In this population, chronotype is weakly associated with depressive symptoms and moderately correlated with self-reported sleep disturbance. Sleep disturbance is significantly associated with depression scores, but objective sleep parameters are not directly correlated with Beck Depression Inventory (BDI-II) scores, with the exceptions of a moderate correlation between social jetlag and depression scores in females and a marginal correlation between sleep duration and depression scores. Multiple regression and path analyses reveal that chronotype effects on depressive symptoms in this population are mediated largely by sleep disturbance. The PER3 VNTR genotype significantly predicts depressive symptoms in a model with objective sleep parameters, but it does not significantly predict depressive symptoms in a model with chronotype or subjective sleep disturbance. Interestingly, PER35,5 genotypes, in males only, are independently related to chronotype and depression scores. Our results support hypotheses linking subjective sleep quality and chronotype and provide a first step in understanding how objective sleep structure may be linked to chronotype and depressive symptoms. Our results also suggest that circadian gene variants may show sex-specific effects linking sleep duration and sleep structure to depression.Depression and its related mood disorders are a major global health issue that disproportionately affects young adults. A number of factors that influence depressive symptoms are particularly relevant to the young adult developmental stage, including sleep loss, poor sleep quality, and the tendency toward eveningness in circadian preferences. However, relatively few studies have examined the relationship between sleep and circadian phenotypes, and their respective influences on mood, or considered potential molecular mechanisms driving these associations. Here, we use a multi-year, cross-sectional study of 806 primarily undergraduates to examine the relationships between sleep-wake chronotype, sleep disturbance, depression and genotypes associated with the PER3 variable number of tandom repeats (VNTR) polymorphism—circadian gene variants associated with both chronotype and sleep homeostatic drive. In addition, we use objective, Fitbit-generated sleep structure data on a subset of these participants ( Depression and related disorders, such as anxiety, affect nearly one-fifth of the global population and disproportionately affect young adults ; studiesvia downstream activation and repression of circadian-controlled genes. Mutations in core regulatory clock genes are associated with both sleep and mood disorders, suggesting that molecular mechanisms affecting sleep and mood are influenced, in part, by the circadian clock , and evening-types and individuals with delayed sleep phase disorder (DSPD) have greater frequencies of PER34/4 homozygotes , and deep sleep] to better understand how features of sleep mediate chronotype effects on mood phenotypes. In addition, we test whether a circadian molecular marker, PER3 VNTR genotype, influences the effect of sleep quality and structure on depressive symptoms.n = 806, 229 males, 577 females, median age: 19, range 17–36 ]. Participants were recruited primarily via an introductory biology course comprised of first or second-year undergraduates during the years 2013–2019; sampling occurred during the fall or spring, depending on the year. Participation rate for each sampling period ranged from 94 to 99%. Participants received laboratory credit and a food token for completing the survey study. A randomly selected subset of this study population provided the data used to measure sleep structure parameters ; these participants were compensated with a small monetary payment for wearing a Fitbit for a week. There was no difference in age, Patient-Reported Outcomes Measurement Information System (PROMIS), mid-sleep point on a free day (MSF), or Beck Depression Inventory (BDI-II) scores between the subset and the larger study population. Participants who did not have at least six nights of sleep data recorded, including two weekend nights, were excluded from the study. All methods adhered to the principles of the Declaration of Helsinki; the Institutional Review Board at Colgate University approved all procedures and consent forms . All participants gave written informed consent.Data for the associations between chronotype, sleep disturbance, and depressive symptoms were collected from a multi-year study of a population of predominantly young adults , but this effect is strongly mediated by sleep disturbance [SD = 0.41 ].A multiple regression combining the effects of chronotype and sleep disturbance on depressive symptoms reveals that the significant effects of chronotype become non-significant on addition of sleep disturbance scores in the model . The modF2,266 = 3.1, p = 0.737), sleep disturbance , or depression scores . In the larger data set, VNTR genotypes are not associated with chronotype categories; PER34,4 genotypes are not more likely to be evening-types . However, ET males are nine times more likely to have a PER34,4 genotype  than MT males. In addition, males with a PER35,5 genotype tend to have lower depression scores than males of other genotypes, but this result was not significant . Overall, VNTR genotypes have marginal effects in predicting depressive symptoms in regressions of MSF chronotype and sleep disturbance .In an exploratory regression analysis on the subset of sleep data, objective sleep parameters  are not significant predictors of depressive symptoms . HoweverOverall, our study reveals complex and, potentially, sex-specific relationships between depressive symptoms and chronotype, sleep disturbance, and sleep structure. Our results confirmed previous reports that chronotype is weakly associated with depression scores , with evInterestingly, chronotype was significantly associated with overall self-reported sleep disturbance scores, with evening-types reporting higher sleep disturbance, but not with objective sleep structure parameters or sleep duration. In addition, the degree of misalignment of circadian rhythms with the social environment, or social jetlag, was associated with only one sleep parameter, sleep duration; individuals with greater social jetlag slept longer on free days. Our results on the influence of eveningness on self-reported sleep disturbance support previous studies showing subjective sleep quality measures are associated with chronotype . For objOur study does provide another potential factor to consider – the gender effect seen in our Fitbit-generated sleep structure data. We find that males have significant relationships between chronotype and sleep variables, with evening-types having significantly greater percentage of REM sleep and a lower percentage of deep sleep. However, the variance in these sleep factors across chronotype is greater in females. If sleep structure is related to depression, one might predict an increase in REM sleep and a decrease in restorative deep sleep in individuals battling with mood disorders . Althougvia effects on subjective sleep disturbance. These results support findings from previous studies in young adults  with depressive symptoms  that improve the quality of life and success of undergraduate students and other vulnerable populations.The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.The studies involving human participants were reviewed and approved by Institutional Review Boards of Colgate and Cornell Universities. The patients/participants provided their written informed consent to participate in this study.KI designed the study. KW organized the study and database. KI, CW, and AF performed the statistical analysis. KI wrote the draft of the manuscript. All authors contributed to manuscript revision and approved the submitted version.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


There are growing concerns that poor dietary behaviors at young ages will increase the future risk of chronic diseases in adulthood. We found that female college students who skipped breakfast had higher incidences of dysmenorrhea and irregular menstruation, suggesting that meal skipping affects ovarian and uterine functions. Since dysmenorrhea is more prevalent in those with a past history of dieting, we proposed a novel concept that inadequate dietary habits in adolescence become a trigger for the subsequent development of organic gynecologic diseases. Since inadequate feeding that was limited during the non-active phase impaired reproductive functions in post-adolescent female rats, we hypothesize that circadian rhythm disorders due to breakfast skipping disrupts the hypothalamic–pituitary–ovarian axis, impairs the reproductive rhythm, and leads to ovarian and uterine dysfunction. To explain how reproductive dysfunction is memorized from adolescence to adulthood, we hypothesize that the peripheral clock system also plays a critical role in the latent progression of reproductive diseases together with the central system, and propose naming this concept “adolescent dietary habit-induced obstetric and gynecologic disease (ADHOGD)”. This theory will contribute to analyzing the etiologies of and developing prophylaxes for female reproductive diseases from novel aspects. In this article, we describe the precise outline of the above hypotheses with the supporting evidence in the literature. Among young women, insufficient energy intake and inadequate timing of dietary intake have become common nutritional issues in the world . There aBased on these backgrounds, we found that female college students who skipped breakfast had a significantly higher incidence of dysmenorrhea than those who ate breakfast, according to a questionnaire-based investigation . In addiFrom these findings, we propose a novel concept that disorders of the central clock system due to breakfast skipping disrupt the hypothalamic-pituitary-ovarian axis, impair the reproductive rhythm, and lead to ovarian and uterine dysfunction. We also hypothesize that the peripheral clock system plays a critical role in the latent progression of reproductive diseases. In this article, we precisely present the outline of the above hypothesis, showing the supporting evidence in the literature.Meal-skipping rates are high during young adulthood . Currentn = 956) [n = 4606) [Our longitudinal questionnaire-based study subsequently confirmed the positive relationship between skipping breakfast and the incidence of dysmenorrhea . Later, n = 956) . Another = 4606) . Althoug = 4606) . On the  = 4606) . Currently, meal skipping is considered to increase future risks of various chronic metabolic diseases ,18,27. SFemale college students who skipped breakfast also showed a tendency to have constipation ,31,32,33In general, body-related teasing is known to induce body dissatisfaction and influence dieting behaviors in adolescents . AdditioIn accordance with previous reports ,39, we oCurrently, the hypothesis of developmental origins of health and disease (DOHaD) is causing marked concern throughout the world . This coPrimary dysmenorrhea is mainly caused by abnormal uterine contraction ,46, whicSkipping breakfast interferes with the start of the active phase during the circadian rhythm that is regulated by the central clock system . Since bbrain and muscle aryl hydrocarbon receptor nuclear translocator like protein 1 (BMAL1), Clock, Periods (Per), and Cryptochrome (CRY) [Recently, the relationship between the circadian rhythm and the reproductive function has become a general concern . Circadime (CRY) . Light ame (CRY) . Currentme (CRY) ,63. In ame (CRY) . ConsideBmal1-knockout mice were also reported to be infertile and show implantation failure with a reduction of ovarian functions [Transgenic mice with clock gene mutations were reported to show disrupted reproductive functions such as in ovarian steroidogenesis , estrousunctions ,68. Accounctions .Recently, we found that pregnant women who had a history of dysmenorrhea in young adulthood showed a high incidence of hypertensive disorders of pregnancy (HDP), whereas dysmenorrhea just prior to pregnancy was not significantly correlated with the onset of HDP. HDP is derived from the abnormal placental formation due to inadequate trophoblastic invasion into the maternal uterus . AccordiPer1-3, CRY1-2, BMAL1, and Clock were demonstrated to be expressed in the uterus and oviduct in rodents [BMAL1 gene expression in human endometrial stromal cells attenuated the in vitro decidualization of stromal cells [BMAL1 silencing also impaired the ability of decidual cells to regulate trophoblast invasion, which can impair adequate placental formation. To suggest the involvement of clock dysfunction in obstetric disorders, the authors additionally demonstrated that BMAL1 gene expression was downregulated in the endometrium of women who experience recurrent spontaneous abortion [Bmal1-deficient mice demonstrated that uterine Bmal1 gene expression was essential to maintain a successful pregnancy. On the other hand, a recent study showed that feeding at an unusual time of day (inactive phase) desynchronizes peripheral clocks, causing obesity and metabolic disorders in adult mice [Peripheral oscillators, which are mainly regulated by daily food intake cycles, operate in most peripheral organs . Circadi rodents ,72. Veryal cells . Notablyabortion . To suppult mice . In addiult mice . We alsoBased on these findings, we hypothesized that the peripheral clock system in the uterus also plays a critical role in ADHOGD. It should be noted that the uterus extensively develops, grows, and matures during adolescence and young adulthood from both morphological and functional aspects ,77. TherYoung women who skip breakfast show significantly higher incidences of dysmenorrhea and irregular menstruation, suggesting that meal skipping affects ovarian and uterine functions. Since dysmenorrhea becomes more manifested in those with a past history of dieting, we proposed a novel concept whereby inadequate dietary habits in adolescence become a trigger for the subsequent development of organic gynecologic diseases. As a possible mechanism, we speculate that confliction of circadian rhythm by meal skipping affects the hypothalamic-pituitary-ovarian axis, impairs the reproductive rhythm, and leads to ovarian and uterine dysfunction. We hypothesize that the peripheral clock system in the uterus also plays a critical role in the latent progression of reproductive diseases and propose naming this concept “adolescent dietary habit-induced obstetric and gynecologic disease (ADHOGD)”. This concept will contribute to analyzing the etiologies of and developing prophylaxes for female reproductive diseases such as polycystic ovary syndrome, hypothalamic amenorrhea, endometriosis, infertility, preterm labor, and so on from novel aspects. This hypothesis may change the focus from therapeutic to prophylactic and from dietary content to dietary timing in the management of gynecologic disorders in young women. Further investigation together with developing new methods is recommended to test the hypothesis in the future.


