Determining the tempo and mode of non-avian dinosaur extinction is one of the most contentious issues in palaeobiology. Extensive disagreements remain over whether their extinction was catastrophic and geologically instantaneous or the culmination of long-term evolutionary trends. These conflicts have arisen due to numerous hierarchical sampling biases in the fossil record and differences in analytical methodology, with some studies identifying long-term declines in dinosaur richness prior to the Cretaceous–Palaeogene (K-Pg) boundary and others proposing continued diversification. Here, we use Bayesian phylogenetic generalized linear mixed models to assess the fit of 12 dinosaur phylogenies to three speciation models . We do not find strong support for the downturn model in our analyses, which suggests that dinosaur speciation rates were not in terminal decline prior to the K-Pg boundary and that the clade was still capable of generating new taxa. Nevertheless, we advocate caution in interpreting the results of such models, as they may not accurately reflect the complexities of the underlying data. Indeed, current phylogenetic methods may not provide the best test for hypotheses of dinosaur extinction; the collection of more dinosaur occurrence data will be essential to test these ideas further. The effet al.  = 614) and Lloyd et al. [n = 420), including two subtly different versions of the Benson et al. [et al. [et al. [et al. [Sakamoto et al.  used the [et al.  , ceratopsians [n = 27 and 30, respectively), hadrosauriforms [n = 62), sauropods [n = 87 and 76, respectively) and theropods [et al. [. Maximum and minimum possible ages came from the Paleobiology Database . Zero-length branches were lengthened by imposing a minimum branch duration of 1 Myr; [et al. [We expanded this comparison by using an additional set of nine recently published non-avian dinosaur phylogenies that include representation of all the major clades present during the late Mesozoic, including thyreophorans –39 (n = topsians  (n = 27 uriforms  (n = 62)auropods ,42 ), (ii) the slowdown to asymptote model, with node count modelled as a function of the square root of time elapsed from root to tip ), and (iii) the downturn model, with node count modelled as a function of time elapsed from root to tip and its quadratic term ). Defining the theoretical value for the node count when no time has elapsed is not straightforward. The trees have no tips at t0, meaning that node count will be zero; however, because the tree has a root node, there is technically a node count of 1. We therefore fitted models where the intercept is estimated , with t5 iterations sampling at every 1000 iterations and discarding the first 5 × 104 iterations as burn-in. We used the default priors for MCMCglmm . All models had a mean effective sample size  of greaet al. [et al. [2 or √time elapsed) to choose between models where the difference in DIC was less than 4 units. We feel that our procedure is a potentially fairer test between models, as quadratic terms are almost always significant and would thus lead to the downturn model (c) being preferentially selected over either the null (a) or slowdown to asymptote models (b) even in cases where model fit was similar. For models with estimated intercepts, we also extracted the posterior means of the intercepts to examine how these varied.We fitted each of the three models to all of our 909 trees , with inet al. ), or wit [et al. , which urn model c being pthe null a or slowe models b even inhttps://github.com/nhcooper123/dino-trees/ [https://dx.doi.org/10.5519/0034257) [All analyses used R v. 3.6  and repro-trees/ ). The da0034257) .3.et al. [et al. [The best model varied between trees, between differently dated versions of the same tree, and on the basis of whether intercepts were estimated, set to zero, or set to 1.0 figures . The Lloet al.  supertre [et al.  supertreet al. [et al. [et al. [Our analyses where intercepts were estimated produced qualitatively identical best model results for the Benson et al.  and Lloy [et al.  trees , indicaet al. [et al. [et al. [et al. [For models where intercepts were set to zero , 207 (23%) of our 900 new trees unambiguously favoured the downturn model, 186 (21%) unambiguously favoured the slowdown to asymptote model and 472 (52%) favoured either the downturn or the slowdown to asymptote model. No trees favoured the null model and 35 (4%) favoured no model at all . All three Lloyd et al.  supertre [et al.  supertre [et al.  supertre [et al.  supertreet al. [et al. [et al. [et al. [For models where intercepts were set to 1.0 , 183 (20.3%) of our 900 new trees unambiguously favoured the downturn model, 209 (23.2%) unambiguously favoured the slowdown to asymptote model and 469 (52.1%) favoured either the downturn or the slowdown to asymptote model. No trees favoured the null model and 39 4.33%) favoured no model at all . All three Lloyd et al.  supertre% favoure4.et al. [et al. [2 or √time elapsed). Because these quadratic terms were generally significant this led to their favouring the downturn model over the other two models in these ambiguous cases, potentially leading them to overstate the success of the downturn model. While this is a valid methodological choice, and differences in opinion about model selection procedures are common, we argue that as their selection of the downturn model as the ‘best’ model was methodologically equivocal it is unfair to say there is ‘overwhelming support’ for a downturn in dinosaur speciation rates prior to the Late Cretaceous [In general, our results agree with those of Sakamoto et al.  but we d [et al.  selectedetaceous , p. 5036c) that would be indicative of a continual downturn in dinosaur speciation rates before the K-Pg boundary. In particular, the sauropod tree of Carballido et al. [contra [Using the various combinations of trees, dating protocols and intercept assumptions listed above, we fitted a total of 2727 speciation models. Of these, only 518 (approx. 19%) unambiguously favoured the downturn model c that woo et al.  consiste [contra ), but suet al. [et al. [et al. [et al. [A recent study by Chiarenza et al.  provideset al. . Their cet al. . In addiet al. ,55. Thes [et al.  and Dean [et al.  rejected [et al.  found a  [et al.  recovereFinally, we posit that although phylogenies can be very useful in resolving long-running evolutionary debates ,57, they


Humans evolved from an ape ancestor that was highly intelligent, moderately social and moderately dependent on cultural adaptations for subsistence technology (tools). By the late Pleistocene, humans had become highly dependent on culture for subsistence and for rules to organize a complex social life. Adaptation by cultural traditions transformed our life history, leading to an extended juvenile period to learn subsistence and social skills, post-reproductive survival to help conserve and transmit skills, a dependence on social support for mothers of large-brained, very dependent and nutrient-demanding offspring, males devoting substantial effort to provisioning rather than mating, and the cultivation of large social networks to tap pools in information unavailable to less social species. One measure of the success of the exploitation of culture is that the minimum inter-birth interval of humans is nearly half that of our ape relatives. Another measure is the wide geographical distribution of humans compared with other apes, based on subsistence systems adapted to fine-scale spatial environmental variation. An important macro-evolutionary question is why our big-brained, culture-intensive life-history strategy evolved so recently and in only our lineage. We suggest that increasing spatial and temporal variation in the Pleistocene favoured cultural adaptations.This article is part of the theme issue ‘Life history and learning: how childhood, caregiving and old age shape cognition and culture in humans and other animals'. Severalet al.'s  classic et al.'s . Thus, t [et al.  The othe [et al. . Kaplan  [et al.  provide  [et al. –8. Walke [et al.  note tha [et al.  have recet al.'s  work on  [et al.  argue thThe reason that our large brain features so prominently in discussions of the evolution of the human life history is that it generates strong life-history trade-offs. Aiello & Wheeler  noted thet al. [Isler & Van Schaik  argue thet al.  suggest et al. . The onlet al. [et al. believed hunter–gatherers could not solve. Hawkes [There is some controversy over who were the most important providers of alloparental care over the course of human evolution. Kristen Hawkes et al.  noted th. Hawkes  proposedet al. [et al.'s [et al. [the human life history in the face of likely large changes over the Pleistocene and considerable ecological and cultural variation around the current central tendency. Another extreme human life-history variant is modern ‘demographic transition' societies that have far fewer children than are economically feasible [Hawkes et al.  went on et al.'s  data are [et al.  label thfeasible . Modernifeasible .et al.'s [contra [One constraint on the small game hypothesis is that human physiology precludes getting a large share of calories from lean protein. Humans cannot process enough nitrogenous waste from using protein as a source of calories above about 35% of total caloric needs, and small animals are generally very low in fat . Kaplan et al.'s  data sug [contra  but cons [contra ). Large  [contra  might coWood & Hill  tested t2.Given that our large brain coevolved with our slow life history and our cooperative breeding adaptation, it is of interest what the brain is adapted to do. Some of the controversy involves whether big brains are for managing our social life ,29 or foet al. [A different controversy involves the roles of culture and individual intelligence in the evolution of our large brain. This is a subtle question. The subtlety is twofold. First, empirically, the comparative biology of brain size suggests that it is correlated with both individual and social learning across a wide range of behaviours and species –34. Readet al.  argue thet al. ,37. It iSecond, Boyd & Richerson  showed tSome prominent evolutionary psychologists deny that culture, in the sense of socially transmitted traditions, plays any significant part in the evolution of humans or other animals (e.g. ). Tooby More recently, Cosmides & Tooby  have procultural variation operating via social selection  acting shments, .Cultural evolutionists argue cultural traditions like technology and social organization typically evolve cumulatively over prolonged periods, often resulting in quite sophisticated adaptations ,55,58–60et al. [et al. [et al. [et al.'s [Homo naledi fossils suggests that Lower Pleistocene brains and their cultures were viable long after more sophisticated cultures and larger brains evolved in other human lineages [In essence, the cultural niche hypothesis defended by Boyd et al.  holds th [et al.  point ou [et al.  to argue [et al. , and by  [et al. . By Ache [et al. . Hill etet al.'s  hypotheslineages . At the lineages . The builineages . Social Thus, in principle, our large and costly brain might be explained by its ability to support more innate modules, better individual intelligence or cumulative culture. That so much of our subsistence is a product of sophisticated technology and social organization that is transmitted culturally suggests that cumulative culture is a major part of the answer.3.et al. [The four life-history characteristics enumerated by Kaplan et al.  could ha(a)et al. [There is a fifth major life-history difference between humans and chimpanzees in addition to the four enumerated by Kaplan et al. . Humans et al. . When thet al. , as is tet al. . High paet al. , would aet al. . Young cThe size of social networks people can use to access cultural variation is important because people can actively bias their acquisition of culture (and their teaching) . Learneret al. [Long-term quantitative studies of hunting and gathering groups demonstrate that they tend to live in fluid bands of 30–50 individuals that regularly exchange members, such that the whole ethnolinguistic tribe of a few hundred to a few thousand people participates in a common social network, a form of social structure that seems to be uniquely human . Hadza aet al.  studied et al.  looked cet al.  give theSheer population size limits the scale of social networks and the level of social complexity that a societies can sustain ,80. DiamIncreasing network sizes engenders trade-offs. Societies living at low population densities require costly investments in travel to maintain large social networks. In the Ju/hoansi (!Kung), a system of gift exchange links people in distant camps . In the Increasing social network size also carries the risk of acquiring maladaptive ideas. Acquiring culture by vertical transmission is relatively safe in that parents and offspring are closely related genetically and tend live in the same ecological circumstances. Even so, parent–offspring conflict is a well-studied problem . HoweverHuman social networks are poly-functional. A person's information network, network of relatives, network of economic partners and network of acquaintances have different costs and payoffs yet they tend to heavily overlap. For example, Thornhill & Fincher  defend t(b)Do humans show any signs of being adapted to social as opposed to individual learning? At least at the margin of time these activities trade-off against one another even if both capacities extensively share cognitive resources. A now voluminous literature documents that children are adept at social learning even as compared with other apes ,99. This4.The evidence we have reviewed suggests that the long, slow human life history coevolved with our large brain. Brains are substantially organs of phenotypic flexibility, and brain size increased in many mammalian lineages in the Cenozoic, with humans holding down the upper tail of the distribution of brain size relative to body weight. This evolution seems to have been driven by increasingly variable environments. The question is how do humans, and by extension other large-brained creatures, pay the high overhead costs of large brains? The general answer seems to be learning and other forms of individual creativity plus social learning. The extraordinarily large modern human brain depends upon high skilled food acquisition strategies that make nutrient-dense foods available by exploiting a great variety of locally available food resources. Cooperative breeding, especially the heavy involvement of men in helping provisioning of mothers with dependent offspring, requires institutions of marriage and kinship. Culturally transmitted subsistence skills and techniques and culturally transmitted social institutions make possible a life history that is simultaneously slower but capable of higher completed family size than in other apes.


