Rep.7, 11046 (doi:10.1038/s41598-017-10669-4)) by using the same ecologically valid paradigm in an independent sample with similar age ranges. We were able to replicate the previously observed results of a preservation or even enhancement in socio-affective processes, but a decline in socio-cognitive processes for older adults. Our findings add to the understanding of how social affect and cognition change across the adult lifespan and may suggest targets for intervention studies aiming to foster successful social interactions and well-being until advanced old age.Anticipating population ageing to reach a historically unprecedented level in this century and considering the public goal of promoting well-being until old age, research in many fields has started to focus on processes and factors that contribute to healthy ageing. Since human interactions have a tremendous impact on our mental and physical well-being, scientists are increasingly investigating the basic processes that enable successful social interactions such as social affect  and social cognition (Theory of Mind). However, regarding the replication crisis in psychological science it is crucial to probe the reproducibility of findings revealed by each specific method. To this end, we aimed to replicate the effect of age on empathy, compassion and Theory of Mind observed in Reiter and colleagues' study (Reiter  Thus, met al. –14. Reseet al. ,16.with somebody [for somebody that is strongly linked to the motivation to help the other person [Socio-affective processes include empathy and compassion. The definition of empathy ranges from a multifaceted construct—comprising affect sharing and empathic concern as an emotional and mentalizing or perspective taking as a cognitive component of empathy –20—to a somebody . More prsomebody ,22. Compr person ,24. Somer person . In the et al. [Socio-affective and socio-cognitive processes are not just conceptualized distinct, there is also evidence from behavioural, neuroimaging and (sub-)clinical studies that supports their independence ,26. Inveet al.  found noet al. –30 mostlet al. , while aet al. ,32,33 moet al. . Furtheret al. –39. Wheret al. .et al. [either the socio-emotional or the socio-cognitive route [et al. [et al. [In view of the conceptual and neural differentiation of social affect and social cognition as well as the findings from (sub-)clinical populations, it is likely that these two subcomponents of social understanding might also be differentially affected by age. Over the last 30 years, a vast body of research emerged on this topic . One of et al. . The autet al. . Howeverve route ,59–61, w [et al. . The aut [et al. ) that en [et al.  observedHowever, with regard to the replication crisis in science in general and in psychological science in particular ,64, it iet al. [et al. [The reproducibility of the results of Reiter et al.  on the e [et al.  by using. 2This study was part of a larger project consisting of three testing sessions. Here, we focus on the behavioural measures of the third session in which participants performed the EmpaToM task ,28,67 an. 2.1N = 45, age range = 18–30; OA: N = 56, age range = 65–78). All participants were recruited via flyers and newspaper announcements in the greater Dresden city region. The OA were additionally recruited from sport, language and university courses as well as choirs for OA and the database of the Lifespan Developmental Neuroscience Lab at Technische Universität Dresden. All participants spoke and understood German fluently, had normal or corrected to normal vision, and were right-handed as well as MRI-suitable individuals . Additionally, participants were not allowed to have participated in the study of Reiter et al. [et al.’s sample [a priori exclusion criteria for the present sample: self-reported psychological or neurological disease currently or within the last 12 month, consumption of more than five cups of coffee a day , smoking of more than five cigarettes per day, drinking of more than 12 g (women)/24 g (men) pure alcohol per day, consumption of illegal drugs more often than two times per month or having a history of drug abuse or addiction, as well as having a prescribed hearing aid device, attested tinnitus or colour blindness. The OA were additionally screened for cognitive impairment (in the first session of the project) via the official German version of the Montreal Cognitive Assessment , problems with hearing or sight during the task (N = 4), termination of the session by the participant or due to technical problems (N = 2) or neurological abnormalities in the brain (N = 4). In contrast to Reiter et al. [N = 2) that performed below chance level (less than 0.33). This did, however, not affect the results in the present study .One hundred and one individuals participated in the third session . With a [et al. , who use [et al. . Hence, r et al. , we did M = 24.00, s.d. = 3.20) and 44 older  individuals. Between the two age groups, the gender distribution was comparable. YA and OA did also not differ significantly in their years of education, relationship status or residence  which verified that the final sample size was nevertheless appropriate to detect the effects with a power of 1-beta = 0.95.The final sample comprised 42 younger  and performed the battery of cognitive functioning tasks afterwards, or vice versa. At the end of the third session, they received 8.50€ per hour for participation. The Technische Universität Dresden ethics committee granted ethical approval in accordance with the Helsinki declaration for the whole project (EK 486 112 015).et al. [et al. [The EmpaToM  is a videt al. , we allo [et al. , we did  [et al.  nor repl [et al.  affect tet al.’s sample [et al.’s sample [et al.’s sample [et al.’s sample [et al. [In addition, participants performed a battery of cognitive functioning tasks. These comprised the Trail Making Test A and B , the Ids sample . Regardis sample  as well s sample . Howevers sample  and finds sample , which as sample  and the s sample ,76, OA w [et al.  with com. 2.3t-tests or χ2-tests using the R package stats [t-test were violated, a non-parametric test (Mann–Whitney U-test) was performed using the R package stats [Age differences in sample characteristics and cognitive measures were examined with independent ge stats . If the ge stats . Regardiet al. [Analogous to Reiter et al.  measureset al. [t-test using the R package stats [U-Test using the R package stats [Coherent with Reiter et al. , the folet al. . If the et al. . To analge stats . Since tge stats  as well.et al. [z-scores of the TMT A (RT) and B (RT), IDP (mean of RT and accuracy), and DSb ) as well as a unit-weighted composite score reflecting a proxy of verbal intelligence (based on the z-scores of the SAW (RT) and SAW (accuracy)) to test for potential effects of these cognitive abilities on the observed age effects. We performed the repeated measures analysis of covariance  separately for the proxy of fluid intelligence and for the proxy of verbal intelligence with the R package rstatix [Also consistent with Reiter et al. , we furt rstatix  and the  rstatix . All dat rstatix  and R 3. rstatix .. 3. 3.1F1,81 = 675.143, p < 0.001, ηp2 = 0.893), suggesting that emotionally negative videos elicited a stronger empathic response than neutral videos in our participants  and no significant interaction effect  in our sample. Thus, YA and OA did not differ significantly in their empathic response to emotionally negative and neutral videos. Since the assumption of normality was violated for the valence ratings of the neutral videos in YA and emotional videos in OA, we additionally performed a robust mixed ANOVA that validated the former analysis and replicated the findings of Reiter et al. [The mixed ANOVA of the valence ratings revealed a significant main effect of emotionality of the video , with OA reporting more compassion than YA across all conditions , with higher accuracies for ToM questions , and a significant main effect of age group , with YA answering more accurately than OA . We also found a significant interaction between question type and age group  that qualified the main effects and is in line with the findings of Reiter et al. [post hoc tests showed that both YA and OA performed significantly better in the ToM condition than in the factual reasoning condition, but the effect was stronger for OA  than YA   . Thus, tF1,84 = 0.570, p = 0.452, ηp2 = 0.007) nor of age group , suggesting that ToM and factual reasoning questions were answered similarly fast by YA and OA in our sample and replicates Reiter et al.’s results [F1,84 = 0.906, p = 0.344, ηp2 = 0.011) ; however, robust post hoc tests on the difference scores  did not reveal a significant difference in RT as a function of age group .The mixed ANOVA for RTs showed neither a significant main effect of question type ( results . The int= 0.011) . This fi results , who fou. 3.3Fs1,83 ≥ 18.123, all ps < 0.001, all ηp2 ≥ 0.179) and compassion . The separate adjustment for the proxy of fluid intelligence or the proxy of verbal intelligence did not alter the absence of an age effect on empathy either . Since the assumption of homogeneity of variances was violated for empathy as well as for compassion, we additionally conducted robust ANCOVAs that validated our former analysis. Thus, adding a proxy of fluid intelligence or verbal intelligence as covariate in the analyses did not alter the effects of age on ToM/factual reasoning accuracy and compassion or the absence of an effect of age on ToM/factual RT and empathy, which is in line with Reiter et al. [To test for potential effects of fluid intelligence and verbal intelligence on the observed age effects on ToM/factual reasoning accuracy and compassion as well as the absence of an age effect on empathy, we conducted separate mixed ANCOVAs, with either the proxy of fluid intelligence or the proxy of verbal intelligence as covariate. Controlling for these covariates separately, yielded the same age effects as observed above on ToM/factual reasoning accuracy  and compassion (as concern for the narrator). For empathy we found a significant main effect of video type on the valence ratings, showing that emotionally negative videos are experienced as more negative than neutral videos in both YA and OA . The maet al. , indicat. 