Alzheimer’s disease (AD) represents an enormous public health challenge currently and with increasing urgency in the coming decades. Our understanding of the etiology and pathogenesis of AD is rather incomplete, which is manifested in stagnated therapeutic developments. Apart from the well-established Amyloid Hypothesis of AD, gaining traction in recent years is the Pathogen Hypothesis, which postulates a causal role of infectious agents in the development of AD. Particularly, infection by viruses, among a diverse range of microorganisms, has been implicated. Recently, we described a prominent antiviral immune response in human AD brains as well as murine amyloid beta models, which has consequential effects on neuropathology. Such findings expectedly allude to the question about viral infections and AD. In this Perspective, we would like to discuss the molecular mechanism underlying the antiviral immune response, highlight how such pathway directly promotes AD pathogenesis, and depict a multilayered connection between antiviral immune response and other agents and factors relevant to AD. By tying together these threads of evidence, we provide a cohesive perspective on the uprising of antiviral immune response in AD. Hallmarked by the deposition of β-amyloid plaques and accumulation of neurofibrillary tangles, Alzheimer’s disease (AD) manifests with complex pathophysiology and its etiology remains elusive. Among the many viewpoints for the underlying mechanisms, the Pathogen Hypothesis was proposed initially based on the clinical discoveries of Herpes Simplex Virus-1 (HSV-1) in association with AD, further expanded to include other microbes, and gained experimental support in recent years . In partMammalian antiviral innate immune defense mechanism utilize an array of nucleic acid innate immune sensors to detect viral genomes or their replication products, which activate a cascade of signaling events to induce rapid gene expression . Among tAlthough normally non-immunogenic, host self-derived nucleic acids can provoke IFN response under pathological conditions such as autoimmune diseases. In systemic lupus erythematosus, immune complexes comprised of a patient’s DNA or RNA and associated autoantibodies activate plasmacytoid dendritic cells (pDCs), a subset of innate immune cells, to stimulate IFN production and flares of systemic inflammation . In psorin vitro. More strikingly, nucleic acid-containing amyloid fibrils are potent inducers of IFN response from pDCs in vitro and in vivo  is protected by a highly complex barrier structure, but is by no means invincible to infections. Many neurotropic viruses can gain access to the brain through blood circulation or peripheral nerves . Acute vAlthough responsible for a common form of acute viral encephalitis, herpes simplex virus mostly carries out latent infection in the general population. TREX1 or ribonuclease RNASEH2 lead to the accumulation of aberrant cytosolic nucleic acid species, IFN production, and encephalopathy in Aicardi-Goutieres syndrome . In normal aging brain, heightened IFN signaling from microglia inside the parenchyma and choroid plexus, an epithelial tissue located within the ventricles, has been shown to be detrimental to neurogenesis and cognitive function . On the in vivo, whereas blocking IFNγ signaling rescued the premature death of trisomy 16 cortical neurons in vitro (Adult Down syndrome (DS) patients, who mostly carry trisomy 21 in their genome, unanimously develop the neuropathological changes of AD . Besidesin vitro . By contin vitro . Since bin vitro , these fIn summary, we have described the IFN pathway activation, a prototypic antiviral immune response, as a major component of CNS neuroinflammatory network in AD and connected IFN response to various endogenous, pathological, infectious and genetic risk factors that have been implicated in AD pathogenesis. The discussion here is largely focused on the molecular events originated within CNS, but peripheral IFN, manifested as a result of viral infection, autoimmune condition or drug administration, nevertheless affects brain functions . TherefoThe original contributions presented in the study are included in the article/supplementary material, further inquiries can be directed to the corresponding author.WC formulated the concepts and wrote the manuscript. ER contributed to the manuscript writing and figure preparation. All authors contributed to the article and approved the submitted version.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Crithidia bombi, which may reflect the ease with which this parasite is transmitted. These results suggest that agri-environment schemes such as flower strips, which are known to enhance the nutrition and survival of bumblebees, may also have negative impacts on pollinators through enhanced disease transmission. Future studies should assess how changing the design of these schemes could minimize disease transmission and thus maximise their health benefits to wild pollinators.Supplemental feeding of wildlife populations can locally increase the density of individuals, which may in turn impact disease dynamics. Flower strips are a widely used intervention in intensive agricultural systems to nutritionally support pollinators such as bees. Using a controlled experimental semi-field design, we asked how density impacts transmission of a virus and a trypanosome parasite in bumblebees. We manipulated bumblebee density by using different numbers of colonies within the same area of floral resource. In high-density compartments, slow bee paralysis virus was transmitted more quickly, resulting in higher prevalence and level of infection in bumblebee hosts. By contrast, there was no impact of density on the transmission of the trypanosome  As spDaphnia, host density influenced the likelihood of infection by protozoan parasites )Our experiment examined within-species transmission, but, as noted above, in the wild bumblebees live in complex multi-species assemblages of floral visitors. An increasing number of studies suggest that between-species transmission, in particular from managed honeybees, may be driving emergent diseases in wild pollinators ,57,63–655.Controlled semi-field experiments demonstrate the importance of density-dependent transmission for viruses in bumblebees. However, current agri-environment schemes designed to support pollinator assemblages ignore their potential role for disease transmission among flower visitors. We suggest that future development of such schemes should take a more holistic, integrated approach that considers both nutrition and disease risk, to design conservation interventions that maximize pollinator health.


