The discovery of the rapid antidepressant effects of the dissociativeanaesthetic ketamine, an uncompetitive N-Methyl-D-Aspartate receptorantagonist, is arguably the most important breakthrough in depressionresearch in the last 50 years. Ketamine remains an off-label treatmentfor treatment-resistant depression with factors that limit widespreaduse including its dissociative effects and abuse potential. Ketamineis a racemic mixture, composed of equal amounts of (S)-ketamine and(R)-ketamine. An (S)-ketamine nasal spray has been developed andapproved for use in treatment-resistant depression in the UnitedStates and Europe; however, some concerns regarding efficacy and sideeffects remain. Although (R)-ketamine is a less potentN-Methyl-D-Aspartate receptor antagonist than (S)-ketamine, increasingpreclinical evidence suggests (R)-ketamine may have more potent andlonger lasting antidepressant effects than (S)-ketamine, alongsidefewer side effects. Furthermore, a recent pilot trial of (R)-ketaminehas demonstrated rapid-acting and sustained antidepressant effects inindividuals with treatment-resistant depression. Research is ongoingto determine the specific cellular and molecular mechanisms underlyingthe antidepressant actions of ketamine and its component enantiomersin an effort to develop future rapid-acting antidepressants that lackundesirable effects. Here, we briefly review findings regarding theantidepressant effects of ketamine and its enantiomers beforeconsidering underlying mechanisms including N-Methyl-D-Aspartatereceptor antagonism, γ-aminobutyric acid-ergic interneuron inhibition,α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic receptor activation,brain-derived neurotrophic factor and tropomyosin kinase B signalling,mammalian target of rapamycin complex 1 and extracellularsignal-regulated kinase signalling, inhibition of glycogen synthasekinase-3 and inhibition of lateral habenula bursting, alongsidepotential roles of the monoaminergic and opioid receptor systems. There are significant limitations to current widely prescribed antidepressanttreatments. These include a significant delay in the onset of therapeuticaction (weeks to months) and approximately one-third of patients with majordepressive disorder (MDD) failing to demonstrate an adequate response .For indIncreasing evidence has revealed that the dissociative anaesthetic ketamine, anuncompetitive N-Methyl-D-Aspartate (NMDA) receptor antagonist, has thepotential to overcome such limitations, demonstrating rapid antidepressantand anti-suicidal effects, even in treatment-resistant patients . It has In this review, we summarise findings on the antidepressant effects of ketamineand its enantiomers. We then discuss underlying therapeutic mechanisms,exploring the case that ketamine’s enantiomers and metabolites may producecomplementary antidepressant effects via distinct mechanisms, beforeconsidering future directions of enquiry.Ketamine is a racemic mixture that consists of equal amounts of twoenantiomers, (S)-ketamine and (R)-ketamine (or esketamine and arketamine).(S)-ket(S)-ketamine and (R)-ketamine both undergo extensive metabolism by cytochromeP450 enzymes to corresponding forms of norketamine, dehydronorketamine(DHNK), hydroxyketamine (HK) and hydroxynorketamines (HNKs)  Figure . (S)-ket-ketamine: In the first double-blind, placebo-controlledstudy of racemic ketamine in MDD, it was demonstrated that a singlesub-anaesthetic intravenous (IV) infusion (0.5 mg/kg over 40 mins) resultedin rapid antidepressant effects, within hours of administration  and therefore may have been underpowered to detect a true differencebetween treatment groups. In addition, all patients were maintained on theirmedication regimes throughout the infusion phase and the impact thatconcomitant medications may have had on ketamine’s effects cannot be ruledout.Unfortunately, the antidepressant effect of a single dose of ketamine is notgenerally sustained beyond 1 week . In theAlthough there are specialist centres around the world and an increasing numberof ketamine clinics in the United States offering ketamine infusions fordepression , the use(S)-ketamine: As NMDA receptor antagonism was understood to play akey role in ketamine’s antidepressant mechanism, (S)-ketamine wasinvestigated as a novel antidepressant candidate by Janssen Research &Development due to its higher affinity for the NMDA receptor. In a firstproof-of-concept trial, IV (S)-ketamine at doses of 0.2 mg/kg and 0.4 mg/kgled to rapid and robust antidepressant effects in individuals with TRD(with TRD. Side efwith TRD.A fixed-dose (S)-ketamine nasal spray has subsequently been developed andtested in TRD. A number of Phase II and III trials have shown thatintranasal (S)-ketamine plus an existing or newly initiated oralantidepressant outperforms placebo plus an oral antidepressant forindividuals with TRD , althougSpravatoTM, for adults with TRD incombination with an oral antidepressant. However, some questions remainregarding uncertainty of efficacy, safety, potential for abuse and need forcareful monitoring, which currently limit wider use -ketaminenasal spray, ider use .(R)-ketamine: Preclinical findings have suggested (R)-ketamine hasthe potential for more potent and longer-lasting antidepressant effects thanboth ketamine and (S)-ketamine and it appears to have less behaviouralside-effects and abuse liability (iability . Giveniiability .Perception Neuroscience to further investigatesafety and tolerability of differing doses of (R)-ketamine in healthyvolunteers before exploring its potential in depression . In addition, a large trial comparing the efficacyand safety of (R)-ketamine with (S)-ketamine and -ketamine in TRD isalready underway in China (ChiCTR1800015879).In the first open-label pilot study of (R)-ketamine, seven subjects with TRDreceived a single IV infusion of (R)-ketamine at a dose of 0.5 mg/kg over40 mins . Mean MoIt has been widely acknowledged that the rapid antidepressant effects ofketamine are mediated through blockade of NMDA receptors located onγ-aminobutyric acid (GABA)-ergic inhibitory interneurons . This inIt has been reported that the metabolism of -ketamine to HNK, isrequired for ketamine’s antidepressant-like effects in rodents . Adminis(S)-ketamine is primarily metabolised to (S)-norketamine and it has beenshown in an animal model that although the antidepressant actions ofthe metabolite are similarly potent to the parent compound,(S)-norketamine appeared to lack the associated side effects . ImportaA receptors (GABA-NAMs) exert rapidantidepressant actions similar to -ketamine in animal models-ketamine at GABAergic interneurons issupported by findings that the NMDA receptor antagonist MK-801initially inhibits the firing of fast-spiking GABAergic interneuronsand, at a delayed rate, increases the firing rate of pyramidal neurons. Widman l models,likely l models.AlthougBDNF and its receptor TrkB have been consistently implicated in theaetiology of depression and mechanism of action of currentantidepressants . BDNF seIn the study by Considering the individual enantiomers, in a chronic social defeat stressand learned helplessness models of depression, a TRkB antagonist wasable to block the antidepressant effects of both (S)-ketamine  and (R)-The mammalian target of rapamycin complex 1 (mTORC1) and extracellularsignal-regulated kinase (ERK) are key signalling molecules in pathwaysthat regulate protein synthesis with roles in synaptic development andplasticity . ThefunFurther preclinical work demonstrated that the antidepressant effects of(S)-ketamine but not (R)-ketamine were blocked by mTORC1 inhibitionand that (S)-ketamine, but not (R)-ketamine, significantly attenuateddecreased phosphorylation of mTOR in the prefrontal cortex of mice ina chronic social defeat stress model . ThissaIn a recent randomised double-blind cross-over study, Activation of mTORC1 signalling has been linked to phosphorylation(deactivation) of glycogen synthase kinase-3 (GSK-3) and inhibition ofGSK-3 has been shown to be necessary for the rapid antidepressant-likeeffects of -ketamine in mice .Furtherd=2.27) than valproate(d=0.79), there was no significant differenceobserved between these two agents -ketamine infusions inindividuals with TRD . Inpatio agents .FurtherAlthough some evidence highlights GSK-3 as an important regulatory targetfor ketamine’s antidepressant effects, clinical studies have not yetconfirmed preclinical findings. Further evaluation of the role ofGSK-3 in the antidepressant effects of ketamine’s individualenantiomers and metabolites is still required.s is a subunit of the G protein GS thatstimulates the generation of cAMP by activating adenylyl cyclase.Localisation of Gαs within lipid raft microdomains in theplasma membrane acts to regulate cellular signalling, and indeedproduction of cAMP is diminished when Gαs is localised tolipid raft microdomains  thatacts to upregulate neurotrophic factors and increase synaptogenesishas been associated with conventional antidepressant action .Gαs is odomains . Anumberessants .s from lipid raft domains to non-raft domains andan increase in cAMP, followed by an increase in BDNF expression after24 hours. The -ketamine induced increase in cAMP was found topersist after knocking out the NMDA receptor indicating an NMDAreceptor-independent mechanism. Further, administration of theketamine metabolite -HNK also resulted in redistribution ofGαs from lipid rafts and an increase cAMP production.These findings suggest the translocation of Gαs from lipidrafts is a reliable hallmark of antidepressant action; however,further research is needed to examine to what degree this mechanismcontributes to the antidepressant effect of the individual enantiomersof ketamine.1a receptor antagonist blocks theantidepressant-like effects of -ketamine in mice and attenuatesketamine-induced increases in phosphorylation of Akt  signalling playsa role in the antidepressant effects of ketamine. Preclinical work hasdemonstrated that the antidepressant-like action of -ketamine isblocked by pre-treatment with a 5-HT-depleting agent .