4.2et al.’s findings [Post hoct-tests on the interaction effect differed slightly from Reiter et al.’s results [et al.’s sample [et al. [et al. [We assessed socio-cognitive processes after the participants had watched a short video of a protagonist talking about an emotionally negative or neutral life-event by measuring ToM (as the reasoning about a narrator's thought). Factual reasoning (as the reasoning about facts included in the story) was used as control condition. Regarding the accuracy for ToM and factual reasoning, we found a significant main effect of age, revealing that YA outperform OA in both conditions. We also found a significant main effect of question type, showing that both YA and OA perform better for ToM versus factual reasoning questions. The main effects as well as a significant interaction of question type and age that qualified the main effects are in line with Reiter findings . Post ho results . OA perfs sample . However [et al. , in the  [et al. . Neverthet al.’s results [et al. [et al. [et al. [et al. [Regarding the response time for the ToM and factual reasoning questions, we found neither a significant main effect of age nor of question type, revealing that both age groups respond equally fast, and both question types are answered equally quickly. This is again in line with Reiter  results . However [et al.  had repo [et al. , OA perf [et al.  had part [et al. , which m. 4.3et al.’s results [To control for potential effects of diverging cognitive abilities between the age groups, we conducted separate ANCOVA analyses for empathy, compassion and ToM with either a proxy of fluid intelligence or a proxy of verbal intelligence as covariate. These control analyses did not qualitatively change the observed results which are in line with Reiter  results . This in results ) as well results ,82). As  results ,84. The . 4.4et al. [et al.’s sample [We were able to replicate the effect of age on empathy and compassion reported by Reiter et al. —revealins sample ). This ss sample ,86 that et al. [et al. [Nevertheless, the current study has three main limitations that would be important to address in future research. First, testing high-functioning individuals in our sample with equal levels of education in YA and OA, can be regarded as both a strength and a limitation. The latter particularly relates to limitations with respect to external validity or the question whether the findings can be generalized towards other populations. In a similar vein, we note that the current sample is, as in most previous research, a ‘western, educated, industrial, rich and democratic’ (WEIRD) sample , which iet al. , as well [et al.  as well  [et al. ,99. ThusDespite these limitations the current study lends more evidence to the view that socio-affective and socio-cognitive processes age differentially across the adult lifespan. Further, in view of the replication crisis in psychological science, reproducing the results of a study with an independent sample in a different setting (e.g. here MRI versus test cabins) has great value not only for science itself, but also for the reliability of the results and thus new ideas, theories or interventions that can be derived from these results. Overall, our findings support the assumption that the socio-affective processes are preserved or even enhanced while socio-cognitive processes decline with age, which is derived from the socio-emotional selective theory  in the aClick here for additional data file.


Methane is an important greenhouse gas, emissions of which have vital consequences for global climate change. Understanding and quantifying the sources (and sinks) of atmospheric methane is integral for climate change mitigation and emission reduction strategies, such as those outlined in the 2015 UN Paris Agreement on Climate Change. There are ongoing international efforts to constrain the global methane budget, using a wide variety of measurement platforms across a range of spatial and temporal scales. The advancements in unmanned aerial vehicle (UAV) technology over the past decade have opened up a new avenue for methane emission quantification. UAVs can be uniquely equipped to monitor natural and anthropogenic emissions at local scales, displaying clear advantages in versatility and manoeuvrability relative to other platforms. Their use is not without challenge, however: further miniaturization of high-performance methane instrumentation is needed to fully use the benefits UAVs afford. Developments in the models used to simulate atmospheric transport and dispersion across small, local scales are also crucial to improved flux accuracy and precision. This paper aims to provide an overview of currently available UAV-based technologies and sampling methodologies which can be used to quantify methane emission fluxes at local scales.This article is part of a discussion meeting issue 'Rising methane: is warming feeding warming? (part 1)'. Both emissions of CH4 to atmosphere and the absolute concentration of CH4 in the atmosphere have increased over the past decade )in situ atmospheric measurements or remote sensing of methane concentrations at the spatial scale of local sources (less than 1 km) and small facilities. UAVs can be equipped to dynamically monitor the lower atmosphere and planetary boundary layer (e.g. )Aside from classifications based on size and payload weight, UAVs can be broadly divided into fixed-wing and rotary-wing aircraft. Fixed-wing UAVs resemble traditional airplanes and the largest of these  can ben. 3Accurate wind speed and wind direction is typically required for flux quantification. These can be measured (or inferred) from a suitable nearby monitoring station, equipped with anemometer instrumentation, but doing so may introduce some measure of uncertainty . Preferaet al. )et al.  tested a ) ) ([CH4]) . U⊥ is tThe mass balance method generally requires that the wind field is constant between upwind and downwind measurements, i.e. that the wind speed and wind direction does not change. However, wind field variability can be implicitly accounted for through robust uncertainty propagation . When uset al. [4 and CO2 to quantify a methane flux from landfill. They used two UAV platforms: a fixed-wing UAV equipped with a high-precision CO2 instrument  and a rotary-wing UAV tethered to an off-axis Integrated Cavity Output Spectrometer  Ultraportable Greenhouse Gas Analyzer [2, while the fixed-wing UAV was used for horizontal measurements of CO2, from which the horizontal distribution of methane was inferred using emission ratios characteristic of landfills (−1) with the uncertainties mainly influenced by the variability in the methane background, and the variability in the wind. Their reported uncertainties were comparable to uncertainties derived from other non-UAV-based landfill flux estimation methods (e.g. [2 can be a poor proxy for landfill methane emissions in the presence of interfering (off-site) sources [Allen et al.  used theAnalyzer ). The roandfills . Methaneds (e.g. ). Howeve sources  and the  sources .Figure et al. [−1. This was larger than the fluxes quantified by two additional, non-UAV-based methods, which yielded methane emission rates of 5.8 g s−1. Nathan et al. [Nathan et al.  used theet al. ,93) was n et al.  noted thn et al. .A variant of the mass balance method was used to detect and quantify natural gas leaks using a UAV equipped with a backscatter tunable diode laser absorption spectrometer . This inistances . The metThe mass balance approach, incorporated alongside algorithms to both detect and localize leaks, was used to quantify natural gas emissions using a UAV-based remote sensing measurement platform . Methane (b)F is the emission flux, U⊥ is the perpendicular wind speed, H is the height of the emission plume source above the ground and yσ and zσ are Gaussian dispersion parameters of the plume in the y  and z  directions, respectively. It should be noted that for the Gaussian plume method, y is conventionally used to refer to the cross-plume coordinate, whereas x is used to denote the same quantity for the mass balance box method.Gaussian plume models can be used to model enhancements in methane concentration downwind of a point source using Gaussian statistics . This meet al. [−1, larger than both the mass balance derived flux (14 ± 8 g s−1), and the two independent non-UAV-derived fluxes (both 5.8 g s−1). Ali et al. [Nathan et al.  used thei et al.  used theThe Gaussian plume method usually requires a large amount of time averaging in order for the instantaneous plumes to resolve into an observed Gaussian plume morphology suited to Gaussian inversion. When sampling in close proximity to an emission source (less than approximately 100 m), the time scale of measurements is unlikely to allow for adequate time averaging, and a non-Gaussian distribution of methane may be observed. The turbulent fluctuation of the wind field combined with the short flight duration of many UAVs may result in the measurement of an instantaneous plume and, therefore, a poorly defined plume morphology. However, repeated flights (or longer duration sampling) in less variable wind conditions, combined with carefully designed flight patterns can mitigate this. Individual case studies would need to be evaluated for the conditions specific to the environment at the time of sampling, by comparing data with fitted Gaussian plume assumptions.yσ and zσ increase linearly with distance from the source location . This method requires adequate