Plasmodium falciparum histidine-rich protein 2 (PfHRP-2), parasite lactate dehydrogenase (pLDH), aldolase, glutamate dehydrogenase (GDH), and the biocrystal hemozoin. The models that demonstrate a potential for field application have been discussed, looking at the fabrication and analytical performance characteristics, including (but not exclusively limited to): response time, sensitivity, detection limit, linear range, and storage stability, which are first summarized in a tabular form and then described in detail. The conclusion summarizes the state-of-the-art technologies applied in the field, the current challenges and the emerging prospects for malaria biosensors.The impact of malaria on global health has continually prompted the need to develop more effective diagnostic strategies that could overcome deficiencies in accurate and early detection. In this review, we examine the various biosensor-based methods for malaria diagnostic biomarkers, namely;  Anopheles mosquitoes. The greatest burden of the disease is in the tropical and subtropical regions of the world ..74].Until date, most malaria biosensors have been principally confined to the established markers. Some other likely candidate targets have been speculated; heat-shock protein 70 (Hsp70), dihydrofolate reductase (DHFR)–thymidylate synthase (TS), heme-detoxification protein (HDP), glutamate-rich protein, hypoxanthine phosphoribosyl transferase, and phosphoglycerate mutase ,104,105.2–108 cells/mL of infected RBCs  antibodies directed against both antigens as low as 6–50 pg/L (concentrations in the range of 10–15 M) by means of EIS. Overall, the method demonstrated a promising approach to real-time probing of antibodies in serum directly without the need for sample preparation . S. SP. vivP. falciparum [P. falciparum and P. vivax [P. falciparum DNA fragments is suitable for automation and integration into portable molecular POCTs [P. falciparum DNA target sequences is then concentrated for SERS measurement. The assay produced a LoD of 100 attomoles DNA (Nucleic acid markers have been exploited as reliable alternatives to proteins and antibodies in malaria diagnostics . A DNA blciparum . The poslciparum  and distP. vivax . Both ofar POCTs . The assoles DNA .Simultaneous analysis of biomarkers maximizes the use of samples and gives a wide range of results that significantly improves test accuracy. Increased throughput, reduced reagents/assay setup and less labor are among features that could decrease assay errors and make parallel testing ideal for healthcare delivery. Moreover, the benefit directly translates to convenient sampling and low cost to providers and patients at the same time ensuring optimum outcome. A combination of antigens and genes have been reported in ICT and nucleic-acid-based methods respectively and are discussed elsewhere ,112,113.The drawbacks of diagnostics in spite of the vast array of tests require more effective alternatives that is crucial in the long-term fight against/and probable elimination of malaria. Taking the tremendous success of glucometers into account, biosensors are a potential driving force in the emerging medical demands for onsite diagnostics in resource-limited settings. The growing interests in biosensor research and the progress made with the prototypes from published data implies that the technology could have a significant impact on malaria diagnosis regardless of being in nascent stages. These concepts, so far have offered ideal and desirable characteristics like high performance, low fabrication costs and easy operation with the aim of practicality in clinical diagnostics. The sensors discussed in this review present with impressively low detection limits that are superior to immune chromatographic dipstick which is usually between 2–16 nM. Overall, it can be argued that the slight increments in LoDs observed in serum samples relative to buffer are attributable to the complexity of serum that can hinder interactions between biorecognition molecules and target biomarkers. Given that a major challenge to malaria diagnosis is the lack of highly sensitive tools POCTs at low parasitemia, the impressive analytical performances that biosensors have demonstrated would allow for ultra-sensitive diagnosis of asymptotic cases and monitoring treatment with antimalarial drugs. To put this in perspective, randomized control trials that apply these technologies to a large number of clinical samples are needed in order to evaluate the practical usefulness. For easy translation of malaria biosensors to the field, aspects related to complexity and instrumentation which minimizes user intervention while ensuring low-cost are key areas which remains unclear. Studies are required to model self-contained devices as most biosensors presented rely on traditional immunoassay principles and involve multiple assay steps. Innovative technologies should take advantage of 3-D printing techniques and microfluidics to integrate prototype sensors into self-contained lab-on-chip models in readiness for field deployment.The increase in preference for probing LDH in blood seems an amelioration in malaria diagnostics since its concentration correlates with parasite density and implies metabolically active parasites. This is in contrast to HRP-2 that either has delayed clearance or is absent in certain mutant strains. Furthermore, considering the advantages of electrochemical systems in POCTs, efforts towards improved system complexity could propose appropriate electrochemical systems for enzyme biomarkers like LDH, GDH, and aldolase that could be easily integrated into enzyme-based biosensors. For instance, substrates that are specific to these enzymes could be incorporated for direct enzymatic reaction detectable through electrochemical methods. Undoubtedly, the future of biosensors is being changed by the growing demands for novel biotechnologies in POC diagnostics. These innovations coupled with the capacity of multiplexing are necessary in fulfilling a major requirement of detection and species differentiation in the clinical diagnosis of malaria.


Over the recent decades there has been tremendous progress in understanding and controlling friction between surfaces in relative motion. However the complex nature of the involved processes has forced most of this work to be of rather empirical nature. Two very distinctive physical systems, hard two-dimensional layered materials and soft microscopic systems, such as optically or topographically trapped colloids, have recently opened novel rationally designed lines of research in the field of tribology, leading to a number of new discoveries. Here, we provide an overview of these emerging directions of research, and discuss how the interplay between hard and soft matter promotes our understanding of frictional phenomena. Structural lubricity is one of the most interesting concepts in modern tribology, which promises to achieve ultra-low friction over a wide range of length-scales. Here the authors highlight novel research lines in this area achievable by combining theoretical and experimental efforts on hard two-dimensional materials and soft colloidal and cold ion systems. Setting aside empiricism and engineering, scientists embracing distinct—yet complementary—experimental, theoretical, and computational approaches, have recently made crucial systematic inroads in the atomic-level physics involved in frictional phenomena. Spanning several decades of the involved scales of length, time, and energy, the ubiquitous nature of friction, together with a remarkable number of material-independent features, suggests indeed potentially fruitful links among physics, chemistry, materials science, soft matter, geology, and other fields of past and current research and technology, making tribology a pretty unique subject.Despite the fundamental and practical relevance of dissipative interface phenomena and its long history in science, the complexity of the highly out-of-equilibrium processes across confined sliding interfaces still leaves several key aspects of tribology not well understoodOn the wave of recent advances and achievements in the field, we aim at providing, via this Perspectives article, a glance at some topics which in our view contain potential seeds for further work, development and new connections. Specifically, out of serendipity, two very distinctive operative frameworks have recently brought about promising avenues of research and new discoveries in tribology: the friction-related physics of hard two-dimensional (2D) layered atomic materials and that of optically and topographically trapped soft microscopic systems, such as cold ions and colloids.At microscopic scales Fig. , where m5. The most common example is lubricated friction, where lubricant additives, polymer brushes, ionic liquids, or hydration layers are used to provide ultralow friction and remarkable durability of interfaces9. Notably, ultralow friction can be achieved also in the absence of such lubricants. Thus, amorphous carbon coatings can provide ultralow friction coefficients13, and they are already used in industrial applications, including razor blades, magnetic hard drives, microelectromechanical systems and more.There are many alternative strategies for achieving ultralow frictionWhen dealing with well-defined mating interfaces, the idea of nearly frictionless sliding, in both soft and hard matter contacts, relies on the possibility to predict the physical response of a system under shear based on interface geometrical features more than upon distinct system-dependent characteristics. Structural incommensurability, arising from surface lattice mismatch or misalignment as shown in Fig. 18 in the physical context of frustrated incommensurate structures and its connection to the renowned Kolmogorov–Arnold–Moser framework of the onset of chaos in dynamical systems20, is cast as one of the deepest achievements in recent theoretical mechanics and dynamical systems theory.Aubry’s pioneering study of the Frenkel–Kontorova (FK) class of models23, can be rigorously drawn in the framework of the standard one-dimensional (1D) FK model, describing a chain of nearest-neighbor interacting particles (the slider) subject to a corrugated periodic potential (the underneath substrate)25. According to Aubry, in the thermodynamic limit, an incommensurate 1D chain/substrate interface undergoes a sharp remarkable transition at a critical value of the substrate potential corrugation between a lubric frictionless dynamics and a pinned frictional sliding. In other words, the static friction, i.e., the minimal force required to initiate motion, should vanish, provided the two mismatched mating surfaces are stiff enough compared to their mutual interaction; in addition, under such conditions, the incommensurate contact geometry promotes also an ultralow dissipative dynamical regime during sliding.Aubry theory leaves nowadays the elegant mathematical realm of number theory and Hamiltonian maps to become practically relevant and cornerstone of the physical understanding of many length-scale competition phenomena in complex systems, as can be found, e.g., in both condensed matter and soft-matter physics. Particularly, the surprising existence of a possible frictionless sliding regime, today a pervasive nano/mesoscale tribological concept called superlubricity and first named by Hirano and ShinjoEven though, at first glance, Aubry’s mathematically rigorously proven scenario may seem quite academic when compared to the intrinsic 2D (or even 3D) complexity of a microscopic sliding interface, structural superlubricity has indeed started to be observed in an increasing number of recently implemented experimental cases.28. In 2004, one groundbreaking experiment in the field demonstrated interlayer registry-dependent friction in a nanoscale contact between a pristine graphite surface and a graphite flake attached to an AFM tip29. There, it was shown that when the atomic lattices of the two contacting surfaces are aligned in registry, high interlayer friction values are measured, while when orientationally misaligned friction practically vanishes. This observation first marked layered materials as promising candidates for dry friction applications in nano/microelectromechanical systems (NEMS/MEMS), as well as improved lubricants or