-kn of Akt . -kapamycin .Finallytagonist . Taken tmulation .Considering the individual enantiomers, an in vivo microdialysis studyhas shown that both (R)- and (S)-ketamine acutely increase 5-HTrelease in the prefrontal cortex, with (R)-ketamine causing a greaterincrease than (S)-ketamine . Althoug2/D3 receptor tracer binding (2/D3 receptor in the monkey striatum andsuggests that unlike (R)-ketamine, (S)-ketamine can cause dopaminerelease in the striatum that may contribute to thepsychotomimetic/dissociative side effects in humans -ketamine administrationis associated with significantly increased dopamine levels in thecortex, striatum and nucleus accumbens in rodents  and ther binding . A furthn humans . Other g release .In a study examining the effects of -ketamine and metabolites onevoked striatal dopamine release and dopamine receptors,-ketamine did not alter the magnitude or kinetics of electricalstimulation-evoked dopamine release in the nucleus accumbens ofanesthetised mice and neither ketamine’s enantiomers nor itsmetabolites had affinity for dopamine receptors or the dopaminetransporter . This su1receptor activity in the mPFC are necessary for the rapidantidepressant actions of -ketamine using optogenetic stimulationin a mouse model (1receptor-dependent induction of mTORC/ERK and inactivation of eEF2kinase resulting in increased protein synthesis (1receptor antagonist did not block the antidepressant effects of(R)-ketamine in a chronic social defeat stress model -ketamine infusion .WhetherKetamine interacts with mu, kappa and, to a lesser extent, delta-opioidreceptors  . TheaffIn rodent models of depression , it was shown that naltrexone pre-treatment didnot block the antidepressant effects of -ketamine . HoweverThere are several potential convergences between opioid signalling andother mechanisms implicated in the antidepressant action of ketamine.For example, administration of endogenous opioids has been shown toupregulate BDNF expression in the frontal cortex, hippocampus andamygdala . MoreoveThe role of the opioid system in the antidepressant effect of ketamineremains controversial and a topic of debate . FurtherSpravatoTM, has been approved in both theUnited States and Europe, although some concerns remain regarding efficacyand side effects. The first pilot study of (R)-ketamine in TRD hasdemonstrated encouraging results and, considering preclinical findings, itappears (R)-ketamine may have a more favourable safety profile than(S)-ketamine. Accumulating preclinical evidence also suggests (R)-ketamineto have more potent and longer-lasting antidepressant effects than both-ketamine and (S)-ketamine. As studies of (R)-ketamine progress throughPhase I and Phase II, results from direct comparison studies of the safetyand efficacy of (R)-ketamine and (S)-ketamine in TRD will be crucial. Otherkey outstanding questions are outlined in The discovery of the rapid antidepressant effects of -ketamine, includingin treatment-resistant patients, has appropriately been hailed ‘the mostimportant discovery in half a century’ in depression research . ThroughAlthough NMDA receptor inhibition and subsequent AMPA receptor activation havea role in the antidepressant effects of ketamine, further mechanistic workis building a more nuanced understanding of the distinct molecular andcellular mechanisms of ketamine, its enantiomers and metabolites, includingBDNF-TrkB, mTORC1 and ERK signalling. Although there may be a role formonoaminergic and opioid receptor systems in the antidepressant effects ordetrimental side effects of ketamine, further work examining the effects ofeach of the component enantiomers on these systems is required. All thewhile, new pieces of the ketamine puzzle are being discovered and otherpotential future directions of enquiry include examining the role of thetransforming growth factor β1 system  and theAs we further our understanding of the similarities and differences in thesignalling pathways associated with (S)-ketamine, (R)-ketamine and theirmetabolites, we should bear in mind potential complementary or synergisticantidepressant effects that might arise via distinct mechanisms. A deeperunderstanding of the precise molecular and cellular mechanisms underlyingthe antidepressant effects and negative side effects of -ketamine,(S)-ketamine and (R)-ketamine will be invaluable as we seek to developfuture rapid-acting antidepressants with favourable safety profiles,alongside treatment strategies to maintain adequate response.


The catecholamine norepinephrine (NE) links hindbrain metabolic-sensory neurons with key glucostatic control structures in the brain, including the ventromedial hypothalamic nucleus (VMN). In the brain, the glycogen reserve is maintained within the astrocyte cell compartment as an alternative energy source to blood-derived glucose. VMN astrocytes are direct targets for metabolic stimulus-driven noradrenergic signaling due to their adrenergic receptor expression (AR). The current review discusses recent affirmative evidence that neuro-metabolic stability in the VMN may be shaped by NE influence on astrocyte glycogen metabolism and glycogen-derived substrate fuel supply. Noradrenergic modulation of estrogen receptor (ER) control of VMN glycogen phosphorylase (GP) isoform expression supports the interaction of catecholamine and estradiol signals in shaping the physiological stimulus-specific control of astrocyte glycogen mobilization. Sex-dimorphic NE control of glycogen synthase and GP brain versus muscle type proteins may be due, in part, to the dissimilar noradrenergic governance of astrocyte AR and ER variant profiles in males versus females. Forthcoming advances in the understanding of the molecular mechanistic framework for catecholamine stimulus integration with other regulatory inputs to VMN astrocytes will undoubtedly reveal useful new molecular targets in each sex for glycogen mediated defense of neuronal metabolic equilibrium during neuro-glucopenia. The brain consumes a disproportionate fraction of bodily energy relative to its size in order to maintain neuron plasma membrane electrical stability and synaptic firing. Inadequate access to the principal substrate fuel glucose from the systemic circulation jeopardizes these vital brain cell functions, potentially leading to neurological dysfunction or nerve cell injury. The brain is continuously appraised of cellular metabolic equilibrium by dedicated interoceptive cells located within and outside the central nervous system. Input from these metabolic sensors is utilized by a distinctive neural glucostatic circuitry to regulate autonomic, neuroendocrine and behavioral outflow that maintains plasma glucose within a narrow physiological range. The final common control of these diverse functions derives from the hypothalamus, the principal autonomic motor center in the brain. The ventromedial hypothalamic nucleus (VMN), a critical component of the glucostatic network, functions to integrate multiple metabolic cues including nutrient profiles and indicators of peripheral (adipose tissue) energy stores to shape glucose counter-regulation. The complex carbohydrate glycogen is stored in brain astrocytes as a crucial metabolic fuel depot ,2,3. An Glucose homeostasis results from the stabilization of plasma glucose levels within the relatively limited range of approximately 3.9–8.3 mmol/L despite dynamic variations in glucose acquisition by dietary delivery/endogenous production versus metabolism . Plasma Neurons that exhibit electrical sensitivity to substrate fuel availability in vitro and in vivo reside in neuroanatomically distinct sensory , motor  and circumventricular (area postrema) divisions of the DVC ,8,12,13.Blood glucose gains primary access to the brain by constitutive glucose transporter (GLUT)-1 and GLUT-3 mediated uptake into astrocytes; specifically, end-feet processes of these glial cells, which function as an important component of the blood-brain barrier . In the In the brain, glycogen is maintained primarily within astrocytes ,28,29. G65/67 (GAD) protein content infer that the diminution of local glycogen disassembly may suppress the gluco-inhibitory neurotransmitter γ-aminobutyric acid (GABA) [Site-targeted delivery of the GP inhibitor 1,4-dideoxy-1,4-imino-d-Arabinitol (DAB) to the VMN augments local expression of neuronal nitric oxide synthase (nNOS), a marker protein for the gluco-stimulatory gaseous transmitter nitric oxide . Meanwhid (GABA) . In lighd (GABA)  by down-d (GABA) , outcome1 (α1), alpha2- (α2) and beta1- (β1) adrenergic receptor (AR) proteins [DVC NE governs hypoglycemic patterns of VMN AMPK activation and neurotransmitter marker protein expression . Noradreproteins , it is rproteins . In an eproteins . OutcomeFurther work utilized the selective catecholamine neurotoxin 6-hydroxydopamine (6OHDA) to address the role of noradrenergic transmission in the hindbrain lactoprivic regulation of the VMN GP variant expression . ResultsIt should be noted that outcomes documenting lactoprivic augmentation of the gluco-inhibitory transmitter GABA likely reflect, to some extent, differences associated with this experimental model  compared with systemic insulin-induced hypoglycemia (which is characterized by body-wide sensor activation). It is possible that under the latter circumstances, noradrenergic stimulation of GAD expression may be attenuated by a neurotransmitter cue(s), as-yet-unidentified, that is provided to the VMN by non-hindbrain energy sensors. Alternatively, up-regulated GABA transmission may be a signal of glycogen conservation due to negligible disassembly caused by down-regulated expression of NE-sensitive GPbb, alongside an absence of metabolic deficit activation of GPbb activity. These findings are consistent with earlier reports that NE depletion reduces glycogenolysis during energy imbalance .In light of proof that ample tissue glycogen levels are maintained in the brainstem , it was The seminal finding that variation in the noradrenergic signal volume, governed by the hindbrain metabolic state, imposes a differential regulation of VMN glycogen metabolic enzyme expression prompts a consideration of molecular mechanisms that may control loss-or-gain or shift in direction of NE effects on these protein targets. In recent work, we used a hypothalamic astrocyte primary cell culture model  to addre1- and β2-AR protein profiles exhibited bi-directional responses to increasing NE doses; female astrocytes exhibited diminished β1-AR content at low dose exposure but enhanced β2-AR expression at high NE dosages. Thus, in each sex, graded variations in noradrenergic stimulation may modulate astrocyte receptivity to NE in vivo albeit with different effects at specific signal strengths. Estradiol imposes sex-dimorphic control of hypothalamic astrocyte AMPK activity and glycogen mass through the regulation of the ER variant expression [Sex differences in the noradrenergic regulation of glia AR and ER expression involved dosage efficacy and direction of regulatory action. In male rats, βpression . Outcomepression ,61. Currpression  and regupression , reinforHypothalamic astrocytes express AMPK and require catecholaminergic input for activation of this sensor during hypoglycemia . In maleResearch discussed here bolsters growing recognition that metabolic signal-initiated patterns of hindbrain noradrenergic input impact VMN neuro-metabolic stability. VMN glycogen mass and/or turnover is a plausibly important metabolic variable that is subject to careful monitoring. As glycogen-derived substrate fuel stream has documented influence on VMN metabolic regulatory transmitter signaling, as indicated by evidence that NE causes 4CIN-reversible changes in the VMN nNOS and GAD protein expression , the astNovel evidence for intrinsic sex differences in hypothalamic astrocyte responses to NE exposure in vitro provides ample justification for research efforts going forward to focus on whether intrinsic functional dimorphism in reactivity to this neurotransmitter occurs in vivo and, if so, potential causal mechanisms.2-AR protein expression, cAMP signaling and glycogenolysis in vitro but does not coincidently modify lactate release [NE stimulates astrocyte synthesis of the chemokine CCL2 ,68. This release . Thus, mEarly reports that NE exposure in vitro only depletes a sub-fraction of cortical astrocyte glycogen content advanced the concept of stimulus-specific control of glycogen metabolism . Our obs


