the succeeding respiratory complexes . There is also, to a lesser extent, a reduction in ATP synthesis. Mitochondrial respiration is driven by redox reactions, organized through mitochondrial electron transport chain complexes. The complex I (a NADH:Ubiquinone oxidoreductase) proton pump is the first step in the respiratory chain and couples electron transfer with proton translocation across the mitochondrial membrane. This complex contains 45 subunits, 7 of which are mtDNA encoded . The 3 momplexes . LHON muCurrently, there is no recommended treatment for those individuals who are known carriers of the LHON mutation . That beAs LHON is a maternally inherited, a male (affected or unaffected) with a known pathogenic mtDNA mutation cannot transmit this to any of his offspring, and a female with a similar mutation will transmit it to all her offspring. As LHON exhibits incomplete penetrance the appearance of disease may vary widely within each family or between families with the same mtDNA mutation. 60% of families will have a history of visual loss affecting maternal relatives. Incomplete penetrance further complicates genetic counseling because it is unknown when or if an LHON mutation carrier will become symptomatic. Unaffected family members should be cautioned that finding the mutation in relatives in maternal line is very likely, and the results will not aide in prognosis. The current international consensus statement released in 2021 is that all maternally related family member do not currently need to be tested. It is generally agreed upon however that they should be screened [If individuals do receive genetic testing, the large majority of positive individuals will be homoplasmic. Heteroplasmy is well documented and genetic testing results documenting heteroplasmy must be carefully interpreted as mtDNA in peripheral blood may not reflect the heteroplasmic load in the RGCs . Along sLastly, haplotype J is now well documented as leading to a higher conversion and therefore increases the penetrance of LHON mutations. The phenomena is felt to be due to lower mtDNA and mtDNA-encoded polypeptides This is currently of less importance in regard to genetic counselling clinically, but if known for a carrier may lead to some prognostic value ,29. The incomplete penetrance seen in LHON lends to the classic hypothesis and well accepted statement that mtDNA mutations are necessary for an individual to be affected but alone are insufficient to cause vision loss. It is the addition of environmental factors which allows for conversion from asymptomatic individuals  to affected individuals . A growing body of data is now available on mitochondrial toxicity, in particular increases in oxidative stress induced by tobacco smoke, other smoke, and alcohol.Multiple studies have shown that alcohol consumption at high levels, especially binge-drinking, and smoking tobacco have a strong association with more severe symptoms and prognosis in LHON carriers ,30. AffeIncreasing in the mitochondrial quantity and mtDNA copy number to increase total ATP production is a theorized method in which one may prevent disease conversion in pre-symptomatic individuals. This compensatory mechanism has been highlighted in LHON and it has been postulated that increased mitochondrial biogenesis may be a responsible driving factor in its incomplete penetrance . AlthougAdditionally, dysfunctional mitochondria are implicated in the triggering of apoptosis. The quality control of mitochondria requires a complex balance of mitobiogenesis and autophagy. Autophagy involves lysosomal degradation and elimination of dysfunctional and damaged proteins and organelles. Autophagy may be significantly compromised in cells with LHON mtDNA mutations. This results in reduced clearance of dysfunctional mitochondria (mitophagy) and is a contributing factor to cellular impairment and death. Pharmacologic activation of mitophagy in LHON cell models has been shown to selectively clear damaged mitochondria and improves overall cellular survival . TransmiMany drugs have been studied, among them metformin, rosiglitazone, resveratrol, vitamin B3 and NAD+ precursors which have shown promising results; but no studies specifically relate to vision loss or optic nerve tissues ,38,39. SIn relation to the section above, estrogens should be discussed during the pre-symptomatic stage of LHON, particularly when it comes to prevention in female patients, as this has one of the largest bodies of literature in relation to potential neuroprotection in LHON. Reduced prevalence of LHON affected women is well established, but there is also an increased incidence of disease seen with patients that may correlate with a decline in estrogens . It has been found that estradiol levels increase mtDNA content, oxygen consumption and ATP levels in human cybrids with LHON mutations . In vitrOxidative stress and its modulation is a focal point in mitochondrial disease. The potential benefit in using antioxidants to help reduce the neurotoxic strain imposed on RGCs by ROS is quite a desirable theoretical avenue of treatment. The hypothesized mechanism of reducing the neurotoxic stress from multiple supplements  and their combinations have been attempted to help reduce this ROS imposed stress. None so far have been shown to have any significant benefit in LHON and currently there is not enough scientific evidence to support their use clinically. Of the various over the counter options, the ubiquinone family seemed the most promising for effective treatment. This coenzyme shuttles electrons between complex I and II to complex III of the ETC. The reduced CoQ10 therefore also has antioxidant properties, minimizing ROS generation. Additionally, CoQ10 might deliver the electron as needed to complex III thus restoring energy production. However, the major hindrance to its effect is its inability to cross cell membranes, the mitochondrial membrane, and blood–brain barrier due to its long lipophilic tail and therefore low bioavailability. It has been shown to be partly effective in other mitochondrial related disorders but has not panned out for LHON ,44. In oIdebenone is a synthetically derived molecule which similarly facilitates electron transfer between ETC complexes allowing for improved ATP production and minimized ROS generation . This soIn 2011 a randomized clinical trial “Rescue of Hereditary Optic