lubrication additives for more macroscopic devices.Back in the nineties, few proximal-probe based studies provided the first experimental evidence of superlubricity40, telescopic dynamics and water flow in nanotubes and confined geometries43, rare-gas island inertial motion46, sliding colloidal layers and ion-crystal simulators53, and transport in quasi-1D Wigner solids55, have recently uncovered signatures of a superlubric behavior.Still focusing on nano- up to mesoscale experimental systems, other works, including nanomanipulation of deposited adsorbates (in the form of nanoclusters and 2D graphite-like patches)59, sustainable against external load, high velocities, ambient conditions and varying contact geometries, and to upscale this phenomenon to microscopic and eventually macroscopic system length scales.With the potential of controlling dissipation and reducing wear in high-performance mechanical devices functioning at various scales, all that has brought about great fundamental and technological interest toward the nature of structural superlubricity, its robustness, the feasibility of scaling it up, and the possible mechanisms leading to its failure and breakdown. The search and design of novel ultralow frictional interfaces is a timely subject of practical importance. Specifically, the main challenges today are to achieve robust superlubricity60, where weak van der Waals (vdW) forces couple extremely in-plane stiff atomic layers, does emerge as an exemplary candidate.As prescribed by Aubry’s theory, dry solid-on-solid contacts displaying vanishing low values of static and kinetic friction, arising from effective cancellation of lateral force components, are characterized by key ingredients, such as interface incommensurability, intrinsic rigidity of the mating surfaces, paired with mutual weak interactions. Within this framework, the class of 2D layered materials61. Prototype examples for mechanical and tribological applications are 2D layered materials such as graphite, hexagonal boron nitride (h-BN), molybdenum disulfide (MoS2), and other dichalcogenides. Structural anisotropy of these elements, characterized by strong intralayer covalent bonding and weaker interlayer dispersive interactions, results in low interlayer friction and wear resistance. This state of affairs makes 2D materials promising candidates for achieving structural superlubricity in realistic mechanical systems. However, realizing this potential still requires a solution of many demanding problems standing in the way of scaling up of superlubricity to macroscopic contacts under ambient conditions.Indeed, the number of materials that can be thinned down to single layers has been widely increased since the first isolation of graphene monolayers Fig. . Mechani64. Importantly, the superlubric behavior was observed to sustain under ambient conditions and at high sliding velocities up to 25 m/s65. Besides, measurements carried out in microscale graphitic junctions of different sizes, showed a power-law scaling of kinetic friction with the contact area, and found an exponent of 0.35 confirming the incommensurability of superlubric contacts63. The next step toward macroscale superlubricity was attained using centimeter-long bichiral double-walled carbon nanotubes that exhibit the intrinsic interwall incommensurability66.Considerable progress in upscaling superlubricity was achieved in 2012 when this phenomenon was found for graphite samples of micrometer size67. To overcome this issue, heterogeneous junctions (heterostacks) formed, e.g., between graphite and hexagonal boron nitride (h-BN) have been suggested theoretically70, predicting that even for aligned contacts the heights of potential barriers toward sliding significantly decrease above a certain contact size. This phenomenon results from the intrinsic mismatch of the interlayer lattice periodicities in hetero contacts, which leads to the formation of moiré superstructures, and to a transition from stick-slip motion to smooth soliton-like sliding with increasing contact size70. Thus, ultralow friction should be observed even for aligned heterojunctions, and dynamical surface reorientations will not eliminate superlubricity. Recent experiments supported these predictions, demonstrating superlubricity in the microscale heterogeneous contacts of graphite and hexagonal boron nitride for all mutual surface orientations71. Furthermore, the heterogeneous contacts may exhibit ultralow friction for significantly higher normal loads, than the homogeneous ones70. These experimental and theoretical findings show that van der Waals heterostructures hold great promises for dry tribological applications, which require ultralow friction and wear.One of the major obstacles in realizing robust superlubricity at homogeneous graphite-like contacts is their possible spontaneous reorientation, locking them back into commensurate high-friction configurations34 and those realized during interwall dynamics in multiwalled nanotubes73, or in sliding and rolling of carbon nanotubes on substrates75, or even involving confined C60 molecules acting as nanoscale bearings76.Less ideal forms of interface geometries may also, under suitable operative conditions, exhibit structural superlubricity. Concepts related to interface incommensurability and mechanical interlocking are still called into play when dealing with more complex mechanical contact configurations, as, for example, contacts between crystalline and amorphous surfaces82. This threshold depends on the ratio between the intra-surface and interfacial stiffness and on the imposed driving procedure. First, experimental verification of this prediction was reported recently for antimony particles sliding atop MoS283. For 2D layered materials the effects of elasticity can become important already in microscale contacts. Undesirable elasticity effects can be diminished using multi-contact interfacial configurations, in which a macroscale junction is divided into a large collection of decoupled or weakly coupled, randomly oriented small-scale patches86. The elasticity effects can be also diminished by depositing the van der Waals material coatings on rigid substrates. This configuration prevents deformation of the contacting sliding layers therefore hampering the onset of local partially commensurate areas and reducing considerably frictional energy dissipation.Another intrinsic hindrance preventing immediate upscaling superlubricity comes from the mechanical contact elasticity at the slider/substrate interface. It has been theoretically anticipated that even for an incommensurate matching there exists a threshold size of contact, above which the elastic deformations may become sufficiently large to locally bring regions into a commensurate mating configuration Fig. , leading46. Due to their low confinement, the edge bonds are more flexible than those located within the bulk of the contact area, and thus may undergo larger distortions during sliding87. Thus, the contact edges may act as pinning sites during sliding, enhancing frictional energy dissipation, and impeding superlubricity. Computational studies revealed that for incommensurate finite contacts, the edge atoms, more compliant to the action of the normal load, become easily pinned to the substrate88. In addition, under ambient conditions, exposure of edges to contaminations is inevitable, which also boosts frictional dissipation. Interestingly, these investigations demonstrated that at heterogeneous interfaces, e.g., between graphene and hexagonal boron nitride, the compliance and pinning of edge atoms under high normal loads is significantly reduced compared to the case of homogeneous graphitic contacts70.Under realistic conditions, where the contacts always have a finite size, the total friction is contributed not only by the surface atoms located within the contact area, but also by those at the contact edges89. The grain boundaries typically induce large out-of-plane surface deformations that may degrade or even completely eliminate the superlubric behavior. Understanding the effects of edges and grain boundaries on the frictional dissipation, and the mechanical response in general, in layered material contacts is crucial for achieving large-scale superlubricity, but this information is still lacking, and it is hard to obtain from measurements with solid contacts and from simulations.Scaling up structural superlubricity toward the macroscopic world is further challenged by the often polycrystalline nature of layered materials at larger scales, where the contacting surfaces often exhibit a mosaic of randomly oriented grains that are separated by grain boundaries91. The contaminant-related friction may be diminished by surface heating, which leads to desorption of adsorbates93. Alternatively, in situ interface cleaning can be achieved by applying mechanical oscillations to the frictional contact, which can significantly reduce the concentration of contaminants and restore superlubric regimes95. Recent experimental and computational studies of structural superlubricity under ambient conditions suggested that contaminants are pushed out from the contact formed by mesoscopic gold islands and graphite, thus providing a robust ultralow friction state36. It is still not clear how general is the observed phenomenon, but the search for self-cleaning incommensurate contacts that repel contaminants may open up a possibility for realization of structural superlubricity under ambient conditions.In addition to the discussed above intrinsic factors, which can suppress superlubricity, there are a number of external factors that can also hinder the superlubric regime. In this respect, it was found that a presence of contaminants, which are inevitable under ambient conditions, often leads to finite static friction and an increase of kinetic friction at the nominally incommensurate interface97. Therefore, it is important to determine the conditions under which the ultralow friction and wear are maintained for loads and velocities used in realistic applications.High normal loads applied to frictional contacts and high sliding velocities constitute other extrinsic factors limiting superlubricity. Normal loads and driving velocities explored in most friction measurements with nano- and microscale junctions of layered materials are significantly smaller than those usually applied in macroscopic tribological contacts. Obviously, increased friction and wear should be observed when the normal load and driving velocity exceed some threshold values that depend on the contact properties98. Most tribological systems show a logarithmic dependence of friction on the sliding velocity, which results from thermally activated crossing of sliding potential barriers3. Although such barriers are significantly suppressed in incommensurate contacts, the logarithmic velocity dependence of friction was also found in some superlubric systems71. This effect may originate from the contributions of edges, grain boundaries, and interfacial contaminants, however, more experimental and simulation studies are required to address this problem, which is important for both fundamental understanding of mechanism of superlubricity and for applications.If contact pressure may in general favor interface locking and friction, particularly acting on the compliant edge boundaries of a finite-size system, on the other hand, an increase in normal load can also suppress out-of-plane surface deformations, especially relevant for 2D layered materials, thereby inhibiting a dissipation channel and leading to a negative friction coefficientThe ubiquity of friction and dissipative related processes, together with central correspondences regardless of the specific system and length scale, has recently stimulated novel experimental approaches and theoretical formulations capable of setting a common framework toward an understanding of these phenomena which is independent of the length scale and the specific choice of the involved materials. Such ongoing quest for the “holy grail” of a unifying approach to friction may undisputedly benefit from investigations at the small microscopic level, with sliding mechanical contacts now well-characterized and operating under well-designed experimental conditions.100, such as the Prandtl–Tomlinson and the Frenkel–Kontorova type of