spatial sampling density in both the y and z directions to resolve a flux [A recently developed adaptation of the Gaussian plume flux inversion, referred to as the near-field Gaussian plume inversion (NGI) technique, was adapted for downwind sampling of turbulent plumes close to the emission source . In the e a flux .et al. [et al. [Shah et al.  tested t [et al.  conclude−1, in good agreement with fluxes derived from nearby ground-based monitoring [The NGI method was used to quantity the methane emission flux resulting from unintended cold-venting during flowback operations at a hydraulic fracturing facility in the UK  figure . In thisnitoring . The rannitoring .Figure et al. [z direction (zσ) was accounted for by using an open-path methane sensor measuring column-integrated methane, and dispersion in the y direction (yσ) was assumed to be small, due to the close proximity to the source. Quantified fluxes were shown to be in agreement with the mass balance approach used in the same study [Another approach related to the Gaussian inversion method was used by Golston et al.  to quantme study . (c)A methane emission flux could feasibly be quantified using UAVs in conjunction with the tracer release method, and a controlled release of a reference tracer gas at a known rate from the source origin e.g. ,101–103)–103101–1Alternative atmospheric inversion approaches could also be applied to UAV-based measurement data to quantify methane fluxes. For example, Lagrangian particle dispersion models simulate the paths of many massless particles as they travel with the local wind field (e.g. ) and havet al. [et al. [Ravikumar et al.  compared [et al.  conclude. 6The literature reviewed here demonstrates the clear capability of small UAV-based measurement platforms for quantifying methane fluxes from a range of sources at local scales (less than 1 km). Such capability has only become available over the past decade, as a result of advances in both UAV technology and the miniaturization of instrumentation for the measurement of methane. Lightweight and high-performance instrumentation, with the capacity to measure methane with a precision on the order of a few ppb, are now becoming commercially available (at the time of writing), and developments continue to be made at pace. Such instrumentation will enable more accurate flux quantification, particularly of smaller emission sources which may not be captured by current methodology. Further developments in UAV technology would also be highly beneficial. Enhanced flight duration, as a result of improved battery capacity, would be extremely advantageous in terms of plume mapping, allowing for extended temporal ranges and refined spatial resolution of methane measurements. A summary of advantages and disadvantages of different UAV platform types, sampling methodologies, and flux quantification techniques is presented in UAVs provide a highly versatile platform, uniquely suited for high-density spatial mapping of methane plumes, and quantifying methane emissions at local scales. Their use could easily be incorporated into policy and regulation concerning monitoring and quantification of methane emissions from polluting industries  and also


Adipose tissue senescence is implicated as a major player in obesity- and ageing-related disorders. There is a growing body of research studying relevant mechanisms in age-related diseases, as well as the use of adipose-derived stem cells in regenerative medicine. The cell banking of tissue by utilising cryopreservation would allow for much greater flexibility of use. Dimethyl sulfoxide (DMSO) is the most commonly used cryopreservative agent but is toxic to cells. Trehalose is a sugar synthesised by lower organisms to withstand extreme cold and drought that has been trialled as a cryopreservative agent. To examine the efficacy of trehalose in the cryopreservation of human adipose tissue, we conducted a systematic review of studies that used trehalose for the cryopreservation of human adipose tissues and adipose-derived stem cells. Thirteen articles, including fourteen studies, were included in the final review. All seven studies that examined DMSO and trehalose showed that they could be combined effectively to cryopreserve adipocytes. Although studies that compared nonpermeable trehalose with DMSO found trehalose to be inferior, studies that devised methods to deliver nonpermeable trehalose into the cell found it comparable to DMSO. Trehalose is only comparable to DMSO when methods are devised to introduce it into the cell. There is some evidence to support using trehalose instead of using no cryopreservative agent. Ageing results in adipose tissue dysfunction, resulting in systemic effects such as peripheral insulin resistance and inflammation. Cellular senescence and progenitor cell dysfunction are also seen in ageing adipose tissues, and these may represent potential therapeutic targets in age-related disease. Adipose tissue comprises many cell types, generally divided into two fractions: the adipocyte fraction (AF), which contains primarily mature adipocytes, and the stromovascular fraction (SVF), which comprises progenitor cells , pericytes, and fibroblasts. Keeping these cells in long-term culture for clinical use or research has inherent limitations, and cell banking of tissue by utilising cryopreservation would allow for much greater flexibility of use .ADSCs also show great promise in regenerative medicine as these cells can differentiate into several different cell types, which can be used for tissue regeneration. For example, ADSCs can differentiate into a large number of cell types, which include chondrogenic, myogenic, osteogenic, angiogenic, and neuronal lineages . With reVarious methods of stromal vascular fraction cryopreservation have been demonstrated , but theDimethyl sulfoxide (DMSO) is the most commonly used CPA in the cryopreservation of ADSCs. DMSO is a small amphipathic molecule with two non-polar methyl groups and a polar sulfoxide group. The solvent activity of DMSO allows it to readily cross cell membranes. DMSO helps prevent the formation of intracellular ice, thereby protecting the tissue from freezing injuries and helping achieve maximum tissue survival during the freezing and thawing process. DMSO is toxic to cells at room temperature and washing and centrifugation are needed after thawing to remove it from the tissue . This isOwing to their low toxicity, sugars have been used as extracellular cryopreservation agents. Trehalose (α-d-glucopyranosyl α-d-glucopyranoside) is a disaccharide of glucose with low toxicity in contrast to DMSO. It is synthesised by species of bacteria, fungi, yeast, insects and plants that are prone to dehydration but is absent from vertebrates . Such orIn an update on the cryopreservation of adipose tissue and ADSCs, Shu et al.  highlighA systematic review was performed using the following databases: PubMed, CINAHL, Web of Science, MEDLINE, Cochrane Library and EMBASE. We included the following keywords and search terms: adipose stem cells, cryopreservation, trehalose, lipoaspirate, autologous fat grafting and cryopreservation. Those keywords were searched in that order in each database. We used a snowballing technique with an arbitrary cut-off of greater than two hundred articles for each database. An initial reviewer performed the search before being checked by a second reviewer. Any disputes about inclusion were resolved by a third reviewer.The following inclusion criteria were applied: (1) interventional studies using trehalose in the cryopreservation of human adipose-derived stem cells, including case series, case–control, cohort studies and randomised controlled trials; (2) studies translated into the English language; and (3) full article accessible.Exclusion criteria: (1) unpublished literature; (2) literature published before 2000; (3) articles not available for free viewing; and (4) studies using animal adipose-derived stem cells.A total of 430 articles were retrieved. A total of thirteen articles with fourteen suitable studies were relevant for this systematic literature review. The studies included and excluded are summarised in Eight studies were included and are described in Pu et al.  tested tIn a later study, Pu et al.  injectedCui et al.  comparedPu et al.  found inFour Level 3 studies were available for analysis . Two stuRao et al.  examinedP < 0.05).Roato et al.  comparedFour studies were included for analysis . Study pP < 0.5). Rao et al. [Cui et al.  examinedo et al.  used nano et al.  and Cui o et al.  reportedThere are several limitations to this review. Firstly, there is an English language bias in our search strategy as only articles published in English were reviewed. However, a study by Juni et al.  found thThe results from eight studies in our systematic review showed that DMSO and trehalose could be combined effectively to cryopreserve adipocytes. The advantage of this combination is that it reduces the dose of DMSO required. All four studies comparing trehalose and DMSO to simple cryotherapy with liquid nitrogen alone found a significant improvement in cryopreservation ,23,25. PP > 0.05). Wolter et al. [To maximise cell viability, a balance is required between reducing intracellular ice formation (by cooling slowly) and cooling quickly enough to minimise the solution effects. Mazur proposed a two-factor hypothesis attributing a lower cell survival rate when the cooling rate is slower than optimal . A faster cooling rate than optimal also decreases cell survival due to intracellular ice formation ,34,35,36r et al.  comparedStorage temperature is thought to significantly influence the viability of cryopreserved adipose tissue. Subzero temperatures result in ice formation damage, but this can be mitigated by the storage of tissue at less than −70 °C. Rapid thawing can improve cell viability by reducing the risk of intracellular ice formation, and subsequent ice crystal growth. Son et al.  performeLong-term cryopreserved ADSCs demonstrate a low risk of tumorigenicity and therefore offer great potential in regenerative medicine . DevelopFinally, there remains some variation in the cryopreservation of tissues and further studies are required to determine optimal protocols. Choudhery et al. demonstrated that adipose tissue could be successfully cryopreserved without compromising cell morphology, as well as subsequent proliferation and differentiation potential , making