Disease Outpatient Study” (RHODOS) presented evidence that idebenone can be beneficial to preserve and/or restore vision . This prThese results then triggered a seminal event in LHON treatment, the European Union’s authorization of the use of Idebenone in patients with LHON in 2015. This was then followed by the 2017 consensus conference to address therapeutic issues in the treatment of LHON. The consensus statement and most current recommendation is that Idebenone should be started as soon as possible in patients with disease onset of less than 1 year. Additionally, in these subacute/dynamic patients, treatment should be continued for at least 1 year to assess the start of therapeutic response or until a plateau in terms of improvement is reached ,53.Since then, the previously reported beneficial effect of idebenone on recovery and preservation of vision has been confirmed by multiple other major studies, including the expanded access program (EAP) , a JapanElamipretide (MTP-131) is an antioxidant peptide which has been found to increase ATP synthesis as well as reduce ROS production . It targAlpha-tocotrienol quinone (EPI-734) is a third generation quinone molecule that has been studied in depth in vitro and needs further investigation to determine benefit in LHON patients. It has been used and studied in other inherited mitochondrial diseases and has been found to be approximately 1000 to 10,000 fold more potent than coenzyme Q10 (because of drug delivery) or idebenone (because of electron delivery) in protecting mitochondria. It is an antioxidant para-benzoquinone that also replenishes glutathione, acting on oxidoreductase enzymes. Case reports or case series support its benefit in LHON; the largest to date being a small open-label trial, in which EPI-743 arrested disease progression and reversed vision loss in 4 of 5 treated patients with LHON ,59.Drugs or molecules developed for use in other pathologic processes which play a role in the modulation of the cellar intrinsic pathway or apoptosis may lead to promising therapy. Cyclosporine A inhibits opening of the mitochondrial permeability transition pore and is one of the anti-apoptotic drugs that has been studied in LHON. Oral cyclosporine (2.5 mg/kg/day) was given to patients in the hope of preventing second-eye involvement in patients with strictly unilateral Leber’s hereditary optic neuropathy. Despite cyclosporine treatment, second-eye involvement occurred in all five patients included in the study, resulting in severe loss of vision, down to 20/200 or less. In addition, there was also a worsening of the visual acuity, the mean visual field defect, and the average thickness of the GC-IPL in the first eye affected . Other mPhotons from Near Infrared Light (NIR) light can penetrate diseased retina, be absorbed by mitochondrial photoreceptors, such as cytochrome c oxidase, and potentially promote mitochondrial energy production. Evidence has suggested that NIR light may restore biological function of damaged mitochondria, upregulate cytoprotective factors and inhibit apoptosis . NIR ligElectrical stimulation techniques may have shown potential benefits for treatment in diseases which affect the retina and optic nerve. Non-invasive electrical stimulation has been evaluated in traumatic non-arteritis optic neuropathies as well as retinal disease . Recent The current therapies discussed thus have focused on prevention and compensation. These pharmacologic mechanisms have been ones which ameliorate mitochondrial dysfunction at the cellular level by reducing, or attempting to reduce, ROS generation and the impact of mtDNA mutations on cellular homeostasis. Gene therapy, which alternatively targets replacement and repair of dysfunctional or damaged cellular pathways, poses an exciting new approach in the field. In order to deliver a gene product to a mitochondrion there are several steps. The vector must endocytose within the cell, internalize within the mitochondria, and affect mitochondrial metabolism. This is very problematic, given that many DNA molecules do not cross mitochondrial membranes unaided. Additionally, mitochondrial genes require allotopic expression. Allotopic expression of mitochondrial genes is the deliberate relocation of mitochondrial gene into the nucleus followed by the importation of the genetically encoded polypeptide from the cytoplasm into mitochondria. The most studied mechanism to date for replacement of dysfunctional mtDNA is via intravitreal (IVT) injection of viral vectors. The currently most established gene therapy consists of a wild-type of ND4 subunit packaged into an adeno-associated-virus 2 (AAV2) which is injected into the vitreous and targets the closest cells which are RGCs near the macula. In theory, upon cellular transfection, the wild-type ND4 gene is allotopically expressed. The AAV2 gene therapy vector carrying the wild-type ND4 gene gets transported to the nucleus where it is transcribed into messenger RNA which is later transcribed by ribosomes. The genetic sequences encoded also optimize translocation into the inner mitochondrial matrix and allow its integration within complex I to restore function . PreclinTo date, there have been several phase I, II and III studies for LHON gene therapy. RESCUE and REVERSE are two randomized, double-blind, sham-controlled, multi-center, phase III clinical studies in which an intravitreal injection administered AAV recombinant wild type ND4 in one eye and an intravitreal administration of sham injection was delivered in the fellow eye . In 2017Currently there is not enough evidence to recommend treatment in patients in the later chronic stage who have experienced vision loss bilaterally, and there is no evidence to recommend treatment beyond 5 years from onset . OverallCurrently, with limited available treatment options, the majority of patients with symptomatic LHON will progress to legal blindness . As previously mentioned, many will have central scotomas with residual peripheral vision and are good candidates for low vision therapy and rehabilitation services. Additionally, those patients with a fenestration through which they have improved acuity, may be able to maximize this through assistive devices to maintain independence. Ample digital devises, such as smart phones or iPads, can be very helpful. Evidence supporting low vision rehabilitation is less robust, and a recent 