models, relying on a firm fundamental background but whose direct experimental testing has been locked away for several decades.In this scenario, the prediction of the system’s universal physical response is primarily based on interface geometrical features and relative interaction considerations, more than upon distinct system-dependent characteristics (such as the specific governing forces and their range). Accordingly, tribological experiments measuring how general parameters, e.g., substrate corrugation versus interlayer stiffness, lattice mismatch, shear velocities, and driving forces impact the frictional behavior, do become crucial. The most promising theoretical tool to understand system-independent general properties of friction are simplified ‘mechanical’ approaches101.In this view, out of serendipity, the field of microscopic tribology can now benefit from the experimental opportunities offered by subjecting nano/microparticles to artificial optical potentials, as shown in Fig. 102, exploiting the versatility of these novel approaches has allowed to cast new light on elemental interface processes at the single-particle level with unprecedented real-time resolution, avoiding interpretative pitfalls arising from indirect or ex situ characterization of contact surfaces.While atomic force microscopy, surface-force apparatus, and quartz-crystal microbalance, workhorse techniques in experimental nanotribology, do measure the system rheological and dissipative response in terms of crucial, but averaged, physical quantities105. Such experiments would allow for the direct comparison with theory and numerical simulations of the Frenkel–Kontorova model which make detailed predictions regarding the motion on a single-particle level. Recently, experiments with laser-cooled ions have been accomplished where one-dimensional chains of ions were driven across periodically modulated potentials52, the latter provided, e.g., by spatially modulated electrical fields or another nearby chain of interacting ions. Apart from resolving the single-particle dynamics, this approach has another important advantage because it allows to vary the interaction between the sliding chain with the substrate in situ. This will be important, e.g., in the context of the Aubry transition as will be discussed further below.In order to resolve the single-particle dynamics of monolayers sliding across corrugated surfaces, it has been suggested to perform experiments with cold ions in electro-optical fields106. Owing to the colloidal length scale and the presence of a viscous bath, however, their trajectories can be conveniently tracked in space and time with optical methods107. Compared to atoms and ions, interactions between colloids can be tailored, e.g., by functionalization of their surfaces with active end groups or by adding magnetic clusters during their fabrication process108. Therefore, colloidal pair interactions can vary between a short-ranged hard-sphere like behavior to more long-ranged forces as screened Coulomb or dipolar pair potentials. Similar to ions, colloids strongly couple to external fields offering the opportunity to taylor artificial substrates with a high degree of flexibility regarding the geometry, lattice spacing and importantly, also the strength of interaction115.Colloidal suspensions, i.e., micron-sized particles suspended in simple fluids, provide another approach to study nanotribological effects with high spatial and temporal resolution. Similar to atoms, they can form crystalline solids or glasses whose properties closely resemble those of their atomic counterparts25 and references therein) stands nowadays at the core of the superlubricity phenomenon in frustrated incommensurate systems. In the framework of the infinite, 1D incommensurate FK model, where it was originally derived, the transition takes place at a critical value of the ratio between the substrate corrugation and the intra-chain stiffness  in Fig. 119, of the adsorbed  layer relative to the underneath  substrate120.While Aubry restricted his prediction to 1D contacts at zero temperature, recent experiments even demonstrate the validity of his ideas when considering extended 2D interfaces at room temperature Fig. , which m121.In addition to colloidal monolayers, the Aubry transition may be also observed in other geometrically well-characterized soft-matter systems. However, it should be noted, that with increasing softness, additional effects such as lubrication may dominate their friction behavior124. Similar observations have been obtained in colloidal systems forced over topographical or optical periodic landscapes. When an isolated colloidal particle is driven across a periodic surface with 4-fold rotational symmetry, its motion becomes strongly locked to the substrate which results in a devil’s staircase structure when plotting the direction of the particle motion vs. the angle of the external force126.When single or weakly coupled (quasi)particles are driven across a substrate potential, they do not necessarily follow the direction of the applied force. Instead their direction of motion becomes strongly locked to the high symmetry axes of the substrate lattice. Evidence of such directional locking is found, e.g., in type II superconductors. When an external magnetic field penetrates the material a hexagonal lattice of localized flux bundles forms. In the presence of a lattice created by defects, the flux bundle motion shows a clear dependence on the direction of the driving force relative to the symmetry directions of the defect grid128. Depending on the particle size and the lattice spacing, this results in corridors along the high-symmetry directions of the substrate and thus explains the above mentioned devil’s staircase hierarchy. Interestingly, the presence of thermal noise does not lead to the smoothing or even disappearance of the plateaus but to statistical jumps between different well-defined locking states129. In general, such locking-effects strongly depend on the interaction of the particles with the substrate. Accordingly, the phenomenon can be exploited for the sorting of colloids131, living cells132, and other macromolecules134.Directional locking can be understood by considering the corridors of lowest energy of a driven particle moving across the substrate potential136. Numerical simulations demonstrate that when planar chiral particles are driven across a periodic substrate with square geometry, objects with opposite chirality move along orthogonal directions. In the presence of a periodic external driving force, even motion into exactly opposite directions can be achieved137.Particle sorting by directional locking is not limited to spherical particles but has been also demonstrated in case of non-spherical objects. In particular for chiral particles, i.e., when the particle shape is not superimposable with its mirror image, efficient sorting mechanisms of both enantiomers  are important to separate, e.g., organic and inorganic chiral molecules whose pharmacological and toxicological effects may strongly depend on their chirality138.Directional locking is not only affecting the direction of motion of single particles but can also affect the structure of a monolayer of interacting particles driven across a patterned surface. This has been demonstrated in case of electrostatically interacting colloidal particles driven across a quasiperiodic substrate potential with decagonal symmetry. Opposed to the situation where no external drive is applied and the particles follow the symmetry of the underlying substrate potential, numerical simulations and experiments demonstrate square, smectic, and also Archimedean-like states in the driven phases139, directional locking is also observed in nanoscale manipulation and numerical simulations of Au-clusters on layered materials141 as well as in mesoscopic experiments with rigid clusters comprised of several tens of colloids143 (Fig. In addition to individual or weakly interacting particles143 Fig. . ContrarIn this bird’s eye Perspective, we hope to have conveyed the flavor of an exciting research field, that of nano and microscopic friction and related out-of-equilibrium dissipative phenomena, which is alive and well.With the interplay of experiments, theory and simulations, recent advances are currently pursued in a multifaceted approach, starting from the fundamental atomic-scale mechanisms at single-asperity level and then scaling up to interface phenomena in extended well-characterized nano and microscale contacts. Simultaneously, the novel exploitation of mesoscale tunable trapped cold ions and colloidal particles under shear is indeed going to shed light with unparalleled accuracy on several key tribological processes. Last but not least, noteworthy developments in synthesis and fabrication procedures allow now to extend the characterization of well-defined tribological interface, especially dealing with layered materials, up to the millimeter range and beyond, with direct possible implementation in the manufacturing of state-of-the-art mechanical devices. Unquestionably, a full understanding and control of frictional processes is now recognized to involve all relevant size and time scales.The two lines of research outlined here, i.e., hard 2D layered materials and soft-matter systems, can indeed help to identify unexplored energy dissipation mechanisms and to provide comprehensive testing grounds for theoretical predictions with the overall aim to deepen our basic knowledge in this important discipline. The possibility to compare system- and scale-independent hard- and soft-matter interfaces brings about the unique chance to disentangle material-specific and geometrical aspects of the interfaces which allows a detailed view on friction-related phenomena and largely extends our basic knowledge in this field. Further studies in this direction can make a significant contribution to solving one of the most challenging problems in the field of tribology, i.e., bridging the scale gap between the microscopic and the macroscopic frictional mechanisms occurring at a sliding interface.With their fundamental relevance toward practical engineering applications, the framework of 2D materials may strongly benefit from the synergic complementarity with the realm of soft tribology emulators, which offers the exceptional possibility to visually resolve the intimate nature of both structural features and dynamical mechanisms of sliding friction down to the individual particle level. Differently, the usual state-of-the-art tribological techniques provide experimental measurements typically in terms of crucial, yet averaged, frictional quantities, which make systematic inroads unfortunate when paired with the recognized difficulty of tuning the relevant physical parameters in condensed matter tribology.Such “soft-hard” experimental synergy, together with modeling and numerical simulations, may thus contribute in drawing the roadmap to tackle the great scientific and practical challenge of addressing some aspects of the complex nature of frictional phenomena.71, most microscopic measurements aiming at understanding basic tribological mechanisms are thus performed over short sliding distances. Wear is expected to be low but not completely eliminated even in the superlubric sliding regime. Thus, in order to address technological challenges, sustainability of superlubricity at long sliding distances, high normal loads, velocities, and temperatures must be experimentally studied.Setting the ground to robustly scale structural superlubricity up, transposing the desired peculiar nanoscale properties to larger tribological scales and recognizing the conditions of its persistence and the mechanisms of its breakdown, surely remains on the list of outstanding issues that have to be addressed thoroughly. With the aim of controlling frictional-induced energy dissipation in real-life sliding devices, inherent contact elasticity, grain boundaries, surface defects, and interface contamination are, and always will be, unavoidable key elements to consider when moving from the nanoworld up to the macroscopic domain. Besides, although some experiments have demonstrated wearless and superlubric motion for sliding distances of a few mmLively progress along these and newer lines is to be expected in the near future.