NPNL. Our simulation of foetal energy requirements demonstrated that this metabolic threshold of 2.1 × BMRNPNL cannot realistically be crossed by the foetus around the time of birth. These findings imply that metabolic constraints are not the main limiting factor dictating gestation length.A hallmark of modern humans is that our newborns are neurologically immature compared to other primates. It is disputed whether this so-called secondary altriciality evolved due to remodelling of the pelvis associated with bipedal locomotion, as suggested by the obstetrical dilemma hypothesis, or from maternal energetic limitations during pregnancy. Specifically, the ‘Energetics of Gestation and Growth’ (EGG) hypothesis posits that birth is initiated when foetal energy requirements exceed the maximum sustained maternal metabolic rate during pregnancy at around 2.1 × basal metabolic rate (BMR) of the non-pregnant, non-lactating condition (NPNL). However, the metabolic threshold argued under the EGG framework is derived from one study with a small sample size of only 12 women from the UK. Accordingly, we performed a meta-analysis of all published studies on metabolic scopes during pregnancy to better account for variability. After excluding 3 studies with methodological issues, a total of 12 studies with 303 women from 5 high- and 3 low-income countries were analysed. On average, pregnancy was found to be less metabolically challenging than previously suggested. The studies revealed substantial variation in metabolic scope during pregnancy, which was not reflected by variation in birth timing. Further, in a third of the studies, the metabolic rates exceeded 2.1 × BMR Newborn modern humans weigh approximately twice as much as those of great apes, both absolutely and relative to adult body mass, yet neonatal brain size is only 28% of the mother’s brain size, whereas it averages 43% in non-human primates . Human nThe obstetrical dilemma hypothesis has recently been criticized from various perspectives. Most of these critiques focus only on single aspects like the energetic consequences of pelvic width  or factoThe most comprehensive critique of the obstetrical dilemma to date is offered by the ‘Energetics of Gestation and Growth (EGG) hypothesis’. It not only questions that difficult birth was caused by pelvic adaptations to bipedal locomotion but also provides an alternative explanation for secondary altriciality, thus countering two of the central pillars of the obstetrical dilemma hypothesis , 19. In The EGG hypothesis is technically an extension of Ellison’s  ‘metabolin vitro and in vivo studies of the placental transport of 14C-labelled fatty acids showed that this process occurs rapidly [However, since the formulation of the metabolic cross-over hypothesis in 2001,  rapidly , 28. It, rapidly . Support rapidly . Moreove rapidly  metaboli rapidly . Neverth rapidly , so that rapidly  metaboliNPNL), which only slightly exceeds the metabolic scope during lactation [et al. [NPNL for a 280-day-long event like pregnancy [NPNL threshold assumed by the EGG hypothesis. However, the energetic requirements of the foetus would need to increase enormously to cross those of the expectant mother at the time of birth if this threshold would be significantly higher than 2.0 to 2.1 × BMRNPNL. It is, therefore, unclear whether metabolic limitations are the sole determinant for gestation length in humans as suggested by the EGG hypothesis.The maternal energetic threshold of the EGG hypothesis, the so-called ‘metabolic ceiling’, was proposed to lie at approximately 2.0–2.1 times the basal metabolic rate of the non-pregnant, non-lactating condition , th [et al.  with onlregnancy . Therefoet al. [Recently, Prado-Novóa et al.  also demet al. .et al. [Given these issues surrounding the EGG hypothesis, the present study aims to expand the data on which the hypothesis of Dunsworth et al.  rests anet al. , the preet al. [et al. [et al. [t-tests, we found the mean 24 h-energy expenditure observed by Poppitt et al.  [et al.  [et al.  [We identified 15 studies reviewed by Butte and King  and Savaet al.  that meaet al.  was omit [et al.  was excl [et al. , 38. Fin [et al.  was excl kcal/d)  was sign < 0.05)  and Hein < 0.05) , althouget al. [et al. [et al. [et al. [This left 12 studies that were included in the present meta-analysis . Some ofet al.  studied  [et al.  measured [et al.  who comp [et al. , perform [et al. , perform [et al. , 48, 49; [et al.  and the  [et al. . Specifimax) is defined as the ratio of TEE  that can be maintained without depleting energy reserves, thus preserving constant body mass, and basal metabolic rate  [et al. [EP) and fat mass (EF) during pregnancy was taken from Table 8 of Butte and King [Energy expenditure generally shows physiological limits during prolonged physical activities , 53, 54. kcal/d) , 53, 54. [et al.  in addinand King . To obtaet al. [For all studies, the same offspring energy requirements were used as by Dunsworth et al.   and no later measurements were provided. The authors indicated, however, that the non-pregnant, non-lactating women and the pregnant women were not the same individuals. In Butte et al. [n = 63), not only the weighted mean metabolic rate of the three subsamples (2.19 × BMRNPNL in pregnancy week 36) exceeded the threshold of 2.1 × BMR but also each subsample itself . Finally, a high mean metabolic rate was also reported by Dufour et al. [NPNL in pregnancy week 35, but no data have been recorded for week 36 or later.In 4 of the 12 studies , 48, 49,h et al.  that equr et al.  for a stPregnant women potentially use different strategies to lower their own energy expenditure. Such strategies could include energy-sparing mechanisms, like the reduction of the maternal BMR , or loweDifferences in the socioeconomic background of the study populations could explain part of the variation as the mean metabolic scope, particularly before pregnancy, was generally higher in socioeconomically weaker countries. Thus, socioeconomic background has been linked to physically more demanding workloads . Additioet al. [et al. [2 consumption and CO2 production and is considered the most accurate method to measure energy expenditure in a clinical setting [It is possible that the measurement methods contributed to the high variation of the metabolic rates . Thus, tet al.  observed [et al.  found th setting . It is t setting , 49. In  setting , 46, the setting . We consIn the free-living context, the doubly labelled water method (DLW) is usually considered the gold standard, as it imposes a minimal burden to the participants and is sufficiently accurate, having a precision of 2–8% compared against respiratory gas exchange . The DLWet al. [et al. [NPNL with indirect calorimetry. Panter-Brick [NPNL was estimated by using an age-specific predictive equation based on individual height and month-specific mass [et al. [Another method to measure TEE during pregnancy is the flex heart rate method, which was used by Dufour et al.  in Columet al. , 63. On  [et al.  assesseder-Brick  relied ofic mass . The stu [et al.  and Pant [et al.  showed tet al. [r = 0.87) [Finally, Abeysekera et al.  based th = 0.87) . Neverthet al. [et al. [et al. [a priori yield different results compared to DLW.An examination of the additional energy required during pregnancy revealed on average a steady increase during the entire course of pregnancy rather than a plateau during the third trimester as suggested by Dunsworth et al. , which w [et al.  study et al. [e [et al.  to estim [et al. . The equ [et al.  and Dufo [et al. . Neverth [et al. . Therefo [et al.  and Dufo [et al.  are plau [et al.  in the N [et al. , 46 showThe Energetics of Gestation and Growth (EGG) hypothesis posits that maternal energy requirements steeply increase during pregnancy and approach a plateau in the third trimester close to 2.1 × BMR of the non-pregnant, non-lactating condition, and that labour starts when the exponentially growing energetic requirements of the foetus cross the maximum sustained maternal metabolic scope .NPNL. (v) Because the EGG hypothesis posits that labour is only triggered when the energetic requirements of the foetus surpass the maximum sustained metabolic capacity of the pregnant woman, a metabolic ceiling during pregnancy exceeding 2.1 × BMRNPNL would imply an unrealistically high mean birth weight of > 4.9 kg. Conversely, (vi) the remarkably high variation in energy expenditure of the pregnant women strongly contrasts with the relatively low variation in gestation length. If birth timing were in fact dependent on energy expenditure during pregnancy, gestation length would likely be equally variable, which questions the conceptual basis of a metabolic ceiling in determining the onset of labour.The present meta-analysis of the 12 studies that measured TEE during pregnancy demonstrated, however, (i) considerable variation in the observed sustained maternal metabolic scope. (ii) On average, pregnancy was found to be less energetically costly than suggested previously, and (iii) in the great majority of the studies maternal metabolic scope did not plateau in the third trimester, implying a pattern that is not suggestive of a metabolic ceiling being approached by the maternal energy requirements during pregnancy. Moreover, (iv) a large percentage of the studies significantly exceeded the presumed metabolic ceiling of the EGG hypothesis of about 2.1 × BMRA metabolic trigger of human birth has also been challenged by a study of obstetric selection pressures in early hominid fossils using FEA birth simulations, demonstrating that cephalopelvic constraints were similarly high in australopithecines as in modern humans . This imIndependent of the question of birth timing and the EGG hypothesis, it remains unclear from an evolutionary point of view why maternal metabolic capacity would be limited given the potential negative consequences. Further, the significantly higher costs of lactation make it more plausible that this activity might be pushing humans closer to the limit of their energetic capacity, wherever such a threshold might lie. Yet, it is well known that even human lactation can well be supported through increased dietary intake rather than substantial mobilization of adipose tissue , 54, 75.