In the beginning was… the genome.Nature offices in Little Essex Street and handed in these giant Figures together with envelopes containing the manuscript and the other Figures, which I had brought down from Manchester. Nature then lost the copies of Figure 1 but, by 27th March 1992, the paper was accepted and appeared in the journal on the 7th of May 1992  to the physical distance (in kb) showed a > 40-fold range for different intervals along the chromosome, being smallest close to the centromere and greatest half-way down each chromosome arm  that did not appear until the following year. The regular issue of Nature that appeared at the same time carried a ‘News & Views’ piece on the yeast genome from Craig Venter and his colleagues  RNAs , it was immediately obvious that the normal course of genetics would need to be turned on its head. Instead of isolating mutants with defective or altered phenotypes and using classical genetic analyses to define functional genes and map their relative locations on chromosomes, now genes would be both identified and mapped by genome sequencing, and it would be necessary to work in the opposite direction to discover their functions (Oliver https://cordis.europa.eu/project/id/BIO4950080), comprising some 145 laboratories from 14 countries, was established at the beginning of 1996 with the aim of determining the role of 1000 S. cerevisiae genes of unknown function that had been identified from the genome sequence  diploids so that all protein-encoding genes should be interrogated. Studies were carried out both in batch  that the ancestor of this and related species underwent during their evolution. This major evolutionary event was first inferred by Ken Wolfe and Dennis Shields  waltii  by carrying out what have come to be known as ‘multi-omic’ experiments  interactions by using the synthetic genetic array (SGA) methodology developed by Charlie Boone, Brenda Andrews, and colleagues in Toronto  and UNICELLSYS . YSBN had a coordinating role in systems biology research using yeast as a model organism and aimed to employ both mathematical analyses and computational tools to integrate experimental data; it set out to develop common resources and standards. UNICELLSYS had the more focussed aim of achieving a quantitative understanding of how cell growth and proliferation are controlled and coordinated by both extracellular and internal signals. These two networks operated in a very synergistic manner.As with Functional Genomics, the advent of Systems Biology saw the establishment of networks of European yeast researchers to pursue major programmes of work—the Yeast Systems Biology Network , perhaps because the transcriptome analyses were not carried out using RNAseq technology . This model (Yeast1) was rapidly updated even during the course of YSBN (Nookaew et al. et al. et al. S. cerevisiae metabolic network is the subject of an excellent recent review (Yu et al. A major contribution of YSBN was to establish a consensus stoichiometric model of the yeast metabolic network (Herrgård et al. (S. cerevisiae genome sequence (Goffeau et al. et al. Other systems of modelling yeast metabolism have been developed. Reiser et al.  construcet al. et al. et al. kcat or km values (Kroll et al. et al. et al. et al. (et al. (E.coli (Edwards and Palsson The most important developments of the genome-scale stoichiometric model of yeast metabolism have involved the exploitation of transcriptomic (Lee . et al.  used mac (et al. . In thiset al. et al. et al. et al. et al. et al. et al. Much of the work described in this article is the result of open-ended research programmes that aimed to produce useful data. In other words, they were hypothesis generating, rather than hypothesis testing. Science has always needed both these kinds of research (Kell and Oliver


R. irregularis genes during co-existence of two R. irregularis isolates in cassava. (3) An adjusted probability of <0.1 led to a substantial number of false positives. (4) There was no evidence that two R. irregularis isolates showed divergence in a putative MAT-locus indicating mating compatibility. We consider that there are serious errors in the assumptions and rationale used for the re-analyses, a mis-interpretation of the original data, as well as the discarding of important data without any justification.The commentary by Malar et al.  raised fAlthough Malar et al. “do not exclude the possibility that the genes identified by Mateus et al. are involved in mating,” they qualify the homology inference between genes differentially expressed in the co-inoculation treatment and genes involved in mating in other fungal species as “spurious evolutionary relationships” or “not the best ortholog”. Those statements imply that they attach no importance to the demonstrated sequence homology relationships identified in Mateus et al. Orthology does not necessarily imply conservation of gene function and genes with equivalent functions are not necessarily orthologs . TherefoMalar et al. have not considered, or have misunderstood, the experimental evidence on gene expression in interpreting their homology search. It is not surprising that their “best homologs” were not upregulated, because we already saw that those genes were not upregulated in the original dataset. Our approach comprised performing an experiment to identify genes that were specifically upregulated when two isolates coexisted in planta. We then identified their putative function by homology. We did not look at whether the genes were the closest orthologs. However, we discussed the limitations of an homology approach to identify gene function . To our R. irregularis). In fact, Malar et al. compared the 18 genes against “all protein gene catalogs of fungal species from the JGI fungal genomic resource” comprising 1318 taxa. The interpretation of the number of hits on a such large dataset is misleading because if a gene is highly conserved across the fungal kingdom, we would expect hundreds of hits in this database. In contrast, if an R. irregularis gene is highly specific to the Glomeromycotina taxa, we would expect very few hits (because there are less Glomeromycotina genomes in the database). Consequently, the number of hits in Table 1 from Malar et al. reflect the size of the database used and how conserved a given gene is, rather than whether a gene is from a large gene family. Malar et al. identified the so-called “closest ortholog” in R. irregularis of fungal mating genes from other fungal species by showing the “best hit” using OrthoMCL. However, differentiating paralogs from orthologs is a complicated task, in very distant species, especially if the organisms are highly paralogous. A more cautious analysis for each gene, including a confirmation that they are located in similar genomic locations, would lend more certitude that a given gene could be an ortholog. Consequently, the evaluation of RNA expression of their “best hit” remains incomplete in terms of the effort to find the best orthologs.Malar et al. claim the identification of hundreds of hits of the 18 genes differentially expressed in Mateus et al. “against the high-quality protein databases from the JGI Mycocosm Rhiir2” , show that they indeed performed a two-way comparison of DAOM197198 vs. co-inoculation with DEseq2 but then a comparison with the three treatments B1, co-inoculation, and DAOM197198 (raw data from GitHub repository: madhubioinfo). The consequence of the three treatments as input in the DEseq2 analysis is that the software compares the first and the third treatments. In their reanalysis, Malar et al. actually compared B1 vs. DAOM197198, which completely invalidates their claims.Malar et al. claim to have found specific upregulation in only two putative orthologous genes in their reanalysis of the RNA-seq data. But they have discarded data without giving any scientific justification. In Supplementary Table 2, Malar et al. appear to have manually edited several cases with: “N.A = cannot be calculated based on mapped reads” even though the analysis should always yield a value, and did in our analysis. There is no explanation given about why they considered expression values could not be calculated, especially for only one comparison of a given gene (DAOM197198 vs. Co-inoculation) but not the other comparison of the same gene (B1 vs. co-inoculation). Consequently, the omission of those values that ensured their reanalysis would provide a different conclusion from that of Mateus et al.As clearly stated in Mateus et al., we used an adjusted probability of <0.1 to maximize the identification of interesting genes. However, the change of probability threshold does not change the main finding that a large number of genes upregulated are homologs of genes involved in fungal mating. Eleven, of the 18 highlighted genes, displayed an adjusted probability <0.05 in both comparisons, and the other 7, an adjusted probability below 0.05 in one comparison and between 0.05 and 0.1 in the other comparison (Supplementary Table 6 ). FurtheThe authors questioned the possibility of mating compatibility between isolates B1 and DAOM197198, because no sequence information at the putative MAT-locus  was provTo conclude, Malar et al. asked “Where is the evidence?” To respond to their question, the evidence of specific gene upregulation during AM fungal co-existence is where Malar et al. chose not to see it.