Tumor vascularization occurs through several distinct biological processes, which not only vary between tumor type and anatomic location, but also occur simultaneously within the same cancer tissue. These processes are orchestrated by a range of secreted factors and signaling pathways and can involve participation of non-endothelial cells, such as progenitors or cancer stem cells. Anti-angiogenic therapies using either antibodies or tyrosine kinase inhibitors have been approved to treat several types of cancer. However, the benefit of treatment has so far been modest, some patients not responding at all and others acquiring resistance. It is becoming increasingly clear that blocking tumors from accessing the circulation is not an easy task to accomplish. Tumor vessel functionality and gene expression often differ vastly when comparing different cancer subtypes, and vessel phenotype can be markedly heterogeneous within a single tumor. Here, we summarize the current understanding of cellular and molecular mechanisms involved in tumor angiogenesis and discuss challenges and opportunities associated with vascular targeting. Malignant cells require oxygen and nutrients to survive and proliferate, and therefore need to reside in close proximity to blood vessels to access the blood circulation system. The early observation that rapidly growing tumors were heavily vascularized, while dormant ones were not, led Judah Folkman to propose that initiation of tumor angiogenesis was required for tumor progression . FurtherSmall dormant tumors that are devoid of active blood vessel formation can frequently be observed in human tissue and in genetically engineered mouse models of multistage carcinoma at early stages of cancer progression. Tumor progression is often accompanied by ingrowth of blood vessels, consistent with a need for malignant cells to have access to the circulation system to thrive. Tumors can be vascularized either through co-option of the pre-existing vasculature , or by iThe blood circulation system is critical in delivering nutrients and chemicals to tissues, removing waste products, and maintaining homeostasis. The vascular system, composed of the aorta, arteries, capillaries and veins transports blood throughout the body. Arteries carry blood away from the heart, transporting oxygenated blood to the tissues. The capillary networks have narrow walls that help in gas exchange between the blood and tissues. Oxygen is released into the tissues and carbon dioxide is absorbed by the blood, and is transported back to the heart through veins. Transmigration of immune cells into tissues is facilitated by post-capillary venules. The capillary wall is made of an endothelial cell layer surrounded by a basement membrane and is supported by pericytes. Angiogenesis is typically initiated from the capillaries and it plays an important part in tumor growth, maintenance and metastasis. Blood vessel formation in tumors can be induced through several cellular processes Fig.  as briefNew capillaries can bud from parental vessels through a multi-step process known as sprouting angiogenesis. Formation of sprouts involves (a) Tip cell selection: a cell from the parent vessel becomes the migratory leading cell and blocks its neighboring cells from adopting a tip cell fate by a lateral inhibition process. (b) Sprout extension: the tip cell migrates along the chemotactic path, followed by trailing stalk cells and (c) Lumen formation: connection of the luminal space of the sprout with the parent vessel. The developing sprout then connects with other vessels through a process called anastomosis.Endothelial cells are normally quiescent, but can be induced to sprout and initiate angiogenesis by pro-angiogenic factors including vascular endothelial growth factor (VEGF). Tip and stalk cell selection is regulated through cross-talk between the VEGF and Dll4/Notch pathways . In respA less studied process of neo-angiogenesis is “intussusception”, where transluminal tissue pillars develop within existing vessels and subsequently fuse to remodel the vascular plexus, first described in remodeling of lung capillaries , 30. The+ EPCs from the bone marrow . H. H245]. The capability MDSC regulating tumor angiogenesis is similar to M2-like TAMs. MDSCs promote and sustain tumor angiogenesis primarily by secretion of MMPs. In particular, MMP-9 is known to boost angiogenesis and stimulate tumor neovasculature by increasing the bioavailability of VEGF . This inNeutrophils are the most abundant leukocyte population, providing the first line of defense against invading pathogens. They are a rich source of soluble factors such as ROS, peptides, cytokines and enzymes that exert antimicrobial activities . NeutropConclusive evidence of neutrophils involvement in tumor angiogenesis came from studies in the RIP1-Tag2 multi-step pancreatic carcinogenesis mouse model. Neutrophil depletion using anti-GR1 antibodies reduced the number of dysplastic islets that were undergoing angiogenesis . In addiThere are three major types of lymphocytes, namely T cells and B cells, which constitute the adaptive immune system and NK cells, which are part of the innate immune system. The contribution of lymphocytes towards tumor angiogenesis is not as well understood, as that of myeloid cell types.brightCD16−KIR+, dNK cells), characterized by poor cytotoxicity and pro-angiogenic capacity have been identified in the decidua during pregnancy. They secrete VEGF, placental growth factor (PlGF), IFNγ, IL10 and CXCL8 that are critical for spiral artery formation and decidual vascularization [A subset of NK cells  . HoweverThe concept of targeting angiogenesis as a means to starve tumors was introduced by Judah Folkman and colleagues 48 years ago . Since tResistance to anti-angiogenic therapy is an important issue that likely explains the variable response in different types of tumors and the limited overall survival benefits. Resistance can be classified into intrinsic resistance, observed from the outset of the therapy, and acquired resistance, observed after an initial positive response to therapy . SeveralAs already mentioned, anti-angiogenic therapy can promote tumor invasion and metastasis in pre-clinical cancer models, which might be triggered by increased hypoxia due to vessel depletion. Indeed, the transcription of HIF-regulated genes controls different steps of tumor invasion and metastasis, including EMT, activation of MET signaling, recruitment of stromal cells, vascular mimicry and vessel co-option . Vessel Recruitment of stromal cells, immune cells and progenitors is another potential mechanism for resistance to anti-angiogenic therapy. In particular, many studies have pointed out an important role of bone marrow derived cells (BMDCs) in this aspect. Recruitment of BMDCs in glioblastoma can cause resistance to vatalanib treatment and the depletion of BMDCs can potentiate the effects of this anti-angiogenic drug . ReleaseIn addition, alternative pro-angiogenic signaling pathways including ANGPT-2, FGF-2, IL-8 can be induced by tumor cells in response to a pharmacological inhibition of the VEGF signaling pathway . In receAlthough anti-angiogenic drugs were initially designed to block blood vessel formation, their ability to control tumor growth may be due to several different mechanism, which are not mutually exclusive. To improve vascular targeting, a thorough understanding of the cellular and molecular mechanisms that hinder tumor progression in response to anti-angiogenic therapy in specific tumors is necessary. The possible mechanism of actions of angiogenesis inhibitors on tumor blood vessels can be broadly classified into three categories: (a) vessel depletion, (b) vessel normalization, and (c) immune activation Fig. .Fig. 3EfThe development of anti-angiogenic drugs was initiated by the hypothesis that starving tumors by blocking angiogenesis would slow tumor progression and improve patient survival . Early pSeveral studies demonstrate that before reaching complete depletion of the vascular bed, anti-VEGF drugs induce an early and transient phase in which vessels assume normal shape and function , 307. ThDespite a high vascular density, tumors are usually hypoxic and nutrient-deprived since the tumor vessels are abnormal, leaky and malfunction. Such abnormal vasculature significantly compromises the efficacy of most anti-cancer therapies by limiting the delivery of drugs as well as promoting resistance to treatment.The vessel normalization hypothesis, introduced by Rakesh Jain in 2001  suggestsEvidence that support the notion that vessel normalization occur in response to anti-angiogenic therapy has also been obtained from clinical studies. The functionality of the tumor vasculature in glioblastoma patients treated with anti-VEGF therapies has been evaluated by magnetic resonance imaging (MRI). MRI analysis of patients treated with cediranib revealed a decrease in vessel diameter, vascular permeability, and edema. More importantly, survival of patients with recurrent glioblastoma following cediranib-treatment was found to correlate with a vascular normalization index . ImprovePro-angiogenic factors in tumors induce down-regulation of adhesion molecules on endothelial cells in the tumor vasculature and induce anergy to inflammatory signals such as TNFα and IL-1. Hereby, tumors with an angiogenic phenotype may escape the infiltration of cytotoxic leukocytes . Using aInhibition of VEGF signaling in the tumor microenvironment may be beneficial not only in terms of improving immune cell recruitment, but can also directly improve immune cell activation. Normalization of the tumor vascular network and decreased hypoxia can promote T cell infiltration and induce polarization of TAM to an M1-like phenotype . Anti-anTumor vessels are often dysfunctional and anergic to inflammatory stimuli, leading to a hostile tumor microenvironment that fuel cancer progression and aggravate therapeutic approaches. Current vascular targeting strategies are based on inhibition of key angiogenic signaling pathways known to promote tumor angiogenesis. Although several anti-angiogenic drugs have been approved, intrinsic and acquired resistance to therapy limit their efficacy. An increased understanding of tumor vessel phenotype and mechanisms involved in treatment response and resistance to therapy is necessary to overcome the hurdles that prevent successful control of the angiogenic response in tumors. Alternatively, vascular targeting should instead be designed to target the tumor vessels in new ways that are conceptually different from inhibition of angiogenesis. This may involve altering the timing and dosing of already existing anti-angiogenic therapy in combination with other drugs, or development of novel therapeutics to either directly target the tumor vessels or optimize their function to fit the cancer therapy at hand.The fact that tumor vessels differ molecularly from their normal counterparts can be used to develop treatment strategies that specifically target malignant cells and tumor vasculature. Therapeutic vaccination strategies to raise endogenous antibodies against antigens specifically expressed by tumor vasculature have shown efficacy in pre-clinical cancer models . Prophylhttp://clinicaltrials.gov) [+ T cells induced HEV development within the tumor after Treg depletion [Going beyond anti-angiogenesis and vascular normalization, strategies that can alter vessel phenotype to optimize specific types of cancer therapy are quickly emerging. It is already established that targeting VEGF/VEGFR signaling can enhance the efficacy of cancer immunotherapy by increasing expression of adhesion molecules and chemokines necessary for capture and transendothelial migration of T-cells , 328. Seals.gov) , 328. Toals.gov) . Depletials.gov) . The preepletion . Consistepletion . Specifiepletion . With reepletion . The obsepletion . This ex


