Developing electrophysiological platforms to capture electrical activities of neurons and exert modulatory stimuli lays the foundation for many neuroscience-related disciplines, including the neuron–machine interface, neuroprosthesis, and mapping of brain circuitry. Intrinsically more advantageous than genetic and chemical neuronal probes, electrical interfaces directly target the fundamental driving force—transmembrane currents—behind the complicated and diverse neuronal signals, allowing for the discovery of neural computational mechanisms of the most accurate extent. Furthermore, establishing electrical access to neurons is so far the most promising solution to integrate large-scale, high-speed modern electronics with neurons that are highly dynamic and adaptive. Over the evolution of electrode-based electrophysiologies, there has long been a trade-off in terms of precision, invasiveness, and parallel access due to limitations in fabrication techniques and insufficient understanding of membrane–electrode interactions. On the one hand, intracellular platforms based on patch clamps and sharp electrodes suffer from acute cellular damage, fluid diffusion, and labor-intensive micromanipulation, with little room for parallel recordings. On the other hand, conventional extracellular microelectrode arrays cannot detect from subcellular compartments or capture subthreshold membrane potentials because of the large electrode size and poor seal resistance, making it impossible to depict a comprehensive picture of a neuron's electrical activities. Recently, the application of nanotechnology on neuronal electrophysiology has brought about a promising solution to mitigate these conflicts on a single chip. In particular, three dimensional nanostructures of 10–100 nm in diameter are naturally fit to achieve the purpose of precise and localized interrogations. Engineering them into vertical nanoprobes bound on planar substrates resulted in excellent membrane–electrode seals and high-density electrode distribution. There is no doubt that 3D vertical nanoelectrodes have achieved a fundamental milestone in terms of high precision, low invasiveness, and parallel recording at the neuron–electrode interface, albeit with there being substantial engineering issues that remain before the potential of nano neural interfaces can be fully exploited. Within this framework, we review the qualitative breakthroughs and opportunities brought about by 3D vertical nanoelectrodes, and discuss the major limitations of current electrode designs with respect to rational and seamless cell-on-chip systems. Developing electrophysiological platforms to capture electrical activities of neurons and exert modulatory stimuli lays the foundation for many neuroscience-related disciplines, including the neuron–machine interface, neuroprosthesis, and mapping of brain circuitry. In the central nervous system, neuronal networks are able to learn adaptively from environmental inputs, form cognitions, and carry memory storage. In the peripheral nervous system, neurons can sense a diversity of mechanical, chemical, and thermal stimuli, while delivering accurate controls through neuromuscular junctions for both long-range, high-strength and short-range, delicate motions. Moreover, the highly efficient transformation from chemical energy to ionic gradients allows a neuron to generate and transmit electric signals. Therefore, it has long been a major pursuit in neuroscience, bioengineering, and electrical engineering to develop seamless neural interfaces for probing, understanding, and modulating neural activities. And, despite advances in engineering large-scale electrophysiological approaches for e.g. patch clamps) and extracellular microelectrode arrays (MEAs) have intrinsically suffered from the trade-off between invasiveness and precision. Patch clamping often leads to severe cell damage within hours of electrode insertion, and is difficult to expand to multisite, parallel recordings. While planar MEAs impose no invasiveness to cells, the large dimension of electrode size  and poor electrode–membrane seal make it difficult to reveal subcellular and subthreshold neuronal activities. Looking at nature, the nanoscale ion channels are the driving forces behind the great diversity of neuronal dynamics, which provide biomimetic inspiration to overcome this trade-off by shrinking the electrode dimensions to the nanoscale. Although a variety of nanostructures, such as planar nanowires and suspended nanoparticles,8–13 have been applied to neuronal recording and stimulation, a revolutionary breakthrough towards an organized and high-resolution neural interface has been enabled by 3D nanofabrication techniques. Vertical nanoelectrode arrays significantly reduce the projection area on the planar substrate to which they are bound without compromising the interfacial area they share with the local cell membrane, allowing for the realization of high-density fabrication and parallel recording from different compartments of single neurons. More importantly, their topography of 3D protrusions provides intimate electrode–membrane seals, drastically improving the reliability and fidelity of recording. In this way, nanoelectrode arrays not only induce sufficient invasiveness to ensure high-quality signals, but also cause minimal cellular damage even after electrical or optical poration. Therefore, these advantages that have arisen from scaling down to the nanoscale have provided new possibilities for solid-state electronics to “talk” to electrogenic neurons.At the implementation level, constructing a bridge between electronic devices and neurons requires the electrodes not only to have appropriate electrical properties for signal detection and/or current injection, but also to be able to adapt to the dynamic and fragile nature of cells. Conventional tools of intracellular micropipettes  to Na15 Unlike the digital all-or-none feature of action potentials, these synaptic inputs are analog signals with different amplitudes and durations  from distal dendrites, either excitatory or inhibitory, are integrated at the neuronal soma that determines if an action potential will be fired or not .15 Unlikurations .16 More Rseal. Moreover, the membrane curvature is spontaneously formed without forced insertions, thus imposing minimum invasiveness and damage to the neuron. Since the junctional area is also at nanoscale, the signal source from the neuronal membrane is highly localized, allowing for the recovery of spatial distribution of the ion channels and membrane properties.The recording/stimulation principle of 3D vertical nanoelectrodes is the same as planar MEAs. However, the small dimensions and vertical protrusions of nanoelectrodes can bring about significant improvements to some critical parameters. The equivalent circuit of a nanoelectrode–neuron interface is shown in C. elegan's nervous system, an invertebrate worm with only 280 neurons.18 As mentioned before, a single neuron is a complex computational device that receives inputs from multiple pre-synaptic terminals followed by neurocomputation to produce action potentials of various frequencies. Although the properties of ion channels have been extensively characterized using patch-clamp electrophysiology,19,20 their non-uniform spatial distribution and time variance make it challenging to decode the computation process. The situation becomes more frustrating for neural networks, where the variance of synapse location and time-dependent synaptic strength are involved. Therefore, to sketch the complete picture of nervous system operations, it is imperative, at the fundamental level, to first develop in vitro sensors with high resolution and long-term robustness to reveal the electrical activities of single neurons.Even for animals with the seemingly simplest behaviors, there might be a rather complicated nervous system functioning in the background. In contrast to the rapid development in solid-state electronics, in which billions of transistors are integrated per chip, it took decades for scientists to resolve the complete connectivity of 24 Such a cleft, filled with highly conductive electrolyte, contributes to most of the current leakage of the entire recording system.25 Although the topographical improvement using mushroom protrusions has greatly enhanced the membrane adhesion for cardiomyocytes and Aplysia neurons,26 the inevitable cell membrane deformation of micrometer size occupies a considerable amount of membrane area, making it difficult to simultaneously extract information from multiple sites of a small mammalian neuron.Fundamentally more advanced than microelectrodes, nanoelectrodes are not only a matter of size-scaling, but a breakthrough regarding both the quantity and quality of information extracted from cells. MEAs, developed for the purpose of large scale and parallel neural recordings, are intrinsically limited by their large dimensions and defects at the neuronal interface. First of all, with nanoscale ion channels and submicron neurites and synapses, it is difficult for MEAs to pinpoint specific cellular compartments and extract localized information from plasma membrane, as the electrode size of 5–30 μm can only characterize the averaged electrical activities of the attached membrane. As a result, subcellular information reflecting the operation of nanoscale ion channels and submicron neurites and synapses is often attenuated or buried in the signals recorded from larger areas. Second, planar microelectrodes suffer from relatively weak electrode-cell coupling that originates from a 70–100 nm cleft between the cell membrane and solid-state probes.27 Nanopillars, with a diameter of about 100 nm, can interface with only a small portion of the cell membrane, allowing for multiple detection at different sites from a cell. Moreover, nanowire field-effect transistors (FETs) have reached the 10 nm scale, approaching the dimensions of single ion channels28 (though this device has not achieved extracellular recordings from single ion channels). Early studies often fabricated multiple nanopillars on the same conductive pad, resulting in a loss of individuality, as they are electrically interconnected , giving rise to a seal resistance of 8029 to 500 MΩ,22 which is a more than two orders of magnitude enhancement. Besides the morphologically tight adhesion, the membrane curvature caused by vertical nano-protrusions may also induce the aggregation of ion channels, increasing the local ionic current.31,32The spontaneous membrane engulfment around nanoelectrodes provides an excellent seal for recording subthreshold membrane potentials. Various studies have characterized the plasma membrane deformation on vertical nanoprotrusions using fluorescence and electron microscopic techniques . Althoug33,34 Despite the increased electrode impedance caused by the reduced surface area, the improvement in the membrane–electrode seal clearly overweighs the sacrifice of electrode impedance, which has been proved through both theoretical study25 and electrophysiological tests.21–23,35,36 More importantly, membrane engulfment was also observed at large neurites that are difficult to access by patch-clamps and microelectrode arrays  or cell penetrating pipettes (CPPs) have enhanced the internalization efficiency to 15%,44,51 it is still not affordable for neuronal recordings, since a nanoelectrode array has much fewer probes than relatively simple drug delivery platforms. In addition, a systematic study using nanopillars, nanocones, and sharp nanopillars has revealed that vertical nanostructures spontaneously penetrate the cellular membrane to form a steady intracellular coupling only in rare cases, and suggested that most spontaneous penetrations might occur only