2020 Cochrane review found that there was low-certainty evidence that some rehabilitation interventions improved vision related quality of life compared to usual care. Particularly, psychological therapy and methods of enhancing vision had the best evidence . OverallThe impact of LHON extends beyond visual and activity related limitations. An important aspect of care for patients with LHON is the psychologic impact it has on both patients and carriers ,75. As LAs stated, the chronic phase has particularly challenging limitations in treatment. Once optic atrophy has occurred there is currently no avenue to reverse this process or regrow damaged fibers. The intention of the previously discussed therapies was maximizing remaining vision and prevention of further loss. Induced pluripotent stem cells (iPSCs) present a potential remarkable and spectacular avenue for reversal of vision loss. This field of research could in theory be effective for such a broad scope of etiologies of vision that it would revolutionize the field. At this time, however, it is an extremely challenging option with many hurdles to overcome before becoming clinically feasible. Unlike other cellular derived lines which are more amenable to stem cell related therapy, the neuronal pathway a RCG must travel the re-establish cerebral connections is exceedingly difficult. In 2006, technology to reprogram somatic cells from patients to human iPSCs was developed . Since tGiven the limitations in current therapeutics in patients with LHON, the concept of mitochondrial genetic engineering presents a novel and exciting focus. Gene editing tools such as zinc-finger nucleases (mitoZFN) and transcription activator-like effector nucleases (MitoTALENs) have been shown to eliminate mutant mtDNA and shift heteroplasmy . Recent In conclusion, the field of therapeutic options for LHON is still limited but has recently shown a dramatic expansion in scientific research and potential. This article reviewed supportive, preventative, and interventional therapeutics, how they are currently applied to clinical disease stages, and future promising areas within the field. The novel insights in molecular mechanism and cellular pathology found in LHON has created multiple new approaches for a deeper understanding of LHON and other mitochondrial optic neuropathies and related diseases. Presently, Idebenone remains the cornerstone of treatment and the only currently licensed therapy; however, the vast arsenal of potential therapies spans everything from repurposing established drugs, creation of novel molecules and gene therapy. The future of the field holds the promise of more efficacious treatment options for the treatment of LHON optic neuropathy.


Cells is devoted to the many aspects of MSC-based therapy and includes original data papers and reviews on preclinical models, clinical studies, as well as MSC manufacturing protocols.The use of mesenchymal stromal cells (MSCs) for clinical application is intensively investigated for a variety of areas, such as bone repair, haematological and autoimmune diseases, and solid organ transplantation. Moreover, increasing insights into their mechanism of actions revealed a lively (paracrine) interaction between MSCs and recipient (immune) cells and identified novel mediators such as extracellular vesicles (EVs). Both paracrine signals and EVs have been shown to play a role in the effects of MSCs and may potentially complement MSC-based therapies or represent therapeutic alternatives. This Special Issue of There are still limited options for functional rehabilitation after ischemic stroke. Early approaches using MSCs have been shown to be unable to replace dead neurons after stroke events in a preclinical setting, but other features of MSCs, such as the secretion of growth factors at the injured site, the promotion of axonal outgrowth, neuro- and angiogenesis, and synaptic remodeling, have been identified in recent years as key mediators of their therapeutic effect. Another treatment option commonly used for stroke recovery is physical exercise (PE). PE ameliorates motor dysfunction, improves neuroplasticity, and decreases cognitive decline in the brain. Nucci et al. investigated the effect of MSC-based cell therapy and physical exercise training in a murine stroke model using noninvasive molecular imaging techniques. The combination of local MSC treatment and PE resulted in the improved execution of complex movements and a faster recovery of symmetry over time . MoreoveSystemically delivered MSCs for ischemic stroke therapy would have to cross the blood–brain barrier (BBB) in order to reach their site of action and induce their therapeutic effects. Yarygin et al. reviewed the available data on intra–arterial MSC administration for the intra-cerebral delivery of MSCs. They concluded that administered MSCs either temporarily attach to the walls of the cerebral vessels and ultimately return to the blood stream or penetrate the BBB, resulting in homing in the perivascular space and deeper migration into the parenchyma .The preconditioning of MSCs prior to in vivo administration is a promising opportunity to enhance their therapeutic efficacy. The preconditioning of MSCs by modifying their physical environment, chemical or pharmaceutical agents, bioactive factors, or specific gene manipulation could prolong survival and improve the function of immunomodulatory effects. The review of Cheng provides insights into different preconditioning strategies aimed at optimizing MSCs for inhibiting allograft rejection. It discusses novel directions, such as the simultaneous transfection of multiple genes or the ex vivo treatment of the allograft before implantation for further improvements .This Special Issue also covers the important topics of MSC manufacturing guidelines and storage. According to European regulation 1394/2007, MSCs are an advanced therapy medicinal product that must be produced following good manufacturing practice (GMP) standards. This requires strictly organized protocols for staff organization; premises/equipment qualification and monitoring; raw material management; starting materials; technical manufacturing processes; quality controls; and the release, thawing and infusion of MSCs . LechantNot only are manufacturing protocols important factors that can affect MSC viability and subsequent efficacy before their actual