Premenopausal women generally have a favorable cardiovascular risk profile, owing to young age and the protective effects of estrogen. Rates of hypertension and more advanced cardiovascular disease (CVD) are low in premenopausal women. A large body of epidemiological evidence has shown that lifestyle behaviors in midlife, i.e., cardiorespiratory fitness, physical activity, and healthy diet, are associated with lower risk of overt CVD and adverse cardiovascular outcomes in the future for men and women. Despite differences in future cardiovascular risk, brachial blood pressures might be similar between premenopausal women with favorable vs. unfavorable levels of lifestyle behaviors in early-to-mid-life. Here we make the case for deeper phenotyping by means of vascular function measurements, such as arterial stiffness, augmentation index, and endothelial function, to identify potential mechanistic pathways linking lifestyle behaviors in early-to-mid-adulthood with lifelong CVD risk in women. We describe considerations for vascular function measurement in premenopausal women and opportunities for investigators to fill in knowledge gaps to further our understanding of CVD risk assessment and CVD progression in premenopausal women. Lifestyle interventions sometimes do not elicit significant improvements in brachial blood pressure (BP) in premenopausal women , 2. ReasWe propose that more sensitive measures of vascular structure and function are needed to understand how lifestyle influences CVD risk and better evaluate the effects of lifestyle behaviors and lifestyle interventions in premenopausal women. Some studies that employed more sensitive vascular measurements pre- and post- exercise intervention, such as central BP, arterial stiffness and distensibility, carotid intima-media thickness, or ventricular-vascular coupling, reported improvements in some of these measures while brachial BP stayed the same , 2, 8–11Ascertainment of hard outcomes, i.e., cardiovascular events, morbidity, and mortality, can guide clinical decision making and are used for evaluation of intervention success . HoweverWith the exception of coronary artery calcium, the value of novel or subclinical CVD risk factors, such as poor endothelial function (measured using flow-mediated dilation), systemic inflammation (c-reactive protein), and ankle-brachial index, beyond traditional CVD risk factors for identifying individuals at high-risk for atherosclerotic CVD endpoints has not been consistently supported , 29. HowImportantly, the atherosclerotic CVD risk prediction models were calibrated to identify patients at risk for atherosclerotic, but not myocardial, CVD . A new r2. Augmentation index has been shown to be lower in the late luteal phase when progesterone is at its highest and estrogen is above the levels found in the early follicular phase  might further complicate vascular assessment in premenopausal women and requires thoughtful consideration. While arterial stiffness was similar in women who do and do not use OCP, brachial and central BPs were higher during the non-placebo phases of the pill cycle in women who use OCP , 18, 19.Premenopausal women may also become pregnant. Pregnancy is associated with profound cardiovascular changes in women, and pregnancy-related pathologies, such as hypertensive disorders of pregnancy, intrauterine growth restriction, and preterm birth, are associated with excess lifelong CVD risk . Given tBest practices for collecting vascular structure and function measurements during reproduction have not been established. The current recommendations for assessment of vascular function in non-pregnant adults require an overnight fast preceding testing and specify that vascular testing should be performed with the participant in the supine position , 39. A lPregnant women are advised to avoid lying supine to prevent the weight of the baby and uterus from compressing the vena cava. Lying supine for a 30–45 min testing session past the first trimester would not be acceptable for fetal health and might cause a drop in maternal BP. We propose the use of a reclining, padded table set at a standard 45° angle and conducting testing 1–2 h after a light meal that is low in vasoactive saturated fat and refined carbohydrates . The repEarly menopause and conditions related to subfertility have also been associated with higher risk of overt CVD in women , 42. AntWomen diagnosed with polycystic ovarian syndrome (PCOS), an endocrine disorder characterized by high AMH levels, hyperinsulinemia, amenorrhea or oligomenorrhea, and hyperandrogenism, also present with unfavorable lipid profiles, hypertension, subfertility, higher fasting insulin levels and arterial stiffness, and have a higher risk for developing CVD , 44. StuGiven their generally healthy traditional CVD risk factor profile, sensitive assessments of vascular structure and function are needed to evaluate the success of lifestyle interventions in premenopausal women. Our understanding of CVD development and progression is lacking in women. Tracking changes in vascular function in response to lifestyle interventions, as well as evaluating associations of vascular structure and function with easily measured sex-specific biomarkers, will critically inform preventive and treatment strategies and risk stratification in women. Cycling hormone levels, OCP use, pregnancy and breastfeeding, and conditions linked to subfertility present challenges to achieving reproducible vascular measurements in premenopausal women and/or caveats for the application of findings of smaller studies. Rather than ignoring potentially important factors related to ovarian hormone levels or excluding women from studies, we suggest that investigators carefully consider these variables when planning research and systematically fill in the knowledge gap. Our specific recommendations for research are as follows:Sensitive measures of vascular structure and function should be included in clinical trials involving different types of lifestyle behaviors, i.e., resistance interval and endurance exercise training, weight loss, in premenopausal women.Studies should account for the menstrual cycle phase and perform vascular data collection during the early follicular phase when possible.Investigators should gather information regarding type and length of oral contraceptive use to use in main analyses or sensitivity analyses.Vascular measurement protocols for data collection during pregnancy and post-partum, including meal timing and body posture, must be developed and standardized.More lifestyle-based clinical trials should be conducted in women with POI and PCOS to better understand the role of lifestyle in mitigating the excess CVD risk observed with these conditions.The original contributions presented in the study are included in the article/supplementary material, further inquiries can be directed to the corresponding author/s.AL-C and SB drafted the manuscript, prepared the table, and critically edited the manuscript. All authors contributed to the article and approved the submitted version.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Sepsis is a major cause of acute kidney injury (AKI) among patients in the intensive care unit. However, the numbers of basic science papers for septic AKI account for only 1% of all publications on AKI. This may be partially attributable to the specific pathophysiology of septic AKI as compared to that of the other types of AKI because it shows only modest histological changes despite functional decline and often requires real-time functional analysis. To increase the scope of research in this field, this article reviews the basic research information that has been reported thus far on the subject of septic AKI, mainly from the viewpoint of functional dysregulation, including some knowledge acquired with multiphoton intravital imaging. Moreover, the efficacy and limitation of the potential novel therapies are discussed. Finally, the author proposes several points that should be considered when designing the study, such as monitoring the long-term effects of the intervention and reflecting the clinical settings for identifying the molecular mechanisms and for challenging the intervention effects. Acute kidney injury (AKI) is an important challenge that negatively impacts patient survival in an intensive care unit (ICU). Although there was a 9% decline in the in-hospital mortality rate among patients treated with renal replacement therapy in the ICU from 2007 (44.9%) to 2016 (36.1%) in Japan, the mortality rate of sepsis patients remains high at > 50% [Escherichia coli infusions ameliorated both oliguria and serum creatinine levels, despite reduced renal blood flow in sheep [Changes in the GFR, from the viewpoint of hemodynamic change, are currently investigated on relatively big animals, such as dogs and sheep, because it is easier to create ICU-like experimental settings for these animals as compared to that for smaller animals, such as rodents, and to perform continuous inulin/inulin-like compound-used clearance analyses. The former may be essential for the replication of a hyperdynamic state-like human septic AKI . Changesin sheep . Studiesin sheep , medullain sheep , and mitin sheep  during sin sheep , therebyIn addition to the afferent/efferent arteriole-dependent intraglomerular pressure changes, Xu et al.  reportedThe prerenal causes of GFR reduction were often resolved by fluid and vasopressor treatment although this might not be the case if sepsis increased the systemic vascular permeability. The optimization of fluid resuscitation for sepsis or septic AKI in clinical studies has been debated in multiple review articles –19. HoweAKI could occur with oliguria and without a rise in the serum creatinine level. The unresponsiveness of serum creatinine could result from the dilution of creatinine by fluids , 26 or vThe renal capillary network was often destroyed during non-septic AKI , and angAs noted above, AKI is currently diagnosed via the GFR or urine output, and it is important to ensure patient survival during the acute phase. The pathological features of AKI differed, depending on the AKI cause. Renal ischemia/reperfusion injury that is widely employed as an AKI model induces dramatic changes in the renal histology. Conditions related to renal ischemia/reperfusion injury include acute tubular necrosis, cell sloughing, cast formation, perivascular inflammation , capillaStandard supportive treatments, such as fluids and vasoconstrictors, are used to restore the GFR; however, they do not address the changes in the tubules in the septic AKI cases. In addition, there are concerns surrounding fluid resuscitation –47. TherIschemic preconditioning is a possible strategy against perioperative end-organ damage, including AKI. Repeated short-time hypoxic/ischemic stress to the kidney has caused a tolerance to AKI in multiple experimental models, including sepsis –51, ischSalmonella abortus equi endotoxin [Endotoxin preconditioning involves the injection of a single low-dose endotoxin, including LPS, to protect the organs from subsequent events. Experiments on pigs have reported that endotoxin preconditioning suppressed the systemic oxygen demand despite unchanged global hemodynamics in response to the ndotoxin . Althougndotoxin . Moreovendotoxin . The benndotoxin , 69 and ndotoxin . Howeverndotoxin  and lupundotoxin . Chen etndotoxin  demonstrndotoxin . For insndotoxin .Metz et al. developed a concept that has been termed the cholinergic anti-inflammatory pathway , mediateAtrial natriuretic peptide (ANP) is secreted from the atrium following reduced cardiac function. Endogenous ANP secretion is increased during human septic shock  and in tPickkers et al. performed several studies using the endotoxin challenge in humans –111 and The current limitations of these therapeutic candidates are summarized in Table As much as possible, studies to find the pathogenesis/molecular mechanism should additionally examine the effect of factors existing in the clinical setting. Young rodent tests might show different results than older mice. Use of rodents might benefit the modification of genes (and are less expensive); however, the hemodynamic pattern does not reflect the clinical contribution of the target molecule that might be less or more when in actual clinical use. Septic patients are usually under antibiotic therapy and fluid administration , and thiIntervention studies must confirm either mortality rate or CKD development because these are the objectives of the therapy. Attenuating the injury only at one time point does not guarantee that the treatment could prevent patient mortality.As far as possible, intervention studies should reflect the clinical setting. Anti-immune therapy might suppress host defenses. Thus, even if it works against sterile inflammation, it might worsen bacteremia and increase mortality.Septic AKI is associated with considerable hospital-based mortality; once a novel treatment method has been developed, it could save thousands of lives. Recent developments using artificial intelligence have enabled the prediction of “new” antibiotics . However