during the initial hours of cell plating with membrane resealing afterwards.53 Therefore, it is important to address the significant issues in terms of both mechanism and application before spontaneous poration can be exploited further for nano neural electrodes.Spontaneous membrane penetration enabled by vertical nanoprotrusions was observed and widely applied in delivering biomolecules into the cytoplasm . Without41 However, since electroporation has to be conducted by the same electrodes as used for the recordings, the switching between pulse injection and recording modes will inevitably result in a blind recording period caused by overcharging. Additionally, the number, size,21,54 and distribution of membrane pores from electroporation are difficult to predict and characterize.55 To address these issues, the combination of vertical nanoelectrodes and plasmonic optoporation was developed for opening transient pores at the tips of nanopillars  into the water conduction band, which induce a chain reaction of photon absorption and the release of more hot electrons. Eventually, water molecules are accelerated by hot electron impact, producing a mechanical shock wave that ruptures the cell membrane  for hot electron excitation42,56 at the solid–water interface. Comparing to electroporation, this approach also has the advantage of localized pore formation, in addition to continuous recording. Because of its essence of mechanical nanoshockwaves, plasmonic optoporation is likely to produce single pores that are highly localized and easier to modulate. Considering that the lipid membrane reseals after poration, a single and localized pore could make the recorded signals better in terms of fidelity and consistency than a group of sparsely distributed pores. This might also be the reason for the prolonged period of intracellular access of up to 80 min,37 as it takes more time for the cell membrane to reseal a large pore than many small pores.On the other hand, artificially induced membrane poration through electrical or optical approaches has been well applied to intracellular recordings. In electroporation, cell membrane can be ruptured under a strong electric field applied at the nanoelectrode tip, allowing for significant improvement in the signal amplitude and intracellular-like shapes. The mechanism of this process was investigated by computer simulations. Specifically, water defects, caused by the interaction of water dipoles with the electric field gradient at the water/lipid interface, are significantly promoted by the external electric field. As a result, water can penetrate the lipid bilayer from both sides, leading to pore formation .41 Howevopillars .37 Upon membrane .42 So faFor both electroporation and optoporation, the recording window is relatively short . However, nanoelectrode penetration is a highly local process that has little impact on the adjacent membrane as well as seal resistance. In fact, the cell membrane spontaneously reseals after the recording window. Thus, the poration could be repeatedly performed  over a long-term cell culture. Although this repeated sampling is not a continuous recording, considering that the neuronal long-term plasticity (learning and memory) is a much slower process, repeated intracellular access with nanoelectrodes still holds great promise for studying long-term neural network dynamics.2, which can then be selectively etched by HF (22 Fabrication of hollow nanotubes exploited the tone-inversion of photoresists by secondary electrons during focused ion-beam (FIB) milling  and noble metals  due to their compatibility with photo- and e-beam lithography as well as chemical and physical etching. The top-down process has been widely used for metal-coated semiconductor nanopillars and nanotubes, in which high-resolution lithography was first applied to define the diameters of electrodes on photoresists, followed by reactive ion etching to remove uncovered material and leave out vertical nanostructures of high aspect-ratios. Additionally, since the top-down approach is a well-established process in dealing with semiconductors, it promotes innovative improvements to achieve various electrode geometries, smaller diameters, and higher densities. For example, the diameter of Si nanowire electrodes was reduced from 600 to 150 nm by thermal oxide thinning, based on that Si can be thermally oxidized to SiOed by HF .22 Fabri milling , resulti etching .2321 In comparison to semiconductor-based processes, bottom-up deposition involves much fewer steps that greatly reduce fabrication failure. Moreover, it is insensitive to substrates so that transparent glass and quartz can conveniently serve as substrates for cell observation under inverted microscopes. However, because of the limited maneuverability of noble metals at the nanoscale, bottom-up fabricated electrodes are often pillar-shaped without sharpened tips (like Si electrodes), which may more or less affect the cell membrane–electrode interface due to their restrained bending ability under the pulling force from the cell membrane and the larger curvature at the electrode tips.57 Another factor that makes semiconductor electrodes more popular is their compatibility with CMOS  technologies that integrate on-chip amplifiers in the vicinity of electrode probes, significantly reducing the amplitude loss by eliminating the stray capacitance.58Noble metals of Pt and Au have excelled in microelectrodes because of their inertness in biological fluids and excellent conductivity, rendering robust neural interfaces for long-term sensing. For nanoelectrodes, however, metals are highly resistant to plasma etching, and it would be expensive and unreliable to use conventional lift-off processes for high-aspect-ratio protrusions that are 1–3 μm in height. Therefore, the bottom-up approach by selective deposition has been mostly adopted for metal nanoelectrodes. Specifically, nanoscale holes can be milled on a layer of photoresist by e-beam lithography, followed by electrochemical deposition or FIB-assisted deposition to create the protrusion structure .21 In co38,59–61 Composed of three terminals: source (S), drain (D), and gate (G), an FET's charge carrier density in the S–D channel is modulated by the voltage applied on the gate, thus resulting in a conductance controlled by the gate. During neural recordings, the alternating electrical potential from a neuron serves as the gate voltage that modulates the S–D current  deposition is also the most critical process in the fabrication of nanowire field-effect transistors (NWFETs). current . The oriRseal, due to high electrode impedance. More importantly, it is imperative to measure the Rseal for each electrode in order to recover a neuron's actual electrical activities. Indeed, the tight adhesion between the cell membrane and vertical nanostructures yields detectable intracellular-like signals after membrane poration. However, interfacing with different neuronal compartments of different dimensions, each electrode will possess a unique Rseal, causing substantial variance across electrodes. Moreover, neuronal migration may also contribute to the time-variation of Rseal. Apparently, if this critical parameter is not available or well characterized, the quantitative information of the recorded data will be lost.Neuronal signals acquired by nanoelectrodes are strongly affected by the seal resistance, 29,30 Alternatively, FIB and SEM (FIB-SEM) can target the biointerface at any desired locations through FIB milling , both the amplitude and shape of recorded signals will be affected substantially by the electrode–membrane seal. Moreover, low-frequency field potentials may have important links to the brain's perception, motion, and memory,73) but not simulate presynaptic spikes to evoke excitatory post-synaptic currents . Although larger current injection can be realized by breaking into the faradaic regime,22 the electrochemical reactions will inevitably cause electrode degradation and even water electrolysis for polarizable electrodes such as Au or Pt. Additionally, the slow recovery/discharging process after stimulation makes it difficult to immediately switch an electrode from stimulation to recording mode.On the other hand, this issue becomes more problematic for neural modulation purposes. The small surface area of nanoelectrodes results in an extremely low charge delivery capacity, which might be able to inject miniature post-synaptic currents . More importantly, combined with non-polarizable Ag/AgCl electrodes and a Wheatstone Bridge circuit, a micropipette electrode in current-clamp mode is able to implement simultaneous stimulation and recording.28,59 With FETs integrated at the bottom of nanotubes, this bioelectronic probe provides high-resolution intracellular mapping of electrogenic cells. Yet the use of FETs as an interface also precluded the compatibility for current injection. Surprisingly, the breakthrough in nanoscale ionic neural interface occurred mainly in the field of drug delivery, where vertical nanostraws (hollow nanotubes) were used to impale cell membrane, allowing for the diffusion of molecules into the cytoplasm  can accurately access individual synapses and monitor the dynamics of neurotransmitter fluxes84  that are critical to information processing. Ideally, neural interfaces should be able to selectively monitor the electrical activities of neurons without inducing acute damage or substantially altering the natural status of the neurons. For 3D vertical nanoelectrodes, scaling down the electrode sizes to less than 200 nm has caused minimal membrane deformation , while enabling precise electrical neuronal interfaces at the subcellular level. Arranging the nanoprotrusions into individually addressable arrays further opens up the possibility of parallel, large-scale neuronal recording without labor-intensive manipulations.The advancement of nano neural electrodes is a joint effort involving nanofabrication, neuroscience, electronic engineering, and biophysics. Behind this development lies the rationale of matching to the nanoscale neuronal compartments (85 and lying-down nanowires86), because the unique topography of substrate-bound vertical nanostructures addresses two significant barriers towards robust neuron-electronic integration: membrane–electrode seal and reliable intracellular access. The spontaneous membrane engulfment greatly reduces the current leakage through the seal, ensuring a good signal-to-noise ratio. And, membrane poration via electrical or optical approaches can be realized with low power injection, high repeatability, and membrane recovery. Overcoming these fundamental issues has made vertical nanoelectrodes a promising platform for neuron-chip communications.Besides the rationale of accurate probing, organized vertical nanoelectrode arrays are more prominent over other nanoscale interfaces (such as dispersed nanoparticles and nanowires,in situ will inevitably make the recorded signals less trustworthy. On the other hand, the high electrode impedance from the EDL capacitance has put an intrinsic limitation on both recording and stimulation, which might be resolved by switching to direct ionic interfaces. Moreover, regarding device functionality, aligning nanoelectrodes with neurites and synapses needs to be addressed in the future to fully exploit the nanoscale advantages.Nevertheless, several key challenges still need to be addressed to push this technology further towards functional cell-on-chip systems. As mentioned in the remaining dilemmas, the prerequisite for understanding and decoding neuronal computation from recorded data is awareness of the recording conditions. The lack of approaches for characterizing the seal resistance There are no conflicts to declare.