A best-evidence topic was written according to a structured protocol. The question addressed was the following: in patient undergoing lung transplantation, are lungs from donors of age >60 years old (yo) associated with equivalent outcomes—including primary graft dysfunction, respiratory function and survival—than lungs from donors ≤60yo? Altogether, >200 papers were found using the reported search, of which 12 represented the best evidence to answer the clinical question. The authors, journals, dates, country of publication, patients group studied, study type, relevant outcomes, and results of these papers were tabulated. Amongst the 12 papers reviewed, survival results were different depending on whether donor age was analysed raw or adjusted for recipients’ age and initial diagnosis. Indeed, recipients with interstitial lung disease (ILD), pulmonary hypertension or cystic fibrosis (CF) had significantly inferior overall survival when receiving grafts from older donors. When older grafts are allocated to younger donors, a significant decrease in survival has been noticed in the case of single lung transplantation. In addition, 3 papers showed worse results regarding peak forced expiratory volume in 1 second (FEV1) in patients receiving older organs, and 4 showed comparable primary graft dysfunction incidence rates. We conclude that when carefully assessed and allocated to the recipient who could benefit most from the transplant , who would not require a prolonged cardiopulmonary bypass (CPB)), lung grafts from donors of >60yo offer comparable results to younger donors. A best-evidence topic was constructed according to a structured protocol. A best-evidence topic was constructed according to a structured protocol. This is fully described in the ICVTS .In [patients undergoing lung transplantation], are lungs from [donors > 60 years old] associated with  than lungs from donors ≤60 years old?As a thoracic surgeon on call for lung procurement, you receive a call from the agency of health care. A graft from a 76-year-old (yo) male patient is offered. The cause of death is a sudden brain Haemorrhage. The clinical record shows no medical and no smoking history. The initial study shows a thoracic diameter similar to your patient’s one with a Pa02/Fi02 ratio evaluated at 420 mmHg. The CMV serology is negative. The CT-scan imaging shows no emphysema or suspicious nodules. The bronchoscopy shows perfect integrity of the airway. Then, you find a potential match for a 64yo man with COPD, but you hesitate to go through the lung transplantation (LTx) based on the donor’s age.Medline search using PubMed interface was performed with the following query: [lung transplantation OR lung transplant] AND [old donors OR age] AND [outcomes], with results only in English and including all types of publications. Afterwards, references from the retrieved studies were looked up manually.209 papers were found using the reported search. From these papers, 12 were selected, providing the best evidence to answer the question. These are presented in Table et al. [De Perrot et al.  analysedet al. [Baldwin et al.  evaluateet al. [Shigemura et al.  publisheet al. [Sommer et al.  studied et al. [P < 0.05). Regarding long-term outcomes, recipients of young, old and very old lungs showed similar 1- and 3-year survival rates.In 2017, Hecker et al.  investiget al. [Holley et al.  publisheet al. [Schultz et al.  publisheet al. [Whited et al.  aimed toet al. [In 2019, Hall et al.  retrospeet al. [Auråen et al.  publisheet al. [Renard et al.  studied et al. [Vanluyten et al.  publisheWe are here chronologically presenting data from cohorts studied between 1986 and 2019. The patients with the oldest data are included in large cohorts and the authors of the studies captured sufficient data for a robust statistical analysis. Considering the level of evidence, all the studies are retrospective non-randomized studies. Nevertheless, we looked for large cohort studies that analysed donor age as a binary variable. The data presented here are showing that when analysed as a binary variable and without adjusting for the presence of other extended donor criteria, old donor age can be associated with worse OS. However, when the recipient's age and underlying diagnosis were considered, these results almost consistently lost their statistical significance. Some authors have sub-stratified patients by single or bilateral LTx. In this way, in younger recipients, the use of older donors was associated with a similar survival when performing double LTx and a significant decrease in survival performing single LTx. Nevertheless, it has been noticed that double lung transplant was the dominant procedure in the younger recipients’ cohorts and the single transplant was the dominant procedure in the older recipients’ cohorts. Regarding respiratory functions, results are showing better short- and long-term postoperative peak FEV1 from younger lungs. The donor age was not associated with PGD risk in the papers presented. Considering CLAD, there are data available. However, some authors described a significant difference in CLAD-free survival when using older donors.Hence, during the allocation process, lungs from donors ≥60yo should not be systematically rejected based on this parameter alone and the recipients’ characteristics should also be considered. From the data collected here, the proper recipients for lungs from old donors appear to be emphysema patients with no pulmonary hypertension and >60 years old. Thus, accepting lung grafts from older donors can be a safe and feasible strategy but donor age should be perceived as a risk factor that needs to be balanced against the urgency for transplantation.


Mast cells and basophils share some phenotypic characteristics, such as the expression of a high-affinity membrane receptor for IgE, FcεRI, and the granule storage of inflammatory mediators including histamine. Mast cells are found to be resident in nearly all vascularized tissues, whereas basophils migrate into the inflamed tissues from the circulation.Sauer et al. verified the involvement of STAT3 in type IIb autoimmune CSU. The roles of histamine in chronic inducible urticaria were reviewed by Kulthanan et al., Takimoto-Ito et al. reported a case in which activated basophils remained in circulation during treatment with omalizumab. Miyake et al. summarized recent findings of human and murine basophils, including their roles in immune tolerance. The regulatory roles of mast cells were also discussed by Zhang et al. and Honda and Honda Keith. Poto et al. and Numata et al. investigated, respectively, the actions of autoantibodies against IgE and sweat antigen-induced chronic activation of basophils in patients with atopic dermatitis. Kamei et al. demonstrated an IgE-dependent murine model of oral allergy syndrome. El Ansari et al. found that allergen-specific IgA could suppress the IgE-mediated activation of mast cells.Accumulating evidence suggests that cutaneous mast cells play critical roles in chronic spontaneous urticaria (CSU) and that there are several promising therapeutic approaches . Sauer eMcNeil). In this topic, signal transduction and regulation of intracellular localization of MRGPRX2 were investigated . West and Bulfone-Paus discussed the heterogeneity of tissue mast cells, with special attention given to the expression of MRGPRX2. Numata et al. summarized the roles of mast cells in cutaneous diseases. In indolent systemic mastocytosis, an increased number of mast cells expressing MRGPRX2 was found in the skin but was not linked to symptom severity .MRGPRX2 has received attention as a novel therapeutic target of chronic urticaria and drug-induced anaphylaxis, because its agonists have been found to have a structural diversity . Novel murine experimental models for the depletion of mast cells and basophils were also introduced here .We hope that this topic encourages and accelerates the research of mast cells and basophils.All authors listed have made a substantial, direct, and intellectual contribution to the work and approved it for publication.