This article provides an overview of both established and innovative applications of femtosecond (fs)-laser-assisted surgical techniques in ophthalmology. Fs-laser technology is unique because it allows cutting tissue at very high precision inside the eye. Fs lasers are mainly used for surgery of the human cornea and lens. New areas of application in ophthalmology are on the horizon. The latest improvement is the high pulse frequency, low-energy concept; by enlarging the numerical aperture of the focusing optics, the pulse energy threshold for optical breakdown decreases, and cutting with practically no side effects is enabled. This effect is called “photoablation”. In ophthalmology, it is applied to change the curvature of the cornea with pulsed UV light from excimer lasers. For shorter pulse durations in the ps to fs range and even higher intensities above 1011 W/cm2, more exotic interactions can be achieved, as will be explained in detail below. A more comprehensive general overview of laser–tissue interaction mechanisms can be found in excellent quality in several text books . Al. Al30]. The stability of the vacuum docking contact during laser emission is of primordial importance. Loss of contact harbors the risk of cutting in wrong planes. Therefore, all lasers are designed to automatically monitor vacuum levels, sometimes complemented with imaging of the eye position (eye tracking), and to immediately stop laser emission upon loss of contact. Of course, the eye surgeons also monitor their patients during the procedure and can manually interrupt or temporarily pause the treatment when they anticipate problems. In case of laser systems with an articulated arm, the surgeons can also use their substantial manual skills to actively stabilize the laser handpiece while in contact with the eye. In any case, after a vacuum loss, the treatment can usually be resumed immediately after a new docking.The human eye functions like the lens of a camera. Images are focused on the retina through a converging system composed mainly of the cornea. If the corneal curvature and thus its refractive power does not precisely match the axial length of the eye, refractive problems like near-sightedness (myopia) or far-sightedness (hyperopia) ensue . Refractive surgery consists of either reducing the refractive power of the cornea (by flattening) or by increasing its power (by steepening) or by modifying its curvature on a determined meridian to correct astigmatism .LASIKIn the laser in situ keratomileusis (LASIK) procedure, a corneal flap is created. The flap is lifted and then excimer- or solid-state UV-laser energy is used to change the cornea’s refractive power by flattening or steepening the stromal bed. Later, the flap is repositioned. Before the advent of fs-laser technology, the flap was created using mechanical devices called microkeratomes. With fs-laser technology, the flap can be completed in various patterns . KeziriaStromal keratophakia (additive refractive surgery)Keratophakia as a means to sculpt corneal curvature by adding tissue has been studied since 1949 by Barraquer . BecauseMultiple refractive surgery methods use fs-laser cuts to create “pocket”-shaped openings in the cornea, from which either material can be removed or implanted. In both cases, the refractive power of the cornea changes.Corneal stromal lenticule extractionWhile many fs-associated surgical interventions in ophthalmology are merely improvements of pre-existing techniques, lenticule extraction is unique to fs-laser technology: the procedure was introduced in 2011 to treat myopia, and later also myopic astigmatism. It became known under the brand name “SMILE”  of the Carl Zeiss Meditec AG. Later, other companies introduced their own laser systems for similar lenticule procedures under different brand names, including “SmartSight” by Schwind and “CLEAR”  by Ziemer Ophthalmic Systems AG.The procedure is a “flapless” laser refractive technique that uses a single femtosecond laser system to create a pocket. The content of the pocket—the lenticule—is removed via a small access tunnel incision, and as a result, the cornea is flattened see . InsteadIntrastromal corneal ring segmentsFs technology allows creating stromal pockets of specific size and shape at specific positions. Corneal ring segments  are placThe concept of corneal cuts for astigmatic correction was established more than 100 years ago  and undeBackgroundKeratoplasty (cornea transplantation) ranks among the oldest and most commonly performed human tissue transplantation types worldwide . A corneTrephinationA perfect trephination system produces a congruent recipient bed and donor buttons and thereby allows well-centered tension-free fitting, and efficiently waterproof-adapting incision edges . DiffereIt is often problematic to ensure proper centration with trephination in the recipient eye. Fs technology allows for perfect limbal oriented centration through OCT-visualization.Another problem with trephination is the mechanism by which the recipient eye and donor button are fixated and stabilized; any mechanical impact on the tissue during trephination causes compression and distortion and will decrease the fit of recipient and donor . Common Different stabilization systems could explain this superiority. While the excimer-assisted keratoplasty does not require applanation of the cornea, it is needed for the fs laser used in the cited studies. The new liquid optics interface assisted fs-Keratoplasty method could solve this problem: Here, cutting both recipient and donor is achieved within a liquid interface, which leaves the curvature of the cornea undisturbed. This reduces shear- and compression artifacts in the tissue and improves congruent fitting of the recipient and donor . It willSidecutsIn femtolaser-assisted keratoplasty (FLAK), different side-cut profiles can be chosen . TheoretBackgroundThe cornea is structured in five parallel layers. Often, not all layers of the cornea are diseased. Scars from trauma or infection commonly involve the anterior layers . In contrast, some inherited corneal dystrophies  affect only the inner most layers In deep anterior lamellar keratoplasty (DALK), approximately 95% of the anterior corneal layers are removed, and only the innermost layers, Descemet’s membrane, and endothelial cell layer stay behind . It is pPosterior lamellar keratoplastyPosterior lamellar keratoplasty has revolutionized corneal transplant surgery in the past two decades: the cornea’s clarity depends on healthy endothelial cells. These cells are thought of as non-regenerating highly specialized cells. In a disease called “Fuchs endothelial dystrophy”, but also after traumatic or multiple surgical interventions, these cells cease to do their job in maintaining corneal clarity and patients eventually become blind. Transplanting these cells, be it with a small amount of corneal stroma, so-called Descemet stripping endothelial keratoplasty “DSEK”, or with Descemet’s membrane alone as the carrier, so-called Descemet’s membrane endothelial keratoplasty “DMEK”, reverses the process of corneal opacification and especially in the case of DMEK causes perfect visual acuity ,65. PartOphthalmic surgeons have been trying to implement laser in cataract surgery for decades. Bille and Schanzlin proposed ultrashort laser pulses for ablation of the cataractous lens in 1992 . Nagy waThe main advantages of fs-assisted cataract surgeries are the precision and repeatability of laser incisions to the tissue; reduction in ultrasound energy used for emulsification (liquification) of the lens nucleus by precutting it into small pieces; perfect sizing of corneal incision with regards to position, length and depth; and predictability in capsulotomy size and position. Despite the aforementioned obvious advantages and numerous studies showing superiority in performing the single surgical steps over the ones manually performed by surgeons, meta-analysis studies could not prove overall outcome advantages of fs-laser-assisted surgery versus the conventional phacoemulsification manual operation ,72. A raThe question remains: Why should a high-precision system not be superior to the manual? Two answers merit consideration. (1) Several studies were done using the first-generation femtosecond laser systems comparing those to the conventional phacoemulsification surgery that has undergone evolution and perfection for several decades. Results of comparative studies using the newest laser devices could give a better comparison. (2) In most comparative studies, the conventional surgery has been performed by outstanding, high-volume, and exceptionally talented cataract surgeons. Comparing the visual outcomes of such surgeons to the machine results is like assessing the abilities and outcomes of first-generation autonomous driving systems to those of Formula-1 drivers. In the following, we describe each step of the cataract surgery taken over by the fs machine.Traditionally, cataract surgeons access the cataractous lens by manually opening the anterior lens capsule by pulling in a continuous curvilinear manner. This maneuver is performed using a needle or forceps and is called capsulorhexis. While the hard inner part of the lens is removed, the outer capsular bag is maintained. This bag is used as a mounting plate to fixate an implanted acrylic intraocular lens at the same position from where the cataractous lens was removed. Size, position, and shape of the capsulorhexis are related to the effective lens position, a determinant of the intraocular lens (IOL) power. The IOL power determines the postoperative refractive error of the eye. Inappropriate sizing of the capsular opening may result in IOL tilt, decentration, and increased posterior lens capsule opacification ,77,78. PWhile a perfect capsular opening is of the highest significance for surgical success, it is one of the most challenging maneuvers in cataract surgery. The learning curve of surgeons in training can be quite flat. At the same time, even in the hands of the most experienced surgeons, the capsulorhexis can be unpredictable and perfect sizing at the submillimeter precision appears only possible with machines performing the step. Furthermore, the manual outcome is dependent on the axial length of the eye, pupil size, image enlargement by the cornea , and inhThe human lens loses transparency and flexibility throughout life. A cataractous lens cannot be removed through a small incision by suction alone; rather, emulsification or fragmentation of the hard lens nucleus is necessary. Conventional cataract surgery uses ultrasound energy within the eye to liquefy the lens nucleus. Femtosecond laser technology allows precutting the nucleus in almost any imaginable shape and reduces the ultrasound energy needed for emulsification. This is an advantage as the ultrasound energy is a cause of oxidative stress, heat, and inflammation, and damage to the tissue . The mosFull-thickness incisionsFull-thickness incisions through the cornea are necessary for the introduction of instruments into the eye. Traditionally, a metal scalpel or diamonds are used for creating them in different sizes. Fs technology allows predictable sizing  of full-thickness corneal incisions. Since these incisions need to be self-sealing, a perfect wound architecture incision is mandatory to prevent wound leakage postoperatively. Incorrect positioning of the wound induces astigmatism and can provoke prolapse of the iris during the surgery. Studies have shown increased repeatability and safety of wound construction using fs technology, resulting in higher stability and water tightness ,90,91.Partial-thickness incisionsPartial-thickness incisions into the cornea help to reduce preoperatively existing corneal astigmatism. Several nomograms have been developed, addressing length, position, and depth of the incisions for different amounts of astigmatism. Fs technology allows higher predictability and repeatability of partial thickness incisions, or even completely intrastromal corneal incisions . Since tProbably the most important evolutionary step in ophthalmic fs-laser devices will be miniaturization, mobility, and versatility. Tools available soon are fs laser-assisted primary posterior capsulotomy and lens capsule marking for positioning of toric IOL. On the horizon, another technology involves changing the IOL power postoperatively through fs-laser energy to achieve emmetropia in all eyes .In summary, fs-laser technology has evolved over the past decades into a precise and reliable tool in ophthalmic surgery. While some of the applications have not finished evolving and require further research and development, fs-laser-assisted cataract and corneal surgery have reached highly standardized levels worldwide. For these surgeries, fs-laser technology has improved patient safety and clinical outcomes and opened gateways to new surgical approaches.