Atherosclerosis (AS) is widely accepted to be a multistep pathophysiological process associated with several other processes such as angiogenesis and inflammatory response. Long non-coding RNAs (lncRNAs) are non-protein coding RNAs (more than 200 nucleotides in length) and can regulate gene expression at the transcriptional and post-transcriptional levels. Recent studies suggest that lncRNA-H19 plays important roles in the regulation of angiogenesis, adipocyte differentiation, lipid metabolism, inflammatory response, cellular proliferation and apoptosis. In this review, we primarily discuss the roles of lncRNA-H19 in atherosclerosis-related pathophysiological processes and the potential mechanisms by which lncRNA-H19 regulates the development of atherosclerosis, to help provide a better understanding of the biological functions of lncRNA-H19 in atherosclerosis. Atherosclerosis (AS) drives cardiovascular disease, which is one of the main causes of mortality in the world  are widely classified as transcripts > 200 nucleotides with limited coding potential  by increasing p53 activity  gene. The H19 gene is 2.3 kb in length and is highly evolutionarily conserved, suggesting that it may have some important biological functions  growth and capillary-like structure formation  gene  gene, thus increasing VASH1 expression and secretion in HAMSCs, and suppressing angiogenesis  is considered as one of the crucial factors associated with angiogenesis , brown adipose tissue (BAT) and perivascular adipose tissue (PVAT) play differential roles in atherosclerosis. WAT, acting as a lipid sink, prevents the accumulation of lipids in circulation but active BAT helps combust lipids. PVAT has properties of both WAT and BAT. Evidence suggests a critical role for PVAT in regulating the focal inflammatory state and vessel homeostasis via pro-atherogenic or anti-atherogenic mechanisms such as releasing inflammatory cytokines and adipokines, depending on the PVAT state in health and disease  are precursor cells of adipocytes  is strongly associated with atherosclerosis  was first discovered in 1986  signaling is a vital regulator of NF-κB-mediated inflammatory responses  and increases the expression of anti-inflammatory factors (IL-4 and IL-10)  expression by downregulating microRNA let-7. IL-6, which is a pro-inflammatory cytokine, activates NF-κB, thereby completing a positive feedback loop of the NF-κB/let-7/IL-6 pathway , which is a critical regulator of vasoconstriction and proliferation of arteries. Through the lncRNA-H19/let-7b/AT1R axis, lncRNA-H19 also promotes the proliferation of VSMCs  proliferation in a miR-675-dependent manner. They identified that phosphatase and tensin homology deleted on chromosome ten (PTEN) is a target of miR-675 and PTEN is a well-known tumor suppressor that mediates VSMC proliferation. Hence, a lncRNA-H19/miR-675/PTEN axis was uncovered, through which lncRNA-H19 promotes the proliferation of VSMCs  induces proliferation of VSMCs. It has been reported that Hcy treatment results in the demethylation of differentially methylated regions between the Atherosclerosis is the major cause of coronary artery disease, leading to myocardial ischemia and infarction  patients, is positively correlated with cardiac biomarkers, such as creatine kinase (CK), suggesting a potential role for lncRNA-H19 in AMI diagnosis   What influences lncRNA-H19 levels in the onset, development and progression of atherosclerosis? (ii) How does lncRNA-H19 strictly regulate multiple targets in vitro and in vivo? (iii) Are there any other lncRNA-H19 targets that can increase the risk of atherosclerosis? (iv) Does lncRNA-H19 influence the process of atherosclerosis by targeting different pathways in various cell types and tissues? (v) Why and when lncRNA-H19 may serve pro-apoptotic vs. anti-apoptotic roles and pro-survival vs. deleterious roles in I/R injury? Note that there are still some technical challenges that make future lncRNA-H19-based therapeutics difficult. Importantly, the potential significance of lncRNA-H19 requires careful studies that complement the development of reliable strategies to specifically target different genes and proteins in atherosclerosis-related cells. Understanding the molecular mechanisms and cellular pathways controlled by lncRNA-H19 will be warranted.


The long-term pandemic of coronavirus disease 2019 (COVID-19) requires sensitive and accurate diagnostic assays to detect severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus and SARS-CoV-2 antibodies in infected individuals. Currently, RNA of SARS-CoV-2 virus is mainly detected by reverse transcription-polymerase chain reaction (RT-PCR)-based nucleic acid assays, while SARS-CoV-2 antigen and antibody are identified by immunological assays. Both nucleic acid assays and immunological assays rely on the luminescence signals of specific luminescence probes for qualitative and quantitative detection. The exploration of novel luminescence probes will play a crucial role in improving the detection sensitivity of the assays. As innate probes, aggregation-induced emission (AIE) luminogens (AIEgens) exhibit negligible luminescence in the free state but enhanced luminescence in the aggregated or restricted states. Moreover, AIEgen-based nanoparticles (AIE dots) offer efficient luminescence, good biocompatibility and water solubility, and superior photostability. Both AIEgens and AIE dots have been widely used for high-performance detection of biomolecules and small molecules, chemical/biological imaging, and medical therapeutics. In this review, the availability of AIEgens and AIE dots in nucleic acid assays and immunological assays are enumerated and discussed. By building a bridge between AIE materials and COVID-19, we hope to inspire researchers to use AIE materials as a powerful weapon against COVID-19. The patRS-CoV-2 . DespiteCurrent detection methods can be divided into two categories: nucleic acid assays and immunological assays –12. NuclAggregation-induced emission (AIE), first proposed by Tang in 2001, is a diametrically opposite phenomenon to ACQ . The lumSince the number of viruses reach the maximum in the first few days of infection, nucleic acid assays can provide the earliest information to determine the presence or absence of the SARS-CoV-2 virus . Althoug1, 3, and 5 as examples  and lateral flow immunoassays.3 to 2.5 × 106 copies/μL  can specifically bind the SARS-CoV-2 viral protein to form antibody–protein complex, which can be detected by an additional tracer antibody. This assay is very fast and results can be obtained within 1–5 h. Various AIEgens with switchable luminescence have been successfully used in ELISA –58, and opies/μL . AdditioE. coli O157:H7 monoclonal antibody, forming the fluorescent immunoprobe (AIEFM-mAb)  was immobilized on the test line (T) of the nitrocellulose membrane while goat anti-mouse antibody (sAb) was immobilized on the control line (C). The target E. coli O157:H7 (if present) can be specifically captured by mixing with AIEFM-mAb to generate bright fluorescence on both the test line and control line. Otherwise, AIEFM-mAb can only be captured by sAb on the control line to emit bright fluorescence. The advantage of using AIE-active TCBPE is that the higher the loading of TCBPE, the stronger the fluorescence signal and the higher the detection sensitivity. As a result, the AIEFM endows the lateral flow immunosensor with a very low detection limit, down to 3.98 × 103 CFU/mL, which is about 10 times lower than that of the two commercial FMs  . Anti-E.cial FMs . Undoubtcial FMs , 63. The2000), poly(ethylene glycol)-b-polystyrene (PEG-b-PS), 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[folate(polyethylene glycol)-5000] (DSPE-PEG5000-folate), etc.), bovine serum albumin (BSA), and biocompatible surfactants . It is worth noting that the internal dispersion of AIEgens throughout the amphiphilic matrix may be stochastic and vary from molecular to aggregate distributions when preparing and using the composite AIE dots. Compared to the pure AIE dots, the composite AIE dots preserve the properties of both materials to offer a number of unique advantages: (1) synthetic efforts are minimized by reducing the hydrophilic groups; (2) luminescence performance are well-tuned by mixing various AIEgens in different concentrations; and (3) high specificity and responsiveness are achieved by using the amphiphilic matrices to conjugate with diverse linkers.To facilitate the understanding and utilization of AIE dots, current AIE dots are divide into two categories: pure AIE dots and composite AIE dots –25. The Global health is facing the most dangerous situation regarding the COVID-19 pandemic. Owing to the large number of infected cases worldwide, the rapid long-term spread, and lack of effective treatments, there is a continuing need for more sensitive, low-cost, rapid, and point-of-care diagnostic kits to identify cases of SARS-CoV-2 infection in communities and individuals without advanced facilities. To date, while RT-PCR assays have become the primary technique for detection of SARS-CoV-2 RNA, other nucleic acid assays including isothermal amplification assays, hybridization microarray assays, amplicon-based metagenomics sequencing, and the cutting-edge CRISPR-related technologies have also been developed . ChallenIn addition, there are some issues that need further improvement when AIE materials are used to COVID-19 detection: (1) development of the sequence-specific DNA probes labeled with AIE dots and quenchers; (2) promotion of AIE dots-based lateral flow immunoassay kits for COVID-19 clinical testing; (3) diversification of AIEgens and AIE dots that can be directly labeled with highly targeted molecules ; and (4) further attempts to use AIE materials for studying immune mechanisms and pharmacological treatment of COVID-19 infection . With thAll authors listed have made a substantial, direct and intellectual contribution to the work, and approved it for publication.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