Sheaffer blue ink is an effective method to stain arbuscular mycorrhizal (AM) fungi in a variety of plant species. It has, however, received criticism for its potential rapid degradation and short-term viability. The long and medium term storage and viability of stained samples has not, to date, been described for this particular staining method. This short communication reports on the viability of 730 samples stained with Sheaffer blue ink stored for the duration of 4 years in microscope slide boxes out of direct sunlight. There was no significant difference in micrograph image quality and presence of stain between years as indicated by the number of AM fungal structures quantified. In conclusion Sheaffer blue ink stain does not deteriorate in the medium term. The present communication evaluates the medium-term viability of Sheaffer blue ink staining for arbuscular mycorrhizal fungi. To date, Sheaffer blue ink stained root materials have not been subject to a further quantification and assessed for the longevity of the original stain during prolonged storage. The samples evaluated in the present communication show that root tissues retain the original stain and maintain the capacity to identify AM fungal structures for a minimum of 4 years.Arbuscular mycorrhizal (AM) fungi are symbiotic biotrophs  that are34H28N6O14S4) or lactophenol cotton blue as a method of detecting mycorrhizal structures such as vesicles, arbuscules and intra-radiating hyphae  dissolved in ethyl acetate [C4H8O2]) and viewed under a light microscope at ×100 magnification. Samples were stored at room temperature in microscope slide boxes avoiding direct sunlight. Images of samples  stained with Sheaffer blue ink n=30 accord samples  were takt-tests of equal variance compared the baseline year (the year that the root samples were stained) with observations of root arbuscules for the same samples in the year 2021 . Statistical significance was determined by P values ≤0.05.Statistical analyses were conducted using the R commander  software package. Arbuscular mycorrhizal fungal root arbuscules were quantified both visually and using CamLabLite version 1.0.8942.20170412 . The mean and standard error was calculated for each set of sample data. A single factor ANOVA tested for differences in winter wheat root mycorrhizal structures between years before further statistical testing was performed. Paired two-tail P=0.76, df: 4, 256, F value: 51.90, F critical: 2.64, Single factor ANOVA). A paired two-tail t-test of equal variance revealed no significant difference .Single factor ANOVA testing revealed no statistical difference between AM fungal root structures between the years quantified  (P=0.76,fference  between et al. [et al. [The present communication evaluates the medium-term viability of Sheaffer blue ink staining for AM fungi structures in winter wheat roots when stored under appropriate conditions. It contests the short-term viability of Sheaffer blue ink staining reported by Vierheilig et al.  and cite [et al. .et al. [et al. [et al. [et al. [Modifications to the ink staining approach, implemented by Wilkes et al. , who use [et al. . Recent  [et al.  identify [et al. . Additio [et al.  staininget al. [et al. [et al. [et al. [et al. [et al. [et al. [et al. [et al. [Vierheilig et al.  did not  [et al. . Kowel e [et al.  also use [et al.  demonstr [et al.  The impa [et al.  fail to  [et al.  and Wilk [et al.  to those [et al.  when comThe present communication has been able to demonstrate that Sheaffer blue ink, in the staining of AM fungal root components, has medium term storage potential when root tissues are fixed in FAA solution. The avoidance of boiling in KOH and using a fixative afterwards preserves AM structures, improves staining clarity and longevity. The continued monitoring of sample clarity and an evaluation of the impact of storage procedure, for example the effects of temperature and sunlight, on the persistence of ink as a mycorrhizal stain will be undertaken as future work.


Tritrichomonas foetus is a venereal trichomonad parasite which causes reproductive issues in cattle. No other trichomonads are known to be urogenital pathogens in cattle, but there are several reports of Tetratrichomonas and Pentatrichomonas isolates of unclear origin from the cattle urogenital tract (UGT) in the Americas. This study reports the first case of a non-T. foetus cattle urogenital trichomonad isolate in Europe. Molecular analysis of the internal transcribed spacer (ITS) 1-5.8S ribosomal RNA-ITS 2 and 18S ribosomal RNA loci suggest that the isolate is a Tetratrichomonas species from a lineage containing other previously described bull preputial isolates. We identified close sequence similarity between published urogenital and gastrointestinal Tetratrichomonas spp., and this is reviewed alongside further evidence regarding the gastrointestinal origin of non-T. foetus isolates. Routine screening for T. foetus is based on culture and identification by microscopy, and so considering other trichomonad parasites of the bovine UGT is important to avoid misdiagnosis. Tritrichomonas foetus is an important bovine venereal parasite which causes reproductive issues including spontaneous abortion, pyometra and infertility  of animals including invertebrates, fish, mammals, reptiles, amphibians and birds , respectively , 5 g L−1 neutralized liver digest (Oxoid) and 20 g L−1 glucose, pH 7.4], supplemented with 200 units mL−1 penicillin, 200 μg mL−1 streptomycin and 1000 units mL−1 polymixin B, overlaid on a solid medium slope prepared by heating 7 mL horse serum at 75°C for 2 h. Cultures were incubated at 37°C for 7 days, and were examined microscopically on days 4 and 7 for motile protozoa. Positive cultures were passaged in fresh media, and 10 mL culture was harvested by centrifugation at 300 × g for 10 min, fixed in 15 mL 100% ethanol and stored at −20°C. No cryopreserved stock of the isolate was generated.Samples were collected from bulls in the UK during routine screening for parasites, in accordance with the principles defined in the European Convention for the Protection of Vertebrate Animals used for Experimental and Other Scientific Purposes. To collect samples, the preputial cavity was washed with 30 mL pre-warmed phosphate buffered saline (PBS) (pH 7.2). Washings were pelleted by centrifugation at 300 × Genomic DNA was extracted from ethanol-fixed parasite isolates using the DNeasy ultraclean microbial kit (Qiagen) according to the manufacturer's instructions. Loci for molecular sequence typing were amplified by polymerase chain reaction (PCR) using generic Taq polymerase (NEB). A region containing the 5.8S ribosomal RNA (rRNA) and flanking internal transcribed spacers (ITS) 1 and 2 was amplified using the trichomonad-specific TFR1 and TFR2 primers, and the 18S rRNA gene was amplified using the generic eukaryotic primers Euk 1700 and Euk 1900. Resulting products were cloned into pCR4TOPO using the TOPO TA cloning kit (ThermoFisher Scientific) according to the manufacturer's instructions. Sequences were generated for the inserts from five independent clones by Sanger sequencing (Eurofins) on both strands using the T7 and T3 promoter primers, and additional internal sequencing primers were designed to cover the full length of the 18S rRNA gene for both strands. All primer sequences are listed in Supplementary Table S1.et al., et al., et al., et al., et al., et al., et al., A reference collection of ITS1-5.8S rRNA-ITS2 and 18S rRNA trichomonad sequences were selected from the NCBI RefSeq database .In order to identify the isolate, sequences were generated for five clones for the 18S rRNA and ITS1-5.8S rRNA-ITS2 loci. For the 18S rRNA locus, three unique clonal sequences were generated which shared 99.5–99.7% identity, suggesting a single-species infection. ML analysis of the 18S rRNA locus placed all clone sequences together within Tetratrichomonas with moderate bootstrap support . The isolates were also placed in a separate lineage from urogenital Tetratrichomonas isolates from cattle originating from a previous study (accession AF342742).For the ITS1-5.8S rRNA-ITS2 locus, there were three unique sequences which shared 95.2–99% identity, suggesting some diversity amongst the parasites present. In agreement with the 18S rRNA locus, ML analysis also placed all the isolates together within Tetratrichomonas lineage of the isolates in more detail, ML analysis was performed on the 18S rRNA sequence from a greater taxonomic sampling of Tetratrichomonas spp., including several bull urogenital isolates from previous studies  amongst Tetratrichomonas 18S rRNA sequences. For the published urogenital Tetratrichomonas isolate 2004–5000 (Dufernez et al., Tetratrichomonas sequence generated in this study. Representative BLAST results for a single sample are shown in Supplementary Table S2, and an alignment between metatranscriptome sequences and urogenital Tetratrichomonas 18S rRNA sequences are shown in Supplementary file S4.In order to investigate the potential gastrointestinal origin of urogenital Tetratrichomonas and Pentatrichomonas spp., which appear to have been isolated in roughly equal proportions. In contrast, there has only been a single report of a Pseudotrichomonas isolate.In addition to the isolates identified in this study, there have been numerous reports of trichomonads isolated from the UGT of cattle, summarized in Tt. foetus trichomonad isolated from the bovine UGT in Europe. In agreement with previous reports of cattle urogenital trichomonads (Walker et al., et al., et al., Tetratrichomonas sp. The phylogenies presented here are largely in agreement with previous studies regarding the interrelationship between trichomonads and the reported lineages of Tetratrichomonas spp. (Cepicka et al., et al., et al., Tetratrichomonas genus.This study reports the first case of a non-Tetratrichomonas lineage 4 as defined by Cepicka et al. (Tetratrichomonas isolates defined by Dufernez et al. (et al., Tetratrichomonas lineage 10 (Cepicka et al., et al. (et al., Tetratrichomonas isolates.Our results support the placement of the new isolates in a et al. , which cz et al. . The new et al., , corresp, et al. , althoug et al., . These oTetratrichomonas lineage 4 was described as mostly limited to the tortoise GIT (Cepicka et al., et al., et al., Tetratrichomonas lineage 10 (Cepicka et al., Te. buttreyi (Dufernez et al., Tetratrichomonas lineage 10 also appears to be mostly restricted to tortoises (Cepicka et al., Tetratrichomonas lineages, P. hominis, and a