Discovered as a contaminant of adenovirus stocks in the 1960s, adeno-associated virus (AAV) is a mono-stranded DNA virus that depends on helper factors to replicate. Even though AAV is endemic in the human population (35–80%), it is remarkable that many issues concerning the natural infection by this virus remain unanswered. In this study, we reflect on the main basic aspects of AAV biology and provide an overview of the studies exploring the impact of AAV infection on human health, focusing on three major research areas including, (i) cervical and (ii) liver cancer, and (iii) reproductive system disorders. Conflicting results have been obtained into the association of AAV infection with the occurrence of adverse reproductive outcomes, such as placental complications, spontaneous abortion, and fertility disorders, or with a protective role in HPV-related cervical carcinogenesis. Noteworthy, recent reports have identified AAV insertional mutagenesis as a novel risk factor for the development of hepatocellular carcinoma. This latest finding raises concern regarding the widespread usage of AAV vectors in liver-targeted gene therapy. Adeno-associated virus (AAV) was discovered as a contaminant in a simian adenovirus type 15 preparation in 1965 . Since tParvoviridae family, AAV belongs to the Dependoparvovirus genus. There are at least 13 naturally occurring serotypes, each with a different tissue tropism . Nevertheless, considering the extensive usage of rAAV vectors in liver-targeted gene therapy and its potential for insertion into the human genome, patients treated with rAAV vectors should be followed longitudinally to monitor long-term consequences and determine the risk of HCC development.


Adipose tissue undergoes significant anatomical and functional changes with aging, leading to an increased risk of metabolic diseases. Age-related changes in adipose tissue include overall defective adipogenesis, dysfunctional adipokine secretion, inflammation, and impaired ability to produce heat by nonshivering thermogenesis. Thermogenesis in adipose tissue is accomplished by brown and beige adipocytes, which also play a role in regulating energy homeostasis. Brown adipocytes develop prenatally, are found in dedicated depots, and involute in early infancy in humans. In contrast, beige adipocytes arise postnatally in white adipose tissue and persist throughout life, despite being lost with aging. In recent years, there have been significant advances in the understanding of age-related reduction in thermogenic adipocyte mass and function. Mechanisms underlying such changes are beginning to be delineated. They comprise diminished adipose precursor cell pool size and adipogenic potential, mitochondrial dysfunction, decreased sympathetic signaling, and altered paracrine and endocrine signals. This review presents current evidence from animal models and human studies for the mechanisms underlying thermogenic adipocyte loss and discusses potential strategies targeting brown and beige adipocytes to increase health span and longevity. Adaptive thermogenesis is the process of heat generation elicited to maintain body temperature when homeothermic organisms are exposed to cold and occurs through non-shivering and shivering mechanisms . Shiveri+2-ATPase 2b (SERCA2b) and ryanodine receptor 2 (RyR2) (Ikeda 2017), (ii) the futile cycle of creatinine metabolism (Kazak 2015), and (iii) the triglyceride-fatty acid futile cycling  activity from calcium transport into the sarcoplasmic reticulum and is mediated by the protein sarcolipin . In adip cycling . Further cycling .Nonshivering thermogenesis in thermogenic adipocytes is long known to play a central role in the successful adaptation of small placental mammals to cold environments by enabling the maintenance of a stable body temperature. Energy dissipation as heat is also acknowledged to be a major regulator of energy homeostasis by affecting energy expenditure, glucose  and lipiEvidence from rodent studies indicates that thermogenic adipocytes affect adipose tissue biology through anti-inflammatory, anti-fibrotic, and pro-angiogenesis actions . They caTwo distinct types of thermogenic adipocytes are recognized in mammals, namely brown and beige/brite (brown-in-white) adipocytes. Both types of thermogenic adipocytes share morphological features, such as multilocular lipid droplets, mitochondria abundance, and expression of the thermogenic machinery, consistent with their ability to execute thermogenesis . HoweverMyf5) precursors in the dermomyotome and are found in distinct brown adipose tissue depots, such as the interscapular and perirenal regions of both rodents and human infants, which are densely innervated with sympathetic fibers. They constitutively express high levels of proteins comprising the thermogenic machinery and produce heat primarily by the UCP1-dependent mechanism  or by white-to-beige conversion (transdifferentiation) of phenotypically mature white adipocytes  was recently described in the adipose tissue of humans and mice. In mice, it was reported that AKG increased the appearance of beige adipocytes within white adipose tissue until late infancy by the interleukin 6/STAT3 pathway. In contrast, lack of AKG intake in infancy led to premature loss of beige adipocytes and increased white adipose tissue mass . TherefoData from recent studies involving cellular lineage determination and single-cell transcriptomics suggest there are different subtypes of brown and beige adipocytes, arising from distinct progenitors and comprising adipocyte populations with specific functional features . In mous+ and PDGFRα+ adipose progenitor cells. It expresses the thermogenic machinery in a β-adrenergic-dependent fashion and uses the energy from fatty acid oxidation to generate heat  as opposed to ‘standard’ conditions  indicate that classical brown adipose tissue exhibits molecular markers of human thermogenic adipose tissue .Despite the controversy regarding the comparative phenotype of thermogenic adipocytes in mice and humans, current evidence indicates a gradual decline in thermogenic adipose tissue mass and activity after puberty . In humaCurrent evidence indicates that a myriad of mechanisms is implicated in the decline of thermogenic adipose tissue mass and function with aging, which affect the number and ability of precursor cells to differentiate into thermogenic adipocytes and their thermogenic capacity, as summarized in Human brown adipose tissue involutes early in postnatal life, differently from mouse tissue. In mice, brown adipocyte progenitor cells (APC) remain in the brown adipose tissue throughout life and are essential for tissue regeneration after damage . They caFSTL1 gene, encoding the follistatin-like one glycoprotein, which could act to maintain their renewal capacity and differentiation competence. In parallel with tissue involution in rabbits, there is decreased brown APC expression of FSTL1 and reduced competence or ability to differentiate into brown adipocytes upon β3-adrenergic activation. Conversely, mice retain FSTL1 constitutive expression at later life stages, and its genetic ablation recapitulates the brown adipocyte involution observed in rabbits and humans , Pax3, Pdgfra, or Pdgfrb (de novo beige biogenesis). Notably, aging is associated with a significantly decreased ability of beige APC to proliferate and differentiate  mice and human stromal vascular cells from the abdomen and hip of more senior individuals were shown to exhibit an overall senescence-like phenotype. Such phenotype includes upregulation of genes related to cell cycle arrest and induction of the senescence-associated secretory program consisting of cytokines, chemokines, proteases, and growth factors  F344 rats were shown to exhibit a comparable proliferative response to serum-supplemented culture medium when compared with their young (6-month-old) counterparts .It is currently unknown whether aging affects the proportion of different subpopulations of thermogenic adipocytes by differential effects on their precursors. It was recently shown that brown adipocytes expressing high levels of adiponectin declined gradually in aged mice, as opposed to those expressing lower levels of adiponectin .Events intrinsically related to thermogenic adipocytes that could mediate the age-related decline in their thermogenic capacity are cellular senescence affecting APC, as previously discussed, and those related to mitochondrial function. Thermogenic adipocytes are rich in cristae-dense mitochondria crucial for UCP1-dependent heat production. They are maintained by constitutive mitochondrial biogenesis in brown adipocytes. Conversely, in beige adipocytes, mitochondrial content increases upon thermogenic stimulation, which signals for mitochondrial biogenesis, and decreases when the latter is withdrawn by autophagy-mediated clearance .Aging is associated with global mitochondrial dysfunction , most liThe transcription factor forkhead box protein A3 (Foxa3) was previously described to mediate high-fat diet-induced expansion of visceral white adipose tissue and was recently implicated in decreasing thermogenic adipose tissue activity with aging. It was shown that Foxa3 is upregulated in mouse brown and inguinal white adipose depots during aging and suppresses cAMP-responsive element binding protein 1-regulated transcription of PGC1α, a master regulator of mitochondrial biogenesis . Foxa3 kMitochondrial proteomic analysis of mouse interscapular brown adipose tissue revealed that aging led to diminished fuel oxidation ability, determined by reduced levels of mitochondrial lipoylation due to reduced mitochondrial iron-sulfur cluster formation required for maturation of lipoate-containing proteins in mitochondria. The latter finding was shown to be directly implicated in the impairment of glucose catabolism and thermogenic activity in this adipose depot .Nfe2l1 led to an impaired cold response, as indicated by lower mitochondrial content, abnormal thermogenic function, and whitening of brown adipose tissue  that is observed in both rodents  and humaIn contrast to the findings from rodent studies, human studies using the radiolabeled norepinephrine analog 123I-metaiodobenzylguanidine combined with PET/CT imaging indicated that older subjects exhibit both reduced thermogenic adipose tissue activity and decreased sympathetic drive when compared with their younger counterparts, independently from body mass . TherefoAging is associated with several endocrine changes that may affect thermogenic adipose tissue, including diminished gonadotropic and thyrotropic axis function. Estrogen and androgen receptors are expressed in human  and mousThe actions of sex steroids on brown adipocytes have been thoroughly reviewed elsewhere . There iLikewise, the effects of progesterone on brown adipocytes and its underlying mechanisms remain to be established and could be mediated by progesterone, glucocorticoid, or even mineralocorticoid receptors . DespiteThe impact of age-related changes in gonadotropic axis function may also be associated with increased circulating levels of the gonadotropin follicle-stimulating hormone (FSH). Blocking FSH action in mice with a polyclonal antibody was shown to reduce adiposity, induce beige adipocyte recruitment, activate brown adipose tissue, and augment thermogenesis. These effects were recapitulated in heterozygous FSH receptor knockout mice .Conversely, corticotropic axis function does not change considerably with age, and glucocorticoid receptor signaling in adipose tissue is well-known for its actions to impair brown adipose tissue  thermogenic function . It is tThyroid hormone is a critical regulator of thermogenic adipocyte recruitment and activity, affecting brown and beige adipocytes in rodent models and humans . Aging iGhrelin receptor (growth hormone secretagogue receptor) is highly expressed in the hypothalamus and at peripheral sites, including adipose tissue . Ghrelinex vivo’ (in vivo could mediate an FGF-resistant state in aged subjects.Fibroblast growth factor 21 (FGF21) is primarily secreted by the liver, acting as an endocrine factor or hepatokine, and has also been shown to be produced in adipose tissue, with paracrine and endocrine actions . FGF21 aex vivo’ . It cannSeveral skeletal muscle-derived factors referred to as myokines (or exerkines when released in response to exercise), may mediate the favorable effects of exercise on health span and longevity . Some myAging is associated with a systemic pro-inflammatory state, so-called ‘inflammaging’, which poses a high risk of metabolic disease, disability, frailty, and premature death . InflammPro-inflammatory cytokines were shown to induce brown adipocyte apoptosis directly , impair Findings from a study involving a murine brown preadipocyte cell line indicated that mature brown adipocytes exhibit increased expression of enzymes related to antioxidative defense when compared with preadipocytes. However, exposure to inflammatory cytokines such as tumor necrosis factor-alpha or interleukin 1β decreased antioxidative enzyme expression, which was accompanied by increased reactive oxygen species production and down-regulation of thermogenesis-related proteins, such as UCP1 and β-Klotho .It was reported that aging upregulated genes related to catecholamine catabolism in mouse visceral adipose macrophages in an NLRP3 inflammasome-dependent fashion, leading to decreased lipolysis . AlthougA recent study investigated the transcriptomic signature of 17 organs and plasma proteomics across mouse lifespan and found widespread activation of immune cells with age, first detectable in white adipose tissue depots at middle age . Such fiOverall changes in adipose tissue mass, distribution, and function with aging have a significant impact on other tissues, given the widespread crosstalk between adipose tissue and other tissues through several pathways, including those involving metabolic fuels, endocrine factors affecting insulin sensitivity, the inflammatory response, and circulating miRNAs .In particular, the implications of age-related changes in thermogenic adipocytes are not fully understood. It is well-acknowledged that brown and beige adipocytes’ diminished mass and activity impairs the response to cold and increases the risk of cold-induced hypothermia in the elderly. Moreover, decreased energy expenditure may contribute to reduced energy consumption in the elderly .There is increasing evidence that loss of thermogenic adipocyte activity with aging affects metabolic health. Brown and beige adipocytes act as a metabolic sink for glucose and lipids, so the age-related impairment in their function may contribute to an insulin-resistant state and metabolic disease risk . It is aMany strategies to improve health span or even extend lifespan have targeted the adipose tissue, with promising results . Most ofThe advances in understanding the mechanisms underlying thermogenic adipose tissue aging have provided promising targets and strategies to improve health span. For example, alpha-lipoic acid supplementation restored mitochondrial lipoylation in adipocytes of aged mice, enhancing the ability of brown adipose tissue to uptake glucose and execute thermogenesis, implying that the age-associated decline in brown adipose tissue function is potentially reversible . It is cAnother essential question to be addressed is the effectiveness of such strategies during adulthood. Overall, the decline in the ability to recruit beige adipocytes is apparent by the fourth decade of life in humans  and is aFSTL1 repression , intermittent fasting, and dietary restriction, have favorable effects on health and increase longevity . CaloricGenetic mouse models have provided invaluable insights not only for understanding thermogenic adipocyte recruitment and function but also for identifying potential targets to improve thermogenic adipocyte function to increase longevity. A classical model of mammalian longevity and increased health span is the growth hormone (GH) receptor knockout mouse , which eTransgenic mice overexpressing phosphatase and tensin homolog deleted on chromoson 10 (PTEN) exhibited increased longevity, which was accompanied by decreased adiposity, improved glucose tolerance, increased energy expenditure, and hyperactive brown adipose tissue activity . SimilarThe interest in understanding thermogenic brown and beige adipocyte biology was fueled in recent years due to their identification in adult humans and delineation of their role in whole-body energy homeostasis. Data from rodent models and human studies have pointed to age-related changes in thermogenic adipocytes and indicated their role in disease development. Increasing understanding of the mechanisms underlying thermogenic adipocyte dysfunction in aging has identified some pathways targeted to increase health span and longevity in rodent models, and many more may be discovered soon. Further studies should address how these findings can be translated into human physiology.


FBN1 gene. Ehlers-Danlos syndrome (EDS) is a group of inherited connective tissue disorders with similar clinical features to MFS that often requires genetic testing to confirm a diagnosis of an EDS subtype. MFS is diagnosed using the Ghent Nosology criteria, which screens for cardiovascular, musculoskeletal, integumentary, ocular, and pulmonary abnormalities. Though genetic testing has recently been increasingly emphasized in diagnosing MFS, it is not currently a mandatory component of the Ghent Nosology. We present the case of a nine-year-old male who presented with joint hypermobility of the shoulders, knees, and thumbs, and a family history of joint hypermobility in his 15-year-old brother. Genetic testing ruled out MFS, and the patient subsequently underwent testing for EDS, which further ruled out classical and hypermobile EDS. This case highlights the importance of supplementing the Ghent Nosology criteria with genetic testing in diagnosing MFS because it can aid in generating a differential diagnosis and optimizing diagnostic accuracy.Marfan syndrome (MFS) is an autosomal dominant connective tissue disorder associated with the mutation of the  FBN1) gene on chromosome 15 .,5,8.1,5,Genetic testing is often vital in diagnosing EDS subtypes and providing a definitive diagnosis for MFS. However, it is not currently a mandatory component of the Ghent Nosology criteria used to diagnose MFS. This case highlights the importance of including genetic testing in optimizing diagnostic accuracy for patients who present with clinical features consistent with MFS and EDS.


Neuroimaging studies have shown gray matter structural and functional alterations in patients with idiopathic blepharospasm (iBSP) but with variations. Here we aimed to investigate the specific and common neurostructural/functional abnormalities in patients with iBSP.A systematic literature search from PubMed, Web of Science and Embase was conducted to identify relevant publications. We conducted separate meta-analysis for whole-brain voxel-based morphometry (VBM) studies and for functional imaging studies, and a multimodal meta-analysis across VBM and functional studies in iBSP, using anisotropic effect size-based signed differential mapping.The structural database comprised 129 patients with iBSP and 144 healthy controls whilst the functional database included 183 patients with iBSP and 253 healthy controls. The meta-analysis of VBM studies showed increased gray matter in bilateral precentral and postcentral gyri, right supplementary motor area and bilateral paracentral lobules, while decreased gray matter in right superior and inferior parietal gyri, left inferior parietal gyrus, left inferior temporal gyrus, left fusiform gyrus and parahippocampal gyrus. The meta-analysis of functional studies revealed hyperactivity in right dorsolateral superior frontal gyrus, left thalamus and right fusiform gyrus, while hypoactivity in left temporal pole, left insula, left precentral gyrus, bilateral precuneus and paracentral lobules, right supplementary motor area and middle frontal gyrus. The multimodal meta-analysis identified conjoint anatomic and functional changes in left precentral gyrus, bilateral supplementary motor areas and paracentral lobules, right inferior occipital gyrus and fusiform gyrus.The patterns of conjoint and dissociated gray matter alterations identified in the meta-analysis may enhance our understanding of the pathophysiological mechanisms underlying iBSP. Idiopathic blepharospasm (iBSP) is an adult-onset focal dystonia that has a peak age at onset in the fifth to sixth decade with a female predominance . IBSP isConventional neuroimaging studies and autopsy studies failed to find structural brain lesions in iBSP. Nevertheless, studies with advanced neuroimaging approaches have revealed structural and functional brain alterations in iBSP, and have advanced our understanding of the brain alterations in patients with iBSP. For example, the voxel-based morphometry (VBM) was used to measure the microstructural changes of brain, and functional magnetic resonance imaging (MRI) and positron emission tomography (PET) have been used to evaluate the functional changes. Using VBM, Etgen et al. reported gray matter (GM) increase in bilateral putamen and GM decrease in the left inferior parietal lobule in patients with iBSP when compared to healthy controls . While sAlthough studies have examined structural or functional brain alterations in iBSP, the pattern for convergence or divergence in regional GM alterations remains unknown. Therefore, we performed separate meta-analysis to identify consistent and reliable GM structural and functional alterations in iBSP by integrating all eligible studies reporting whole-brain GM abnormalities and brain activity, separately. Then we undertook multimodal meta-analysis of GM structural and functional studies to detect converging findings that may be indicated as important brain nodes across different neuroimaging modalities in iBSP, using anisotropic effect size-based signed differential mapping (AES-SDM), a coordinate-based meta-analytic tool that has been widely applied in neuroimaging studies of neurological disorders . We alsoSystematic literature search was conducted for peer-reviewed human studies in PubMed, Embase and Web of Science published until 16 August 2021. The following English search terms were used: “blepharospasm” and . The language was restricted to English. In addition, manual searches were conducted within review articles and via the reference lists of individual studies. After duplicate removal, 739 studies were identified .Studies were considered for inclusion if they met the following criteria: (a) published in an original English paper with peer review, (b) comparison of patients with iBSP and healthy controls, (c) employed whole-brain VBM to detect GM structural changes (volume or density) or functional imaging  to explore GM functional alterations; to minimize the heterogeneity of the functional imaging paradigms, only functional studies in the resting state were included; (d) reported whole-brain results of changes in three-dimensional coordinates  [Talairach or Montreal Neurological Institute (MNI)]. Studies only reporting regions of interests (ROI) findings were excluded. Studies were independently ascertained and checked by the two researchers. To achieve a high standard of reporting we have adopted “Preferred Reporting Items for Systematic Reviews and Meta-Analyses” (PRISMA) guidelines. The study selection procedures were summarized in t-values, z-scores, or p-values) with significant differences between patients with iBSP and healthy controls in GM changes or brain functional activity were extracted from each study according to the SDM tutorial. Two authors conducted the study selection and extracted the data that needed to perform the meta-analyses independently.For each included study, we recorded the following demographic and clinical characteristics: sample size, sex, mean age, mean age at disease onset, disease duration, mean score of motor symptom and medication. Peak coordinates and their effect sizes . The procedures have been described in detail elsewhere , as previous simulations indicated that this threshold provided an optimal balance between sensitivity and false-positive rate  and functional (PF) alterations to identify regions with alterations in each modality using separate meta-analysis. The multimodal analysis combined the two probabilities maps, incorporating the P-values to identify a union of alterations in both modalities (U). The estimation of U is straightforward as U = PGM + PF- PGM × PF. However, the U statistic in its raw form is overly conservative, and to reduce the imbalance between the false-positive and negative rates, U was adjusted according to P = U+(1-U) × ln(1 − U) (p < 0.0025) than that used in unimodal meta-analyses. It should be noted that this analysis did not aim to detect correlations between structural and functional abnormalities, but to localize brain regions in which iBSP is associated with both structural and functional alterations.Areas of overlapping functional and structural abnormalities between patients with iBSP and healthy controls were assessed by conjunction analysis using the multimodal meta-analysis in AES-SDM. This multimodal meta-analysis approach aims to ensure that the false-positive rate is not increased compared with that in studies of any single modality . We obtan(1 − U) . A more Robustness of the significant results was assessed by means of exploration of the residual heterogeneity, as well as by jackknife sensitivity analyses. Specifically, we examined the funnel plots of the peaks of maximum heterogeneity in order to check whether findings might have been driven by few or small noisy studies, or to detect any gross abnormality such as studies reporting opposite results . As regaP = 0.0005 and cluster extent = 10 voxels was used to minimize spurious findings  and bilateral paracentral lobules, while decreased GM in right superior and inferior parietal gyri, left inferior parietal gyrus, left inferior temporal gyrus, left fusiform and parahippocampal gyrus .Patients with iBSP showed increased activity compared to healthy controls in the right dorsolateral superior frontal gyrus, left thalamus and right fusiform gyrus, while decreased activity in left temporal pole and left insula, left precentral gyrus, bilateral precuneus and paracentral lobules, left superior parietal gyrus, right SMA and middle frontal gyrus .Multimodal analysis in iBSP showed increased GM and hypoactivity relative to healthy controls in bilateral SMA and bilateral paracentral lobules, and left precentral gyrus. Right inferior occipital gyrus and fusiform gyrus showed GM decrease with hyperactivity .In the linear regression analysis, age did not have any significant effects on the observed between-group GM or brain activity differences.The whole-brain jackknife sensitivity analysis showed that the findings of GM increase in the bilateral precentral and postcentral gyri were highly replicable, being preserved in all combinations of the datasets. The GM increase in right SMA and bilateral paracentral lobules were significant in all but two combinations, all the other GM alterations were significant in all but one combination. The brain functional alterations in left temporal pole, left insula, left middle temporal gyrus, left precuneus and left precentral gyrus were significant in all combinations of the datasets. Similarly, the results in right superior frontal gyrus, left thalamus, right lingual gyrus and right SMA were significant in all but two combinations. The results of the meta-analyses thus showed high replicability and reliability in these regions. Egger's tests were non-significant, suggesting there was no evidence of publication bias for the reported clusters , 4. The To the best of our knowledge, this is the first multimodal neuroimaging meta-analysis that combines information from studies investigating whole-brain gray matter and studies investigating the functional brain on iBSP. The main findings were conjoint increased GM and hypoactivity in left precentral gyrus, bilateral SMA and paracentral lobules, and decreased GM and hyperactivity in right inferior occipital gyrus and fusiform in patients with patients with iBSP compared to controls. In addition, patients with iBSP showed decreased GM in bilateral inferior parietal and left superior parietal gyri, left inferior temporal gyri, left fusiform gyrus and left parahippocampal gyrus, while hyperactivation in right dorsolateral superior frontal gyrus, left thalamus and hypoactivity in left temporal pole and insula and bilateral precuneus. The specific and common structural and functional abnormalities revealed in the present meta-analysis may give new insight into the neuropathology of iBSP.GM increase with hypoactivity was observed in the left precentral gyrus, bilateral SMA and paracentral lobules, regions that are functionally belong to the motor network. These results were consistent with the abnormal excitability of the primary motor cortex found in iBSP using transcranial magnetic stimulation (TMS) , 29. TheWe found regions in the right inferior occipital gyrus and fusiform gyrus had GM atrophy and hyperactivity, and the left fusiform had GM atrophy. Abnormalities of structure and functional activity in these regions were also reported in other forms of dystonia , 34. FusSeveral regions showed only structural or functional alterations in iBSP. The structural alterations included increased GM in bilateral postcentral gyri, decreased GM in bilateral inferior parietal gyri, left superior parietal gyrus, left inferior temporal gyrus and left parahippocampal gyrus. Although iBSP is a movement disorder, sensory symptoms including burning sensation and grittiness in the eye, dry eye and photophobia are frequently reported , 40. In An additional involvement of GM atrophy was found in the left inferior temporal gyrus and left parahippocampal gyrus in patients with iBSP. Inferior temporal gyrus is known to be a visual association area that receives inputs from the occipital lobe through reciprocal projection and links comprehensive information toward the prefrontal cortex for higher-order functions . NeuroimThe increased functional activities in left thalamus and right dorsolateral superior frontal gyrus, and decreased functional activities in bilateral precuneus, left temporal pole and insula were found in iBSP and not accompanied by structural alterations. Glucose hypermetabolism in the thalamus was observed in patients with iBSP, which may be related to the primary cause of compensatory mechanism of iBSP sharing the common pathophysiological mechanism to other types of focal dystonia . It is aBoth the temporal pole and insula are complex structures with different anatomical and functional subregions, and have wide connections with other cortical and subcortical areas. The temporal pole is associated with several high-order brain functions such as language and semantic processing, socio-emotional processing and autobiographic memory . AbnormaSome limitations of this study should be noted. First, our multimodal analysis found regions showing both structural and functional alterations, but whether the relationships between structural and functional changes are causal cannot be addressed. Future studies are encouraged to detect the spatial and temporal relationships between the structure and function of these regions identified in current meta-analysis. Second, the differences in medical treatments, age of onset, disease duration, sex and age of included studies, and the difference in the preprocessing pipelines of included studies may influence the results of meta-analyses. Third, this meta-analysis was conducted based on the reported coordinates with significant differences rather than raw data from individual cases, which would affect the accuracy of the results. Fourth, we cannot determine whether these alterations are parts of the pathophysiology of iBSP or a consequence of the disease. And lastly, the GM volume or density was not specified in the results as we included studies examining GM density or volume as previous studies did , 63, altOur meta-analyses found that GM abnormalities in iBSP were characterized by conjoint alterations of regional structure and function mainly in the motor cortex and visual cortex, and separate GM structural changes in sensorimotor cortex and limbic lobe, and widespread altered function in the thalamus, insula, temporal pole, and regions involved in DMN and cognitive executive network. These results give us a description about underlying brain alterations that may be involved in motor and non-motor symptoms of iBSP, which may help provide new insight into the neuropathology of this disease.The original contributions presented in the study are included in the article/MZ: execution, data acquisition, statistical analysis, and manuscript preparation. XH and BYL: execution, data acquisition, manuscript review, and critique. HFS: manuscript review and critique. JY: conception, organization, manuscript review and critique, and responsible for the overall content as the guarantor. All authors contributed to the article and approved the submitted version.This work was supported by the National Natural Science Foundation of China (grant number 81971071), the Applied Basic Research Programs of Science and Technology Department of Sichuan Province (grant number 2021YJ0447), and 1.3.5 project for disciplines of excellence-Clinical Research Incubation Project, West China Hospital, Sichuan University (grant number 2021HXFH044).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.


Targeting calcitonin gene-related peptide (CGRP) and its receptor by antibodies and antagonists was a breakthrough in migraine prevention and treatment. However, not all migraine patients respond to CGRP-based therapy and a fraction of those who respond complain of aliments mainly in the gastrointestinal tract. In addition, CGRP and migraine are associated with obesity and metabolic diseases, including diabetes. Therefore, CGRP may play an important role in the functioning of the gut-brain-microflora axis. CGRP secretion may be modulated by dietary compounds associated with the disruption of calcium signaling and upregulation of mitogen-activated kinase phosphatases 1 and 3. CGRP may display anorexigenic properties through induction of anorexigenic neuropeptides, such as cholecystokinin and/or inhibit orexigenic neuropeptides, such as neuropeptide Y and melanin-concentrating hormone CH, resulting in the suppression of food intake, functionally coupled to the activation of the hypothalamic 3′,5′-cyclic adenosine monophosphate. The anorexigenic action of CGRP observed in animal studies may reflect its general potential to control appetite/satiety or general food intake. Therefore, dietary nutrients may modulate CGRP, and CGRP may modulate their intake. Therefore, anti-CGRP therapy should consider this mutual dependence to increase the efficacy of the therapy and reduce its unwanted side effects. This narrative review presents information on molecular aspects of the interaction between dietary nutrients and CGRP and their reported and prospective use to improve anti-CGRP therapy in migraine. Migraine is a common disabling disorder affecting mainly individuals in productive age and consequently resulting in a high burden for societies. Despite its high prevalence and serious consequences, it is still an undertreated disorder. The main reason for this may be poor knowledge of molecular mechanisms underlying migraine induction and progression.Calcitonin gene-related peptide (CGRP) is a neuropeptide, which, along with its receptor, is involved in pain transmission and released from trigeminal termini during migraine attacks . HoweverDietary nutrients may modulate the action of many therapeutic drugs. Appropriate diet and hydration are essential in ameliorating the side effects of anticancer and antiviral drugs . This maIn this narrative review, we present and update information on molecular aspects of the interaction between dietary nutrients and CGRP, their reported and putative clinical implications and perspectives on improving anti-CGRP therapy in migraine with appropriate nutrition. We also discuss some nutritional issues of metabolic disorders, including obesity and diabetes, that may associate with migraine and the CGRP system. Some basic information on CGRP and migraine pathogenesis is also provided.Migraine is a neurological disorder involving sensory abnormalities occurring prior to, during and following a headache . HistoriIn fact, the pathogenesis of migraine is poorly known, and many factors, both genetic/epigenetic and environmental/lifestyle, can be involved (reviewed in ). MigraiComprehensive diets regulate the right proportion between the core components of foods, including minerals, vitamins, proteins, carbohydrates and fats. However, some individuals report decreases in the frequency of headache attacks and/or their intensity after modifications to such a comprehensive diet . In clinSeveral kinds of diet were postulated to prevent migraine or mitigate its effects , but theNot only the diet content but also the meal schedule was associated with migraine. A higher prevalence of migraine was shown in individuals who had not a regular diet and regular diet schedule and who had fewer than three meals per day . Diet coNutraceuticals, food or dietary supplements exerting beneficial health effects were studied in migraine prevention and therapy (reviewed in ). They aIt is not the purpose of this work to present general information on associations between dietary nutrients and migraine. For this work, such information is important in the context of the CGRP system and CGRP-targeted antimigraine therapy. It should be underlined that migraine may be a diet-sensitive disease, and this should be considered when antimigraine treatment is to be modified by dietary compounds.CALCA) gene located at 11p5a. Alternative processing of the CALCA gene produces calcitonin (CT) and CGRP-alpha (CGRP1), one of the two isoforms of CGRP [CALCB gene, but CGRP-alpha is a predominant CGRP form in the trigeminal ganglia and will be further referred to as CGRP unless otherwise stated. CGRP is a 37-aa neuropeptide involved in many phenomena and effects, including pain transmission and migraine [Human CGRP is encoded by the calcitonin-related polypeptide alpha . HoweveThe treatment of migraine remains challenging, and a possible causative role of CGRP in migraine stimulated studies on its therapeutic utilization in this disease, which resulted in the development of the first migraine-specific preventive treatment drugs. Three classes of therapeutics targeting the CGRP system were approved for both chronic and episodic migraine treatment and prevention. These are (1) monoclonal antibodies against CGRP; (2) a monoclonal antibody targeting the canonical CGRP receptor, and (3) small molecular weight antagonists of CGRP receptors (gepants) (reviewed in ). These The canonical CGRP receptor is a complex of a G protein-coupled receptor (GPCR), calcitonin-like receptor (CLR) and receptor activity-modifying protein 1 (RAMP1), but the second receptor, AMY-1, consisting of calcitonin receptor (CR) and RAMP1 may also be bound by CGRP  Figure . CGRP coAlthough the precise mechanism of the involvement of CGRP in migraine is still not fully understood, its vasodilatory effect on cranial arteries as a link to migraine was established . As suggInteraction between drugs and dietary nutrients may result in changes in their metabolism and transport . HoweverSome studies show that certain kinds of diet affect circulating CGRP levels, suggesting that certain components of the diet may influence migraine outcomes (reviewed in ). SecretGrape seed extract was shown to suppress basal expression of CGRP in spinal neurons in rats . This ef2+ and Cu+, and modulating pro- and antioxidant enzymes.Cocoa bean preparations have a long history of health-related beneficial effects . They diOn the other hand, the epicatechin content of cocoa is associated with its beneficial effects on vascular endothelium through its impact on both acute and chronic upregulation of nitric oxide (NO) production . Cocoa pLower levels of CGRP in serum during episodic migraine were observed after dietary supplementation with vitamin D (2000 IU per day) as compared with individuals who received a placebo . This efBeneficial effects in migraine were observed for some synthetic formulations of natural compounds—in general, they are moderately effective and generally safe (reviewed in ). GinkgoCinnamon is reported to display anti-inflammatory and neuroprotective properties and modulate some hippocampal functions . It was The regulation of appetite and feeding behavior involves both central and peripheral neurohormonal mechanisms that are not completely known . In CNS,It was shown in several studies that CGRP exhibited central anorexigenic effects in animals, but these studies may not reflect general CGRP properties in appetite/satiety and food intake regulation ,77. It wMechanisms underlying the anorexigenic potential of CGRP were investigated in rats . IntrapeAnorexigenic behavior in rodents and primates is regulated by the lateral parabrachial nucleus, which is a duct for visceral signals from the caudal hindbrain to forebrain areas linked with appetite control ,81. It wAlthough those studies were conducted in rats, there are fundamental regulatory mechanisms that have exerted strong evolutionary pressure . As a coAlthough the cause of obesity is a long-term energy imbalance featured by increased energy intake and reduced energy expenditure, and so it can be avoided by increased physical activity, and dietary nutrients may play an important role in obesity pathogenesis .−/− mice were characterized by higher core temperatures, increased energy expenditures, and a relative daytime (nonactive) depression in respiratory quotients, suggesting an increased β-oxidation [−/− mice showed improved glucose handling and insulin sensitivity. These results clearly showed the role of CGRP as a mediator of energy metabolism and a target in obesity-reducing treatment.It is generally accepted that migraine and obesity are related to each other as obesity exacerbates migraine by increasing the risk of the disease and higher frequency and severity of headaches (reviewed in ). Althouxidation . In respType 2 diabetes mellitus (T2DM) occurs more frequently in obese individuals than in persons with normal weight and is associated with impairment in insulin secretion and activity (reviewed in ). AlthouIt was proposed that the early development of insulin resistance, impaired glucose tolerance and T2DM could be modulated by the increased activity of sensory nerves in Zucker diabetic fatty rats ,92. SmalIt was postulated that CGRP might affect adipocytes to promote lipid utilization and pancreatic β-cells to modulate insulin secretion . AnalysiThe studies on the role of CGRP and CGRP-related therapy in obese and diabetic animals were translated into two randomized, double-blind, placebo-controlled trials to assess the safety and metabolic effects of eptinezumab, an anti-CGRP antibody, in non-migraine overweight/obese patients and patients with T1DM . No chanMigraine patients are at a higher risk of GI disorders, and this relationship is bidirectional, in agreement with the conception of the gut-brain-microbiota axis . The preCGRP and its receptor were localized in enteric neurons in various human GI segments ,75. In tConstipation is frequently mentioned as a gut-related adverse effect of anti-CGRP migraine therapeutics as the post-approval real-world surveys showed that it might occur in more than 50% of patients treated with erenumab, an antibody targeting the CGRP receptor, fremanezumab, or galcanezumab, antibodies targeting CGRP (reviewed in ). CGRP iFDA Adverse Event Reporting System (FAERS) shows that the prevalence of adverse, GI-related syndromes associated with the administration of CGRP-based antibodies ranges from 0.3% (diarrhea) to 7.6% (nausea) (reviewed in ). Gepant2) patients, and the 100-mg dose did not exceed a 10% separation for certain subpopulations of patients (obesity classes I (30 to <35 kg/m2) and II, patients with migraine diagnosed between 20 and 30 years of age, and patients diagnosed less than 9 years), the fraction of migraineurs with ≥50% migraine responder rate was higher for eptinezumab- than placebo-treated patients. Therefore, these studies point to BMI as a factor that can modulate anti-CGRP therapy, but this issue should be addressed in further research also in the context of nutrition. An explorative, prospective, questionnaire-based study showed that most chronic migraine patients treated with CGRP antibodies displayed complete prevention of migraine symptoms without indication of initial onset followed by attack abortion [p = 0.007). Altogether, these studies show that gastrointestinal homeostasis may be associated with the efficiency of anti-CGRP therapy. Therefore, nutrition may play a role in this fundamental anti-migraine therapy.Recent randomized controlled trials and real-life studies provided important information on GI homeostasis and anti-CGRP therapy. The multicenter prospective cohort GARLIT real-life study enrolled patients with high-frequency episodic migraine and chronic migraine ,102. It abortion . About oabortion . FurtherIn summary, CGRP is an important element of the gut-brain axis, and its modulation may lead to GI functional disorders. Such disorders are reported to be side effects of anti-CGRP therapy in migraine. However, this association may be featured by several factors, including intake of dietary nutrients, as it may be involved in both migraine and GI tract disorders. Dietary nutrients improving functions of the GI, in particular preventing or inhibiting diarrhea and constipation as well as nausea, may be useful in reducing unwanted side effects of anti-CGRP migraine therapy.Dietary nutrients are of concern to migraine patients as some of them are migraine triggers and may exacerbate migraine headaches. On the other hand, CGRP, a central neuropeptide in migraine pathogenesis, is sensitive to many dietary nutrients. This sensitivity may influence CGRP properties that could contribute to migraine pathogenesis, such as light aversion, neurogenic inflammation, peripheral and central sensitization of nociceptive pathways, cortical spreading depression, and regulation of nitric NO production . FurtherMany studies associate the diet with migraine pathogenesis, first of all recommending dietary components to avoid so as not to induce headaches or exacerbate existing ones. Much fewer studies deal with dietary components that are recommended to be added to the standard diet. As with any kind of diet, migraine preventive diets should be adjusted to individual needs and constitutions, understood as the set of genetic/epigenetic and phenotypic features.The introduction of antibodies against CGRP, along with antibodies and antagonists to its receptor, revolutionized the prevention and treatment of migraine, but several outstanding questions remain . Some ofAnother problem associated with anti-CGRP is the unwanted side effects of the therapy. Generally, such therapy is considered safe, but some patients complain of ailments, mainly from the GI tract . TherefoThe anorexigenic potential of CGRP is important in any dietary intervention in migraine patients and obviously impedes such intervention. Therefore, dietary nutrients recommended in migraine and in anti-CGRP therapy should be provided in the form of diet supplements rather than natural products. Additionally, obesity and metabolic diseases, including diabetes, that may be influenced by CGRP make this problem even more complex.The fundamental outstanding question is whether dietary nutrients may be used to improve anti-CGRP migraine therapy making it more effective and/or safe. To address this question, many elements should be taken into consideration, including two fundamental ones. Firstly, dietary nutrients that are migraine triggers should be excluded as such compounds, even if they may improve the mechanistic action of ant-CGRP drugs. Secondly, nutritional strategy in obese and/or diabetic patients in anti-CGRP therapy should be adjusted to the anorexigenic potential of CGRP.This review does not answer important questions on the use of nutrients in anti-CGRP therapy. Such questions can be addressed when outcomes of clinical trials and results of laboratory studies are available. This review aimed to underline the importance of dietary nutrients in anti-CGRP therapy of migraine and justify the need for further research as the currently available data are scarce.


P1-HCPro bi-cistron of a plum pox virus construct (PPV-GFP) was replaced by that of potato virus Y (PVY) spread slowly systemically in Nicotiana benthamiana plants and accumulated to levels that were 5−10% those of parental PPV-GFP. We tested whether consecutive mechanical passages could increase its virulence, and found that after several passages, chimera titers rose and symptoms increased. We sequenced over half the genome of passaged chimera lineages infecting two plants. The regions sequenced were 5′NCR-P1-HCPro-P3; Vpg/NIa; GFP-CP, because of being potential sites for mutations/deletions leading to adaptation. We found few substitutions, all non-synonymous: two in one chimera , and three in the other . HCPro substitutions 2053 AUU(Ile)→ACU(Thr), and 2359 CUG(Leu)→CGG(Arg) occurred at positions where single nucleotide polymorphisms were observed in NGS libraries of sRNA reads from agroinfiltrated plants (generation 1). Remarkably, position 2053 was the only one in the sequenced protein-encoding genome in which polymorphisms were common to the four libraries, suggesting that selective pressure existed to alter that specific nucleotide, previous to any passage. Mutations 5729 and 5733 in the Vpg by contrast did not correlate with polymorphisms in generation 1 libraries. Reverse genetics showed that substitution 2053 alone increased several-fold viral local accumulation, speed of systemic spread, and systemic titers.A viral chimera in which the  Potyviruses that among its functions performs two that are essential to the virus infection cycle: it suppresses gene silencing, neutralizing selectively plant antiviral defenses based on small RNAs (sRNAs) of viral sequence ..P1-HCProt of PVY b and [3]Potyviruses indicate that mutations leading to adaptation do not distribute uniformly along the viral genome but target instead specific variable regions of some of the encoded viral proteins [On the other hand, how plant viruses adapt and evolve in response to new hosts and environments, and vectors, is currently a very interesting topic of research, as climate change and increased global trade multiply the opportunities for those situations to occur . Studiesproteins ,11.N. benthamiana) of an intrinsically un-adapted, artificial virus lacking any starting sequence diversity.We wanted therefore to test whether we could increase the virulence of our chimera for practical purposes, but also to study how an un-adapted virus could evolve towards becoming better adapted to its host. The fundamentals of adaptation of natural viruses to new contexts may or may not differ from those of adaptation to a compatible experimental host (We agroinfiltrated plants with the chimera constructs (generation 1 plants) and after several consecutive mechanical passages that lasted around four months a we seleP1HCPro bi-cistron (in fragment 1) is a heterologous sequence that originates from another potyvirus, PVY; that the Vpg-NIa (in fragment 2) contains hyper-variable regions , where non-synonymous substitutions have been shown to occur under positive selection, facilitating the genetic adaptation of viral isolates [GFP cistron is a non-viral sequence that could be lost, and the second one that the CP has been also reported as a potential target for viral adaptation and evolution [We sequenced three fragments covering together over half of the genomes of the chimeras infecting plants A and D a. The reisolates ,11; and volution .GFP cistron had been maintained through the months of passages, suggesting that its fitness cost to the virus in this host is low, or lower than the cost of deleting it. Overall, we could find only a few non-synonymous substitutions, but different among the two chimeras, indicating that divergent chimera evolution in different plants had occurred . In the case of the mutation in HCPro of the chimera infecting plant D, restoration of the intact PVY HCPro sequence may have led to a slightly more efficient polyprotein processing, although western blot data shows that the original unmodified chimera processes HCPro properly (not shown). In the case of mutation of HCPro amino acid 352, we have no indication of why it could be selected.These data indicate that mutations in specific sites of the viral genome are occurring repeatedly, prior to any passage bottlenecks, and that selective pressures are already operating in generation 1 infections that will lead to some becoming eventually prevalent, as happened to those in positions 2053 and 2359 in the To test the effect on virulence of the nucleotide 2053 mutation leading to HCPro amino acid substitution 352 Ile→Thr, we incorporated it into the original chimera, creating chimera 352Ile→Thr, and compared both viruses. We found that chimera 352Ile→Thr titers were several-fold higher in agroinfiltrated tissues as well as in systemic tissues than those of the unmodified chimera, and also that the virus moved much faster . TherefoPrunus). With regard to experimental plants, PVY infects N. benthamiana and tobacco, but not arabidopsis Col 0 plants. PPV-GFP by contrast infects N. benthamiana, arabidopsis Col 0, and the agroinfiltrated leaves of tobacco leaves but does not spread systemically [in vitro [in vivo is a matter for further research.We finally tested the host range of our chimera relative to those of the parental viruses. PVY infects many solanaceous plants of economic importance such as potato, pepper or tomato. PPV has also devastating effects on bone fruit trees . We show that in generation 1 infections, selective pressures created genomic variability at specific nucleotides, which with regard at least to those in yviruses . This co


Oral submucous fibrosis (OSMF) is a chronic oral potentially malignant disorder (OPMD). It is described as a scarring disease of the oral mucosa associated with excess oxidants and insufficient antioxidants. While it is becoming increasingly accepted that oxidative stress results in excessive accumulation of collagen and progressive fibrosis of the submucosal tissues, there is limited data regarding the moderation of oxidative stress to initiate or prevent OSMF. To assess the scope for mechanism-based approaches to prevent or reverse OSMF, we systematically evaluated the existing literature and investigated the role of oxidative stress in the pathogenesis and chemoprevention of OSMF. A search for relevant articles on PubMed and Scopus was undertaken using pre-defined inclusion and exclusion criteria. A total of 78 articles were selected in accordance with Preferred Reporting Items for Systematic Reviews and Meta-Analyses extension for Scoping Reviews (PRISMA-ScR) guidelines. The articles eligible for assessment investigated both OSMF and/or oxidative stress biomarkers or specific antioxidants. Both in vitro and human studies consistently demonstrated variations in oxidative stress biomarker levels in OSMF and revealed an increase in oxidative stress, paralleling the development of the disease. Furthermore, the use of antioxidant supplements was overall associated with an improvement in clinical outcomes. Having identified the significance of oxidative stress in OSMF and the therapeutic potential of antioxidant supplements, this scoping review highlights the need for further well-designed studies in the development of mechanism-based interventions for managing OSMF. Oral submucous fibrosis (OSMF) is a chronic and subtle oral potentially malignant disorder (OPMD) affecting the oral cavity and oropharynx, in mainly the South-Asian populations . ClinicaAlthough OSMF is multifactorial in origin, there is overwhelming evidence that long-term areca-nut chewing ), is considered the main etiological factor. Constituents of BQ have in fact been shown to generate substantial amounts of reactive oxygen species (ROS) , which mThe aim of this study was to evaluate the existing literature systematically to assess the role of oxidative stress in OSMF. It was concluded that excess oxidants may drive the pathogenesis, while an excess of antioxidants may play a fundamental role in the chemoprevention of OSMF and the potential reversal of its debilitating effects.This scoping review was done according to Preferred Reporting Items for Systematic Reviews and Meta-Analyses extension for Scoping Reviews (PRISMA-ScR) guidelines. A search for relevant articles on Pubmed and Scopus published up to June 2021 was completed using the search string: ‘ AND ’. Inclusion criteria included studies with OSMF human subjects, in vivo or in vitro models of OSMF, as well as studies assessing oxidative stress biomarkers or molecules or using antioxidants in models of fibrosis. Exclusion criteria included non-English language and non-peer-reviewed studies, systematic reviews, meta-analyses, and books/book chapters.The selection process involved three stages ; the seaData extracted from each article were the name of the first author, publication year, study type , PICO, oxidative stress biomarkers (if present), study design or model, assay methodology, details of treatment/intervention, if any. n = 1038) articles were retrieved from Scopus and PubMed, respectively. After removing 300 duplicates, the datasets were combined, giving a total of 738 manuscripts. A further 614 papers were excluded due to inconsistent titles and abstracts, leaving a total of 124 articles. After reading the full text, a further 45 manuscripts were excluded. The remaining 78 articles were deemed to be eligible for detailed assessment, of which 52 were laboratory-based studies  have been shown to have cytotoxic effects on hamster cheek pouch , normal 2O2) and sodium, compared to healthy controls  r2−) and H2O2 by converting them into water, are copper/zinc, iron/manganese, and nickel type [Glutathione S-transferase (GST), a family of Phase II detoxification enzymes that function to protect cellular macromolecules from attack by reactive electrophiles, are long-time known to protect cells from oxidative stress . Howeverkel type , and werkel type ,30,39, bkel type . Lower akel type  demonstrIn summary, there is sound evidence that oxidative stress biomarkers are altered in OSMF tissues as well as in patients’ blood. Multiple compounds with antioxidant and anti-cariogenic properties were investigated to assess their effectiveness in managing or preventing OSMF. The mechanism of action of antioxidants may involve immune system stimulation and breakage of the free radical chain reactions .Six articles investigated the efficacy of aloe vera in managing OSMF with substantial antioxidant vitamins and enzymes, topical aloe vera was effective in improving at least one of the following parameters: mouth burning and opening ,74,76,77Lycopene is a carotene, carotenoid pigment, and phytochemical with potent antioxidant and anti-carcinogenic properties. Documented as a non-invasive treatment option, it yields significant improvements in OSMF signs and symptoms . Its admCurcumin, the main natural polyphenol found in Curcuma species , has beeEpigallocatechin gallate (EGCG) is a plant-based potent antioxidant protecting against cellular damage caused by free radicals , with poLastly, other compounds such as alpha-lipoic acid (ALA) are worth noting due to their effects in alleviating burning sensation and improving mouth opening. Along with conventional intralesional steroid injections, ALA was indeed able to reverse higher clinical stages . Jiang eEight papers compared the chemopreventive effects of nutrient antioxidants against each other. When compared to standard antioxidant capsules, aloe vera produced greater improvements in burning sensation and mouth opening , while pIn summary, results of clinical studies show that there is clear scope for investigating the preventive and therapeutic effects of antioxidants in betel nut chewers and patients with OSMF through well-designed, large, controlled studies.By providing a broad overview of relevant studies, this scoping review has established the current understanding of the topic of interest while identifying gaps in the literature. However, we must acknowledge the possibility that we may have missed some relevant studies including non-English articles and articles with no full text. The balance of breadth and depth of the data reported in this article was also challenging due to the volume of articles identified and time constraints. This present review was not aimed at providing a comprehensive pathogenic model of OSMF. While oxidative stress is indeed associated with OSMF, tissue inflammation is crucial for the induction of tissue fibrosis. The involvement of prostanoids and other inflammatory mediators ,91 as weThe current literature is in favour of antioxidants being used for OSMF prevention and management because it is relatively non-toxic and can be easily supplemented in the diet. Our findings have important public health implications for the management and reversal of OSMF’s debilitating effects. Additionally, our results can be used as a helpful precursor for future systematic reviews to deepen knowledge of pathogenesis and chemoprevention of OSMF. This review may also serve as a framework for future studies to help inform the development of mechanism-based interventions and a clear guidelines for managing patients with OSMF using antioxidants.Understanding the molecular profile of distinct BQ components and how these mediate the pathogenesis of OSMF is a key challenge if we are to develop mechanism-based preventive or therapeutic strategies for this potentially malignant disease ,94. This


We review and bridge the literature on the internationalization of state-owned firms and sovereign wealth funds to provide a novel understanding of how government ownership affects foreign investments in three ways. First, we explain how state-owned firms and funds behave differently from private ones because they need to balance governments’ nonbusiness objectives and firms’ business goals. This results in competing predictions on whether government ownership helps or hinders internationalization due to particular nonbusiness objectives. Second, building on the review, we provide suggestions on how to extend research topics and theories of the firm by incorporating these nonbusiness objectives in the internationalization decisions in four areas: home government’s endowments, characteristics, and attitudes; host-country expansion’s support, influence, and impact; home- and host-country relationship conflicts, mediation, and disguising; and management’s orientation, opacity, and arbitrage. Third, we capture how governments may use state-owned multinationals and sovereign wealth funds to nudge host-country governments by introducing the concept of discreet power and the use of four strategies  to achieve it. This helps to outline the beginning of a unified approach to how governments use their foreign investments to achieve nonbusiness goals.The online version contains supplementary material available at 10.1057/s41267-022-00522-w. There has been a resurgence of state capitalism in international business, through which governments are increasingly becoming foreign investors. The 1980s and 1990s were a period of retrenchment for state capitalism as communist countries transitioned towards capitalism, and advanced and developing economies underwent deregulation and privatization , but lower-level governments also own them, like the Alaska Permanent Fund Corporation.There are significant differences in goals between state and private investors. At the core of the theory of the firm lies its objective function, which is maximizing profits or market returns  have politically influenced nonbusiness motives; (2) select more challenging host countries and use more acquisitions, even if the market reactions are negative; and (3) show conflicting behavior on the level of internationalization, with some studies proposing a higher level and others a lower one  sovereign wealth funds invest abroad in search of diversification and behave differently from private funds, and (2) they invest in better firms and countries with stronger investor protection.The sparse literature on the internationalization of sovereign wealth funds can be grouped into two sets: studies on their Sovereign wealth funds invest abroad to diversify their macroeconomic and political exposures by investing in global equities and debt, especially those of developed economy firms. This stream of research on portfolio selection unanimously finds that sovereign wealth funds behave differently, as they invest in firms in strategic industries , risk appetite, and returns. Particular attention is given to sovereign wealth funds’ transparency and their target firms’ performance  and agents (managers) to achieve desired goals . State-owned firms suffer from unique agency problems in comparison to private firms. These firms are viewed as suffering from multilevel principal-agent problems which result in reduced competitiveness and performance relative to private firms  capital and sovereign wealth fund home-country markets Rose, . The staThe resource-based view explains how private firms develop and use resources and capabilities to satisfy customers’ needs better than rivals Barney, . From thThe literature on sovereign wealth funds does not explicitly build on the resource-based view. Instead, it discusses how sovereign wealth funds mitigate the resource curse by reducing the negative consequences of natural resource wealth, reducing corruption from the misuse of wealth in emerging economies Below is the link to the electronic supplementary material.


PLoS Medicine used movement readings from participants’ wrists to assess their sleep and then investigated associations with several psychiatric disorders.People with psychiatric disorders often experience sleep problems. A recent post hoc cross-sectional study in  Polysomnography, which records brain waves, blood oxygen, heart rate, breathing and movement, has traditionally been used to monitor sleep. However, this requires specialist equipment and is therefore not amenable to simultaneously monitoring the sleep of large numbers of people at home. Self-reporting of the quality and duration of sleep is often used instead, but is known to be less accurate. Recently wrist-worn devices have enabled movement patterns to be quantified whilst normal activities are being undertaken, and such accelerometer readings can be used to monitor sleep.Wainberg et al.1 investigated associations between sleep and diagnoses of schizophrenia spectrum disorders, bipolar disorder/mania, major depressive disorder and anxiety disorders. In contrast to many prior studies, participants were not selected for study based on known inpatient psychiatric diagnoses but were representative of the wider population. Over 80,000 participants from the UK Biobank were provided with an accelerometer and asked to wear it on their dominant wrist for 7 days. Ten sleep measures were assessed, including bedtime, wake-up time, sleep duration, sleep efficiency, number of sleep periods, naps and variability in sleep patterns.Whilst it has been shown that poor sleep is both a causal risk factor and common in people with many psychiatric disorders, most studies have focused on people experiencing particular disorders. Similar sleep pattern differences were present across all the disorders studied, generally with the disorders associated with reduced sleep quality rather than changes in sleep duration. Reduced sleep quality was also associated with polygenic risk scores derived from genome-wide association studies for major depression, bipolar disorder and schizophrenia. Whilst the effects tended to be small, participants were not necessarily experiencing symptoms of the disorder when their sleep was assessed, since these were lifetime diagnoses.A secondary analysis considered 6 sleep properties self-reported at baseline assessment. Some associations were seen between these sleep properties and the inpatient psychiatric diagnoses and psychiatric polygenic risk scores, despite the different assessment methods and the baseline assessment having occurred 5 years earlier. Comparisons were also made between self-reported and accelerometry-derived sleep measurements, with associations seen between sleep duration, naps and wake up times. Self-reported chronotype was also associated both with accelerometry-derived bedtime and wake-up times.The initial analysis was undertaken on data from self-reported white participants, who are the majority of the participants in the UK biobank. However, a replication analysis of a cohort of 2692 self-reported non-white participants found similar associations. Males and females were also confirmed to have similar sleep alterations across lifetime psychopathologies.Whilst relatively high median sleep efficiency, low wake time after sleep onset, and long sleep bout durations were obtained in this study compared to prior polysomnography-based studies, it was nevertheless able to assess the normal sleep patterns of vast numbers of participants, using a potentially less disruptive measure. Such wearable devices could provide a scalable way to objectively measure sleep across many thousands of individuals undertaking their normal behaviours, facilitating studies investigating the impact of sleep on development of other disorders.