Hypoparathyroidism requires management with both calcium supplementation and active vitamin D to avert a state of hypocalcemia. During late gestation and the postpartum period , there is an under-recognized, yet intriguing occurrence of apparent ‘pseudohyperparathyroidism’, whereby supplementation dosages may need to either be reduced or discontinued, to prevent hypercalcemia. The explanation for this apparent phenomenon of improved parathyroid status (‘remission’ or ‘resolution’) is incompletely understood; the purpose of this review is to analyze the case reports of this enigma within the medical (and grey) literature, providing an overall pathophysiological explanation and recommendation for the management of such patients. A literature search was conducted through PubMed/Medline, CINAHL, Cochrane Library Database, Scopus, UpToDate, Google Scholar, and the grey literature without a time-restricted period, analyzing all available articles within the literature describing an apparent improvement in parathyroid status in late-gestation and postpartum (lactating) females. Non-hypoparathyroid case reports were also included to further analyze and synthesize an overall likely pathophysiological explanation. Through the literature search, 24 papers were identified covering such a phenomenon in patients with hypoparathyroidism, alongside multiple additional reports of a similar occurrence in patients without underlying hypoparathyroidism. The pathophysiology is believed to occur due to the placental production of parathyroid hormone-related peptide (PTHrP) during gestation, with further production from the lactating mammary glands during the postpartum period. A typical pattern is observed, with increased PTHrP and suppressed PTH throughout both gestation and lactation . The concept of PTHrP-induced hypercalcemia is further demonstrated in patients without hypoparathyroidism, including subjects with placental hypersecretion and mammary gland enlargement. It is evident that patients with hypoparathyroidism may require a dosage reduction during late gestation and lactation, due to the risk for hypercalcemia. In addition to patients with hypoparathyroidism, this pathophysiological phenomenon occurs in unsuspecting patients, demonstrating the need for all clinicians in contact with pregnant females to be aware of this uncommon - yet perilous - occurrence. Initially described in the great Indian rhinoceros by Sir Richard Owen in 1852, the parathyroid glands were an addition to the knowledgebase in the ever-evolving field of endocrinology ..3].Hypoparathyroidism has a prevalence of up to 6.6% in the general population, encompassing all age groups, but lacks a specific gender preponderance ,5. The pDuring the postpartum period, an apparent phenomenon is noted, whereby lactating females (with hypoparathyroidism) may demonstrate an apparent state of calcitriol toxicity  requiring a temporary reduction (or discontinuation) of their medications. This is characterized by subversion into an apparent temporary remission (or improvement) of hypoparathyroidism and has also been documented during late gestation. As this occurrence is infrequently recorded, the focus of this article is to reinforce this pivotal concept, as well as to provide an overall recommendation for the management of such patients.Outside the field of endocrinology, the quandary for calcitriol toxicity (temporary hypoparathyroidism resolution) during lactation is unlikely to be common knowledge. Parous women with a diagnosis of hypoparathyroidism are likely to attend appointments with specialties other than endocrinology - examples include obstetrics, midwifery, and general practitioners. This demonstrates multiple occasions for potential proactive maternal care. Due to the limited understanding of this phenomenon, coupled with the potential for preventable maternal harm, the purpose of this review is to broaden the awareness of the potential for hypercalcemia in postpartum, lactating females, with an established diagnosis of hypoparathyroidism.The primary objective of this review is, therefore, to1. Provide a scoping review of the medical literature pertaining to the phenomenon of an ‘improvement’ or ‘remission’ of hypoparathyroidism during late gestation and lactation, with the intent of enhancing awareness of this potential (harmful) outcome across further medical specialties (and afflicted patients).The secondary objectives of this review are to1. Provide an overview of the proposed pathophysiological hypotheses pertaining to the apparent improvement in hypoparathyroidism .2. Review the recommendations for the management of hypoparathyroidism in late gestation and the postpartum period.Whilst the temporary resolution of hypoparathyroidism is described in the medical literature, this is mainly limited to case reports and case series. Due to the paucity of data, this review is justified, with the intent to provide awareness to the greater medical community of the potential for calcitriol toxicity (requiring dosage revision) during both late gestation and postpartum (lactation). Ensuring that a broader group of healthcare professionals (and motivated patients) are aware of this potential outcome will safeguard proactive patient care and result in a reduction in preventable feto-maternal harm.Materials and methodsThis review is delivered as a scoping review, encompassing the broad medical literature, with a subset focus on relevant case reports/series from the ‘grey’ literature, which further substantiates the apparent (under-studied) phenomenon of hypoparathyroidism improvement during late gestation and in postpartum, lactating females. Providing a scoping review allows for a multi-faceted approach in the deciphering of this apparent endocrine anomaly, covering multiple case documentation, reviewing (and synthesizing) the proposed pathophysiology, providing rationales for the recommended management of this unique subset of patients, and highlighting areas of uncertainty.Search StrategyWhilst, in research, the highest-regarded bodies of evidence  are randomized controlled trials, meta-analyses, and systematic reviews, infrequent  medical occurrences  are often limited to less-rigorous bodies of evidence. For this reason, this review encompasses all forms of articles published . In addition to the variable article types available, to prevent further exclusion of informative data, the electronic search conducted did not have a time-restricted period. The search strategy entailed Cochrane Library Database, Scopus, PubMed/MEDLINE, the Cumulative Index to Nursing and Allied Health Literature (CINAHL), UpToDate, and Google Scholar (in addition to the grey literature). The following keywords were used in the retrieval of available literature pertaining to the principal topic of this professional project: “hypoparathyroid” AND “lactation” OR “hypoparathyroidism” AND “lactation” OR “hypoparathyroidism” AND “breastfeed” OR “hypoparathyroidism” AND “remission” AND “lactation” OR “hypoparathyroid” AND “remission” AND “postpartum” OR “hypoparathyroidism” AND “resolution” AND “lactation” OR “calcitriol treatment decreased” AND “hypoparathyroid” AND “lactation”.Publications in languages other than English were included . The search included reports of this occurrence without hypoparathyroidism to further support the pathophysiological hypotheses. Male subjects were excluded from the search strategy; animal studies were included in the search strategy, to be incorporated under the pathophysiological section of this professional project . The focus of inclusion is postpartum (lactating) females. However, to further reiterate the pathophysiological hypotheses, non-lactating, postpartum females (and those during gestation) were also included. Further, articles were included from the reference list of an initial article identified (when relevant to the aforementioned inclusion criteria). Both full-text articles and abstracts (whereby the case description was apparent and data could be deciphered) were included . A further n = 49 citations were identified from primary reference lists and used to further the pathophysiological explanation of this professional project Figure .Quality of EvidenceAs the purpose of this scoping review is to synthesize the cases relating to the apparent improvement in hypoparathyroidism and to analyze areas of uncertainty in current knowledge, a quality appraisal was not performed for this review.Risk of Bias AssessmentScoping reviews are prone to bias, especially when considering a rare documented medical occurrence whereby only patients experiencing the phenomenon are likely to be depicted in the literature (an uneventful postpartum period is unlikely to be reported in the literature). There is no data to suggest the percentage of pregnant and lactating postpartum females with hypoparathyroidism who do not experience the enigma of temporary resolution or remission. As this is a scoping review, a risk of bias assessment was not performed.ResultsWright et al.  posed anTwo years later, the same patient delivered her second child; again, during her pregnancy, serum calcium was unremarkable, and she was adherent to 100,000 IU of calciferol and 76 mEq of calcium daily. The authors record that, for the second time, she did not breastfeed and note that, within 24 hours postpartum, calcium rose to 6.0 mEq/L, at which point all medications were discontinued. Serum calcium subsequently decreased to 5.1 mEq/L over the following six days postpartum. Calciferol  and 38-57 mEq/L of calcium were restarted; however, symptomatic hypercalcemia reappeared (6.0 mEq/L), requiring discontinuation on postpartum day 20. Roughly one week following the discontinuation of her medications, the serum calcium decreased to 4.8 mEq/L, at which point half her usual dosage was reintroduced for the succeeding 12 days.A second patient was described by Wright et al. , who wasIn contrast to Wright et al. , SadeghiMarkestad et al.  presenteRude et al.  presenteCundy et al.  describeCaplan et al.  presenteHöper et al.  presenteCathébras et al.  further Shomali et al.  presenteAnai et al.  presenteMather et al.  presenteYasumatsu et al.  presenteSweeney et al.  presenteKrysiak et al.  presenteSegal et al.  depictedAl-Nozha et al.  reportedHatswell et al.  analyzedShah et al.  demonstrDixon et al.  posed a DiscussionA pattern is noted in the above case series, whereby calcium requirements are rather labile during pregnancy, but tend to decrease postpartum. Marcucci et al.  includedIn a case series performed by Hartogsohn et al. , 17 pregWang et al.  presenteProposed PathophysiologyThe apparent improvement in hypoparathyroidism during late gestation and breastfeeding is an incompletely understood, yet potentially harmful occurrence  to the untrained clinician. As noted by Winter et al. , this apDuring pregnancy, PTHrP is predominantly released by the placenta . However, there may be concurrent release from enlarged mammary glands during this time period, with a ‘transition’ to release by the mammary glands during the postpartum (lactating) period ; these aPTHrPPTHrP is a hormone responsible for humoral hypercalcemia of malignancy and paraneoplastic syndromes (notably squamous cell carcinoma of the lung). It is important to note that, during lactation, PTHrP is released into the breast milk at 1,000-100,000 times the serum concentration that would be present in humoral hypercalcemia of malignancy . During As noted by Kalkwarf et al. , a lactaGrill et al.  demonstrSimilar findings emerged from the study conducted by Kalkwarf et al. , noting In a longitudinal study performed by Ardawi et al. , levels In a study of non-hypoparathyroid, lactating women by Dobnig et al. , at two-While Grill et al.  noted elAt two to three days postpartum, Dobnig et al.  demonstrAs demonstrated in the study by Segal et al. , there iSowers et al.  demonstrIn the first six months of the study performed by Kalkwarf et al. , the autHypercalcemiaIn the cases reported under the ‘results’ section of the review, it is important to note that not all authors were able to measure serum PTHrP. In the report posed by Shomali et al. , howeverAnai et al.  demonstrIn the first patient described by Yasumatsu et al. , it is nThe phenomenon of apparent PTH-independent hypercalcemia in pregnant and lactating females (as noted in the above series) has furthermore been replicated across numerous case reports consisting of patients without a diagnosis of hypoparathyroidism. Marx et al.  presenteVan Heerden et al.  parallelSiskind et al.  presenteSato  describeLepre et al.  describeIn a similar manner to the case report by Segal et al. , KebapciBreslau et al.  describeProlactinProlactin is a hormone involved in lactation and is released from the lactotrophs in the anterior pituitary in response to a myriad of stimuli (predominantly nipple suckling). As demonstrated by Sowers et al. , there iIt should be noted that elevated PTHrP has been documented in other cases of hyperprolactinemia, such as that with prolactinomas  . In rat Kovacs et al.  analyzedOf further peculiarity, serum prolactin appears to not only be intricately associated with serum PTHrP but also 1,25-hydroxyvitamin D3 (active vitamin D). Following the work by Cundy et al. , the appVitamin DCundy et al.  attempteThe correlation between the prolongation of active vitamin D and prolactin is ill-defined, albeit three strong hypotheses are currently understood to be partly correct: delayed clearance, PTHrP-mediated, and prolactin-induced. The most straightforward explanation for a prolongation of the half-life of active vitamin D and prolactin would be that there is a reduction in the clearance of the former (as opposed to enhanced endogenous production). This hypothesis, while simple, is not accepted by many, as it is evident that serum 1,25-hydroxyvitamin D3 levels may continue to rise following discontinuation of calcitriol supplementation .PTHrP demonstrates similarity in structure to PTH (the first 34 amino acids of PTHrP are similar in structure to PTH), acting upon the same cell receptors . AnecdotProlactin has similarly been suggested to be able to stimulate the enzyme 1-α-hydroxylase (CYP27B1) within the maternal kidney. In both chick and rat studies, this theory has been demonstrated on multiple occasions -69. As rMarkestad et al.  presenteHypocalcemiaOne should be aware that the converse has been described during both gestation and the postpartum periods, whereby there is an apparent ‘worsening’ of serum calcium . The case report posed by Sweeney et al.  differs Al-Nozha et al.  further Eller-Vainicher et al.  presenteDurst et al.  describeBernstein et al.  depictedHarrad et al.  presenteCallies et al.  presenteJabbar et al.  discusseKurzel et al.  presenteRecommendationsBoth the hypoparathyroid mother and her infant should be carefully monitored to detect abnormal calcium levels ,13. The Strengths and limitationsA major strength of this article includes the enhanced awareness of a niche endocrine phenomenon, for which access will be available to all healthcare professionals who may encounter a female patient with hypoparathyroidism. Moreover, this article provides an up-to-date review of the available literature depicted by various authors globally. As with all articles, however, limitations must be highlighted; in this case, it is important to note that a great proportion of females with hypoparathyroidism may not require dosage adjustments during either gestation or lactation, with these apparently ‘normal’ cases unlikely to be reported upon . Further risks include selection bias and misinterpretation of data from articles published in German, Japanese, and French . Moreover, as there are multiple underlying causes of hypoparathyroidism . For this reason, an individualized approach with close monitoring is essential when treating hypoparathyroid females during gestation and the postpartum period.