Pseudotrichomonas sp. have been isolated from the cattle UGT (Walker et al., et al., Tt. foetus, which shows a remarkable degree of genetic homogeneity suggestive of a recent founder event (Kleina et al., et al., et al., et al., Tetratrichomonas 18S rRNA (Dufernez et al., et al., Tt. foetus trichomonads in a backdrop of frequent Tt. foetus monitoring (Yao, Tt. foetus is also possible.At least two ing Yao,  suggestsTetratrichomonas spp. (Zhang et al., Te. buttreyi (Dufernez et al., P. hominis is known to occupy the GIT of a very wide range of animals (Li et al., et al., Pseudotrichomonas keilini (Dufernez et al., Monoceromonas ruminantium is also relatively closely related (Dufernez et al., et al., P. keilini is an exception or mislabelled as free-living. The presence of gastrointestinal-associated bacteria at the same site as urogenital trichomonad isolation provides further evidence for their gut origin (Cobo et al., et al., There are plausible sources for the isolated urogenital trichomonads from the bovine GIT; Tetratrichomonas preputial isolates in the cattle UGT, results have ranged from no persistence (Cobo et al., et al., et al., Pentatrichomonas hominis derived from the cattle gut also failed to persist in the UGT of heifers (Cobo et al., Tetratrichomonas to regularly establish colonization during experimental infection (Cobo et al., Tt. foetus trichomonads (Cobo et al., During experimental inoculation of Tt. foetus trichomonad isolates obtained from the cattle UGT do not represent an emerging urogenital inhabitant but rather a sporadic transmission from another source, most likely the gut. However, transmission of parasites between these mucosal sites may increase the possibility of new pathogens emerging, as has been documented for other trichomonads (Maritz et al., Together, the evidence supports the hypothesis that non-Tt. foetus trichomonads in the cattle UGT is most likely misdiagnosis through confusion with Tt. foetus, which could cause unnecessary culling of suspected infected animals (Campero et al., et al., Tt. foetus monitoring is culture-based isolation and morphological identification by light microscopy (Parker et al., et al., et al., et al., et al., Tt. foetus detection (Oyhenart et al., Tetratrichomonas sp. are short-lived in the bull UGT (Cobo et al., et al., et al., The most significant concern associated with non-


Most research exploring the link between traumatic events and psychotic experiences has focused on either Australia, Europe or North America. In this study, we expand the existing knowledge to Thailand and investigate the impact of the type and the number of traumatic events on psychotic experiences in Thailand.We used data from the nationally representative 2013 Thai National Mental Health Survey (TNMHS), including questions on traumatic events and psychotic experiences. We regressed the lifetime experience of hallucinations or delusions against the following independent variables: the experience of any traumatic event during lifetime ; the experience of either no traumatic event, one interpersonal, one unintentional or both interpersonal and unintentional traumatic events  and the number of traumatic events experienced during lifetime . We adjusted the regression models for sociodemographic indicators and psychiatric disorders, and considered survey weights.About 6%  of the respondents stated that they had either hallucinatory or delusional experiences during their lifetime. The risk of reporting such experiences was more than doubled as high among respondents who had experienced at least one traumatic event during their lifetime than among those who had not yet experienced one, with higher risks for interpersonal or multiple traumatic events. Our results further indicated an increase in the risk of psychotic experiences as the number of traumatic events increased, with up to an eight-fold higher risk for people exposed to five or more traumatic events in their lifetime, compared to those with no traumatic events.Individuals reporting interpersonal or multiple traumatic events face much higher risk of psychotic experiences. Effective and widely accessible secondary prevention programmes for people having experienced interpersonal or multiple traumatic events constitute a key intervention option. The objective of the current study was to address this research gap using data from the 2013 Thai National Mental Health Survey (TNMHS). According to the existing literature, we investigated the following hypotheses:et al., Data from the TNMHS in 2013 were used, a sub-nationally representative cross-sectional survey that adopted the methodology of the World Mental Health Survey Initiative , a fully standardised interview on mental disorders in accordance with the ICD-10  or auditory hallucinations (‘Have you ever heard any voices that other people said did not exist?’), or delusions such as thought insertion and/or withdrawal, mind control and/or passivity, ideas of references, plot to harm and/or being followed during lifetime, after excluding the possibility of dreaming or the influence of substances . Furthermore, respondents were asked whether they had ever experienced one or more traumatic events . Survey weights were applied in all regression analyses. All analyses were carried out using Stata 15.1 , socioeconomic indicators  and finally psychiatric disorders . Additionally, an unadjusted model was used (model 0). Respondents with missing information on either the dependent, independent or control variables were excluded from analyses  constituted the analytic sample (<1% missing). An overview of sociodemographic characteristics of the sample, the prevalence of psychotic experiences, the prevalence of reporting at least one traumatic event and the weighted percentage of persons fulfilling diagnostic criteria for mental disorders  is displayed in A total of 4727 adults completed the interview (response rate: 74.3%), of which 4715 aged 44.1 years on average (standard deviation [s.d.: 19.5 years), with almost two-fifths reporting having been under the age of 18 . Almost 1%  of the respondents met the criteria for PTSD at any time during their lifetime.About 6% (95% CI: 4.9–7.0) of the respondents reported a psychotic experience during their lifetime, with hallucinatory experiences being more prevalent than delusional experiences. One in two respondents reported at least one traumatic event during their lifetime, with 12.8% (95% CI: 11.5–14.2) reporting two, 5.3% (95% CI: 4.5–6.4) three, 1.9% (95% CI: 1.5–2.5) four and 1.7% (95% CI: 1.3–2.3) five traumatic events. Unintentional, non-interpersonal traumatic event were reported more often than interpersonal traumatic events. On average, the first traumatic event was reported at the age of 26.1 years (Results of the Poisson regression models conducted for hypotheses testing are displayed in p = 0.103). Beginning with two traumatic events, the risk of psychotic experiences increases with each additional event, with risk ratios of 2.5 (95% CI: 1.5–4.2), 3.7 (95% CI: 2.0–6.7) and 5.9 (95% CI: 3.3–10.7) for two, three and four traumatic events, respectively. Compared with respondents without traumatic event, those who reported five or more traumatic events had 7.7 (95% CI: 4.0–14.8) times higher risk of having at least one psychotic experience during their lifetime.The risk of reporting psychotic experiences as a function of the number of traumatic events is shown in et al., et al., et al., et al., et al., et al., et al., To the best of our knowledge, this is the first study investigating the link between traumatic events and psychotic experiences in the Thai population. In line with previous research, we found an increased risk for psychotic experiences in individuals who have suffered from traumatic events (Scott et al., et al., et al., et al., et al., The risk of psychotic experiences increased with the number of traumatic events, pointing to a dose–response relationship which is in line with evidence from previous research (Scott et al., et al., Some limitations have to be taken into account. First, the cross-sectional design of our study does not allow for conclusions about causality. Second, since the TNMHS was conducted in 2013, more research is needed using more recent databases, including other countries in this region, and taking into account longitudinal study designs. Third, psychotic experiences constitute only one symptom of schizophrenia spectrum and other psychotic disorders and may not in all cases lead to clinically meaningful impairment of functioning. Additionally, the TNMHS did not include questions on family history of psychotic experiences, which would have been a key variable given the large genetic component in schizophrenia and other psychotic disorders (e.g. Chou 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., et al., Based on the strong link between traumatic events and psychotic experiences, some researchers call for a distinction to be made between trauma-related psychotic experiences and such experiences in people who were not exposed to trauma (Hardy, et al., et al., et al., et al., et al., et al., Our findings emphasise the close link between traumatic events and psychotic experiences and extend the existing knowledge to the region of Southeast Asia. Experiencing either an interpersonal trauma or from multiple traumatic events were particularly associated with reporting psychotic experiences during lifetime. In Thailand, almost one in five individuals reported having experienced either an interpersonal trauma or multiple traumatic events in their lifetime, and two in five were minors at the time of first trauma exposure. With a total population of 69.8 million inhabitants in Thailand in 2020 (United Nations, Department of Economic and Social Affairs, Population Division, This study was the first to investigate the association between traumatic events and psychotic experiences in a Thai population. We found the experience of interpersonal or multiple traumatic events to be associated with a substantially elevated likelihood of reporting hallucinations or delusions during lifetime. Within our sample, a total of 6% reported having had psychotic experiences at least once in their lives, and one in five respondents stated having experienced interpersonal or multiple traumatic events. The high lifetime prevalence of traumatic and psychotic experiences in the Thai population highlights the need for effective and widely accessible secondary prevention programmes to reduce post-traumatic stress in affected people.