We aimed to analyse clinical characteristics and find potential factors predicting poor prognosis in patients with coronavirus disease 2019 (COVID-19).RNA− or URNA+).We analyzed the demographic and clinical data of COVID-19 patients and detected SARS-CoV-2 RNA in urine sediments collected from 53 COVID-19 patients enrolled in Renmin Hospital of Wuhan University from January 31, 2020 to February 18, 2020 with qRT-PCR analysis, and then classified those patients based on clinical conditions (severe or non-severe syndrome) and urinary SARS-CoV-2 RNA  showed significantly higher positive rate  of urinary SARS-CoV-2 RNA than non-severe patients . UThis study indicated that urinary SARS-CoV-2 RNA detected in urine specimens can be used to predict the progression and prognosis of COVID-19 severity. COVID-19 is a highly contagious disease caused by a newly emerging coronavirus called severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), which belongs to the β-coronavirus cluster that comprises 7 members, including SARS and Middle East Respiratory Syndrome (MERS) .The main methods used for screening and diagnosis of SARS-CoV-2 infection include the detection of SARS-CoV-2 nucleic acid and SARS-CoV-2-specific antibody. The test for SARS-CoV-2 nucleic acid in nasopharyngeal swab specimens with reverse transcription-polymerase chain reaction (RT-PCR) was almost the only pathogen detection method used at the early stage of COVID-19 outbreak in Wuhan, China, and later this test was applied to other body fluid samples, such as anal swab, stool, blood and urine, to avoid false negative results. It has been demonstrated that the positive rates of SARS-CoV-2 nucleic acid in different body fluids are variable, indicating a distinct pattern of persistence and clearance of viral RNA in body fluids in COVID-19 patients -6. The pRNA−, 38 cases) and positive group . We then conducted a retrospective study on those patients’ clinical characteristics, basic diseases and laboratory tests (th edition) published by the National Health Commission of China [From January 31, 2020 to February 18, 2020, a total of 53 patients who were diagnosed with COVID-19 at Renmin Hospital of Wuhan University were tested for SARS-CoV-2 nucleic acid in urine samples with quantitative reverse transcription-polymerase chain reaction (qRT-PCR) analysis. To reduce false negative results, we collected the urine sediment samples from those patients at the admission day case by case. Based on the results of urine SARS-CoV-2 nucleic acid testing, we divided those patients into two groups, including the urinary SARS-CoV-2 negative group (Ury tests . The diaof China . Our stu2)/fraction of inspired oxygen (FiO2) ratio is less than 300 mmHg. We identified 30 cases as non-severe patients and 23 as severe patients  that was examined and confirmed in a designated pathology laboratory.Immunohistochemical staining was performed on kidney specimens from autopsy for thrombomodulin (TM), and von Willebrand factor (vWF) as previously described . Brieflyt-tests was used when the data were normally distributed, otherwise, wilcoxon rank-sum test was used. Chi-square tests and Fisher’s exact tests were applied to categorical variables as appropriate. The cumulative rate of in-hospital survival was investigated using the Kaplan-Meier method. All statistical analyses were performed using SPSS 22.0. p < 0.05 was considered as statistically significant.Continuous variables were expressed using the mean ± standard deviation  or medians and interquartile (IQR) values as appropriate . Categorical variables were shown as the percentages and counts. Two-independent group RNA−) and 15 of them were urinary SARS-CoV-2 positive (URNA+). The URNA+ patients were older and more likely to experience chest tightness and shortness of breath than URNA− patients, but showed no significant differences in male/female distribution, fever, cough, sputum production, fatigue, radiological appearance, hypertension, diabetes, cardiovascular diseases, chronic renal disease  and oxygen saturation (SaO2) than URNA− patients as examined with arterial blood gas analysis . Immune profile evaluation identified a more frequent increase of serum CRP and IgE in URNA+ patients . In addition, we found that URNA+ patients had higher prevalence of increased serum levels of ALT , higher percentage of decreased serum AST , higher case percentage of increased serum myoglobin, ultra-TnI , LDH , urea , and decreased eGFR  than URNA− patients. These data indicated that URNA+ patients had more severe lesions on organs of liver, heart, and kidney. We further found that URNA+ patients showed significantly lower levels of T cells and T helper (Th) cells  in peripheral blood mononuclear cells, and higher levels of serum CRP , ALT , but lower serum AST , and higher levels of DBIL , LDH , urea , and significantly lower eGFR  than URNA− patients. These results suggested that URNA+ patients might develop severe clinical manifestations characterized by the increase of the levels of inflammatory markers and tissue injury-related enzymes.A total of 53 hospitalized COVID-19 patients were enrolled in this study. The characteristics of those patients were detailed in  disease . In addianalysis . The leu2)/fraction of inspired oxygen (FiO2) ratio (less than 300 mmHg). Within the 23 severe patients, we found that 12 of those 23 server patients were urinary SARS-CoV-2 negative (SURNA−) and 11 of them were URNA+ (SURNA+). The positive rate of urine SARS-CoV-2 RNA were significantly higher in sever patients (11/23 severe patients = 47.8%) than that in non-severe patients (4/30 non-severe patients = 13.3%) . This result suggested that urine shedding SARS-CoV-2 correlated with the severity of the disease. To support this notion, we found that SURNA+ patients experienced more comorbidities, including higher prevalence of hypertension and cardiovascular diseases , and renal function impairment . SURNA+ patients also showed significantly lower levers of eGFR  but higher levels of IgE and IgG  than SURNA− patients. In addition, the URNA+ patients among 53 patients had a significantly higher risk of death than the URNA− patients . Furthermore, the severe patients with URNA+ also demonstrated a higher risk of death than the severe patients with URNA−, although the difference did not reach statistical significance between URNA− and URNA+ groups of severe patients . These results suggested that URNA+ patients exhibited significantly higher risk of cardiovascular abnormality, liver injury, renal disfunction and death, and urinary SARS-CoV-2 RNA might serve as a predictive biomarker for determining the prognosis and outcome of the disease.Above results implied a correlation of the urinary SARS-CoV-2 RNA with COVID-19 disease severity and the underlying conditions in COVID-19 patients. We hypothesized that urinary SARS-CoV-2 RNA may serve as a biomarker for predicting the clinical outcomes of severe COVID-19 patients. In the total of 53 patients, we identified 30 non-severe patients and 23 severe patients  based onEndothelial dysfunction is prevalent in chronical cardiovascular disease and infectious or inflammatory diseases (such as that in virus-infected patients) -26. TM aSARS-CoV-2 mainly infects host through respiratory tract and spread to other organs or tissues through blood circulation. Damaged vascular endothelial, which either preexisted or incurred by virus infection, allows virus infiltration between blood stream and tissue mesenchyme by passing through endothelium. Virus-related renal insult and urine shedding are determined by multifactor, such as the virus load of inflow, the vascular endothelial integrity, and the intensity of anti-virus response. In this study, we adopt a model of renal Inflow-Infiltration-Injury-Into urine (“4I” model) to illustrate the course and outcome of SARS-CoV-2 infection in COVID-19 patient kidney . In this model, the kidney is simplified as an effector to response to the virus invasion (input) with the effects of renal damages and virus shedding (output). Output effects are determined by the input strength (amount of loading virus) and the intrinsic nature of the kidney, such as the integrity of vascular endothelium and local immunity . Although the compromised integrity of glomeruli and interstitial vascular vessels both allow the virus infiltration into kidney parenchyma, the infiltrating route through interstitial vessels is ignored, since the exuded virus is unlikely to infect tubular epithelial cells in which the ACE2 enriched in the brush border . In thatWe conducted a comprehensive study on the clinical characteristics, basic diseases and laboratory tests in a cohort of 53 COVID-19 patients, where 30 patients had non-severe symptoms, and 23 patients suffered from severe symptoms. Our results suggested an intriguing association of the urinary SARS-CoV-2 RNA with the clinical manifestation of COVID-19, pointing to a high viral load, new or preexisting vascular endothelial damage in severe COVID-19 patients. This retrospective study indicated for the first time that the positive of SARS-CoV-2 RNA in urine specimens maybe used to predict the progression and prognosis of COVID-19.RNA+)  . Our anaRNA+)  [5, 7, 8RNA+ patients or severe URNA+ subgroup showed higher prevalence of inflammation and immune discord, cardiovascular diseases, liver damage and renal disfunction, and higher risk of death than URNA− patients. The reason for the observed higher prevalence of developing severe clinical manifestations and higher risk of death in URNA+ patients might be caused by a high