Molecularly imprinted polymers (MIPs), due to their unique recognition properties, have found various applications, mainly in extraction and separation techniques; however, their implementation in other research areas, such as sensor construction and drug delivery, has also been substantial. These advances could not be achieved without developing new polymers and monomers that can be successfully applied in MIP synthesis and improve the range of their applications. Although much progress has been made in MIP development, more investigation must be conducted to obtain materials that can fully deserve the name of artificial antibodies.The “Advance in Molecularly Imprinted Polymers” Special Issue connects original research papers and reviews presenting recent advances in the design, synthesis, and broad applications of molecularly imprinted polymers. The Special Issue content covers various topics related to MIP chemistry, which include their molecular modeling, application of new monomers and synthesis techniques for MIP preparation, synthesis, and application of ion-imprinted materials, selective extraction of organic molecules, sensor preparation, and MIP applications in medicine. The presented collection of scientific papers shows that research in MIP chemistry is diversified, ultimately improving the desired properties and creating new potential applications for these materials.−1. The findings of this study indicate the potential of the obtained ion-imprinted polymers for the selective extraction of heavy metal ions from polluted waters.Bivián-Castro et al.  obtainedIon-imprinted polymers were studied by Kondaurov et al. . They obZaharia et al.  obtainedMegahed et al.  obtainedN-(2-arylethyl)-2-methylprop-2-enamides with various substituents in the benzene ring were obtained by Sobiech et al. [Six aromatic h et al.  using 2-Piletsky et al.  presenteThe synthesis of 5-fluorouracil-imprinted microparticles and their application in prolonged drug delivery was reported by Cegłowski et al. . The autJumadilov et al.  studied Chien et al.  developeMIPs possessing dual functional monomers (methacrylic acid and 2-vinylpyridine) were synthesized by Thach et al.  to yieldIn their review article, Lusina and Cegłowski  exploredThe other review article, prepared by Liu et al. , describPolymers. In addition, I would like to express my gratitude to the Editorial Team who helped prepare the “Advance in Molecularly Imprinted Polymers” Special Issue.This Special Issue has brought together experts that have studied and explored various aspects of MIPs. I want to thank all researchers who have contributed to the production of this Special Issue of


Green et al.  apply ne2 injection (e.g. ref. 3 reservoir meant for perturbations on timescales of LIP volcanism surface ocean carbonate saturation (2 release at the onset of Deccan emplacement drove the Late Maastrichtian Warming Event, but carbonate dissolution buffered surface ocean Earth system scientists have long recognized that the rise of pelagic marine calcifiers fundamentally changed the way Earth responds to CO ocean Ω  and mari ocean Ω  and 6.2 release was superimposed relative to earlier Phanerozoic LIPs, Green et al. (2 (Having overlooked the fundamentally different biogeochemical boundary conditions upon which Deccan COn et al.  cite putn et al. . Furthert al. (2  and globt al. (2  returnint al. (2  and is it al. (2  and palet al. (2  and 6 evIn sum, we implore the community to move beyond implicating causality from temporal coincidence, and instead, to focus on how, mechanistically, individual LIPs interacted with their specific boundary conditions to drive specific environmental and macroevolutionary responses. While meaningful research questions about the rate and scale of perturbation required to prompt a mass extinction remain, ultimately, a causal link between any geological phenomenon and mass extinction cannot be made by comparing unconnected events hundreds of millions of years apart. Instead, feasible, model-supported mechanisms for how a given event could precipitate ecological collapse are required—mechanisms that for the Chicxulub impact exist, in the form of ocean acidification  and impa


Urothelial carcinoma (UC) of the bladder is the tenth most diagnosed cancer worldwide and represents a significant cause of morbidity and mortality . UC is ain situ that are thus particularly challenging. For such cases urine cytology is still the most used non-invasive test detecting bladder cancer during follow-up. Despite its high specificity of approximately 86%, the limitation lies in low sensitivity of only 50% .There is a great need for further investigations of molecular markers for bladder cancer in the context of risk stratification and for developing combined targeted therapy options to prevent progression and cancer-related death. According to newer publications, FGFR3 mutation analysis study on bladder cancer patients should be performed for all stages and grades . There aIt is mandatory perceiving that marker systems are playing an important role in all fields of bladder cancer: as an alternative or additional tool for cystoscopy during follow-up, as predictor and prognostic instrument during decisions for systemic therapies or as screening tool for detecting bladder cancer in high-risk groups.This Research Topic has the main aim of offering possibilities to publish new research results in the basic and translational research field of bladder cancer. During the editorial process of this Research Topic we have appreciated that significant progress has been recently done in bladder cancer research and that efforts should be pursued by fostering extensive cooperation between the scientific and medical communities to translate evidence-based research into clinical practice. The identification and validation of bladder cancer markers for predicting recurrence and progression will contribute to establish better treatments for the individual patient based on its predicted response and their specific genetic and molecular characteristics, and molecular staging will allow selection of tumors that will require systemic treatment.The studies in this Research Topic cover the major areas of developing interest in bladder cancer research.Wang et al. studied ferroptosis regulators and identified GCLM as a tumor promotor and immunological biomarker in bladder cancer. Feng et al. evaluated the prognostic significance basement membrane-associated lncRNA in Bladder Cancer.There were several novel markers evaluated in this Research Topic. Lee et al. evaluating alpha-2-macroglobulin in urinary extracellular vesicles and a study by Bian et al. evaluating urinary exosomal long non-coding RNAs as noninvasive biomarkers for diagnosis of bladder cancer. Su et al. analyzed exosome-derived long non-coding RNAs as non-invasive biomarkers of bladder cancer.There were several articles evaluating new diagnostic markers for bladder cancer including a study by Hao et al. study near-infrared targeted imaging using ICG-anti-CD47.To improve imaging of disease, Wang et al. review the impact of cuproptosis-related genes on bladder cancer prognosis, tumor microenvironment invasion, and drug sensitivity. Castaneda et al. identify Novel Biomarkers associated with Bladder Cancer Treatment Outcomes. Wang et al. use proteomics analyses to identify CLIC1 as a predictive biomarker for bladder cancer staging and prognosis. Li et al. study endothelial-related molecular subtypes for bladder cancer patients, and Xiong et al. study inflammation-related lncRNAs in bladder cancer. You et al. study novel pyroptosis-related gene signatures and Xiao et al. evaluate impact of fatty acid metabolism, inflammation and hypoxia on bladder cancer.There were several studies evaluating bladder cancer prognosis. Chen et al. study the tumor microenvironment to develop a nomogram to predict lymph node metastases. Bieri et al. use a modified Immunoscore to improve prediction of progression-free survival in patients with non-muscle-invasive bladder cancer. Luo et al. develop a novel prognostic model based on cellular senescence-related gene signatures for bladder cancerSeveral studies developed prediction models for bladder cancer. Song et al. evaluated aliphatic acid metabolism in bladder cancer with the goal of guiding therapeutic treatment. A study by Wang et al. studied the mechanism by which RAC3 inhibition induces autophagy to impair metastasis in bladder cancer cells via the PI3K/AKT/mTOR pathway.A study by All editors of this Research Topic thank all submitting authors for their work. The lead editors would like to thank all editors and reviewers for the time spent in assigning reviews, commenting on submitted manuscripts as well as reviewing. As editorial team, we hope that this special issue will prove useful in planning bladder cancer research in next future.All authors listed have made a substantial, direct, and intellectual contribution to the work and approved it for publication.