Hereditary deficiency of plasma prekallikrein (PPK) is a rare autosomal recessive disease. The affected patients are often asymptomatic and diagnosed incidentally during preoperative investigations or during hospitalization by isolated prolongation of activated partial thromboplastin time (aPTT). In this article, we report, a 46-year-old woman who was candidate for two invasive procedures (thyroid FNA and hysterectomy) and underwent preoperative evaluation. Due to prolonged aPTT with normal PT she was referred to the IBTO reference coagulation laboratory for specific coagulation assays. Ultimately, the examinations revealed severe PPK deficiency (<1%) with partial deficiency of factor XII level (25%). It was called as "Fletcher factor". In 1973, Wuepper et al. further investigated this unknown factor and they called it as “Prekallikrein” (PK). The gene related to PK is KLKB1, which is located on the q34-q35 region of the long arm of chromosome 4  for all the patients who have prolonged aPTT but normal intrinsic coagulation factors and without evidence of coagulation inhibitors. During this period of time this patient was the second case of PPK deficiency that was diagnosed.6/UL (3.3-5.8), WBC: 6.6 × 103/UL (3.5-11), platelet: 323 × 103/UL, aPTT: 206 seconds, and PT: 10 seconds (see A 46-year-old woman with a history of hypothyroidism has been a candidate for thyroid FNA (fine needle aspiration) to evaluate thyroid cold nodules and hysterectomy for uterine fibroids. During preoperative work up, an isolated prolongation of aPTT was noted and the patient was referred for further evaluation to the special coagulation laboratory of IBTO with suspicion to Lupus Anticoagulant (LA). The patient had a history of easy bruising and also bleeding after one tooth extraction . She also had two previous surgeries (appendectomy and cesarean section) without any abnormal bleeding. According to the ISTH-SSC Bleeding Assessment Tool the total bleeding score was 5 . The patient’s parents did not have any consanguinity. She had two sisters and three brothers, as well as a 16-year-old girl who were rejected for any history of bleeding or thrombotic events. She had a history of long-term hypothyroidism and resistant to routine doses of thyroid hormone. Initial laboratory tests in IBTO showed: HGB: 12.7 g/dL .et al. the deficiency of contact factors is the second cause of unknown prolonged aPTT after antiphospholipid antibodies (et al. in 2017 (Prekallikrein is a glycoprotein mainly synthetized in the liver and secreted into the plasma as a single-chain peptide with a molecular weight of 88,000 Daltons, and normal concentration of 40 mg/mL. About 75-90% of this coagulation factor is combined with HMWK. The activation of prekallikrein results in production of kallikrein, and activation of factor XI and factor XII. Patients with PPK deficiency, which is a rare autosomal recessive disorder , 6, typi in 2017 .et al. (et al. (et al. (The occurrence of bleeding events in the patients with PPK deficiency is not a common phenomenon and the reported studies about bleeding complications after surgical procedures show a wide range of bleeding tendency from no bleeding to major bleeding events -7, 11. Ret al.  in their (et al.  in 2019 et al. (et al. (Thrombotic events among patients with PPK deficiency were reported by several studies , 4, 6. B (et al. . Our caset al. (et al. (The first case of concurrent PPK and factor XII deficiencies was reported in 1981 by Man-Chiu Poon et al. . They de (et al.  one quarThere are some sporadic reports about PPK deficiency with some comorbidities e.g. autoimmune diseases including Graves' disease and systemic lupus erythematous. In the adults with multiple comorbidities or systemic inflammation, the effect of PPK deficiency remains controversial and further researches are needed to confirm the causative association between them -16. Our The data on the preoperative management of patients with PPK deficiency are also limited. Unal and his colleagues in their literature review on the surgical management of PPK deficient patients observed that most of these patients needed no FFP transfusion and FFP transfusion may be used only in patients with PPK deficiency undergoing invasive procedures . Our patIn this study, we presented a case of severe PPK deficiency with concurrent partial factor FXII deficiency that experienced occasional non-significant hemorrhages without any thrombotic events. It seems that PPK deficiency cases are under-diagnosed due to the lack of significant hemorrhagic or thrombotic symptoms in general and low availability of its specific laboratory test in our country. The most significant presentation of these cases is incidental finding of isolated prolongation of aPTT especially in preoperative hemostatic evaluations. Physicians should be aware of this rare congenital deficiency by conducting special coagulation assays and excluding other causes of prolongation of aPTT in order to prevent unnecessary interventions like transfusion of blood products in these patients.