SARS-CoV-2 viral load, which was shown to to be highly associated with in-hospital mortality in COVID-19 patients [We also observed that Upatients . The newStudies on cellular mechanism have confirmed that SARS-CoV-2 shares the same membrane-bound angiotensinconverting enzyme 2 (ACE2) as SARS-CoV to gain access to its target cells -32. In pOur findings support a significant role of vascular endothelial lesion in COVID-19 patients in the development of renal damage and urine shedding. To further illustrate the course and outcome of SARS-CoV-2 infection in COVID-19 patient kidneys, we raised a model of renal Inflow-Infiltration-Injury-Into urine (“4I” model) . This model may help us to understand the role and function of kidneys in SARS-CoV-2 infection. Kidneys are simplified as an effector to response to the virus invasion (input) with the effects of renal damages and virus shedding (output). SARS-CoV-2-related renal insult and urine shedding may determine by multifactor, including the virus load of inflow, the vascular endothelial integrity, as well as the intensity of anti-virus response. Our analysis on the clinical features of the cluster showed that the positive test rate of urine SARS-CoV-2 RNA is remarkably higher in the severe patients than in non-severe patients, and renal vascular endothelial lesion associate with severity of COVID-19 patients. Based on these finding, we propose a function equation and curve to solve the urinary excretion of viral nucleic acid, condition of vascular endothelial integrity and the circulating viral load . The function equation and curve may provide another strategy to calculate and predict the progression and prognosis of the disease.RNA+ in the disease progression of COVID-19. In addition, due to the lacking of clinical data of patients after discharge, we could not assess the association of urinary SARS-CoV-2 RNA with disease recovery. The precise relationship between urinary SARS-CoV-2 RNA and endothelial dysfunction and multiple organ disfunction in these patients require further investigation. Furthermore, our hypothesis about function equation and curve to solve the urinary excretion of viral nucleic acid, condition of vascular endothelial integrity and the circulating viral load should be tested in future studies.However, due to the limited sample size, our model might not be powered sufficiently to reflect the overall complexity of the general population. Therefore, large-scale prospective cohort studies are required in ethnically and geographically diverse cohorts to better understand the association and importance of UWe optimized the method of urine SARS-CoV-2 RNA detection and significantly increased the positive detection rates. We analyzed the clinical characteristics of patients with urinary nucleic acid positive SARS-CoV-2 RNA, and revealed a potential association of vascular endothelial damage with virus urine shedding. Furthermore, we established a model to analyze the relationship between virus urine excretion and the underlying disease condition. This study suggests that the detection of SARS-CoV-2 RNA in urine sediments can provide a robust biomarker for evaluation and prognosis for patients with COVID-19.Supplement


Sepsis is a life-threatening condition that is characterized by multiple organ dysfunction due to abnormal host response to various pathogens, like bacteria, fungi and virus. The differences between viral and bacterial sepsis are indeed of great significance to deepen the understanding of the pathogenesis of sepsis, especially under pandemics of SARS-CoV-2 infection. Dear editor,The pandemics of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection have become a world health crisis that cause significant loss of life. The dysregulated host response to SARS-CoV-2 appears to be associated with the severity and poor outcomes of coronavirus disease 2019 (COVID-19) patients . The devIn this study, we obtained the clinical data from two cohorts up to May 13, 2020, including 41 critically ill COVID-19 patients from the Third People’s Hospital of Shenzhen and 194 non-COVID-19 patients admitted to ICU of the Second People’s Hospital of Shenzhen, China. Demographic characteristics, comorbidities, and laboratory findings on ICU admission and clinical outcomes were collected. Sequential Organ Failure Assessment (SOFA) and Acute Physiology and Chronic Health Evaluation II (APACHE II) scores were calculated within the first 24 h since ICU admission. The bacterial and viral sepsis were identified by blood culture and metagenomic next-generation sequencing.P = 0.01] and APACHE II  were consistently higher among patients with bacterial sepsis than those with SARS-CoV-2-induced sepsis. Meanwhile, ICU mortality rates were significantly higher in patients with bacterial sepsis than those with viral sepsis .Twenty-one patients with SARS-CoV-2-induced sepsis and 46 patients with bacterial sepsis were finally recruited (Additional file As presented in Table In this study, ICU patients with SARS-CoV-2-induced sepsis and those with bacterial sepsis revealed comparable demographic characteristics, like age, gender distribution, and comorbidities, after rigorous screening processes. However, patients with bacterial sepsis were found with more severe organ dysfunction and poor outcomes when compared with those caused by SARS-CoV-2-induced sepsis, including higher values in SOFA and APACHE II, as well as more ICU deaths. The different patterns of immune responses might be the major cause of the divergent outcomes between viral and bacterial sepsis. We further found that failed homeostasis was characterized in both bacterial and viral sepsis but triggered by different pathogens. In the development of viral sepsis, the loss of T lymphocytes and their subsets was the dominant characteristics of dysregulated immune response, thereby contributing to the imbalance between innate and adaptive immune systems; while excessive inflammatory activation was the main feature of bacterial sepsis, which further resulted in intractable immune suppression and multiple organ dysfunction. This is the first report that compared clinical features and host responses between bacterial and SARS-CoV-2-induced viral sepsis. These findings might not only suggest divergent host responses to bacteria and virus but also provide novel insights into further researches on the development of sepsis with underlying etiology of various pathogens.Additional file 1. Appendix Table 1. Baseline characteristics of critically ill patients with SARS-CoV-2- and bacteria-induced sepsis.Additional file 2. Appendix Figure 1. Flow diagram of patient inclusion. COVID-19: Coronavirus disease 2019; SARS-CoV-2: Severe acute respiratory syndrome coronavirus 2; SOFA: Sequential organ failure assessment; ICU: Intensive care unit; LOS: Length of stay; CAP: Community-acquired pneumonia.


With the increasing demand of achieving comprehensive perception in every aspect of life, optical fibers have shown great potential in various applications due to their highly-sensitive, highly-integrated, flexible and real-time sensing capabilities. Among various sensing mechanisms, plasmonics based fiber-optic sensors provide remarkable sensitivity benefiting from their outstanding plasmon–matter interaction. Therefore, surface plasmon resonance (SPR) and localized SPR (LSPR)-based hybrid fiber-optic sensors have captured intensive research attention. Conventionally, SPR- or LSPR-based hybrid fiber-optic sensors rely on the resonant electron oscillations of thin metallic films or metallic nanoparticles functionalized on fiber surfaces. Coupled with the new advances in functional nanomaterials as well as fiber structure design and fabrication in recent years, new solutions continue to emerge to further improve the fiber-optic plasmonic sensors’ performances in terms of sensitivity, specificity and biocompatibility. For instance, 2D materials like graphene can enhance the surface plasmon intensity at the metallic film surface due to the plasmon–matter interaction. Two-dimensional (2D) morphology of transition metal oxides can be doped with abundant free electrons to facilitate intrinsic plasmonics in visible or near-infrared frequencies, realizing exceptional field confinement and high sensitivity detection of analyte molecules. Gold nanoparticles capped with macrocyclic supramolecules show excellent selectivity to target biomolecules and ultralow limits of detection. Moreover, specially designed microstructured optical fibers are able to achieve high birefringence that can suppress the output inaccuracy induced by polarization crosstalk and meanwhile deliver promising sensitivity. This review aims to reveal and explore the frontiers of such hybrid plasmonic fiber-optic platforms in various sensing applications. In addition, macrocyclic supramolecules have been recently proven to be excellent surface functionalization candidates for metallic nanoparticles, contributing to a simple functionalization process, selective target molecules recognition and improved biocompatibility.Plasmonic fiber-optic sensors have captured intensive research attention in recent years due to their high degree of integration, high sensitivity, flexibility and remote-sensing capability ,2,3. Fib3 and achieve highly integrated microfiber based plasmonic sensing in NIR optical frequencies; how to synthesize cyclodextrin-capped gold nanoparticles (AuNPs) in a one-step process and realize microfiber based highly selective detection of cholesterol in human serum, etc. will be demonstrated.In this review, the background and the state-of-the-art of SPR/LSPR fiber-optic sensors will be reviewed. More importantly, the aforementioned emerging hybrid fiber-optic plasmonic sensing solutions will be illustrated in detail. For instance, the exploration of how the highly birefringent MOF based SPR sensor can suppress polarization crosstalk and improve sensitivity in the meantime; how to integrate graphene-on-gold hybrid structure on the fiber-optic platform to strengthen the surface plasmon intensity and to effectively adsorb biomolecules; how to dope abundant free electrons in 2D MoOω, c and ε are the frequency of the TM-polarized incident light, light velocity and dielectric constant, independently. The propagation constant of the evanescent wave at the interface is:θ is the angle of light incidence. SPR can be excited by the TM-polarized total reflected light at the silica-metal interface when the phase matches, that the propagation constant of reflected light equals to the propagation constant of SPP:A surface plasmon polariton (SPP) is an electromagnetic wave that propagates in parallel at the interface between the metal film and dielectric medium. The SPP is TM-polarized. As illustrated in β of the right expression denotes the effects of finite metal layer thickness and high refractive index of prism in real situation. In most fiber-optic SPR sensors, the wavelength interrogation method is employed. The refractive index sensitivity is defined as: sδn is the change in the refractive index of the analyte and δresλ is the shift of resonant wavelength .,81.80,81de model :(6)εmetafound in . Since Lmetalε’ = –2dielectricε. The LSPR resonant frequency is thereby expressed as:Based on Equation (5), the resonance is satisfied  when dielectricε = 2dielectricn. Therefore, the refractive index of the ambient dielectric medium of the nanoparticle impacts the LSPR resonant wavelength:Furthermore, for dielectric medium, Similar to SPR-based fiber-optic sensing platforms, LSPR-based fiber-optic devices have also captured intensive research attention ,87,88,894, NaOH have to be introduced, which violates the purpose of achieving biocompatibility in many LSPR biosensing applications [Benefiting from their macrocyclic cavities, macrocyclic supramolecules like cyclodextrins (CDs), cucurbiturils, pillararenes, calixarenes, etc. show excellent molecular recognition capability by the host-guest interaction. The host–guest interaction is noncovalent interaction between macrocyclic supramolecules and corresponding guest molecules to form inclusion complexations . Encouraications ,106,107.ications , where C1H NMR) and Fourier transform infrared (FTIR) spectra are performed to further illustrate that the hydroxyl groups in β-CDs mainly contribute to reducing Au3+ ions to Au0 atoms [The as-synthesized β-CD-capped AuNPs show good uniformity in particle size with diameters range from ~18 nm to ~21 nm. The dynamic light scattering (DLS) characterization of the AuNPs further verifies the observation . The resu0 atoms .The as-synthesized AuNPs are negatively charged. Hence it can be stably immobilized on positively charged microfiber surface  via electrostatic attraction. As shown in The selectivity of the proposed biosensor to cholesterol is validated by an interference study, where common interfering substances in human serum such as glutamic acid, cysteine, ascorbic acid, dopamine and human serum albumin (HSA) are introduced. Another polysaccharide, chitosan, has also been used for AuNPs synthesis as a reducing and stabilizing agent . Sadani The sensitivity and selectivity of four sensors: BSA attaching to citrate capped AuNPs (BSA on GnP), polyanionic poly(sodium 4-styrenesulfonate) (PSS) immobilized ChGnP (ChGnP on PSS), fluorescent BSA-Au nanoclusters (BSA-AuNC) and BSA immobilized ChGnP (ChGnP on BSA) are compared. As shown in Lee et al. fabricate a fiber-optic LSPR sensor for the detection of ochratoxin A (OTA) utilizing aptamer-modified gold nanorods (GNRs) . The GNRAs discussed in this review, the proper design of MOFs with high birefringence provides wide possibilities in highly integrated microfluidic sensing devices with improved measurement accuracy and stability. Profiting from 2D material-based hybrid plasmonic structures, fiber-optic plasmonic sensors can deliver more promising sensing capability. The exceptional surface to volume ratio, near-field confinement and in situ plasmonic properties tunability of 2D materials facilitate the further enhancement of plasmon-matter interaction so as to enhance the sensitivity and LOD. In addition, the development of supramolecular chemistry brings new solutions in LSPR nanoparticles surface functionalization, leading to excellent target molecule selectivity via host–guest interaction. Given the numerous possibilities in optical design and hybrid plasmonic architectures construction, fiber-optic hybrid plasmonic sensors possess vast potential in various sensing scenarios with distinct advantages of high sensitivity, flexibility, miniaturization and high degree of integration. To further integrate the advances of specialty fibers and various nanomaterials, one major direction in the future is to achieve advanced functional fiber-based sensing by integrating multi-functional materials inside a single fiber or large-scale fabrics ,134,135.


Presurgical motor mapping of cortical muscle representations by single-pulse navigated transcranial magnetic stimulation (sp-nTMS) is used to localize and enclose motor function in patients harboring motor-eloquent lesions. This study investigates paired-pulse nTMS (pp-nTMS) with biphasic pulses for motor mapping of lower extremity (lE) muscle representations, showing that significantly lower stimulation intensities are needed while reliable motor maps can be achieved even in the most demanding patient cases in whom conventionally used sp-nTMS fails. Thus, novel pp-nTMS with biphasic pulses may have potential to considerably facilitate improvements of currently used nTMS-based mapping procedures in the preoperative workup of neurooncological patients.p < 0.0001), and it provided reliable motor maps even in the most demanding cases where sp-nTMS failed . Fiber volumes of the tracked CST were slightly higher when motor maps of pp-nTMS were used, and CST tracking using pp-nTMS data was also possible in the five patients in whom sp-nTMS failed. In conclusion, application of pp-nTMS with biphasic pulses enables preoperative motor mapping of lE muscle representations even in the most challenging patients in whom the motor system is at high risk due to lesion location or resection.Navigated transcranial magnetic stimulation (nTMS) is increasingly used for mapping of motor function prior to surgery in patients harboring motor-eloquent brain lesions. To date, single-pulse nTMS (sp-nTMS) has been predominantly used for this purpose, but novel paired-pulse nTMS (pp-nTMS) with biphasic pulse application has been made available recently. The purpose of this study was to systematically evaluate pp-nTMS with biphasic pulses in comparison to conventionally used sp-nTMS for preoperative motor mapping of lower extremity (lE) muscle representations. Thirty-nine patients  harboring motor-eloquent brain lesions of different entity underwent motor mapping of lE muscle representations in lesion-affected hemispheres and nTMS-based tractography of the corticospinal tract (CST) using data from sp-nTMS and pp-nTMS with biphasic pulses, respectively. Compared to sp-nTMS, pp-nTMS enabled motor mapping with lower stimulation intensities (61.8 ± 13.8% versus 50.7 ± 11.6% of maximum stimulator output,  Navigated transcranial magnetic stimulation (nTMS) has developed into a reliable preoperative tool to map the motor cortex in patients harboring motor-eloquent brain lesions ,2,3,4. SUtility of motor maps and CST tractography for the purpose of valid preoperative surgery planning and intraoperative tumor resection guidance stands and falls with the precision of the method of motor mapping. Consequently, it is essential that presurgical nTMS motor mapping delivers motor maps that come ultimately close to reality and the results of intraoperative DES mapping. While the current standard approach of single-pulse nTMS (sp-nTMS) using an electric-field-navigated system for motor mapping largely fulfills these requirements for uE representations, assessment of lE representations seems more demanding, given the location of lE muscle representations within mainly the dorsal precentral gyrus and reaching down along the interhemispheric gap, thus requiring higher stimulation intensity than uE motor mapping to sufficiently elicit motor-evoked potentials (MEPs).Just recently, paired-pulse nTMS (pp-nTMS) with biphasic pulse wave application to induce short-interval intracortical facilitation (SICF) and combined with electric-field neuronavigation has been made available ,17,18,19To date, existing literature on the matter has exclusively investigated the representations of uE muscles by pp-nTMS with biphasic pulses in small cohorts of healthy subjects ,17,18,28In the present study, we investigate whether novel pp-nTMS with pulses of biphasic wave form could also improve motor mapping of lE muscle representations in patients with brain lesions in close comparison to sp-nTMS procedures. Specifically, we hypothesize that pp-nTMS with biphasic pulses (1) enables more potent lE motor mapping with lower stimulation intensities, and that it (2) provides reliable motor maps even in the most challenging cases where sp-nTMS may fail.This prospective study was conducted at a single university hospital and was approved by the local institutional review board (registration number: 171/19 S). All procedures were performed in accordance with the Declaration of Helsinki and its later amendments. Written informed consent was obtained from all patients prior to study enrollment.The period for study enrollment was September 2019 to July 2020. The following inclusion criteria were considered: (1) age above 18 years, (2) written informed consent, (3) motor-eloquent lesion location according to structural magnetic resonance imaging , and (4) clinical indication for preoperative motor mapping by nTMS and nTMS-based tractography of the CST. Pregnancy and metallic implants  were considered as exclusion criteria.Within the week prior to scheduled surgery for resection or biopsy, patients underwent clinical examinations, cranial MRI, and motor mapping by sp-nTMS as well as nTMS-based DTI fiber tracking (DTI FT) as part of the clinical routine. For study purposes, motor mapping by pp-nTMS with biphasic pulses and derived tractography were performed during the same one-day visit or during immediate preoperative inpatient stay.As part of the routine, patients underwent clinical examinations including assessments of sensory function, coordination, and cranial nerve function. Additionally, the Karnofsky performance status (KPS) scale was determined during the pre- and postoperative as well as 3-months follow-up (FU) assessments. Clinical examinations were supplemented by detailed evaluations of muscle function and strength according to the British Medical Research Council (BMRC) scale, which were performed preoperatively as well as during the first postoperative days and during FU examinations three months after surgery. Additional later examinations were performed depending on clinical course and tumor entity, but were not considered for this study’s purpose.Depending on the comparisons between pre- and postoperative as well as 3-months FU examination results, the following surgery-related motor deficit grades were defined, similar to previously published categories ,13,14,153 isovoxel covering the whole head), DTI sequence , and 3D T1-weighted gradient echo sequence  without and with application of an intravenous contrast agent  were acquired. Further sequences not used for this study’s purposes were acquired according to clinical needs and as part of a standardized multi-sequence imaging protocol for neurooncological patients.Cranial MRI was performed using a 3-Tesla MRI scanner equipped with a 32-channel head coil . A three-dimensional (3D) fluid attenuated inversion recovery (FLAIR) sequence . In all patients, sp-nTMS was carried out first , followed by pp-nTMS after a break of 60 to 120 min . The two investigators were strictly blinded to each other’s motor mappings, used stimulation adjustments, and the analyses of mapping data. The single procedures during motor mapping and their chronological course were identical for sp-nTMS and pp-nTMS mapping sessions (except for the application of single pulses in sp-nTMS and paired-pulses with biphasic wave forms in pp-nTMS), and were followed in close consideration of current nTMS mapping guidelines ,29. AfteImmediately following definition of the rMT, motor mapping was performed using a focal figure-of-eight stimulation coil. The coil was manually moved in all directions until the field of motor-positive stimulation spots was surrounded by at least two rows of motor-negative stimulation points ,32. In tDuring post-hoc analysis of motor mapping data, only stimulation points with MEP amplitudes of at least 50 µV and latencies in the typical ranges were defined as motor-positive spots ,32,37,38https://afni.nimh.nih.gov/), based on the spatial information taken from the contrast-enhanced 3D T1-weighted gradient echo sequence the mapping was conducted with [3 by default [https://afni.nimh.nih.gov/) [The motor hotspot coordinates and motor map extents were used to create heat maps of distribution using Analysis of Functional NeuroImages (AFNI) software  ,19. SpecDeterministic tractography of the CST was performed purely based on nTMS motor maps using Brainlab Elements . As for motor mapping, the two investigators performed nTMS-based DTI FT independently and were blinded to each other’s tractography results. The motor-positive stimulation points were fused with the MRI dataset of the respective patient, followed by definition of the motor-positive stimulation points as one object, which was transferred into a region of interest (ROI) by adding rims of 2 mm to each point ,9,10. InThe overall CST volume was extracted for nTMS-based DTI FT performed with each of the two different tracking adjustments . FurtherSurgery for tumor resection or biopsy was supported by intraoperative neuromonitoring in all cases (and supplemented by intraoperative DES to perform motor mapping in cases where only pp-nTMS provided motor maps and sp-nTMS motor mapping failed). For intraoperative neuromonitoring, a strip electrode  was placed over the precentral gyrus, and an amplitude reduction of 50% in potentials was determined as the stop criterion to avoid lasting motor deficits after surgery ,42,43.In case that intraoperative DES was performed, motor mapping was carried out with a monopolar stimulation electrode  after determination of the motor threshold and using square-waved pulses  in a train-of-five fashion, starting with an intensity of 4 mA that was increased continuously until potentials were detected ,45,46. Sp < 0.05.GraphPad Prism  was used for statistics. Shapiro-Wilk normality tests were conducted to assess data distribution. The level of statistical significance was set at t-tests (Gaussian data distribution) were used to assess differences for continuous variables.Descriptive statistics were calculated for patient, mapping, and tractography characteristics. Results are reported as mean ± standard deviation (SD) with ranges, or as absolute or relative frequencies. Chi-Squared tests were used to assess differences in contingency variables, Wilcoxon matched-pairs signed rank tests (non-Gaussian data distribution) or paired This study enrolled 39 patients suffering from unilateral motor-eloquent brain lesions. Preoperatively, 30 patients (76.9%) showed motor strength impairments of lE muscles according to the BMRC scale. In 9 patients (23.1%), a seizure was the main initial symptom. p = 0.5851).No technical difficulties or side effects were observed during motor mapping with either technique. Furthermore, no breaks during single motor mapping sessions occurred. The average coil-to-cortex distances were similar for motor mapping by sp-nTMS (mean ± SD: 22.6 ± 2.7 mm) and pp-nTMS (mean ± SD: 22.7 ± 2.9 mm), showing no significant difference (p < 0.0001). Discomfort during stimulation according to the VAS scale was on average slightly higher for sp-nTMS  when compared to pp-nTMS , yet without a significant difference between methods (p = 0.2288).Mapping by pp-nTMS was able to elicit MEPs of lE muscles and, thus, provided motor maps in all enrolled patients (100%). In contrast, sp-nTMS mapping failed to elicit MEPs in five patients (12.8%), thus not providing a motor map in these patients. Due to this failure of providing motor maps during sp-nTMS mapping, data of these five patients were not included in the further analyses. The stimulation intensity for motor mapping was significantly higher for sp-nTMS  compared to pp-nTMS . Regarding the CoG, there was a significant difference between sp-nTMS and pp-nTMS mapping for the x- and z-axis . The results of pp-nTMS mapping were highly comparable to those of sp-nTMS regarding the location and coordinates of the motor hotspot (p = 0.0003). Regarding the aspect ratio, significantly higher values were found for pp-nTMS when compared to sp-nTMS (p = 0.0261), indicating that the motor map was more elongated in the direction of the electric field for pp-nTMS. The volume of the motor area was significantly higher for pp-nTMS when compared to sp-nTMS  as compared to tractography using sp-nTMS data , but the differences were not significant . Furthermore, no aberrant fiber courses were detected. The total volume of the CST was higher when motor maps derived from pp-nTMS were used (FA = 0.1: mean ± SD: 34.6 ± 16.1 cmIn five patients (12.8%), solely pp-nTMS was able to provide a motor map, whereas sp-nTMS lacked eliciting MEPs. In four of those patients, intraoperative DES confirmed pp-nTMS results during surgery, while in the remaining patient intraoperative DES was also not able to elicit MEPs. Hence, pp-nTMS remained the only modality to provide information about the motor system in this patient, and the surgery was performed purely based on data derived from pp-nTMS in this case. The CST was reliably tracked in all of those five patients using pp-nTMS motor maps for seeding during tractography. The present study investigated pp-nTMS with biphasic pulses for motor mapping of lE muscle representations and related nTMS-based DTI FT of the CST. As hypothesized based on previous experience with pp-nTMS in motor mapping of uE muscle representations, the novel approach enabled motor mapping with lower stimulation intensities, and provided reliable motor maps even in the most challenging cases where sp-nTMS as the conventionally used method failed.In the investigated cohort of patients with motor-eloquent lesions, pp-nTMS was the only modality to provide motor maps and information about the motor system in one patient. Furthermore, it was able to generate motor maps in good correlation with intraoperative DES results in four patients in whom sp-nTMS failed to elicit MEPs. This finding emphasizes the potential of pp-nTMS with biphasic pulses to further improve the standard of care regarding preoperative motor mapping, which is currently achieved using sp-nTMS by default ,2,3,4. HDuring motor mapping by pp-nTMS, the applied stimulation intensity was lower compared to sp-nTMS, which is in agreement with previous publications investigating motor mapping of uE muscle representations by the same device in healthy subjects or patients with brain lesions ,17,19. WAnalogously to a previous publication on motor mapping of uE muscle representations in patients with brain lesions, there was no statistically significant difference in the location of the motor hotspot when using pp-nTMS compared to sp-nTMS . HoweverFurthermore, according to the aspect ratio, a motor map that was more elongated in the direction of the electric field was present for pp-nTMS compared to sp-nTMS. In contrast, previous work among healthy subjects—yet investigating the motor representations of uE muscles only—revealed comparatively similar aspect ratios for sp-nTMS and pp-nTMS, with the enclosed motor maps appearing to be wider in the anterior–posterior compared to the lateral–medial direction . This diWhen interpreting the results of this study several limitations have to be acknowledged. First, the difference in motor map volumes, CoG location, and aspect ratios between motor mapping using sp-nTMS or pp-nTMS needs to be investigated in more detail. Thus, ideally intraoperative DES from a higher number of patients would be needed to exclude any potential overestimation of the motor cortex by using pp-nTMS for preoperative motor mapping. In this context, intraoperative DES was only applied in patients in whom conventionally used sp-nTMS motor mapping failed, whereas it was not utilized in patients with motor maps generated by both sp-nTMS and pp-nTMS. This is related to the repeated finding of high concordance between preoperative sp-nTMS and intraoperative DES for motor mapping ,2,3,4; hThis study used novel paired-pulse application with pulses of biphasic wave form for preoperative electric-field-navigated motor mapping in patients suffering from motor-eloquent brain lesions. In direct comparison to conventionally used sp-nTMS, pp-nTMS motor mapping with pulses of biphasic wave form for lE muscle representations delivered reliable motor maps even in highly challenging patient cases where sp-nTMS failed. Further, pp-nTMS required lower stimulation intensities than sp-nTMS for successful motor mapping. Yet, differences in motor map volumes and CoG location require further investigation, and specific validation of motor maps derived from pp-nTMS needs to be achieved before the technique might have merit to replace standard sp-nTMS motor mapping in patients with brain tumors.


Read coverage of RNA sequencing data reflects gene expression and RNA processing events. Single-cell RNA sequencing (scRNA-seq) methods, particularly “full-length” ones, provide read coverage of many individual cells and have the potential to reveal cellular heterogeneity in RNA transcription and processing. However, visualization tools suited to highlighting cell-to-cell heterogeneity in read coverage are still lacking.′ UTR lengths, and enhancer RNA transcription.Here, we have developed Millefy, a tool for visualizing read coverage of scRNA-seq data in genomic contexts. Millefy is designed to show read coverage of all individual cells at once in genomic contexts and to highlight cell-to-cell heterogeneity in read coverage. By visualizing read coverage of all cells as a heat map and dynamically reordering cells based on diffusion maps, Millefy facilitates discovery of “local” region-specific, cell-to-cell heterogeneity in read coverage. We applied Millefy to scRNA-seq data sets of mouse embryonic stem cells and triple-negative breast cancers and showed variability of transcribed regions including antisense RNAs, 3 https://github.com/yuifu/millefy) and as a Docker image for use with Jupyter Notebook (https://hub.docker.com/r/yuifu/datascience-notebook-millefy).Millefy simplifies the examination of cellular heterogeneity in RNA transcription and processing events using scRNA-seq data. Millefy is available as an R package ( Single-cell RNA sequencing (scRNA-seq) has been increasingly important in many areas, including developmental biology and cancer biology. In scRNA-seq data analyses, visualization is crucial for quality control (QC) as well as exploratory data analyses. For example, dimensionality reduction techniques such as principal component analysis (PCA)  or t-disIn contrast to visualization of gene expression matrices, visualization of read coverage, which is the distribution of mapped reads along genomic coordinates, helps reveal diverse aspects of RNA sequencing data and thus RNA biology and functional genomics. For example, read coverage reflects transcribed gene structures  , RNA pro′-tag sequencing” scRNA-seq methods, which sequence only the 3 ′ ends of RNAs and cannot be used to extract rich information from read coverage [Given that scRNA-seq has revealed cellular heterogeneity in gene  and splicoverage , 14. Des2– 103) cells and frequently includes latent heterogeneity that is masked by the summation of expression across cells. Second, the tool must associate read coverage with genomic contexts, such as gene structures and epigenomic features, because read coverage data can be interpreted only when it is displayed simultaneously with their genomic contexts. Third, the tool must highlight the cell-to-cell heterogeneity of read coverage within focal regions. This is because there should be “local” region-specific cell-to-cell heterogeneity in read coverage at transcriptional  and post-transcriptional  levels, and such heterogeneity is difficult to notice in advance by cell groupings defined according to global similarity among cells.To explore cell-to-cell heterogeneity in read coverage, we propose several requirements of a tool for visualization of read coverage in scRNA-seq data Table . First, Genome browsers and heat maps are two major tools for read coverage visualization. However, they are insufficient for fulfilling the above requirements.Genome browsers, such as IGV  and JBroTools for heat maps combined with clustering algorithms have been used in the analysis of scRNA-seq data. Thus, heat maps can be used to visualize read coverage of all cells at once and reveal heterogeneity in read coverage. However, tools for generating heat maps are unsuited for visualizing read coverage of scRNA-seq data in genomic contexts, or they lack functionality to directly extract read coverage from standard NGS data formats.Here, we have developed Millefy, which combines genome-browser-like visualization, heat maps, and dynamic reordering of single-cell read coverage and thus facilitates the examination of local heterogeneity within scRNA-seq data. Millefy extracts and organizes various types of useful information from read coverage of scRNA-seq data.Millefy visualizes read coverage from each individual cell as a heat map in which rows represent cells and columns represent genomic bins within a focal region. The heat map is aligned with tracks for gene annotations, genomic features, and bulk NGS data, enabling comparisons of single-cell read coverage with genomic contexts. To highlight latent cell-to-cell heterogeneity in read coverage, the heat map rows  are automatically and dynamically reordered by ’local’ pseudo-time, which is calculated using diffusion maps , a nonliEffective visualization requires iterative adjustment of various aspects of plots by visual inspection. Millefy supports iterative adjustment of plots by the millefy_adjust function, which reuses the read coverage matrices of the last plot, enabling faster adjustment than simply replotting. For example, after viewing plots, users can easily adjust the maximal value of the color scale, which is essential in cases with exceptionally high read coverages in the focal region.Millefy visualizations consist of five types of tracks Fig. : (1) scRMillefy was implemented in R and can import scRNA-seq data without the need for format conversion. For scRNA-seq data, Millefy accepts BAM and BigWig formats, which are standard file formats for NGS data analysis. Millefy is dependent on the rtracklayer package  and Rsamhttps://github.com/jupyter/docker-stacks) for use with Jupyter Notebook.We provide Millefy as an R package and as a Docker image based on the Jupyter Notebook Data Science stack  read coverage cannot capture heterogeneity in read coverage. For example, a change in the merged read coverage cannot indicate whether the number of cells expressing a gene increased or the expression level of that gene increased across all cells. In contrast, Millefy visualizes read coverage of all individual cells in a scRNA-seq dataset as a heat map and thereby provides detailed information on cellular heterogeneity in read coverage.To demonstrate the usefulness of Millefy’s ability to visualize read coverage in scRNA-seq data, we used a time-course RamDA-seq dataset derived from mouse embryonic stem cells (mESCs) upon induction of cell differentiation to primitive endoderm cells  [Sox17, a differentiation marker gene. Cells were reordered according to the first diffusion component values calculated by a diffusion map of read coverage data for the locus, either within user-defined cell groups  patients: the Smart-seq2 data of single sorted cells from six tumors collected from six women with primary, non-metastatic triple-negative invasive ductal carcinomas . The dat′ UTR shortening is observed in the endothelial cell clusters. The Neuroblastoma RAS viral (v-Ras) oncogene homolog (NRAS) and the Jun proto-oncogene (c-JUN) are known to show alternative polyadenylation (APA)-dependent 3 ′ UTR shortening in TNBC cells [′ UTRs of c-JUN and NRAS  with mESC enhancer regions. We downloaded H3K4me1 and H3K4me3 ChIP-seq peak regions for mESCs from the ENCODE project  and defiMyc locus, with the positions of enhancers active in mESCs. The Myc gene models and read coverage reveal that Myc was transcribed in mESCs. In addition, Millefy showed that some of the intergenic regions with transcribed RNA overlapped with the Myc downstream enhancer regions  and C1-SMART-Seq V4 (n=95) data from a dilution of 10 pg of mESC RNA. Figure Mdn1, a gene with a long transcript  consisting of 102 exons. C1-RamDA-seq detected all exons in most samples, while C1-SMART-seq V4 failed to detect a fraction of known exons. Interestingly, the patterns of missing exons in C1-SMART-seq V4 seemed to vary among the samples. The lower reproducibility in read coverage of C1-SMART-seq V4 relative to C1-RamDA-seq is likely owing to technical noise because the samples were prepared not from living cells but from a dilution of 10 pg of RNA. We note that mean read coverage cannot provide such detailed information on reproducibility in read coverage , https://hub.docker.com/r/yuifu/datascience-notebook-millefy(Docker image)Archived version: DOI:10.5281/zenodo.3591109Operating system(s): Platform independentProgramming language: ROther requirements: R version 3.2.2 or higherLicense: MITAny restrictions to use by non-academics: No


The use of granulocyte colony-stimulating factor (G-CSF) in patients with haematological malignancies is associated with less febrile neutropenia episodes. But in the presence of COVID-19 infection, the administration of G-CSF is challenging as it may trigger a robust inflammatory reaction resulting in cytokine storm, respiratory failure and severe outcomes. Additionally, the use of G-CSF after allogeneic HCT is not well established, with heightened risk of graft-versus-host disease.4 Therefore, more HCT patients are at greater risk of infections and the immunopathogenesis of COVID-19 implicates a virus-driven organ damage followed by a robust inflammatory reaction involving the overexpression of several proinflammatory mediators such as IL-6, TNF and host G-CSF resulting in a cytokine storm that contributes to further vital organ compromise, multisystem dysfunction syndrome and death.5This raises concerns for worrisome prognosis among HCT recipients who are contracting severe acute respiratory syndrome coronavirus 2 following HCT primarily during the pre-engraftment period. Given the rapid spread of novel coronavirus 2019 pandemic (COVID-19), confirmed COVID-19 cases have reached more than 31,000,000 worldwide with 960,000 deaths.All in all, concurrent COVID-19 infection in HCT recipients receiving G-CSF therapy can potentially lead to deleterious results and poor clinical outcomes. Further studies and clinical data are timely needed to evaluate the safety and benefits of using G-CSF following HCT during COVID-19 pandemic, as controlling the hyperinflammation response secondary to COVID-19 infection is a cornerstone in the management.


Nematode flavin-containing monooxygenase-2 (fmo-2) is induced by dietary restriction and hypoxia, and is required for the longevity and health benefits of these pathways. It is also sufficient to confer these benefits when overexpressed. As FMOs are well-conserved across taxa, the fmo-2 mechanism has high translational potential. To determine the changes that occur following fmo-2 induction, we performed RNA-seq and untargeted metabolomics analyses. Our data reveal that one-carbon metabolism (OCM) is significantly altered by fmo-2 overexpression. OCM is a nexus for essential metabolic pathways, including transmethylation, transsulfuration, nucleotide synthesis, and amino acid metabolism. We hypothesized that fmo-2 confers longevity benefits by altering key metabolic processes within or downstream of OCM. To test this, we asked whether fmo-2 and OCM interact to regulate longevity by knocking down expression of genes involved with OCM and measuring lifespan and oxidative stress resistance. To understand the biological implications of these interactions, we generated a computational model using qPCR data of key OCM-related genes to predict changes in OCM metabolic flux. Our model predicts significant changes in OCM flux that are regulated by fmo-2 expression levels and are consistent with our RNAi and multi-omics results. We are now testing this model by knocking down genes downstream of OCM to determine their role in fmo-2-mediated benefits. Preliminary results support our hypothesis that changes in metabolic flux through OCM are involved downstream of fmo-2 expression, and may also implicate the UPRER in this pathway. Our future work will elucidate this mechanism and link stress perception and fmo-2-mediated longevity.