Circadian rhythms play a fundamental role in our daily lives, orchestrating a wide range of physiological processes, including energy metabolism, immune function, cellular rejuvenation, and the intricate interplay with the gut microbiota. This Research Topic delves into the profound influence of circadian rhythms on various aspects of metabolism and endocrinology and explores the intricate connections between circadian physiology and metabolic disorders. By elucidating these relationships, this Research Topic aims to provide insights into the development of novel therapeutic strategies for the management of metabolic and endocrine diseases.Li et al. conducted a study revealing an independent association between rest-activity rhythm and obesity phenotypes. Their findings highlight the impact of aberrant rest-activity rhythm on anthropometric and imaging measures of general and abdominal obesity.At the population level, Huang et al. explore the effects of mistimed feeding on circadian reprogramming and the unfolded protein response. Their study demonstrates the cumulative impact of mistimed food intake over multiple generations, resulting in altered diurnal rhythmic transcriptomes and impaired endoplasmic reticulum stress response.Examining the intergenerational impact of circadian disruptions, Sun et al. conducted a bidirectional Mendelian randomization study providing genetic evidence that supports a bidirectional causal relationship between sleep traits and nonalcoholic fatty liver disease (NAFLD). Their findings emphasize the importance of considering sleep disturbances in managing NAFLD.Cortés-Ríos et al. used mathematical modeling to investigate the dosing-time-dependent antihypertensive effect of valsartan and aspirin. Their study sheds light on the optimal dosing time for these medications, underscoring the importance of chronopharmacology in hypertension management.Shifting the focus to pharmacology, Wild et al. conducted a case-control study evaluating sleep patterns in patients treated for non-secreting intra- and parasellar tumors. Their findings revealed altered sleep patterns in these patients compared to healthy controls, highlighting the need for clinicians to address sleep disturbances in this population.Expanding the scope to sleep patterns, Bao et al. performed a prospective clinical trial investigating the effect of melatonin on the quality of repeated-poor and frozen-thawed embryos in humans. Their study demonstrates that melatonin supplementation in the culture medium improves the rate of high-quality embryos, offering a potential rescue strategy for in vitro fertilization (IVF) failures.Moving to a more specific endocrine system, Yin et al. reviewed the use of melatonin for premenstrual syndrome (PMS). They discuss the role of melatonin in modulating sleep disturbance, mood changes, and cognitive impairment associated with PMS, suggesting its potential as a safe and effective treatment option.Exploring the potential therapeutic application of melatonin, Watanabe et al. explored the insulin-independent action of nocturnal melatonin in increasing glucose uptake in the goldfish brain. Their study highlights the role of melatonin in regulating glucose homeostasis and its potential as a circadian regulator of glucose dynamics.Lastly, Li et al., Huang et al., Sun et al., Cortés-Ríos et al., Wild et al., Bao et al., Yin et al., and Watanabe et al. emphasize the importance of circadian regulation and sleep patterns in maintaining metabolic homeostasis and offer potential avenues for therapeutic interventions in metabolic and endocrine disorders. The comprehensive understanding of the ties between circadian rhythms and metabolism presented in this Research Topic will undoubtedly pave the way for innovative approaches in the management of these conditions.The articles included in this Research Topic provide valuable insights into the intricate relationship between circadian rhythm, metabolism, and endocrinology. From the impact of mistimed feeding on circadian reprogramming to the bidirectional relationship between sleep traits and NAFLD, the findings from MR-F: Writing – original draft, Writing – review & editing


Age‐associated reduction of nuclear shape dynamics in excitatory neurons of the visual cortex by Tanita Frey et al., https://doi.org/10.1111/acel.13925Cover legend: The cover image is based on the Research Article


Cryptovaranoides microlanius from the Late Triassic of the United Kingdom was described as the oldest crown squamate. If true, this result would push back the origin of all major lizard clades by 30–65 Myr and suggest that divergence times for reptile clades estimated using genomic and morphological data are grossly inaccurate. Here, we use computed tomography scans and expanded phylogenetic datasets to re-evaluate the phylogenetic affinities of †Cryptovaranoides and other putative early squamates. We robustly reject the crown squamate affinities of †Cryptovaranoides, and instead resolve †Cryptovaranoides as a potential member of the bird and crocodylian total clade, Archosauromorpha. Bayesian total evidence dating supports a Jurassic origin of crown squamates, not Triassic as recently suggested. We highlight how features traditionally linked to lepidosaurs are in fact widespread across Triassic reptiles. Our study reaffirms the importance of critically choosing and constructing morphological datasets and appropriate taxon sampling to test the phylogenetic affinities of problematic fossils and calibrate the Tree of Life.Most living reptile diversity is concentrated in Squamata , which have poorly known origins in space and time. Recently, † Of these, the more than 11 000 living species of squamates represent the most speciose modern tetrapod clade . Living Recent additions and revisions to the early squamate and rhynchocephalian fossil records made possible by the use of new imaging and analytical tools have substantially improved our understanding of squamate origins and the rise of squamates to ecological dominance as rhynchocephalian diversity declined –10. Recoet al. [Cryptovaranoides microlanius  scan data available online [C. microlanius, as well as a wealth of comparative CT data and personal observations. We re-examined the original diagnosis relative to the referred specimens and holotype and re-assessed the primary homology concepts applied to create character scorings from the original study. Our detailed re-assessment of †C. microlanius reveals that the concept of the taxon was based on multiple fossil specimens that were not discovered in association with each other, save for the elements in the holotype block, and that there is little to no anatomical justification for the referral of these morphologically disparate skeletal remains to the same species . FurtheC. microlanius and divergence times for the major groups of squamates using three radically different phylogenetic matrices  under various optimality criteria. We find no support for the placement of †C. microlanius within Anguimorpha or even crown Squamata, with a broader reptile dataset suggesting that it is in fact an archosauromorph reptile. Divergence time estimates support previous estimates for a Mid-Late Jurassic origin of anguimorphs, highlighting how misinterpretations of the fossil record can highly impact our understanding of the origin of major branches of the Tree of Life.We tested the phylogenetic affinities of †. 2. 2.1et al. [C. microlanius are not ideal, as nearly all the non-lepidosaur species included in the original versions of those matrices were excluded. This approach potentially compromises the ability of the matrix to provide a strong test of whether †C. microlanius falls outside crown Squamata. Therefore, we included †C. microlanius in the largest available dataset to infer relationships among the major groups of reptiles [C. microlanius, we further expanded the taxonomic sampling of dataset 1 by adding three taxa with relatively unstable relationships, but which have historically been linked to Lepidosauria: †Fraxinisaura rozynekae from the Middle Triassic of Germany [Palaeagama vielhaueri [Paliguana whitei [Pali. whitei [The data matrices used by Whiteside et al.  to assesreptiles , as it ereptiles  for a de Germany , and †Paelhaueri  and †Pala whitei  from the. whitei  .  [et al. ,33)) is  [et al.  for theiat study . To thisat study –38 into e et al.  in datas [et al.  suggeste [et al.  proposedreserved .Table 1et al. [et al. [C. microlanius being deeply nested within squamates, regardless of the results presented by dataset 1.Finally, dataset 3 is a recent update of Gauthier et al.  by Brown [et al. , that sa [et al. ), taxono. 2.2. 2.2.1All maximum-parsimony (MP) analyses were conducted in T.N.T. v. 1.5  which al. 2.2.2rBayes v. 3.2.7a [Bayesian analysis of the morphological dataset was performed in M. 3.2.7a  using thWe used the Mkv + gamma substitution model  .Stationarity was assessed using standard measures, such as average standard deviation of split frequencies (ASDSF < 0.05) and potential scale reduction factors . Effective sample size values were assessed using Ter v.1.6 , reachin. 2.3rBayes v. 3.2.7a [Divergence times were calculated using relaxed clock Bayesian inference analyses of the morphological dataset for dataset 1 and the combined molecular and morphological data for dataset 2. We implemented total-evidence-dating (TED) using the fossilized birth-death tree model, under relaxed clock models in M. 3.2.7a .. 2.3.1ρ = 1). The age of the root also follows [The tree model and its calibration priors follow the previous analyses of this dataset in . Namely, follows , sampled follows ) and a s follows ). The ra follows . Specifi follows , which i follows .. 2.3.2ρ = 1) where all fossils are considered to be tips only.Sampling strategy among extant taxa was set to ‘diversity’, which is more appropriate when sampling maximizes extant diversity (as performed herein) and fossils are assumed to be sampled randomly ,46. AccoYoungina and crown reptiles) was sampled from an offset exponential distribution with a hard bound for the minimum age  and a soft maximum age, with the mean of the exponential distribution based on a recent TED analysis [The age of the root  [Diphydontosaurus  [The vast majority of our calibrations were based on tip-dating, which accounts for the uncertainty in the placement of extinct taxa and avoids the issue of constraining priors on taxon relationships when implementing bound estimates for node-based age calibrations ,49. The ssic—UK)  → 168.3–ssic—UK) ; SphenodGermany) → 241.5–2Germany) .We provided an informative prior to the base of the clock rate based on the previous non-clock analysis: the median value for tree height in substitutions per character from posterior trees divided by the age of the tree based on the median of the distribution for the root prior: 13.1/280 = 0.0464, in natural log scale = −3.07, and a wide standard deviation (1.0). We employed the uncorrelated independent gamma rate clock model  as in pr. 3. 3.1Reptilia Laurenti, 1768Cryptovaranoides Whiteside et al. [†e et al. .Cryptovaranoides microlanius Whiteside et al. [†e et al. .. 3.1.1Clevosaurus sp.; Whiteside et al. [Cryptovaranoides type block. This observation motivated us to critically re-evaluate the referral of additional material described by Whiteside et al. [Cryptovaranoides .NHMUK PV R36822, a partially articulated skeleton of a single, small reptile preserved in matrix. The presence of a large, isolated interclavicle in the block including the holotype demonstrates that additional reptiles  are repe et al.  to †Cryp. 3.1.2a); coronoid bone that is 40% of the anteroposterior length of the dentary and forms a low, gently rising coronoid process (b); surangular as long as dentary ; absence of incipient or developed rugosities and osteoderms on cranial bones (d); seven cervical vertebrae (c).Pterygoid anterior process considerably longer than posterior process a; corono process b; surang dentary b,c; abseal bones d; seven ertebrae c.. 3.1.3C. microlanius provided by Whiteside et al. [et al. [The diagnosis of †e et al.  cited see et al. ,6,7. Fur [et al. .. 