To investigate the patients transferred by helicopters, as well as an emergent medical services that were performed for them.In this retrospective cross-sectional study, all patients who were transferred by Fars province of Helicopter Emergency Medical Services (HEMS) to Shiraz hospitals, southern Iran (March 2017-March 2019) were investigated. Patients’ information was collected and analyzed includes age, gender, dispatch reason, trauma mechanisms, take hold of emergent medical services, as well as the air transportation time, time between dispatch from the origin hospital and starting the procedures, and patients’ outcome.p<0.0001). Mental status deterioration (25.3%) was the most dispatched indications. The mortality rate was 13.25% totally (11.11% in traumatic vs. 10% in non-traumatic). The mean±SD of air transportation time was significantly lower than ground transportation in both traumatic (p=0.0013) and non-traumatic (p<0.0001) patients. Also, the mean±SD of time between dispatch from the origin hospital and starting the procedures was statistically lower in air transportation in both traumatic (p=0.0028) and non-traumatic (p=0.0017) patients.Eighty-three patients were enrolled with the mean±SD age of 36.9±19.47 years that 75.9% had trauma ( Most of the patients transferred by HEMS were traumatic. The air transportation time as well as the time between dispatches from the origin hospital to the starting of the procedures were significantly lower in HEMS in comparison with ground transportation for both traumatic and non-traumatic patients. Helicopter Emergency Medical Services' (HEM) purpose is to provide specialized services for patient’s triage, treatment, and rapid transfer directly to the trauma center for providing definitive treatment , 2. An aWhen deciding about HEMS using, many factors are important include access to higher-level pre-hospital care that HEMS staff provide, faster access to the main trauma center , simultaneous extract of several injured patients need, and access of local communities to ground emergency medical services (GEMS) centers. In many systems, HEMS staff receive Advanced Life Support (ALS) training that may not be available in rural EMS systems .et al., [4] compared the HEMS with other conventional means of emergency transportations. They reported that it’s an affordable method of transportation in cases which helicopter transmission increases the survival rate. Ringburg et al., [It is proved that HEMS is economical for various clinical conditions if administered carefully. Gearhart  et al.,  showed t et al., .The introduction of HEMS impacts was controversial on the health outcomes of trauma patients . BesidesAccording to our knowledge, few studies investigated the HEMS in Iran -16 and tStudy Design and PopulationThe current retrospective cross-sectional study (Aug-Dec 2019) was conducted on all patients’ medical records who were transferred by Fars province HEMS to Shiraz hospitals , the center of Fars province, southern Iran, from March 2017 to March 2019. Patients who died at the time of arrival to the destination hospitals were excluded.Study ProtocolThe patients’ names and their initial data such as demographic variables , the city and the reason of dispatch (traumatic or non-traumatic), the mechanisms of trauma, the indication of dispatch, and the taken pre-hospital and in-hospital emergent medical services less than six hours were collected from Fars province HEMS center. Then, the started time of the procedures as well as the patients’ outcome (discharge or death) were extracted from the patients’ medical files in the destination hospitals and recorded in a data collection form.To compare the duration of helicopter and ground transportations, the land distance information between the dispatched cities (patients’ locations) and the destination hospitals as well as land estimated times dispatches were obtained from the https://www.google.com/maps website.A variable was defined: “the estimated time interval of dispatch to the start of the procedure, if the patient was dispatched by ground emergency services” in comparing the air and ground emergency services. To calculate this variable, “air transfer to the hospital duration” was reduced from “total duration of dispatch to start of procedures”. This is the duration of patients’ entrance to the emergency department of the hospital and initiating the procedures. Then, it was summed up by the “ground transfer time of the patient to the destination hospital”. Therefore, if patients were transferred by ground emergency vehicles, the time interval between dispatch and starting of procedures was estimated. It should be noted that 30 minutes was considered for boarding and disembarking patients from the ambulance in ground transportation but this time is hidden for helicopter transportation.Statistical Analysisp-value less than 0.05 and confidence interval (CI) of 95% was considered to be statistically significant.The statistical package for social sciences  version 25 and Medcalc software for Windows were used for statistical analysis through descriptive and analytical tests such as independent t-test, Chi Square, and nonparametric tests. Results are presented as mean±standard deviation (SD) for continues variables and were summarized in number (percentage) for categorical variables. Two-sided Ethical ConsiderationsThe current study was supported by Shiraz University of Medical Sciences (grant No. 17434), which was approved by the vice-chancellor of research and technology as well as the local ethics committee (IR.sums.med.rec.1397.344) of Shiraz University of Medical Sciences.p=0.550), that 56 (67.5%) of them were men (p<0.0001). Sixty-three (75.9%) patients had trauma (p<0.0001) which was significantly higher in men . The mortality rate was 13.25% totally . It should be considered that there were no air accidents during the study period and none of the patients died on the way. Totally, as shown in p=0.009). Car-car accident (CCA) and car turn over (CTO) were the most mechanisms of trauma (43.9% vs. 39.02%). In general, 130 indications were recorded for dispatch in traumatic patients which mental status deterioration and long bone open fracture were the most of them (33.33% vs. 31.75%). For each patient, at least one emergent medical services were performed  which blood transfusion was the most of them (44.44%), and 85.71% of the procedures were performed before 202.09 minutes  , that this difference was statistically significant (=0.0028) .As shown in p<0.0001). The mean±SD of time between dispatch from the origin hospital and starting the procedures was 131.03±80.60 minutes in air transportation which was statistically higher than the estimated time in ground transportation (251.40±65.03 minutes) (p=0.0017)  .et al., [et al., [et al., [In the current study, all patients transferred by HEMS of Fars province to hospitals of the Shiraz from 2017 to 2019 and the emergent medical services were investigated. As mentioned before, most of the patients were men which is consistent with previous studies , 17. Beset al., 's study [et al.,  reported[et al.,  also staet al., [et al., [In the current study, detailed investigation of trauma causes revealed that CCA had the highest frequency followed by CTO and motor-car accident. Andruszkow et al.,  reported[et al.,  reported[et al., , they reet al., [et al., [et al., [In the present study, the overall mortality rate was 13.25%, and 11.11% for traumatic patients which was lower than the results reported by Andruszkow et al.,  that rep[et al.,  reported[et al.,  reportedComparison of traumatic patients’ data in the current study with other studies which is used as a reference for evaluating the quality of services provided in trauma, showed that although the mortality rate of traumatic patients dispatched by helicopter was slightly higher in the current study, this difference was not statistically significant.et al., [et al., [In the current study, the dispatch’s indications of non-traumatic patients were ACS, cardiac arrest, abortion, and respiratory arrest. In line with the current study, McQueen et al.,  showed t[et al.,  reportedet al., [et al., [Some studies compared the efficiency of HEMS and GEMS. For example, Davis et al.,  investig[et al.,  in a proet al., [et al., [Schiller et al.,  study onet al., . In anotet al., . Moreove[et al.,  showed tet al., [et al., [Brown et al.,  found th[et al.,  reported[et al., . AccordiOne of the strengths of the current study was that Fars province has one of the best equipped and busiest HEMS in the country due to its vastness and mountainous climate. Therefore, investigating the patients who transferred in this province will provided valuable information about characteristics of the patients and performed emergent medical services. On the other hand, the study also had limitations including a low number of patients who were transferred during the study period. Air Emergency Center reported 109 patients but only 83 patients were analyzed due to the incompleteness of data. To obtain comprehensive results, future studies should collect the number of HEMS all around the country, therefore, such services can be used more effectively.According to the results, most of the patients transferred by HEMS in Fars province were traumatic patients and the main causes of traumatic injuries were CCA, CTO, and motor-car accidents. The overall mortality rate was 13.25% (11.11% for traumatic and 20% for non-traumatic patients). The time between dispatches from the origin hospital to the starting of the procedures was significantly lower in HEMS in comparison with GEMS for both traumatic and non-traumatic patients. Regarding that most of the patients were traumatic, it is necessary to increase the quality of the triage system in HEMS. However, prospective studies with larger sample sizes are necessary.


During Earth's 4.6-billion-year history, its surface has experienced environmental changes that drastically impacted habitability. The changes have been mostly attributed to near-surface processes or astronomical events with little consideration of Earth's deep interior. Recent progresses in high-pressure geochemistry and geophysics, however, indicated that the deep Earth processes may have played a dominant role in the surface habitability . This Sp2O is the key compound that acts as a strong oxidant and oxygen transporter. In this Special Topic, Katsura and Fei [2O in the asthenosphere, and Walter [2O transport all the way down to the CMB. It has been established that when H2O meets Fe at the CMB, Fe can be oxidized to produce superoxide FeO2 with pyrite structure. At the middle of the lower mantle , Hu et al. [2O can also react with ferropericlase O, a major lower mantle mineral, to form pyrite-structured superoxide. At shallower depth of 1000 km (40 GPa), Liu et al. [2O induced oxidation of ferropericlase to form Fe2O3+δ with a new hexagonal structure. Both reactions result in further oxygen enrichment beyond the already highly oxidized slab materials in the deep mantle environment. The subducting process may compile substantial oxygen-rich materials to form oxygen reservoirs at the CMB.Oxygen, the most abundant element in Earth, is the key element that enables lifeforms to thrive and creates the unique living planet as we know. Recent discoveries on the pressure-altered chemistry revealed that the oxygen cycles through Earth's deep interior could also be the origin of the dynamic mantle processes that dictate the disruptive evolution of the life on surface. The Earth shows a vertical gradient of oxygen fugacity over 10 orders of magnitude from the highly oxidized crust at ambient conditions to the highly reduced core–mantle boundary (CMB) at ∼2900 km depth and 130 GPa. The dynamic process of subducting slab, on the other hand, injects the oxidized crust down into the reduced deep lower mantle. Here, Hu et al.  show thainsitu at high pressure–temperature conditions of the deep interior. Mao et al. [Recognition of novel materials in deep Earth relies on seismological observations of their characteristic elastic signatures that must be determined o et al.  present o et al.  uses lono et al.  that couo et al.  highligh