application, but cell storage media and MSC preservation are too. Ścieżyńska et al. reviewed the influence of hypothermic storage fluids on MSC stability. They conclude that MSCs need to recover after cryopreservation, prior to administration, to optimize in vivo survival as well as immunomodulatory and therapeutic properties . HoweverThe review presented by Ścieżyńska et al. summarizes a problem of the MSC field in general. Today, there are more than 75,000 publications on MSCs and this ever-expanding flood of articles makes it hard to extract findings that move the field further towards the development of MSC therapy with clinical efficacy. It will be helpful to encourage the publication of negative studies in the field to form a balanced view of the therapeutic possibilities and impossibilities of MSCs. In addition, it will be necessary to re-think the evaluation of published work solely via impact factors by adding a reproducibility factor as a quality indicator . In such


Abdominal aortic aneurysm (AAA) is a chronic, life-threatening vascular disease whose only therapeutic option is a surgical repair to prevent vessel rupture. The lack of medical therapy results from an inadequate understanding of the etiopathogenesis of AAA. Many studies in animal and human models indicate a ‘short-circuiting’ of the regulation of the inflammatory-immune response as a major player in the AAA chronic process. In this regard, perivascular adipose tissue (PVAT) has received increasing interest because its dysfunction affects large arteries primarily through immune cell infiltration. Consistently, we have recently produced evidence that innate and adaptive immune cells present in the PVAT of AAAs contribute to sustaining a damaging inflammatory loop. However, it is still unclear how the complex crosstalk between adaptive and innate immunity can be self-sustaining. From our perspective, trained immunity may play a role in this crosstalk. Trained immunity is defined as a form of innate immune memory resulting in enhanced responsiveness to repeated triggers. Specific innate stimuli and epigenetic and metabolic reprogramming events induce and shape trained immunity in myeloid progenitor cells improving host defense, but also contributing to the progression of immune-mediated and chronic inflammatory diseases. Here we present this hypothesis with data from the literature and our observations to support it. Abdominal aortic aneurysm (AAA) is a chronic and life-threatening vascular disease with a degenerative course leading to rupture of the aorta with fatal consequences in more than 80% of cases. Epidemiologic studies have identified older age, male sex, smoking, family history of AAA, cardiovascular comorbidities , and to lesser extent hypertension and dyslipidemia as the major risk factors associated with the development of AAA . PatientAlthough historically it was believed that the aneurysm was a final stage of an atherosclerotic process, it is now clear that atherosclerosis and AAA are distinct entities with very specific traits . They shA growing number of studies over the past 2 decades support the idea that immune-mediated inflammation plays an important role in AAA development and progression. A chronic immune-mediated response is followed by the destruction of the aortic media through extracellular matrix (ECM)-mediated degradation by the release of proteolytic enzymes , production of reactive oxygen species (ROS), cytokine action, and vascular smooth muscle cell death, which are typical hallmarks of AAA . The conAll the above implies the involvement of additional as yet unidentified critical factors that do not respond to immunosuppressive chemotherapy and the need to clarify uncertainties about the precise mechanism by which the complex crosstalk between adaptive and innate immunity can be self-sustaining in AAA progression.Cells of innate immunity play a key role in AAA because 1) they can perform pathogenically important effector functions on their own, such as responding to stimuli due to tissue damage and secreting soluble factors that degrade the vessel ECM, and 2) they act as mediators of the adaptive response, acting as antigen-presenting cells (exogenous or endogenous) and activating cells of adaptive immunity. Chronic AAA inflammation would thus be a continuous source of stimuli for cells of innate immunity. This would provide the basis for applying a new immunological concept related to innate immunity, so-called ‘trained immunity’, to AAA pathogenesis.Trained immunity is a term introduced fairly recently in immunology and defines a form of ‘innate immune memory’, which produces an enhanced inflammatory response following secondary stimuli. Stimulation of an innate immune cell (or its progenitors) triggers an enhanced nonspecific immunological reaction as a result of changes in gene transcription caused by modifications in chromatin configuration . An intee.g., by accelerating chronic inflammation and altering the physiological functions of PVAT  and levels of metalloproteinase and TNFα observed in both the aortic wall and PVAT  transcripts, including 84 and 184 unique genes overexpressed in dilated-PVAT of small and large AAAs, respectively  has been established based on publicly available data. For histone modification enrichment, CRRs are scored and ranked according to ChIP-seq features from the ENCODE  were related to the transcriptionally active euchromatin of myeloid cells  stimulation, the latter is typical of trained immunity (restimulation)    .To search for possible biological processes associated with gene signatures of acute (early) or trained immunity, we performed a functional enrichment analysis and identified statistically enriched GO-BP and TF terms. To reduce redundancy, terms were grouped based on similarity and an overview term was used for clustering . InteresAAA is still a disease of unknown etiology that requires new ideas to improve our ability to understand its pathogenetic mechanisms so that effective therapeutic approaches can be developed. In this perspective article, we aimed to highlight possible links between the development and progression of AAA and innate immunity. The existence of innate immunological memory has only recently emerged as a new and rapidly growing immunological concept in several areas, including cardiovascular disease, although its association with AAA has not yet been appreciated. We pointed out that trained immunity is an immunological mechanism capable of generating an excess response when dysregulated, which could contribute to chronic inflammatory disease processes in AAA.A large number of studies have unequivocally demonstrated that innate and adaptive immunity participate together in the evolution of AAA . Furthervice versa is, however, a mere speculative disquisition because both arms of the immune system exert their function through close crosstalk. Consistent with this view, a local environment created by trained innate immune cells during secondary stimulation may indeed influence T-cell responses in AAA, for example by altering the differentiation, polarization, and function of T-cell subtypes, suggesting an important link between trained immunity and an antigen-specific immune response (Whether elements of innate immunity may be more central than adaptive immunity or response .In this scenario, therefore, it may make sense to test whether trained immunity is a sensitive target that could interrupt this “short circuit” and influence disease evolution. The precise definition of the molecular mechanisms underlying trained immunity in AAA could lead to the identification of possible modulators capable of inhibiting inflammation and vessel wall degeneration, opening new therapeutic solutions for AAA . To this


Hyperglycaemia, hypoglycaemia and higher glucose variability during the Early Brain Injury (EBI) period of aneurysmal subarachnoid hemorrhage (aSAH) have been associated with poor clinical outcome. However, it is unclear whether these associations are due to direct glucose-driven injury or if hyperglycaemia simply acts as a marker of initial severity. Actually, strict glucose control with intensive insulin therapy has not been demonstrated as an effective strategy for improving clinical outcomes after aSAH. Currently published studies describing an association between hyperglycaemia and prognosis in aSAH patients have been based on isolated glucose measurements and did not incorporate comprehensive dynamic evaluations, such as those derived from subcutaneous continuous glucose monitoring devices (CMG). Arguably, a more accurate knowledge on glycaemic patterns during the acute phase of aSAH could increase our understanding of the relevance of glycaemia as a prognostic factor in this disease as well as to underpin its contribution to secondary focal and diffuse brain injury. Herein, we have summarized the available evidence on the diagnostic and prognostic relevance of glucose metrics during the acute phase of cerebrovascular diseases, focusing in the EBI period after aSAH. Overall, obtaining a more precise scope of acute longitudinal glucose profiles could eventually be useful for improving glucose management protocols in the setting of acute aSAH and to advance toward a more personalized management of aSAH patients during the EBI phase. Hyperglycaemia occurs in about three out of four aSAH patients within the first 72 h after the initial bleeding and it has been associated with poor clinical outcomes and secondary brain injury –7. HowevGlycaemic dysregulation decisively affects vital and functional prognosis of patients across several cerebrovascular diseases, although it is still unclear how to effectively address the glycaemic status in the acute phase of critical diseases such as aSAH. Traditionally, glycaemic evaluation has relied on static parameters, like glycaemia on admission or premorbid glycaemic status including known-diabetes or glycated hemoglobin (HbA1c) on admission. Nonetheless, glycaemia is a rapidly changing parameter due to both disease evolution and acute management-related factors that are only partially understood. Therefore, static markers may be inadequate to address the true relevance of glycaemic status and to guide insulin therapy in the acute setting. To overcome this limitation, several studies addressed glycaemia as a dynamic variable by means of evaluating the glycaemic variability (GV) , 18. In The exhaustive information obtained through CGM includes static measures such as mean, maximum and minimum glucose levels, the amount of time spent above or below a predefined threshold, and also several dynamic features accounting for GV. CGM also allows the identification of different longitudinal glucose profiles, as well as performing complex analyses of glucose homeostasis, which would be not feasible with conventional testing. The main advantages of CGM, compared to conventional finger prick testing, are the ability to comprehensively analyse a multitude of parameters and to obtain their longitudinal trajectory profiles. Reports on CGM in acute cerebrovascular diseases showed similar-to-better detection of dysglycaemic events, mainly hypoglycaemia, compared to the standard of care. Interestingly, some of these events show circadian variability and nocturnal preference , 24, beiHowever, the information derived from CGM devices should be interpreted carefully, since definitions and thresholds of the novel glycaemic parameters could be complex and heterogeneous. Beyond the extraction of the aforementioned predefined key metrics, the comprehensive analysis of CGM-derived repeated measurements may require sophisticated analytic tools and pre-planned analytical strategies , 20. In The ability of CGM to guide intensive insulin treatments (IIT) is also a matter of debate. Although the use of CGM has spread in the outpatient setting for certain indications, its utility in the inpatient setting remains to be demonstrated. Thus, interventional clinical trials based on CGM are scarce in comparison with accuracy and safety studies. A recent report in a cardiac ICU demonstrated good patient and caregivers' acceptance, accuracy and reliability of subcutaneous CGM measurements . Indeed,In summary, CGM technology seems safe, accurate and might be cost-effective in the acute inpatient setting, although its clinical value for guiding IIT or for prognostic purposes in patients who need tight glycaemic control, as those affected by acute brain lesions, remains to be demonstrated. Importantly, the benefit of strict glycaemic control with IIT for improving clinical outcomes in aSAH remains to be validated in clinical trials before an eventual implementation of CGM for guiding IIT , 10–13.via acute ischemic lesions or microstructural diffuse lesions, is associated with an increased risk of systemic complications and with poorer clinical, cognitive and affective outcomes at long-term  and have not considered including comprehensive dynamic evaluations, such as those derived from CGM devices. Arguably, using CGM to obtain accurate glycaemic profiles during the acute phase of aSAH could help define the relevance of glycaemia as a prognostic factor in this disease, as well as underpin its contribution to secondary focal and diffuse brain injury.Studies focused on hyperacute glucose monitoring in cerebrovascular diseases have included mainly patients affected by acute ischemic stroke (AIS). In this setting, both admission and acute-phase hyperglycaemia have been associated with mid-term and long-term morbidity and mortality following a J-shaped fashion, as well as with higher incidence of infarct growth, ischemic recurrence risk and post-stroke cognitive impairment –76. AfteNonetheless, studies using CGM in stroke are still scarce and have been based in modest population sizes. Disglycaemia measured by CGM is frequent after acute cerebrovascular events and has been linked to infarct growth in AIS , 90 and Hyperglycaemia, hypoglycemia and glucose variability during the acute phase of aSAH have been associated with an increased risk of in hospital complications and with poor long-term functional recovery. However, it is unclear whether these associations are due to direct deleterious effects induced by glucose dysregulation or if hyperglycaemia is an epiphenomenon related to the initial bleeding severity. In addition, it remains inconclusive whether there are specific longitudinal glycaemic profiles during the EBI period with special prognostic implications and/or more prone to lead to secondary brain injury. Arguably, the use of comprehensive dynamic evaluations, such as those derived from CGM devices, could aid to obtain a more precise understanding of such a highly dynamic process that is conditioned by both systemic complications and specific therapeutic approaches. A more accurate knowledge on glycaemic patterns during the acute phase of aSAH could increase our understanding of the relevance of glycaemia as a prognostic factor in this disease and to underpin its contribution to secondary focal and diffuse brain injury. In this context, the evaluation of biomarkers related with brain microvascular, metabolic and microstructural integrity using quantitative advanced MRI along with the quantification of circulating and cerebrospinal fluid (CSF) molecules related with brain injury could be highly informative for identifying glucose profiles associated with higher brain damage after aSAH. More research is needed to understand the most appropriate and informative timing for glucose monitoring, and how to combine glucose metrics with surrogate markers of acute brain injury. Eventually this information may lead to improve glucose management protocols in the setting of acute aSAH, to optimize the design of clinical trials aimed to modulate glucose levels and to advance toward a more personalized management of aSAH sufferers during the EBI period.DS, AM, LP, LL, RT, and SA participated in writing the manuscript and editing critical parts of this review. LL, RT, and SA gave the final approval for this submission. All authors contributed to the article and approved the submitted version.We thank the Spanish Ministry of Economy and Competitiveness for the grant given to SA and RT [project PI19/00936 funded by Instituto de Salud Carlos III and co-funded by the European Regional Development Fund (ERDF)], the Fundació la Marató de TV3 for the grant given to SA (grant number 17/C/2017). DS is supported by a grant from Hospital Clinic de Barcelona (Contracte Clínic de Recerca Emili Letang-Josep Font).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.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


These results suggest that PsyMetRiC is viable for implementation in clinical sites across Western Europe to improve early identification of cardiometabolic risk and inform personalised treatment decisions for young people with psychosis.Patients with a psychotic disorder are more likely to develop a cardiometabolic disorder compared to the general population, resulting in a reduction in life expectancy of around 15 years.Patients with severe mental illness have an increased risk of cardiovascular disease morbidity and mortality.The utility of precision psychiatry tools is ultimately dependent on the quality, relevance and generalisability of the data on which it is developed. However, generalisability of precision psychiatry tools is currently poor. Meta-analytical evidence has shown that while over 600 individualised clinical prediction models have been published in psychiatry, only 5% have been externally validated (performance tested in an independent sample) and less than 1% tested for implementation.For precision psychiatry to meaningful impact real-world clinical care, more research teams need to similarly prioritise robust external, international validation studies to test performance in new settings with corresponding differences in patient sociodemographics and service configuration. Advances in this area are reliant on fostering collaborations between research teams, particularly internationally, to allow for model performance to be tested on varied, representative and information governance-compliant datasets. A model that is resistant to sample differences is suited to implementation in a variety of settings and resistant to changes in population characteristics over time.While PyMetRiC performs well in Western European settings, future work should investigate its performance in other non-European and more ethnically/racially diverse settings. This is particularly important due to ethnic and racial differences in cardiovascular riskBarriers and facilitators for future implementation of PsyMetRiC are also important to consider. Currently, the evidence suggests that the most important facilitators for effective implementation involve employing appropriately-skilled staff and optimising service configuration.Precision psychiatry is a quickly advancing field. To make a positive and equitable impact on real-world mental healthcare, more external validation studies and considerations for prospective clinical use need to be published.None to declare.


Currently, female researchers represent merely a minority, accounting for an estimated 29.3% who end up covering this position worldwide, with a great variability according to each country . SpecifiIn response to such a large gender gap in the scientific research community, the UNESCO Institute for Statistics (UIS) is in the midst of developing new indicators in order to better comprehend the reasons behind women’s decisions to pursue one career over another. Several could be the reasons implicated in limiting and discouraging women’s access to the scientific community, including ancient biases and gender stereotypes. By further understanding such issues, the UIS project concurrently aims at reducing the gender inequality in science, technology, engineering and mathematics (STEM) fields, by possibly promoting reforms in policies and implementing changes in favor of gender equality in all countries with the ultimate goal of empowering women .The present Research Topic spans through various fields of surgical oncology, including breast, ovarian, colorectal, gastric, and esophageal cancer and includes research papers by women involved in oncological surgery.Li et al. focused on retrospectively analyzing the outcomes of three surgical options on disease-free and overall survival rates of young women with breast cancer below the age of 35 years. They found that both survival rates were significantly improved in those patients who underwent breast-conserving surgery compared to mastectomy. On the other hand, Wang et al. used the Surveillance, Epidemiology, and End-results (SEER) database of the US National Cancer Institute registry for the determination of differences in patient survival of each different treatment modality for invasive micropapillary carcinoma (IMPC) of the breast. Authors show how in women with early-stage IMPC, breast-conserving therapy is equivalent to mastectomy in terms of survival outcomes. Xu et al. also assessed breast cancer from a molecular point of view and found that Ferroptosis-related prognostic signature could be proposed as novel biomarkers for the prediction of breast cancer prognosis as they seem connected to the immune microenvironment. Molecular and cellular mechanisms of pyroptosis in patients with breast cancer were looked into also by Ren et al.. The inflammation-dependent programmed cell death mediated by inflammasomes, known as pyroptosis, plays a substantial role in the progression of breast cancer and authors suggest how pyroptosis-related genes could be used as new prognostic biomarkers or even targets for breast cancer treatment.Breast cancer represents, at present, the most common malignancy among women of all ages. Gao et al. retrospectively evaluated the prognostic impact of retroperitoneal lymphadenectomy in patients with ovarian clear cell cancer. Authors found no survival benefit in patients undergoing retroperitoneal lymphadenectomy and was not an independent predictor of tumor recurrence. In their study, Jiang et al. introduce a new, minimally invasive surgical approach for the treatment of giant ovarian cysts > 20 cm in diameter. All patients successfully underwent single-port laparoscopic surgery for the removal of serous or mucinous cystadenomas.Epithelial ovarian cancer is one of the most aggressive gynecologic cancers. Wong et al. determined predictors of morbidity and mortality after palliative surgery in patients with peritoneal carcinomatosis due to various primary malignancies. Authors found elevated preoperative albumin levels and a good Eastern Cooperative Oncology Group (ECOG) performance status were independently associated with better short term outcomes following palliative gastrointestinal surgery, supplying a simplified model to predict superior responders to surgical treatment.Hashimoto et al. analyzed immunohistochemical data in order to identify protein expression patterns in stages II-III colorectal cancer that could somehow predict patient outcomes. A high expression of Tenascin-C was identified as a single prognostic marker and was correlated with a worst prognosis in both stages of colorectal cancer.Colorectal cancer is another one of the most common cancers, ranking third as the leading cause of death in both men and women. Xiang et al. aimed at establishing a reference value for the creation of treatment strategies by identifying current methods for predicting and preventing peritoneal recurrence following surgical resection. Authors highlight how an early gastric cancer diagnosis and a limited loco-regional extension of the primary tumor reduce the risk of peritoneal recurrence, together with intraperitoneal chemotherapy and adjuvant chemotherapy.Although radical resection for gastric cancer is currently the only curative treatment option, recurrence after surgery is most commonly peritoneal. Weis et al.). This study aims to develop patient-centered trial information material for this randomized controlled trial and to increase patient acceptance and compliance with randomized treatment strategies and trials.Finally, this Research Topic also includes a Study Protocol of a prospective multicenter study on patient participation for clinical trials (Promoting gender equality, dismantling stereotypes, and encouraging women to pursue STEM jobs are all necessary to shift entrenched beliefs. Therefore, Frontiers in Oncology is pleased to provide this Research Topic to highlight the contributions of female researchers in all areas of oncology. The work provided here demonstrates the range of oncology research across the board and shows new developments in theory, experimentation, and methodology with applications to interesting and current issues.All authors contributed to manuscript revision, read, and approved the submitted version.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Plant transformation provides a key tool for much basic research, such as the study of gene functions and interactions, protein–protein interactions, developmental processes, as well as applications for crop improvement and the development of plant bioreactors to produce vaccines. Efficient and reproducible transformation technologies are not only essential for the development of transgenic plants but also critical for other applications like transient gene expression studies and gene editing.Agrobacterium tumefaciens was first identified in 1907 as the etiological agent of crown gall disease. The bacterial factor responsible for tumor induction was described in the ‘70s: a DNA plasmid called the Ti plasmid by Zaenen et al.  did not evolve as fast as its domesticated relative. In this context, Zobrist et al. report an innovative protocol by using whorl segments of seedlings germinated from mature seeds of Zea parviglumis.Even though the first transgenic plants were produced in the early ‘80s, not all plant species are transformed as easily as model species, particularly when it comes to some economically important crop species. Some plants are still considered as difficult or recalcitrant to transformation. Almost every plant species has a specific transformation protocol that slowly evolved over the years and has not been updated in the last two decades except for the methodology section of published papers. Protocols were modified to facilitate new breeding techniques like gene editing, and some of the latest methodological improvements include breakthrough advancements like the use of developmental regulator genes and tissue-culture independent gene editing protocols. This Research Topic provides a collection of reviews and original research articles on the recent advancement of plant transformation and gene editing of different crops. Below we briefly describe the original papers are reviews that integrate this Research Topic. Maize, perhaps the crop species with most transgenic commercial traits incorporated so far and for which many new approaches to improve its engineering, including genome editing, is critically summarized by Kok et al. report that supplementation with specific PF-68 concentrations enhances callus proliferation in recalcitrant rice cultivars. Chinese cabbage is another example of a recalcitrant crop. Sivanandhan et al. report stable Agrobacterium tumefaciens-mediated transformation for Chinese cabbage cv. Kenshin by employing antioxidants in the co-cultivation and subsequent regeneration media. The Asteraceae family is the largest and most diversified family of the Angiosperms and includes economically important crops such as lettuce and sunflower. There is a sharp contrast between lettuce that has easily adapted to tissue culture and transformation and sunflower, which is much trickier and difficult to transform. The peculiarities of their genetic transformation protocols are described by Darqui et al..As mentioned above, efficient genetic transformation is currently performed routinely for many plant species, but there are still many recalcitrant species. Even in species where transformation was successfully achieved, some agronomically important genotypes remain reluctant, thus slowing down breeding programs since the obtention of new engineered varieties normally requires the introgression of transgenic or gene-edited traits into elite germplasm. Several reports in this Research Topic deal with this issue by improving tissue culture conditions. For example, pluronic F-68 (PF-68) is a non-ionic surfactant used in plant tissue culture as a growth additive. Campos et al.. Switchgrass is a grass of importance for biofuel production. Upland switchgrass cultivars are recalcitrant to genetic transformation. As for the case of the rice cultivar below, Xu et al. report improved transformation methods for two important upland cultivars. Speaking of forage crops and grasses, dallisgrass is a very important apomictic pasture in South America and other temperate warm regions of the world. In the paper authored by Schrauf et al., the transformation methodology status of cultivars of Paspalum dilatatum transformation is reviewed, and the authors propose this species as a model for molecular breeding in C4 perennial forage species. Although apomixis  is a pursued trait in grasses because the introduced transgene becomes immediately fixed in a highly adapted genetic background capable of large-scale clonal propagation, it may present a serious constraint for conventional breeding. Another review  addresses this paradox by describing the potential molecular pathways involved in apomixis determination and how these pathways could be used for both conventional and molecular breeding.Grapevine has been considered a recalcitrant crop to produce transgenic plants as several other woody perennials. The ability to regenerate plants from transformed explants is considered the main obstacle in the process. State-of-the-art technologies to improve grapevine transformation and regeneration are reviewed by Jozefkowicz et al., proposes the use of the Tnt1 element as a candidate to identify dominant mutations in allogamous tetraploid cultivated alfalfa. They illustrate this potential by showing that a single allelic mutation in the MsNAC39 gene produces multifoliol leaf plants in alfalfa. Sugarcane is another complex polyploid (and aneuploid) species. Modern sugarcane cultivars have extremely large genomes originating from artificial interspecific crosses making traditional breeding extremely difficult. The review authored by Budeguer et al. summarizes the current techniques and state of the art in sugarcane transformation. Potato is another well-studied polyploid crop of global importance that is not apomictic but clonally propagated like apomictic grasses, sharing the difficulties in conventional breeding. Thus, genetic engineering provides the opportunity to introduce/switch-off genes of interest without altering the allelic combination that characterizes successful commercial cultivars or to induce targeted sequence modifications by genome editing. The review by Nahirñak et al. summarizes the latest developments in the field.Many crops belong to polyploid species and require the search of systems to identify new dominant markers. A perspective article from Conti et al. provides a detailed overview of the currently used strategies for the development of genetically modified citruses.Citruses are among the most prevailing fruit crops produced worldwide. As for the examples mentioned above, conventional methods are difficult because of prolonged juvenile periods, complex reproductive stages, occasional low fertility, self-incompatibility, parthenocarpy, or polyembryony. Genetic engineering technologies offer alternative approaches for overcoming these difficulties. The review by Yin et al., the later advances in transgenic technology of forest trees and their application for trait improvement are discussed. Some forest species are widely used in agroforestry plantations for soil stabilization, ecosystem rehabilitation, etc. This is the case of Casuarina equisetifolia. In the study authored by Ren et al., more efficient and rapid regeneration systems based on stem segment explants are reported.Forest-tree breeding using transgenic technology is still in its early stages compared to annual plants except for a few model species. In the review by Razzaq et al. present an example of gene editing to improve fiber quality parameters appreciated by the textile industry.Another important aspect of molecular breeding is the identification and characterization of complex industrial traits. This is the case of fiber length, strength, and other fiber quality parameters in cotton. Cannabis sativa produces unique phytocannabinoids and there are scarce if any reports of in vivo engineering targeting the cannabinoid biosynthesis genes. Matchett-Oates et al. report the successful modulation of cannabinoid biosynthesis genes using RNAi via agroinfiltration, enabling functional genomics of targeted cannabinoid biosynthesis genes.Bacillus thuringiensis toxin gene Cry10Aa to induce coffee berry borer (CBB) resistance, as reported by Valencia-Lozano et al.. Bioassays using transgenic fruits challenged with CBB larvae and adults significantly reduced seed damage to less than 9%, providing a powerful CBB control tool for coffee production.Finally, another woody plant was successfully transformed with HH was a guest associate editor of the Research Topic and wrote the paper text. GS and LH-E were guest associate editors of the Research Topic and edited the text. All authors contributed to the article and approved the submitted version.HH was supported by by the Agencia Nacional de Promoción Científica y Tecnológica, Argentina (ANPCYT: PICT 2018-02926).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.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.