3.2C. microlanius and note issues with the description presented in Whiteside et al. [C. microlanius were found in isolation and lack diagnostic features that would make it referable to the same species of the holotype. Besides the anatomical reinterpretations below, we also provide a list of all specimens referred to †C. microlanius by Whiteside et al. [C. microlanius in the data matrices used to infer its phylogenetic position.In this section, we review the anatomy of †e et al.  based one et al. , noting . 3.2.1Fusion of the premaxillae. Whiteside et al. [C. microlanius as fused, with a median tooth placed centrally. The presence of fused premaxillae is historically considered to be a squamate synapomorphy [C. microlanius would strongly support a squamate identity for this species. However, our re-evaluation of the CT scan data available for the holotype shows that the premaxillae are clearly unfused, and no median tooth is identifiable (c). The isolated premaxilla (NHMUK PV R37378) referred to †C. microlanius by Whiteside et al. [b,c) cannot be directly compared to the holotype specimen because all the teeth of the referred premaxilla are broken. Furthermore, although this referred specimen does appear to show some degree of fusion of the premaxillae near the tooth row margin, the premaxillae are separated throughout most of their extension, and the apparent fusion could be an artefact of preservation (e.g. suture infilling by surrounding sedimentary matrix).e et al.  describepomorphy ,6 and mopomorphy ,9. As sutifiable c. The ise et al.  .anius by , who argagmented a,b.Absence of a jugal posterior process. This widely cited character is related to the partial or complete loss of the lower temporal bar ancestrally in squamates, although it also appears in some stem lepidosaurs and rhynchocephalians [et al. [C. microlanius. However, as noted, this condition is also found elsewhere in lepidosaurs and in many other neodiapsids, such as kuehneosaurids, sauropterygians and ichthyosaurs [C. microlanius with Squamata over other clades of neodiapsids. Furthermore, the posterior region of the jugal is broken on the holotype of †C. microlanius , and so it is possible that a posteroventral process was present but not preserved.phalians ,7,29,33. [et al.  suggestehyosaurs ,53–55. Arolanius a,b, and Peg-in-notch articulation with rod-shaped squamosal. This feature was cited as a potential squamate synapomorphy of †C. microlanius [a–c). The long, thin bone identified as the squamosal by Whiteside et al. [et al. [C. microlanius.rolanius . Howevere et al.  is orien [et al. , but it Quadratojugal not present as separate element. This feature was also listed as a squamate synapomorphy of †C. microlanius by Whiteside et al. [e et al. . We disaFrontal underlaps parietal laterally on frontoparietal suture. This articulation was described as an anguimorph synapomorphy that is present in †C. microlanius based on the inferred articulation of the frontals with the parietals [a,b), and the referred frontals were found in isolation and cannot be anatomically connected to any of the other preserved elements in the skull without ambiguity  foramen and recessus scala tympani. Whiteside et al. [C. microlanius. Whiteside et al. [C. microlanius—see also figure 1d. Despite this, Whiteside et al. [e et al.  describee et al.  acknowlee et al.  still ine et al. ,7,9) or Enclosed vidian canal exiting anteriorly at base of each basipterygoid process. This braincase feature was described as a squamate synapomorphy of †C. microlanius [et al. [rolanius . However [et al.  noted, cFusion of exoccipitals and opisthotics forming an otoccipital. This feature was referred to as a squamate synapomorphy of †C. microlanius [d), we note that braincase fusion is quite variable within Squamata [rolanius . AlthougSquamata  and othe. 3.2.3Septomaxilla probably contacts dorsal surface of palatal shelf of maxilla (septomaxillary facet on maxilla). This feature was described by Whiteside et al. [C. microlanius. However, these are completely disarticulated and the septomaxilla is not preserved (a–c).e et al.  as a squreserved a–c.Long ventral longitudinal ridges converging toward midline of vomer. This feature was described by Whiteside et al. [C. microlanius. The vomers of †C. microlanius are large, flattened, and subrectangular, and are more similar to the vomers of non-squamate lepidosaurs  than toridensis ,9. The auimorphs , such ass apodus  and Elgaria spp. ,59, wherProminent choanal fossa on palatine. This feature was described as an unambiguous synapomorphy of Squamata present in †C. microlanius [C. microlanius is deep and anteroposteriorly restricted, matching the condition in gekkotans [Megachirella watchleri [Bellairsia gracilis [Oculudentavis spp. [rolanius . Howeverrolanius ,6,7,9. Tekkotans ,61 and tgracilis  and †Ocuvis spp. .Gephyrosaurus bridensis [Marmoretta oxoniensis [Taytalura alcoberi [Sphenodon punctatus [Navajosphenodon sani [C. microlanius is mediolaterally restricted so that it barely fills half of the mediolateral length of the anterior margin of the palatine [Tanystropheus hydroides [Macrocnemus bassanii [Stem and crown group members of the other major squamate clades display deeper and more posteriorly extensive choanal fossae ,9,63, whridensis , †Marmoroniensis , †Taytalalcoberi , and sphdon sani . Intrigupalatine . This copalatine  is preseydroides  and †Macbassanii .Short overlap in quadrate-pterygoid contact and the absence of the pterygoid process on the quadrate. This feature was described as a synapomorphy of Squamata found in †C. microlanius by Whiteside et al. [e et al. . However. 3.2.4Angular does not extend posteriorly to reach articular condyle. Although the angular was suggested to terminate before the mandible articular condyle in †C. microlanius as in the squamate total clade [a,b).al clade , the posArticulars and prearticulars medial process present. This feature was scored as present in †C. microlanius by Whiteside et al. [a,c). The absence of a medial process of the articular and prearticular can also be noticed in [figure 6g,h.e et al. . Howeverticed in : figure . 3.2.5Atlas pleurocentrum fused to axis pleurocentrum. The fusion of these elements cannot be assessed because their pleurocentra are not preserved in the holotype. Only the neural arches and neural spine of the atlas and axis are preserved, and their intercentra are missing from the holotype .holotype a,b.Cervical ribs double-headed. Whiteside et al. [C. microlanius, acknowledging this state was unusual for a squamate. However, inspection of the CT data indicates that the cervical ribs of †C. microlanius are in fact single-headed and possess an expanded endpoint for articulation with the vertebral centra . This differs from the condition observed in all known squamates and resembles the rib morphology observed in other reptile clades, including protorosaurs such as †Protorosaurus speneri . What was originally interpreted as the second rib head in †C. microlanius by Whiteside et al. [figure 2b,c—Ant.Pr.) commonly observed on the cervical ribs of several archosauromorphs, including †Protorosaurus speneri (b), †Prolacerta broomi (c), †Mesosuchus browni [Azendhosaurus madagascariensis [Euparkeria capensis [e et al.  is reint speneri b, †Prolaa broomi c, †Mesoss browni , †Azendhariensis  and seveariensis  and †Eupcapensis Cervical and dorsal vertebral intercentra present. Based on the number of preserved, articulated vertebrae, the presence of intercentra on the trunk vertebrae was described by Whiteside et al. [C. microlanius. However, the presence of intercentra was based on the presence of a single isolated bone fragment, suggested as a displaced intercentrum. Upon inspection of the cervical region using CT scan data (c), we observed that cervical centra are in close articulation without any evidence for intercentra or articulatory facets for them, which should be clearly visible in this particularly well-preserved specimen, as they are in extant squamates . TherefCervical vertebrae midventral crest absent. A midventral crest or keel on each caudal centrum was scored as absent in †C. microlanius by Whiteside et al. [b).e et al. . HoweverAnterior dorsal vertebrae, diapophysis fuses to parapophysis. Whiteside et al. [C. microlanius. However, the few preserved dorsals have unfused neural arches and pleurocentra (c), thus logically having their diapophyses  and parapophyses (located on the pleurocentrum) also unfused. Given the juvenile condition of the holotype specimen , it is possible that later during ontogeny those elements could have been fused together, forming a synapophysis. However, there is no evidence to support this given the material available. Secondly, even if synapophyses occur later in the ontogeny of †C. microlanius, these are observed across several groups of reptiles, including all other non-squamate lepidosaurs [e et al.  suggesterocentra c, thus lidosaurs ,68,71,72Zygosphene-zygantra in dorsal vertebrae. Incipient zygosphene-zygantra articulations were mentioned to be present in a set of vertebrae present in the block containing the holotype but separate from the holotype specimen of †C. microlanius [C. microlanius  is quite different from the coracoid emarginations (or ‘foramina’ as labelled by [a). We reinterpret them as an instance of incomplete mineralization of the central region of the coracoid, which is common among juvenile reptiles —e.g. a similar mode of preservation occurs in the coracoid of the Early Cretaceous South American lizard †Tijubina pontei (b). We note the holotype specimen appears to be a juvenile based on unfused neural arches and centra .anius by . We note figures a and 5c elled by ) observea pontei b. We notEntepicondylar and ectepicondylar foramen of humerus present. The ectepicondylar foramen is nearly universally present in Lepidosauria, whereas the entepicondylar foramen is lost in squamates but retained in sphenodontians [et al. [C. microlanius; however, we were unable to observe any foramina in their only illustration . This feature was not discussed by Whiteside et al. [Cryptovaranoides microlanius was scored as having the expanded radial condyle present in the phylogenetic dataset used by the authors, but our inspection of the CT scan data on both humeri of the holotype from different angles clearly shows this is absent in this taxon . This feature was not discussed by Whiteside et al. [C. microlanius suggests the ulnar patella is absent in this taxon  the goois taxon .. 3.3Cryptovaranoides within reptiles, as discussed above. In all of our results using dataset 1, †Paliguana whitei acted as a rogue taxon that contributed to poor resolution across the generated consensus topologies . Removing that species substantially improved the resolution . Under the most robust phylogenetic hypothesis , †C. microlanius is inferred to be the sister to Allokotosauria within Archosauromorpha, albeit with weak support .Dataset 1 was the matrix most suited for testing the broader affinities of †ccuracy, ; figure C. microlanius to lepidosauromorphs are challenged by important reinterpretations of the postcranial skeleton of the holotype, including: absence of ectepicondylar and entepicondylar foramina , thus m (contra ). The op [et al.  is reintrolanius .C. microlanius is supported by the following characters: strong anterior emargination of the maxillary nasal process, which is rarely observed in squamates but is a hallmark feature of archosauromorphs, where it contributes to the formation of the antorbital fenestra . Using relaxed clock Bayesian inference, †C. microlanius is inferred to occupy a similar position but with lower support, less than 50% PP . These results suggest that, when †C. microlanius is included in a lepidosaur-specific dataset, the several character states it shares with some early lepidosauromorphs and archosauromorphs place it close to the root of the tree. We note that only four archosauromorphs are used as an outgroup in this dataset, and so it does not provide an adequate test of the non-lepidosaurian affinities of †C. microlanius . The analyses of datasets 2 and 3 provide strong evidence that the anatomy of †squamate , well ouet al. dataset in [B. gracilis, †Oculudentavis khaungraae, and †Oculudentavis naja. Undated parsimony and tip-dated Bayesian analyses failed to recover †C. microlanius within the squamate crown, again placing this species as a stem squamate, here one node stemward of the clade formed by †B. gracilis, †Huehuecuetzpalli mixteca, †O. khaungraae, and †O. naja .Dataset 3 was analysed with and without the three species of stem-squamates added to the Gauthier taset in : †B. gra†O. naja a,b.Figu. 3.4C. microlanius and its inferred placement within archosauromorphs (C. microlanius (as an archosauromorph) is largely compatible with the specimen age  are estimated to occur during the Middle and Late Jurassic (C. microlanius is again more consistent with its age.Divergence times among the major groups of reptiles are largely unaffected by the inclusion of †pothesis ). Furthe. 4C. microlanius was originally interpreted as nested within Anguimorpha [C. microlanius is accurate, it would radically alter all previous hypotheses on the timing of squamate diversification, and potentially suggest widespread bias towards younger age estimates for squamates and other reptiles in timetrees produced using a wide variety of methods and both morphological and molecular data [C. microlanius posited by Whiteside et al. [The Late Triassic reptile †uimorpha , which iuimorpha ,25,76–78lar data ,24,25,78C. microlanius. First, we find no evidence for referring most of the other specimens noted by Whiteside et al. [C. microlanius to squamates and anguimorphs are in fact not observable (e.g. not preserved), poorly preserved and of ambiguous interpretation, or incorrectly described . We provide a thorough redescription of the holotype and provide, for the first time to our knowledge, detailed images of key anatomical traits that highlight several traits seen in †C. microlanius that are incompatible with a lepidosaur hypothesis, and instead support its affinity to archosauromorphs. Finally, phylogenetic analyses of three separate datasets with radically different taxonomic composition and criteria of character construction, under multiple optimality criteria, consistently reject the hypothesis that †C. microlanius is a crown squamate. Instead, our analyses find that †C. microlanius is a neodiapsid of unclear placement with potential affinities to early archosauromorphs.Reinterpretation of the original data and analyses strongly reject a crown squamate identity for †e et al.  to this Crown reptiles  underwent extensive radiation and diversification during the Early to Middle Triassic but have roots dating back into the Permian . While nC. microlanius shares with crown squamates an elongated (rod-like) squamosal and a large coronoid bone with a prominent dorsal process. Protorosaurian archosauromorphs [C. microlanius). Even when considering highly derived members of the archosauromorph tree, we can find features of interest that have been historically linked to lepidosaurs in systematic studies. For instance, hatchlings of Alligator mississippiensis have their first tooth generation not attached in a socket made of alveolar bone, but rather to the labial face of the medial wall of the tooth bearing element; that is, hatchling A. mississippiensis are  pleurodont [C. microlanius.For example, †romorphs ,28,48,82romorphs –88. Squaromorphs ,86,87 . As such, †C. microlanius and †F.corami are not likely to be conspecific; however, presence of two broadly similar taxa highlights the need for caution when referring isolated elements.Recently, Triassic formations in England have produced several diapsids that may be early diverging lepidosaurs, as well as plentiful examples of early rhynchocephalians ,24,91,92C. microlanius has larger implications for the interpretation of lepidosaur-like skeletons from the Triassic reptile assemblages of the UK and other Triassic faunas—see further discussion on the difficult interpretation on other Triassic fissure fill deposits previously linked to lepidosaurians in  that haC. microlanius remain relatively unclear, this species shares several features with archosauromorphs, the total clade of birds and crocodilians [Cryptovaranoides microlanius is placed as the sister of Allokotosauria, a diverse Triassic archosauromorph clade outside the crown group, with moderate support in analyses of dataset 1 that exclude †Paliguana. However, several anatomical features of †C. microlanius, especially the morphology of its dentition, markedly differ from the conditions observed in allokotosaurs. It appears that †C. microlanius is part of a poorly known radiation of early small-bodied archosauromorphs, but the phylogenetic affinities of this species will only be resolved by future fossil discoveries. †Cryptovaranoides highlights a potential new branch in the exceptional Triassic radiation of crown reptiles and demonstrates the probability that key small-bodied clades might still await discovery .This work did not require ethical approval from a human subject or animal welfare committee.


These advances include novel strategies for reducing CO2 per metric ton of clinker. This approach has the potential to significantly reduce the carbon footprint of cement production, a major source of greenhouse gas emissions. Sim et al. [2 sink to lower CO2 emissions from the cement industry. Their research demonstrated that complete carbonation can be achieved within 10 min at specific CSW ratios (5–25%), with the ability to reduce CO2 emissions from the cement industry.Wojtacha-Rychter et al.  exploredAmin et al. ,4 investAlternative materials and innovative mix designs are being explored to improve concrete performance. Kumar et al.  assessedNiu et al.  examinedThe findings presented in this SI are useful in the ongoing efforts to enhance the sustainability, durability, and performance of cement and concrete products. Moreover, such advances may help address the challenges of evaluating the sustainability of concrete materials and climate change factors that could influence the design of such materials.


Within the nonequilibrium environment of living cells, the transport behaviours are far from the traditional motion in liquid but are more complex and active. With the advantage of high spatial and temporal resolution, the single-particle tracking (SPT) method is widely utilized and has achieved great advances in revealing intracellular transport dynamics. This review describes intracellular transport from a physical perspective and classifies it into two modes: diffusive motion and directed motion. The biological functions and physical mechanisms for these two transport modes are introduced. Next, we review the principle of SPT and its advances in two aspects of intracellular transport. Finally, we discuss the prospect of near infrared SPT in exploring the  Microtubules, the structural backbone of the cytoskeleton, are long hollow tubes with a diameter of 22–25 nm composed of 13 parallel protofilaments. Microtubules are highly dynamic and polarized, alternating between phases of growth and shrinkage . In FRAP experiments, fluorescent molecules in a small given area are first photobleached by a focused laser beam with high intensity. Then, surrounding unbleached fluorescent molecules diffuse into the photobleached area resulting in fluorescence recovery , the laser penetrates the cell vertically. Since all probes across the cell are excited, the overall fluorescence within the cell imposes a strong background on the single probes of interest, making the signal-to-noise ratio relatively low. et al. et al. et al. et al. et al. et al. et al.To improve the signal-to-noise ratio, total internal reflection fluorescence (TIRF) microscopy was invented to selectively excite fluorescent molecules close to the cover glass (<200 nm) Axelrod . TIRF is et al..Due to the observation depth, TIRF is limited to studying the cell membrane. To observe fluorescent molecules in cells, researchers further invented highly inclined and laminated optical sheet microscopy (HILO)  due to the diffraction limit. We obtain an accurate position of the probe through Gaussian fitting. By linking the same particle’s different positions in consecutive images, particle trajectories are constructed . Both tiTo analyse the motion, the MSD of the trajectories is generally calculated:r(t) is the displacement, Δt is the time interval, and < ··· > is the average. Time-averaged MSD is:where M is the total time length of the trajectory. The ensemble-averaged MSD is:where i is the ID of each particle, and N is the total number of all particles. The two MSDs help us understand the motion of target particles, but their results are not always consistent. With increasing Δt, the MSD tends to show an upward trend, which can be described by a remarkable power-law curve MSD ~Δtα. The value of α depends on the motion type of the particle: α = 1 corresponds to Brownian motion, α < 1 refers to subdiffusion, and α > 1 refers to superdiffusion  on cell membranes  on cell membranes and artificial lipid bilayers using SPT   , 3B  technology is an effective method for observing deep tissue (Li et al. et al. et al. et al. et al. et al. et al. et al. et al. et al. et al. et al. et al.in vivo confocal 3D imaging of tumour vasculatures in mice at a depth of 1.2 mm (Zhang et al.in vivo, with promising applications in biophysical studies and biomedical diagnosis.SPT has promoted the study of intracellular transport dynamics, but there are still some challenges. To date, most SPT studies on intracellular dynamics have been carried out at the cellular level Ming-Li Zhang, Hui-Ying Ti, Peng-Ye Wang and Hui Li declare that they have no conflict of interest.


At one text position, the level of significance stated as “A correction has been made to Paragraph 5. This sentence previously stated:vaccinated) and 14 (placebo) dead during eleven weeks, then evidently giving numbers for complete trial groups . If our German-based estimate  is assumed to be the expected value of a binomial probability distribution then the corresponding standard deviation is quite exactly 5. A corresponding binomial test reveals that the 14 dead in the placebo group are significantly different from prognosticated 25 deaths, at a p-value of 0.0012. Hence, counting just 14 dead in Thomas et al. (7) is already utterly unlikely to be explainable by chance; and the 4 dead reported in Polack et al. (1) are an entirely impossible count, which can only reflect some preliminary data analysis. In stark disaccord, the data reported in Polack et al. (1) should without doubt stand on their own and not rely on additional publications, as this was a public dissemination of both safety and efficacy probed by a pivotal vaccine trial. Publishing another, later (6-month) safety data set as in Thomas et al. (7), or even secondary reports like, e.g., by the USA's “Food and Drug Administration,” should not be required when disseminating a primary endpoint assessment of safety .”“In continuation of Polack et al. (1), interestingly, the authors of (7) finally counted 15 (The corrected sentence appears below:vaccinated) and 14 (placebo) dead during 11 weeks, then evidently giving numbers for complete trial groups . If our German-based estimate  is assumed to be the expected value of a binomial probability distribution then the corresponding standard deviation is quite exactly 5. A corresponding binomial test reveals that the 14 dead in the placebo group are significantly different from prognosticated 25 deaths, at a p-value of 0.012. Hence, counting just 14 dead in Thomas et al. (7) is already utterly unlikely to be explainable by chance; and the four dead reported in Polack et al. (1) are an entirely impossible count, which can only reflect some preliminary data analysis. In stark disaccord, the data reported in Polack et al. (1) should without doubt stand on their own and not rely on additional publications, as this was a public dissemination of both safety and efficacy probed by a pivotal vaccine trial. Publishing another, later (6-month) safety data set as in Thomas et al. (7), or even secondary reports like, e.g., by the USA's “Food and Drug Administration,” should not be required when disseminating a primary endpoint assessment of safety .”“In continuation of Polack et al. (1), interestingly, the authors of (7) finally counted 15 (The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated.