C. albicans , unremoved CVC , and a high βDG value (>108.2 pg/mL) . Continuous ophthalmologic examination is recommended in cases of candidemia with the above risk factors with an initial examination within 7 days of onset and a second examination 7–14 days after onset.Ocular candidiasis is a critical and challenging complication of candidemia. The purpose of this study was to investigate the appropriate timing for ophthalmologic examinations, risk factors for complications of ocular lesions, and their association with mortality. This retrospective cohort study applied, using multiple logistic regression analysis and Cox regression models, to cases of candidemia (age ≥ 18 years) for patients who underwent ophthalmologic consultation. Of the 108 candidemia patients who underwent ophthalmologic examination, 27 (25%) contracted patients had ocular candidiasis, and 7 experienced the more severe condition of endophthalmitis, which included subjective ocular symptoms. In most cases, the initial ophthalmologic examination was performed within one week of the onset of candidiasis with a diagnosis of ocular candidiasis, but in three cases, the findings became apparent only after a second examination within 7–14 days after onset of candidiasis. The independent risk factor extracted for the development of ocular candidiasis was the isolation of  Candidemia is a nosocomial infection that is a major cause of morbidity and mortality ,2. More All the preventive guidelines, however, include a few detailed recommendations about when and how often ophthalmological examination should be performed. Although both the necessity and utility of repeated ophthalmological examination has previously been reported, specific guidelines are yet to be published. Furthermore, there have been few reports of the risk factors associated with ocular candidiasis. One factor is that the relationship between ocular candidiasis and prognosis in patients with candidemia is not well known.Here, we describe a retrospective cohort study to determine the risk factors related to ocular candidiasis and identify the relationships between mortality and eye involvement. In addition, we analyze the clinical characteristics of patients with ocular involvement in order to establish appropriate intervention methods via ophthalmological examination.This retrospective cohort study was conducted from April 2013 to March 2020 at the Kurume University Hospital in Japan, which is a tertiary-care university hospital with more than 1000 beds. We used a microbiology database to identify all patients (age ≥ 18 years) with candidemia using microbiology database and collected clinical information from their medical records. This study was approved by the Medical Ethics Committee of the Kurume university hospital (No 21123) and was implemented in accordance with the Declaration of Helsinki. Patient consent was waived owing to the nature of a retrospective study.Candida spp. from at least one blood culture in a patient with clinical sign of infection. Subsequent positive cultures from the same patient were considered as a new episode if an interval of more than 30 days had transpired the two episodes. Exclusion criteria included patients who were less than 18 years of age, patients considered contaminated and untreated, and patients who had received no ophthalmological consultation. The onset of candidemia was defined as the day the initial positive blood sample was drawn, which subsequently formed a culture positive for Candida spp. To classify ocular candidiasis, we referred to criteria established in previous studies [Candidemia was defined as isolation of  studies ,11,12,13 studies .Candida spp., persistent blood stream infection , and a prognosis of 30-day mortality. The ophthalmological clinical course was also collected from the medical records. Ophthalmological clinical information includes the following: when was the ophthalmologic examination performed, counting from the onset of candidemia; how many ophthalmologic examinations resulted in a diagnosis of ocular candidiasis; what are the ophthalmoscopy findings; is there a presence of subjective symptoms of the eyes; and what is the ophthalmology treatment history. The -β-D-glucan (βDG) test values were evaluated via the WAKO β-glucan test , and the highest value measured within 7 days of the onset of candidemia was used.The predisposing factors and clinical information acquired from medical records included age, sex, underlying diseases , immunocompromised status , use of immunosuppressive agents, exposure to chemotherapy, exposure to radiation therapy, HIV infection, recipients of stem cell transplantation), intensive care unit (ICU) admission, history of surgery, prior antibiotics exposure, prior antifungal exposure, presence of a central venous catheter (CVC), removal of the CVC, interval onset of candidemia and removal of CVC, total parenteral nutrition (TPN), mechanical ventilation, hemodialysis, septic shock, initial antifungal drugs, interval onset and administration of antifungal drugs, causative t-test or a Mann−Whitney U test. For the values of βDG, logistic single-regression analysis was used for comparison. Categorical variables were presented as numbers and percentages and compared using either the χ2 test or Fisher’s exact test. We compared demographic characteristics, clinical factors, and outcomes between episodes with and without ocular candidiasis by using univariate analysis, and multivariate analysis was conducted using items that were statistically significant  in univariate analysis. Multivariate statistical analysis was conducted by using logistic regression multivariate analysis with odds ratios (OR) and 95% confidence intervals (CI). A more detailed statistical analysis was performed for βDG. Receiver operating characteristic (ROC) curves for βDG values were described, and their cut-off values were determined with the maximum Youden index. All cases were divided into groups with either high or low βDG values based on the cut-off values, and the relationship between βDG values and ocular candidiasis was illustrated using a Kaplan−Meier estimator. Cox regression models were used to calculate the adjusted hazard ratios (HRs) with a 95% confidence interval. The association between 30-day mortality and ocular candidiasis also was studied using the same methods for statistical analysis. Two-tailed p-values of <0.05 were considered statistically significant. Statistical analyses were performed using R program language, version 4.1.3, and JMP software, version 15.Continuous variables were presented as the mean ± standard deviation (SD) or interquartile range (IQR), and these were compared using either a Student’s The process for selecting target cases appears in Candida spp. was isolated. When one of these two cases had an episode of fever, their blood culture was not submitted, and a central venous catheter was removed, which could have led to a delay in the diagnosis of candidemia. Of the seven patients with probable Candida endophthalmitis, all presented with vitreous opacity, and of them, six patients had bilateral lesions. No cases required surgical treatment of the eye, but one case was treated by direct administration of Voriconazole (VRCZ) into the vitreous. Twenty patients with probable Candida chorioretinitis did not complain of subjective eye symptoms. Fundus examination identified 18 patients with abnormal bilateral findings, and 2 patients had abnormal findings only on one side. All cases presented with cotton wool spots, and some showed petechial hemorrhage.Patient backgrounds, ocular symptoms, and timing of ophthalmologic consultations for 27 patients with ocular candidiasis appear in Primary ophthalmological examinations were conducted an average of 5.6 ± 3.7 days following onset of candidemia, and the median was 5 days. A second examination was performed an average of 10.1 ± 4.8 days following the first, and the median was 8.5 days. Of the 27 patients, 6 died before the second ophthalmologic examination, and the remaining 21 patients all had two or more repeated ophthalmologic examinations. Most cases showed improvement in fundus findings in the second and in subsequent ophthalmologic examinations, but three cases showed a tendency toward exacerbation. In these three cases, second ophthalmological examinations were conducted in 8, 9, and 12 days, respectively, after candidemia had occurred. It is important to emphasize that in these three cases, no abnormalities in the ocular examination were evident during the first examination, but the abnormal findings became apparent during the second examination. In particular, in the patient with AML and severe neutropenia, the ocular lesions gradually worsened with each ophthalmologic examination despite the administration of appropriate antifungal medications, which eventually led to treatment with intravitreal injections.Three of all patients showed no abnormal findings during the first ophthalmologic examination but were diagnosed with ocular candidiasis during the second examination. One of these three cases presented with severe neutropenia and was eventually treated by vitreous injection due to a gradual exacerbation of eye symptoms.Candida albicans was the most commonly isolated species (42.6%), followed by Candida parapsilosis (25.0%), Candida glabrata (14.8%), C. famata (5.6%), C. tropicalis (4.6%), and C. krusei (4.6%). Of the strains detected in this analysis, none with known drug susceptibility were suspected to be highly resistant to antifungal drugs such as fluconazole or echinocandin. Initial antifungal drugs were as follows: azole (59.3%), echinocandin (37.0%), and liposomal amphotericin B (2.8%). Persistent blood stream infection accounted for 36.2%. Among the cases with persistent candidiasis, there was one case each with suspected complications of infectious endocarditis and vertebral osteomyelitis. The rate for 30-day mortality was 20.4%.The characteristics of the patients appear in C. albicans, βDG value were statistically significant, and multivariate analysis was performed according to the method defined. The results indicated that isolation of C. albicans , unremoved CVC , and a high βDG value  were detected as independent risk factors for ocular candidiasis  in the univariate analysis. All items were statistically significant  also in multivariate analysis.Multivariate analysis was performed using items that ware statistically significant  and 95% confidence intervals. High βDG values (>108.2 pg/mL) were statistically significant for the risk factors of ocular candidiasis .As mentioned above, we determined that a high βDG level is an independent risk factor in the development of ocular candidiasis. Therefore, we further analyzed the relationships between βDG levels and ocular candidiasis. Receiver operating characteristic (ROC) curves for βDG values were developed, and their cut-off values were determined using the maximum Youden index of 108.2. The area under curve (AUC) was calculated to be 0.68. We divided the patients into two groups: βDG > 108.2 pg/mL and <108.2 pg/mL. The distribution of time to diagnose ocular candidiasis by ophthalmological examination was estimated using the Kaplan−Meier estimator and analyzed by log-rank testing . The logWe also investigated the association between complications of ocular candidiasis and 30-day mortality. Patients were divided according to their 30-day mortality rates and for a comparison of backgrounds, which included complications from ocular candidiasis . In univp value < 0.05) in the univariate analysis. Unremoved CVC was statistically significant  in multivariate analysis.Multivariate analysis was performed using items that ware statistically significant , and indwelling CV catheters are one of the most important risk factors . TherefoβDG is an adjunct diagnosis for fungal infections and is sometimes used to determine treatment efficacy . The relThough complications of ocular candidiasis were not extracted as a statistically independent risk factor for 30-day mortality, patients with ocular candidiasis experienced a higher mortality rate compared with uncomplicated cases. Some of the patients who were excluded from the study because of no ophthalmologic examination included early death cases in the course of the disease, so the correlation between ocular involvement and prognosis may not have been fully evaluated. In fact, the 30-day mortality rate in the 138 cases was 32.3%, which included patients with no ophthalmologic examination. On the other hand, the 30-day mortality rate for the 108 cases that were ultimately included in the study was 20.4%. There were some reports that the mortality rate was higher in cases with ocular lesions, and although the ocular lesions themselves may not affect the prognosis, the intensity of the disease caused by disseminated lesions is expected to have an effect on the prognosis. Careful management of patients with ocular candidiasis is desirable due to the risk of deterioration in their general condition.We were unable to extract any factors that would make ocular candidiasis less likely to occur in this analysis. Therefore, we recommend that ophthalmological examination be performed in all cases presenting with candidemia. Even after a negative blood culture is confirmed, ophthalmological examination should be repeated in cases of candidemia and ocular candidiasis, and if new lesions have appeared or the lesions do not improve, the antifungal drug should be changed based on tissue migration and drug sensitivity, and treatment should be continued until the fundus findings improve.C. albicans, unremoved CVC, and a high level of βDG. Since the prognosis may be poor in cases with ocular involvement, we also recommend careful systemic management of those patients.Repeated searches for ocular lesions by ophthalmological examination is an essential strategy in cases of candidiasis, and we recommend the first session should occur within 7 days of onset, while a second session should be scheduled for 7–14 days following onset. Furthermore, it is also necessary to constantly check for the presence of subjective eye symptoms. Particular attention should be paid to complications of ocular lesions in cases with the following factors: isolation of


In inverse probability weighted panel models terms for cannabis metrics were significant for chromosomal disorders, trisomies 21 and 13 and Klinefelters syndrome from p < 2.2 × 10−16. In spatiotemporal models cannabis terms were positive and significant for chromosomal disorders, genetic disorders, trisomies 21, 18 and 13, Turners and Klinefelters syndromes from 4.28 × 10−6, 5.79 × 10−12, 1.26 × 10−11, 1.12 × 10−7, 7.52 × 10−9, 7.19 × 10−7 and 7.27 × 10−7. 83.7% of E-value estimates and 74.4% of minimum E-values (mEV) > 9 including four values each at infinity. Considering E-values: the sensitivity of the individual disorders was trisomy 13 > trisomy 21 > Klinefelters > chromosomal disorders > Turners > genetic syndromes > trisomy 18 with mEV’s 1.91 × 1025 to 59.31; and daily cannabis use was the most powerful covariate (median mEV = 1.91 × 1025). Conclusions: Data indicate that, consistent with reports from Hawaii, Canada, Colorado, Australia and USA, CCARs are causally and spatiotemporally related to metrics and intensity of cannabis exposure, directly impact 645 MB (21.5%) of the human genome and may implicate epigenomic-centrosomal mechanisms.Introduction: Laboratory data link cannabinoid exposure to chromosomal mis-segregation errors. Recent epidemiological reports confirm this link and raise concern that elevated chromosomal congenital anomaly rates (CCAR) may be occurring in Europe which is experiencing increased cannabis use, daily intensity of use and cannabinoid potency. Methods: CCAR data from Eurocat. Drug use data from the European Monitoring Centre for Drugs and Drug Addiction. Income from World Bank. Bivariate, multivariate, panel and geotemporospatial regressions analyzed. Inverse probability weighting of panel models and E-values used as major quantitative causal inferential methodologies. Results: In countries where daily cannabis use was rising the trend for CCA’s was upwards whereas in those where daily use was declining it was usually downwards ( Chromosomal congenital anomalies (CCA’s) have been an important feature of several recent epidemiological studies of the associations and causal links of various prenatal drug exposures especially cannabinoids. In reports from Hawaii, Colorado, Canada, Australia and USA congenital chromosomal anomalies have featured prominently on long lists of congenital anomalies elevated after prenatal or community cannabinoid exposure ,6,7,8,9.Particularly concerning in this regard is the well documented exponential dose response of cannabis genotoxicity ,16,17,18Many mechanisms of genotoxicity from cannabinoids are described including altered sperm morphology ,27, disrConsiderable strides have been made in the epigenomic understanding of drug addition and drug dependency in recent years with several multigenerational animal studies appearing ,44,45,46The subject of this study is the seven chromosomal disorders tracked by the European Network of Population-Based Registries for the Epidemiological Surveillance of Congenital Anomalies (Eurocat) namely trisomies 13, 18, 21  along with Klinefelters syndrome , genetic disorders including deletions and chromosomal disorders as a group . One of One of the most intriguing things about the pattern of CCA disease to be discussed is the cellular mechanisms which might be responsible. Whilst various disruptions of normal chromosomal physiology were described several decades ago as noted above ,27,66,67The objective of the present study was to examine the epidemiology of congenital anomalies across Europe with regard to population substance exposure. Use of inverse probability weighting transformed the analysis from a merely observational study to a pseudo-randomized study from which it is proper and appropriate to draw causal inferences. Space-time regression was also employed to examine these changes accounting for effects such as serial correlation, spatial correlation, spatial autocorrelation and random effects. The study builds on an earlier study  and explData on all available congenital anomaly rates was downloaded by each individual year for each of 14 nations from the European Network of Population-Based Registries for the Epidemiological Surveillance of Congenital Anomalies (EUROCAT) website  and analNations were categorized as being either high and rising daily cannabis use or low and/or falling daily cannabis use based on a recent European epidemiological study . Thus BThe availability of several metrics of cannabis use, exposure and consumption made it possible to calculate various derived metrics. Hence, last month cannabis use prevalence data was multiplied by the THC content of cannabis herb and resin to derive compound metrics. These metrics were also multiplied by imputed daily cannabis use prevalence rates to derive further compound metrics for both cannabis herb and resin.Missing data was completed by linear interpolation. This was particularly the case for daily cannabis use. 59 data points on daily cannabis use from EMCDDA were available for these 14 nations across this period. Linear interpolation expanded this dataset to 129 datapoints (further details provided in Results section). Data on cannabis resin THC concentration were not available for Sweden. However, it was noted that the resin to herb THC concentration was almost constant in nearby Norway at 17.7 so this ratio was applied to the Swedish cannabis herb THC concentration data to derive estimates of Swedish cannabis resin THC concentration. Similarly data for the cannabis resin THC concentration in Poland were not available. The resin to herb THC concentration ratio of nearby Germany was used to estimate the resin THC content in Poland from the known Polish herb THC concentrations. Since geospatial analytical techniques do not tolerate missing data the dataset was completed by the last observation carried forward or backwards for Croatia in 2018 and 2019 and The Netherlands in 2010. It was not appropriate to use multiple imputation methods for this dataset as multiple imputation cannot be applied in panel or spatial multivariable regression techniques.Data was processed in R Studio version 1.4.1717 based on R version 4.1.1 from the Comprehensive R Archive Network and the R Foundation for Statistical Computing . The anaBivariate maps were drawn with package colorplaner . All illThe presence of multiple different metrics for cannabis consumption and exposure created a problem for analysis as it was not clear which was the most appropriate metric to employ for any particular model. Indiscriminate use of excessive covariates in a multivariable model would unnecessarily consume degrees of freedom and thereby restrict ability to assess interactions. This issue was formally addressed by the use of random forest regression using the R package ranger  with varPanel analysis was conducted using R package plm  across bThe formal tools of causal inference were used in this analysis. Inverse probability weighting (ipw) is the technique of choice to convert a purely observational study into a pseudo-randomized study from which it is appropriate to make causal inference . All mul10.17632/mvwvxk756z.1 and 10.17632/vd6mt5r5jm.1 (accessed on 22 March 2022).Raw datasets including 3800 lines of computation code in R has been made freely available through the Mendeley data repository at the following URL’s: Ethical approval for this study was provided from the Human Research Ethics Committee of the University of Western Australia number RA/4/20/4724 on 24 September 2021.As shown in As shown in p = 2.29 × 10−4; model AIC = 1811.6, LogLik. = −899.798). p = 0.0005; model F = 12.1, df = 1, 852, p = 0.0005; E-value estimate = 1.86, minimum E-value = 1.46).The time trends for these seven chromosomal anomalies can also be considered dividing the nations into those where daily cannabis use is low or decreasing and those where it is rising based on recent published reports  Figure 8Figure 8.The slopes of the regression lines show in When those slopes which were positive and statistically significant were extracted 51 bivariate relationships remained and these are shown in The next question therefore is how would these various covariates behave in a multivariable analytical context. However, this question is not straightforward to answer due to the multiplicity of covariates listed. This issue was addressed with the use of random Forrest regression using the R package ranger together with the package vip to assess variable importance. These variable importance tables are shown as p = 3.61 × 10−5 to p < 2.2 × 10−16) along with other cannabis exposure metrics.Data was also considered in a formal space-time framework that formally accounts for random effects, serial correlation, spatial correlation and spatial autocorrelation. The case of Klinefelter syndrome shown in E-Values may be extracted from these panel  and geosThese groups may be compared formally using Wilcoxson non-parametric tests. The results of these tests are shown in All seven chromosomal disorders studied, both as a class and individually, are closely related to various metrics of cannabis exposure. This strong statistical signal assessed by various regression models is in close accord with recently reported results from other jurisdictions including Hawaii, Colorado, Canada, Australia and USA ,6,7,8,9,p = 0.0002)  .p < 2.2 × 10−16 for chromosomal disorders at 1 and 2 temporal lags, Down syndrome in additive model, Patau syndrome in additive model, and Klinefelters syndrome at 2 lags . Second, genetic deletions and microdeletions which clearly involve DNA breaks and chromosomal breakage lesions. If one adds together the lengths of the chromosomes involved, here chromosomes 13, 18, 21 and X it is noted that this accounts for (113 + 76 + 46 + 153 = 388) megabases (12.9%) of the 3000 megabases in the human genome.To this landscape of major chromosomal anomalies can be added two cancers with which cannabis is known to be associated namely testicular cancer and acute lymphoid leukaemia. Both of these malignancies are known to be associated with chromosomal breaks, chromosomal translocations and malignant chromosomal fusion events which give rise to oncogenes with constitutive function and conferring a growth advantage on the malignant clone/s. Chromosomes 1, 7, 8, 11, 12, 13, 18, 21, X and Y are implicated in testicular cancer  and togeIf the length of all of the chromosomes implicated in cannabis-related genotoxic disease is added together (excluding duplicates) it totals and impressive 1765 megabases (58.8%) of the human genome which by any measure is a sizeable segment to be registering direct genetic damage.The case of the chromosomal pathologies in testicular cancer is particularly intriguing and its rapidity opens up special insights into chromosomal pathological mechanisms. Metanalysis has demonstrated that the incidence of testicular cancer is 2.6 times greater after cannabis exposure than without it . MoreoveMoreover, the biogenesis of testicular cancer is known to involve genome demethylation, one or two rounds of whole genome doubling events followed by the loss of  50–70 chromosomal arms . This onIt was noted in the Introduction that chromosomal breaks, fusion and bridges have been identified in cells following cannabis exposure. In 1938 the breakage-fusion-bridge cycle was identified by which broken ends of chromosomes became joined to other broken ends as fusions which became disrupted with cell division thereby perpetuating the cycle .Indeed cannabis has been linked with inhibition of telomerase potentially leading to uncapped and unprotected chromosomal ends, and accelerated aging syndromes ,114.If the breakage-fusion-bridge cycle was operating in the testicular germ cell niche this would explain both the relatively rapid appearance of testicular carcinogenesis after young adult exposure and its requirement to take an average of 13 years to occur whilst the cycle turns, builds up genetically modified clones which become selected out and expanded and eventually achieve a growth advantage and become clinically revealed as overt malignant disease.Whilst such oncogenic mechanisms are not relevant to congenital chromosome inherited disorders they do powerfully inform the broader subject of cannabis-induced chromosomal pathology more generally and expand the potential repertoire of human chromosomal pathophysiology which is of relevance to the broader subject.In considering the spectrum of chromosomal pathology in relation to cannabis it should also be noted that cannabis has long been known to test positively in the micronucleus assay . MicronuGiven that chromosomal mis-segregation is so a major part of cannabinoid teratogenesis a major outstanding question must relate to the mechanism whereby chromosomes detach from the mitotic half spindle. Why do the chromosomes detach? This is clearly the source of mis-segregations which lead to trisomies, monosomies, micronucleus formation and chromothripsis.Whilst the exact answer is not known there are many possibilities owing to the impressive complexity of the system of cell division. Tubulin proteins are a complex family of proteins. Microtubules are made of α- and β- tubulin dimers which polymerize into high order elongated structures which form the mitotic spindle. There are 25–30 microtubules in each ray of the spindle. Cannabis was shown in a proteomic screen to reduce tubulin synthesis  so this As mentioned there are many different types of tubulins. They are all subject to a complex system of post-translational modifications (PTM’s) which control subcellular function, address and reactivities. These PTM’s include methylation, acetylation, tyrosination, removal or addition of terminal glutamates including polyglutamylation, glycation or polyglycation, polyamination, phosphorylation, ubiquinylation and palmitoylation . AcetylaMicrotubules of the spindle pointing to the cell equator are enriched for detyrosination and polyglutamylation. Unaligned chromosomes are carried to the metaphase plate by the kinetochore associated protein kinesin-7 motor protein CENPE (centromere protein E) in a manner which is dependent on the detyrosination status of the microtubules implying that the motor protein can read the tyrosination state of the microtubules . Such stPTM’s have interesting effects, as those near the centrioles encourage nucleation and the formation of the poles of the mitotic spindle by polyglutamylation. Importantly phosphorylation of serine 172 on β-tubulin by CDK1 (cyclin dependent kinase 1) prevents incorporation of the tubulin dimer into the growing microtubule and reduces the pool of tubulin available for polymeric assembly. As noted below CDK1 is perturbed epigenomically by cannabinoid withdrawal .p = 0.000033; and again on page 352, p = 0.00445) [Microtubule dynamics were shown to be affected epigenomically by differential DNA methylation in cannabis withdrawal on functional annotation of a recent Ingenuity Pathway Analysis longitudinal whole epigenome screen  .The microtubules are bound to the spindle by a complex multiprotein assembly called the kinetochore which is a giant molecular complex comprising 90 proteins all highly regulated and highly post-translationally modified . It has Clearly this would be a very fruitful area for active investigation.An important recent whole genome DNA methylation screen identified 163 differentially methylated regions (DMR’s) affecting hundreds of genes across the genome during cannabis dependence, and then 127 DMR’s which appeared after 11 weeks of cannabis withdrawal . Three fMSH5, MutS homolog 5 is involved in DNA mismatch repair and facilitates DNA crossing over during meiosis . AbnormaCDK1 is the serine/threonine cyclin dependent kinase I which stands at the very threshold of cell cycle control . CDK1 phCD3D (T-cell surface glycoprotein CD3) is a cell surface glycoprotein on T-cells encoding the CD3 receptor . EPHA2 iThe epigenomewide screen by Schrott and colleagues also identified 256 pathway hits on terms including DNA mainly during cannabis withdrawal . CentralInterestingly it was recently shown that neural progenitor cells derived from induced pluripotent stem cells from patients with Down syndrome had global suppression of transcription related to major alterations in the epigenomic state including chromosomal “introversion”, destruction of the nuclear lamina, abrogation of lamina-associated chromosomal domains, and a reduction in topologically active transcriptional domains which collectively closely recapitulated changes of neural stem cell senescence . This glA fascinating recent study described the manner in which centromeric chromatin is a hotspot for DNA breaks and chromosomal translocations . Indeed Investigators revealed that the modified centrosomal histone 3 variant CENP-A together with its chaperone HJURP and dimethylated lysine 4 of histone 4 (H3K4me2) enable a succession of events to allow the recruitment of the RAD51-BRCA1-BRCA2 complex of the homologous recombination pathway to induce formation of a DNA-RNA hybrid and high fidelity end repair. Moreover, inhibition of RAD51 led to the involvement of RAD52 and the activation of the low fidelity mutagenic microhomology end joining pathway leading to centromeric instability and chromosomal translocations .In this context some of the findings of the recent epigenomic screen of Schrott and colleagues is particularly relevant . It has p = 7.73 × 10−20. Indeed this group of cancer linked mutations is amongst the most highly significant of all the DNA methylation hits identified. Laboratory studies of CENP -A, -C and -N have shown that interference with all three key CENP proteins leads to chromosomal mis-segregation errors, aneuploidy, accelerated aging and cancer [Considering the CENP proteins 105 hits were identified  includinIndeed it has been demonstrated that as many as 90% of solid organ and 50% of haemopoietic cancers are aneuploid which directly implicates epigenomic centromeric/kinetochore processes in tumourigensis .p-values from 0.0009 to 0.0262) and only one for RAD52 indicating that cannabis blocks RAD51 and homologous recombination much more severely than lower fidelity pathways such as microhomology-mediated repair which forces errors into centrosomal replication and downstream chromosomal mis-segregation events [As shown in n events .The patients described in this report were born non-mosaically aneuploid, that is uniformly aneuploid in all body cells. This important finding localizes the genotoxic lesion to either the pre-fertilization male or female gametes or the first cell of the embryo, the bipronuclear fertilized zygote.A fascinating paper recently appeared which provided details on gene activation from the transcriptome analysis of the earliest stages of human life from the oocyte held at the second meiotic division (MII) across fertilization and into the first three zygotic cell divisions to the eight cell stage . An intr6 of the 15 (40%) genes identified in the kinetochore metaphase signalling pathway were also shown to be differentially methylated in the EWAS sperm study of Schrott  includin6 of 9 (66.7%) of the genes identified in the cell cycle control of chromosomal replication pathways were similarly identified in the EWAS analysis of Schrott namely CDK1, MCM6 , ORC2 , PCNA , POLA1 , and TOP2A  .trans as enhancers for multi-chromosomal transcription [The implication of cannabis in the formation of double minute chromosomes, chromosomal rings and micronuclei was noted above ,128. It cription . Such elcription .Such mechanisms may also support the implication of cannabis in various cancer types ,164,165.Numerous laboratory studies document that cannabis genotoxicity and the processes on which genetic stability rests, exhibit an exponential dose–response relationship. Assays of cell growth and cell viability, mutagenesis, inhibition of tubulin, action, DNA and RNA synthesis, and inhibition of DNA methylation resulting in morphine self-administration in offspring all demonstrate exponential dose response relationships ,16,17,18Moreover, this clear exponentiation of dose–response genetic damage in the laboratory is reflected in similar changes epidemiologically as has been well demonstrated in USA ,22,23,24This now well documented exponentiation becomes a major issue downstream of the rising triple convergence of cannabis use prevalence, daily dosing intensity and cannabinoid potency which is widely reported on both sides of the Atlantic ,166,167 A number of factors provide confidence that study results are robust and widely generalizable: 1. Data is based on one of the largest most comprehensive datasets globally; 2. Findings are strongly concordant with all of the other peer reviewed published data in the field internationally ,6,7,8,9;The strengths of the present study are that it used one of the world’s largest most comprehensive datasets, used the database with information on the most number of genetic pathologies, employed both inverse probability weighting and E-values ,169,170,Data present strong evidence demonstrating in inverse probability weighted and causal inferential models that various metrics of cannabis use, especially daily use, are strongly associated with all seven of the chromosomal disorders studied. These findings confirm both many decades of in vitro laboratory work and also several recent epidemiological series from Hawaii, Colorado, Australia, Canada and the USA, as well as an earlier simpler analysis of this same dataset. Naturally these findings are of great concern in their own right. However, when considered in terms of the known exponential dose–response relationship of cannabis genotoxicity and the increasing contamination of the food chain in Europe and parts of the USA, public health concerns must be heightened. The present findings form part of the interface of cannabinoids with the broader spectrum of congenital anomalies which is also an area of great concern. Together with cannabinoid-related carcinogenesis and cannabinoid-potentiated aging syndromes cannabis-related congenital chromosomal anomalies, findings form one pivotal plank of overall cannabinoid genotoxicity. As has been shown elsewhere and also in the present results that cannabis is a much more potent genotoxic than tobacco and alcohol combined see  it is cl


Recent studies have shown the prognostic value of capillary refill time (CRT) and suggested that resuscitation management guided by CRT may reduce morbidity and mortality in patients with septic shock. However, little is known about the current use of CRT in routine clinical practice. This study aimed to assess the modalities of CRT use among French adult and pediatric intensivists.A cross-sectional survey exploring CRT practices in acute circulatory failure was performed. The targeted population was French adult and pediatric intensivists (SFAR and GFRUP networks). An individual invitation letter including a survey of 32 questions was emailed twice. Descriptive and analytical statistics were performed.Among the 6071 physicians who received the letter, 418 (7%) completed the survey. Among all respondents, 82% reported using CRT in routine clinical practice, mainly to diagnose acute circulatory failure, but 45% did not think CRT had any prognostic value. Perfusion goal-directed therapy based on CRT was viewed as likely to improve patient outcome by 37% of respondents. The measurement of CRT was not standardized as the use of a chronometer was rare (3%) and the average of multiple measurements rarely performed (46%). Compared to adult intensivists, pediatric intensivists used CRT more frequently (99% versus 76%) and were more confident in its diagnostic value and its ability to guide treatment.CRT measurement is widely used by intensivists in patients with acute circulatory failure but most often in a non-standardized way. This may lead to a misunderstanding of CRT reliability and clinical usefulness.The online version contains supplementary material available at 10.1186/s12873-022-00681-x. Recent cohort studies have emphasized the prognostic value of capillary refill time (CRT) –3 and a This cross-sectional survey aimed at describing and analyzing the practices in terms of CRT use among French adult and pediatric intensivists, in the setting of acute circulatory failure.The networks from the Société Française d’Anesthésie et de Réanimation (SFAR) and the Groupe Francophone de Réanimation et Urgences Pédiatriques (GFRUP) were solicited in September and October 2021 and asked to email, twice, an invitation letter for survey participation at each physician member of the networks, who are mainly adult/pediatric anesthesiologists and/or intensivists. The members of these learned societies are mainly from France, but also from Belgium, Algeria, Tunisia, Morocco, Luxembourg, and Canada. Responses of different physicians from the same institution were allowed. The invitation letter explained the purposes and benefits of the survey and gave a link to the dedicated web page .The survey included a total of 32 questions, XX of which were multiple choice questions, and is available in Additional files The demographic characteristics of the physicians were collected. The reasons why the physicians perform CRT and how they consider and interpret it were explored. The approach used to perform CRT in routine clinical practice was also assessed and the potential leads to improve clinical practice when using CRT were examined.p-value < 0.05 was considered statistically significant. The descriptive and analytical statistical analyses were computed using the Free Software Foundation’s CRAN R, R version 4.0.4.Categorical variables were expressed as frequencies and percentages, and continuous variables as mean ± standard deviation. Chi-squared tests were used to perform between-group comparisons. All tests were two-sided, and a The 15 physicians who initially answered the survey test reported no issues in terms of feasibility, and the overall quality was rated as good. The comments were used to refine the final version of the survey.Among the 6071 physicians who received and opened the email in September and October, 549 (9%) clicked on the survey link. The survey was then completed by 418 (7%) practitioners. The mean ± SD age of the respondents was 39 ± 10 years and the mean ± SD time of clinical experience and practice was 11 ± 9 years. Overall, 60% of respondents worked in an adult intensive care unit (ICU), 30% in a pediatric ICU, and 7% worked in an adult or pediatric emergency department. The respondents practiced mainly in tertiary teaching hospitals (63%), public hospitals (27%), and private hospitals (9%). Among all respondents, 55% worked in units with 11 to 20 beds, and 28% worked in units with more than 20 beds. Overall, 48% of respondents managed 1 to 5 patients with acute circulatory failure per week  or reproducible (9%) enough for daily use. Furthermore, 308(74%) respondents declared that 3 seconds was the threshold to define abnormal CRT, 29% were convinced CRT could be used to diagnose acute circulatory failure, and 35% thought it could probably have a diagnostic value Table . ConversWhen performing CRT in patients with acute circulatory failure, 42% of respondents declared assessing CRT 3 to 6 times per day and 50% reported doing it 1 to 2 times per day. CRT is measured at the thorax level for 58% of respondents, at the finger level for 51%, and at the knee level for 40%. To control the pressure applied to the skin, 25% use a specific technique and 75% just apply a firm pressure. The skin is compressed during less than 4 seconds according to 60% of respondents, and 98% assess the compression duration without a chronometer. Forty-six percent averaged at least two consecutive CRT measurements  compared to adult ones (76%). The former more frequently reported that CRT is a reliable and reproducible measure and were more confident in the diagnostic value of CRT in the setting of acute circulatory failure. They also more strongly believed in the effectiveness of CRT-based perfusion goal-directed therapy, 63% of them being convinced that CRT is useful at the bedside compared to 37% of adult intensivists. In terms of the technical approach used for CRT measurement, pediatric intensivists used longer compression times and more often averaged multiple measurements Tables  and 3.This survey found that the majority of physicians use CRT, more likely for its diagnostic than its prognostic value. Less than half of them think that a CRT-based perfusion goal-directed therapy can reduce mortality during acute circulatory failure. The standardized approach used in clinical studies does not seem to be applied during routine practice, as almost none of the respondents reported using a chronometer to measure CRT. Pediatric intensivists were more trustful of CRT in terms of reliability, reproducibility, and diagnostic ability, and their approach was closer to the required standards when compared to adult intensivists.CRT has been correlated to microcirculatory and perfusion variables such as NIRS , video mIn the Hernandez et al. clinical trial, CRT was measured by applying firm pressure to the ventral surface of distal phalanx of the right index finger with a glass microscope slide. Compression time was 10 seconds and the time for normal skin color to return was registered using a chronometer . In the Although the absence of link between cardiac output and CRT has been shown , its linThe present study has some limitations. With only a 7% response rate, one could argue the results are not representative of the French intensivist population. This rate is however comparable to other studies using the SFAR , 31or thNevertheless, in the absence of a previous evaluation of CRT use in routine practice, these results provide a relevant picture of current practices in France.CRT is widely used by physicians managing acute circulatory failure, but most often in a sub-optimal way. Moreover, a gap remains between available data and the degree of confidence in CRT to predict patient outcome or to help clinical decision-making and resuscitation strategies. This may lead to a misunderstanding of the reliability and clinical usefulness of CRT. A simple way to increase the reliability of CRT would be the use of a chronometer, reducing the variation in compression level, and averaging several measurements. Nurse training programs and/or the use of specific devices should help standardize CRT measurements and render CRT-targeted therapy effective in routine clinical practice.Additional file 1. Survey letter.Additional file 2. Questionnaire.


In conclusion, the mechanism of pulmonary hypertension induced by portal hypertension may be related to telomere dysfunction and P53 overactivation, and lipid metabolism and intestinal inflammation are also involved in this process.Portal hypertension (PH) is an important cause of pulmonary arterial hypertension(PAH), but its mechanism is still unclear. We used genetic data analysis to explore the shared genes and molecular mechanisms of PH and PAH. We downloaded the PH and PAH data from the GEO database, and used the weighted gene coexpression network analysis method (WGCNA) to analyze the coexpression modules of idiopathic noncirrhotic portal hypertension (INCPH) and cirrhotic portal hypertension (CPH) and pulmonary hypertension, respectively. Enrichment analysis was performed on the common genes, and differential gene expressions (DEGs) were used for verification. The target genes of INCPH and PAH were obtained by string and cytoscape software, and the miRNAs of target genes were predicted by miRwalk, miRDB, and TargetScan and their biological functions were analyzed; finally, we used PanglaoDB to predict the expression of target genes in cells. In WGCNA, gene modules significantly related to PAH, CPH, and INCPH were identified, and enrichment function analysis showed that the common pathway of PAH and CPH were “P53 signaling pathway,” “synthesis of neutral lipids”; PAH and INCPH are “terminal,” “Maintenance Regulation of Granules,” and “Toxin Transport.” DEGs confirmed the results of WGCNA; the common miRNA functions of PAH and cirrhosis were enriched for “P53 signaling pathway,” “TGF- Pulmonary arterial hypertension (PAH) is by a variety of causes pathological physiological state of the abnormally elevated pulmonary artery pressure. It can be divided into 5 types: 1) arterial pulmonary hypertension, 2) pulmonary hypertension due to left heart disease, 3) pulmonary hypertension due to pulmonary disease and/or hypoxia, 4) chronic thromboembolic pulmonary hypertension, and 5) pulmonary hypertension due to unknown multifactorial mechanisms. Portopulmonary hypertension (PPHTN) belongs to the first type . PPHTN iWith the rapid development of biological genome sequencing, engineering, and molecular biotechnology, researchers could explore and acquire disease-related genetic information so as to explain the pathogenesis of diseases at the gene level and develop effective drugs. To explore the molecular mechanisms and common pathways of portal pulmonary hypertension, therefore, we used the gene expression data and weighted gene coexpression network to analyze PAH and PH-related coexpression gene clusters, explore common functions, and map miRNA and mRNA networks.“Portal Hypertension” and “Pulmonary arterial hypertension” were used as keywords to search for Gene expression profiles in NCBI GEO datasets. Inclusion criteria were as follows: 1) Gene expression profile data analyzed by WGCNA should be greater than 15 to ensure accuracy; 2) gene expression profile data should include experimental and control groups; and 3) original data should be available. Finally, GSE77627 and GSE113439 were selected for WGCNA analysis, and GSE77627 and GSE53408 were selected for GEO2R analysis.β (ranging from 1 to 20). Next, soft threshold beta was calculated using Pearson's analysis, and the correlation between genes and gene matrix to construct adjacency matrix, calculated by the formula: aij = | Sij | beta . Then, the adjacency matrix transforms the topological overlap matrix (TOM) and the corresponding dissimilarity (1−TOM). The hierarchical clustering tree was further constructed to divide the similar gene expression into different modules. Finally, the expression profile of each module was summarized using a modular characteristic gene (ME), and the correlation between ME and clinical features was calculated. Modules with high correlation coefficients with clinical features were analyzed (p < 0.001), and genes in these modules were selected for subsequent analysis. In this study, scale independence was set at 0.85, and the soft threshold β in WGCNA analysis of PAH was the value estimated by WGCNA . The other parameters are: network Type = “unsigned”, min Module Size = 30, merge Cut Height = 0.25.The data of GSE77627 are divided into cirrhotic portal hypertension (CPH) and idiopathic noncirrhotic portal hypertension (INCPH). Firstly, the top 25% genes were selected according to their variance (CPH and PH took the top 5000 genes). The matrix was transposed and converted into a matrix with sample rows and genes in columns for WGCNA analysis (using the WGCNA software package in R.4.0.5.). The Hclust function of the R language was used for hierarchical clustering analysis to exclude outlier samples. Then, according to the standard of the scale-free network, the “pickSoftThreshold” function in WGCNA software package is used to select the appropriate soft threshold p < 0.001) and selected genes in the correlation module based on GS and MM values (ABS (GS) > .2& ABS (datKME$MM. Module) > .8). At the same time, the genes with significant correlation between PH and PAH groups were overlapped by Venn diagram to find the shared genes. Finally, Cytoscape's ClueGo function was used for GO enrichment analysis and biological analysis of shared genes.We selected gene modules that were significantly positively associated with the trait . 4. Deleted data with both high and low expression. GO functional enrichment analysis and biological analysis were performed for high and low expression of PAH and PH. In DEGs, we still divided GSE77627 data into two groups for GEO2R analysis, that is, CPH compared with the normal group (G1), INCPH compared with the normal group (G2), and PH compared with the normal group (G3). Finally, the differential gene analysis with PAH was performed in G1, G2, and G3 groups, respectively . Cytoscape's ClueGo function was used for GO functional enrichment analysis and biological analysis of shared genes.miRNA is a noncoding RNA that can inhibit mRNA expression and degrade mRNA to regulate gene expression, which is of key significance for the occurrence and development of diseases. Therefore, we explored the regulation of comiRNA on PH and CPH. The upregulated miRNAs of PAH were searched in the Human miRNA Disease Database (HMDD). Since the HMDD did not record miRNA of PH, CPH is a late complication of cirrhosis. So the miRNAs of cirrhosis were used to replace CPH, and the shared miRNAs between them were identified, and their biological functions and target genes were predicted in miRpath. Finally, the target genes of miRpath, the common genes of WGCNA, and DEGs were screened, and the miRNA-mRNA network map was drawn.String software was used to analyze the shared downregulated genes of WGCNA and DGEs in PH and INCPH, and then Cytoscape was used to identify target genes. Finally, miRNA prediction of target genes was performed by miRwalk, TargetScan, and miRDB. Its biological function was predicted using miEAA 2.0  and miRpPanglaoDB software(PanglaoDB - A Single Cell Sequencing Resource For Gene Expression Data), a database that analyzes the expression of genes in those cell subsets, was used to predict the expression of target genes in cells.In WGCNA, GSE113439 was divided into 17 modules in total, among which two modules  were significantly positively correlated with PAH, including 7 and 1982 screened genes according to GS value and MM value. One module (blue) was negatively correlated with the PAH group, including 551 screened genes . There wIn CPH and PAH positive correlation module, there were 71 common genes , GO analWe also analyzed GSE77627 and GSE53408 data by DEGs, and the results showed that CPH and PAH had 146 downregulated common genes . GO analysis showed “Positive regulation of the inflammatory response”, “Sister chromatid separation,” and “P53 signaling pathway”. There wβ signaling pathway,” and “TNF signaling pathway.”Two miRNAs were upregulated  and two were downregulated . miRpath showed “fatty acid metabolism,” “fatty acid synthesis,” “endoplasmic reticulum protein processing,” “P53 signaling pathway,” “TGF-The common gene of the target gene of upregulated miRpath, the shared gene of WGCNA and downregulated DEGs were screened, and there was a total of one gene, MDM2, which was regulated by has-miR-22a-3p. The common gene of the target gene of downregulated miRpath, the shared gene of WGCNA, and upregulated DEGs were screened, and there was one gene in total, PRDX4, which was regulated by has-miR-34a-5p.MiRwalk, TargetScan, and miRDB were performed on the screened target genes, respectively, for miRNA intersection . Among them, miAEE showed that miRNA of EIF5B, HNRNPA2B1, HSP90B1, METAP2, NARS, SACM1L, and HSPA4 were related to telomere function. In addition, we selected the genes that derived from the intersection of target genes with genes enriched for telomeres by WGCNA and DEGs, and miRpath showed that they were related to cell circle, P53, and DNA repair.MDM2 is expressed in hepatocytes, fibroblasts, and adipocytes in basal cell size; PRDX4 is expressed in hepatic stellate cells, fibroblasts, endothelial cells, and adipocytes. EIF5B, HNRNPA2B1, HSP90B1 METAP2, NARS, SACM1L, HSPA4, GNL3, UTP20, and RARS are expressed in endothelial cells, smooth muscle cells, and fibroblasts.Through WGCNA, DEGs analysis, and miRNA identification, it was concluded that the common pathways of CPH and PAH were “lipid (accumulation) metabolism,” “inflammatory response,” and “P53 signaling pathway.” The common pathways of INCPH and PAH were “telomere,” “intestinal inflammation,” and “toxin transport.” The common pathways of PH (including CPH and INCPH) and PAH included the P53 signaling pathway and telomere.Liver cirrhosis is the precondition for CPH, and the development of HC is inseparable from the activation of hepatic stellate cells (HSCs). The phenotype changes from static to myofibroblasts with contraction and secretion of extracellular matrix, causing liver fibrosis. The formation of portal hypertension is also related to liver fibrosis and the contractility phenotype of HSCs. In addition, portal hypertension is closely related to hepatic sinusoidal endothelial cells and platelets/thrombocyte . INCPH iα (HIF-2α) increased. Pulmonary hypertension is due to vascular remodeling caused by increased proliferation of pulmonary smooth muscle cells and apoptosis of pulmonary endothelial cells, resulting in endothelial cell dysfunction and continuous vasoconstriction [α induces endothelial cell transformation into mesenchymal cells leading to severe pulmonary hypertension [κB pathway [κB [The liver is involved in fat metabolism, including triglyceride metabolism, production, and transportation of very low density lipoprotein. Neutral lipids are inert lipids whose synthesis is the first step of lipid droplet (LD) formation . LD is atriction . HIF-2 αrtension . Endothe pathway . P53 canhway [κB . Surprishway [κB . In the hway [κB . PRDX4 ihway [κB , 37. Somhway [κB . We hypoα and 1β (PGC-1α and PGC-1β) by inducing P53 activation, ultimately leading to mitochondrial dysfunction and promoting cell senescence. With the deepening of the research on PAH, scholars found that cell aging phenotype in irreversible PAH plays a decisive role in apoptosis and senescence, TNF-α induced pulmonary cells to the aging phenotype lead to vascular dysfunction and proved that the aging of vascular smooth muscle cell proliferation of cells can be induced in vitro [The common pathways of INCPH and PAH are telomeres,  inflammation, and toxin transport. Maintenance of telomeres ensures the correct replication of genetic material and protects DNA from damage to ensure genomic stability and accuracy. Telomere dysfunction can lead to cell cycle arrest, aging, and genomic instability, and is associated with bone marrow failure syndrome and cancer . Studiesin vitro . Telomerin vitro . GNL3 anin vitro . In addiin vitro . HNRNPA2in vitro . Under tin vitro .We also analyzed PH (including CPH and INCPH) and PAH by DEGs and WGCNA. Their common pathways include the P53 signaling pathway or telomere, and we thought of it as a potential mechanism. Portal hypertension can lead to oxidative stress, bacterial translocation, intestinal lipid peroxidation, sepsis, etc. Oxidative stress and inflammation will lead to telomere dysfunction and then induce the activation of P53, while overactivated P53 and telomere dysfunction can also induce oxidative stress and inflammation, forming a vicious cycle that promotes senescence and dysfunction of pulmonary vascular cells, reduces the secretion of NO, causes vascular dilatation dysfunction, and promotes or accelerates the formation of pulmonary hypertension.Portal pulmonary hypertension is a rare complication of portal hypertension. At present, there are few studies on the mechanism of the disease. This is the first study to use the WGCNA method to construct a coexpression network to explore the relationship between PH (including INCPH and CPH) and pulmonary hypertension. Our findings reveal several key genes and a common pathway that play an important role in the etiology of portal pulmonary hypertension, which may enhance our understanding of the molecular mechanisms of this disease. In this study, we believe that P53 signaling pathway/telomere dysfunction may be the pathogenesis of the disease, and the mechanism  may be tWe summarized that the mechanism of pulmonary hypertension induced by portal hypertension may be related to telomere dysfunction and P53 overactivation. In addition, lipid metabolism and intestinal inflammation are also involved in this process.


Novel crosslinking bio polyurethane based polymeric solid–solid phase change materials (SSPCM) were synthesized using castor oil (CO) based hyperbranched polyols as crosslinkers. CO-based hyperbranched polyols were synthesized by grafting 1-mercaptoethanol or α–thioglycerol via a thiol-ene click reaction method . Subsequently, the three SSPCMs were synthesized by a two-step prepolymer method. Polyethylene glycol was used as the phase change material in the SSPCMs, while the CO-based hyperbranched polyols and two types of diisocyanate (hexamethylene diisocyanate (HDI) and 4,4'-diphenylmethane diisocyanate) served as the molecular frameworks. Fourier transform infrared spectroscopy indicated the successful synthesis of the SSPCMs. The solid–solid transition of the prepared SSPCMs was confirmed by X-ray diffraction analysis and polarized optical microscopy. The thermal transition properties of the SSPCMs were analyzed by differential scanning microscopy. The isocyanate and crosslinker types had a significant influence on the phase transition properties. The SSPCM samples prepared using HDI and COT exhibited the highest phase transition enthalpy of 126.5 J/g. The thermal cycling test and thermogravimetric analysis revealed that SSPCMs exhibit outstanding thermal durability. Thus, the novel SSPCMs based on hyperbranched polyols have great potential for application as thermal energy storage materials. From this perspective, thermal energy storage (TES) has become an important part of renewable energy systems5. To meet the growing demand for cheaper and more efficient TES, phase change materials (PCM) have recently gained attention for their applicability in the development of TES devices6. PCMs exhibit a large latent heat capacity and uniquely maintain a constant temperature during latent heat transfer, which make them suitable for thermal energy management in various applications such as solar energy storage9, solar desalination12, building materials15, and waste heat recovery18. In the past decade, studies have focused on the suitability of polyethylene glycol (PEG)19, paraffin waxes20, fatty acids21, inorganic salt hydrates22, and the eutectics of these compounds23 as PCMs. However, the development of TES materials using pristine PCMs is challenging, it requires special packaging technology to support the solid–liquid phase transition. Compared to the pristine PCM, solid–solid PCMs (SSPCM) are economical and convenient to produce, face no risk of leakage, and exhibit shape stability26. Thus, various methods have been studied to fabricate SSPCMs, such as grafting, crosslinking polymerization, and introduction of carbon materials, titanium oxide, black phosphorus, and their hybrids35.The continuous increase in fuel prices and efforts toward achieving carbon neutrality in recent years has necessitated the development of sustainable energy sources39. PU is a polymer synthesized through the addition polymerization between a multi-functional alcohol and diisocyanate. As only few types of commercial diisocyanate, the polyols used play a critical role in determining the properties of PUs. Recently, numerous studies on PU-based SSPCMs have been reported in which PEG was used as the PCM material42. A PU-based SSPCM typically has a crosslinked structure, which limits the dissolution and flow of the PEG soft segment. Several researchers have developed improved polymeric SSPCMs using multifunctional polyol as a crosslinking agent45.Polyurethane (PU) is one of the most important polymeric materials that has been widely applied in industries such as automotive, furniture, thermal shielding materials, and sound insulating materials, because it is economical and convenient to produce46. CO is primarily composed of ricinoleic acid, with a double bond on C–9 and a hydroxyl group on C–12; therefore, it has been frequently applied in studies on bio PU (BPU). However, the applications of CO-based PU have been limited because of its low crosslinking density, which leads to low mechanical properties and low productivity. To overcome such drawbacks, many studies have conducted to facilitate the use of CO by epoxidation47, transesterification48, ozonolysis49, and thiol-ene click reactions51. Previously, we have reported the use of the thiol-ene click reaction for grafting 1-mercaptoethanol and α-thioglycerol onto CO53.The use of castor oil (CO) as a bio-based polyol, which can replace conventional petrochemical polyols, has attracted considerable attention because of its biodegradability, abundance, nontoxic nature, cost competitiveness, and biodegradabilityIn this study, two types of CO-based hyperbranched polyols were prepared, which were subsequently used in the production of BPU-based SSPCMs. The structural properties of SSPCM were analyzed by Attenuated total reflection (ATR)-Fourier transform infrared (FTIR) spectroscopy, while X-ray diffraction (XRD) analysis and polarized optical microscopy (POM) were conducted to determine the degree of crystallinity. The phase change properties of the SSPCMs were thoroughly examined by differential scanning calorimetry (DSC). After the thermal cycling test, DSC and FT-IR analysis were performed again to verify the thermal reliability of the SSPCMs. Additionally, the thermal stability of the SSPCMs was verified by thermogravimetric analysis (TGA).-1) was supplied from Kanto Chemical Co., Inc. Ethyl acetate (EA), N, N-dimethylformamide (DMF), anhydrous MgSO4 and NaCl were supplied from Daejung Chemical (Korea). 4, 4'-diphenylmethane diisocyanate (MDI) was purchased from Tokyo Chemical Industry Co., Ltd. Hexamethylene diisocyanate (HDI) was supplied from Wako Chemicals. All reagents were used as received without further purification.CO was supplied from Yakuri Pure Chemical Co., Ltd. Mercaptoethanol, α-thioglycerol, 2, 2-dimethoxy-2-phenylacetophenone (DMPA), acetic anhydride, pyridine, and dibutyltin dilaurate (DBTDL) were purchased from Sigma-Aldrich (USA). PEG , and diffractograms were scanned in a 2θ range of 5°‒70° at scan rate of 3°/min. The spherulite morphologies were observed through POM (Olympus BX-51 TX) equipped with a CCD camera. The thermal behavior of the SSPCMs was investigated by DSC . An accelerated thermal cycling test was performed for 100 cycles in the temperature range of 0‒100 °C using a thermocycler (MJ research). All samples were tested for 5 consecutive heating–cooling scan from 0 to 100 °C at 5 °C/min under nitrogen atmosphere. TGA of the SSPCMs was performed with nitrogen purging from 30 to 800 °C at 10 °C/min.ATR-FTIR spectrometer  was used to determine the structure of the SSPCMs over the range of 500–4000 cm53. The FTIR spectra of the newly prepared polyols are shown in Fig. 53. The hydroxyl values of CO, COM, and COT measured according to the ASTM D1957-86 standard were 160, 270, and 380 mg KOH g−1, respectively.COM and COT were successfully synthesized under the optimized reaction conditions obtained from our previous studies−1 in all the SSPCM spectra indicates that the PUs were successfully synthesized. The newly formed absorbance bands at 3341 and 1539 cm−1 correspond to the N–H stretching and amide vibration within the PU bond, respectively. The FTIR analysis did not show a significant difference in the spectra in terms of the functionality of the crosslinker or the type of isocyanate.The chemical structures of the synthesized SSPCMs were analyzed by FTIR spectroscopy, and the results are presented in Fig. 54. This was more prominent for the SSPCMs prepared using MDI because the rigid benzene ring further confines the PEG chain41.The crystal structures of the SSPCMs were analyzed by XRD analysis, and the results are presented in Fig. For further analysis of the crystal structure of the synthesized SSPCMs, the crystal morphologies of the pristine PEG and the SSPCMs were recorded through POM, and the results are shown in Fig. DSC analysis was performed to examine the phase transition properties of the synthesized SSPCMs. The five consecutive DSC scans after heating and cooling of the pristine PEG and SSPCMs are presented in Figs. Regarding the influence of the functionality of the crosslinker on the changes in the SSPCMs endothermic enthalpy, the degree of latent heat for both MDI- and HDI-based SSPCM followed the order; CO < COM < COT. Studies have revealed several factors that influence the SSPCM phase transition properties, including soft segment content by weight, crystalline state of the soft segments, and steric hindrance of the crosslinking points. With an increase in the functionality of the crosslinker, the synthesized SSPCM can accommodate more PEG, thereby increasing the latent heat storage capacity. For better understanding of the phase transition properties of the SSPCMs on the basis of the functionality of the crosslinker, the relative latent heat efficiency was calculated using the following equation:entclass1pt{minimaConsidering the influence of the isocyanate type on the endothermic enthalpy of the SSPCMs, the latent heat of the HDI-based SSPCM was higher than that of the MDI-based SSPCM when an identical crosslinker was used. The The analysis of the phase transition properties of the SSPCMs by DSC demonstrated that using hyperbranched polyols led to an increase in latent heat efficiency, although the relative latent heat efficiency decreased due to an increase in the crosslinking density. Steric hindrance arising from the structural characteristics of isocyanate is another factor influencing the latent heat efficiency of the SSPCMs.64.The results of the crystalline structure and phase transition property analyses confirmed that the synthesized SSPCMs exhibited repetitive heat storage and release characteristics without any leakage. The phase change mechanisms of the SSPCMs crosslinked with CO and COT are shown in Fig. To verify the thermal stability of the SSPCMs, an accelerated thermal cycling test (100 cycles from 0 to 100 °C) was performed. Even after the 100th thermal cycle, no weight reduction was observed in the SSPCMs, which indicates that they exhibit thermal reliability even after repeated use within the phase transition temperature range. This result verified the applicability of the SSPCMs as PCM materials.Figure Figure 65. Also, the maximum decomposition temperature of all the SSPCMs was higher than that of PEG, exhibiting the outstanding thermal stability of the SSPCMs.TGA was performed to evaluate the thermal stability of the SSPCMs; the results are presented in Fig. In this study, two types of hyperbranched polyols, COM and COT, with an increased hydroxyl value, were prepared via the thiol-ene click reaction with CO. A new SSPCM series for TES was successfully prepared using PEG as the PCM in the SSPCMs, while CO, COM, and COT provided the molecular framework. FTIR analysis of the SSPCMs revealed the formation of PU structures, which confirmed the successful synthesis of the SSPCMs. Moreover, the results of the XRD analysis and POM indicated the solid–solid phase transition. The results of the DSC analysis revealed that the isocyanate and crosslinker types had a significant influence on the phase transition properties. The H.COT exhibited the highest phase transition enthalpy at 126.5 J/g. Furthermore, the results of the thermal cycling test and TGA demonstrated the outstanding durability of the SSPCMs. Thus, the novel SSPCMs based on hyperbranched polyols has great potential to be applied in TES materials.Supplementary Information.


To safely manage this unique alpha-particle-emitting radionuclide, we investigated the permeability of four types of plastic films and two types of rubber gloves against 211At and identified suitable materials that prevent contamination by 211At.211At was placed on them. The radioactivity of the pieces of filter paper under the materials was evaluated by measuring counts using a gamma counter and obtaining autoradiograms 3.5 h later. These experiments were also performed using 225Ac, 125I, 111In, 201Tl, and 99mTc.Four types of plastic films, polyethylene, polyvinylidene chloride, polyvinyl chloride, and a laminated film, and two types of rubber gloves, latex and nitrile, were examined. Small pieces of filter paper were covered with these materials, and a drop containing 100 kBq of 211At solution easily penetrated polyethylene, polyvinyl chloride, and latex rubber. Similar results were obtained for 125I, while other radionuclides did not penetrate films or gloves. These results suggest that halogenic radionuclides under anionic conditions are likely to penetrate plastic films and rubber gloves.211At solution is used, the protection by polyvinylidene chloride, a laminated film, or nitrile rubber would be more effective than that by polyethylene, polyvinyl chloride, or latex rubber.Our evaluation revealed that, when The online version contains supplementary material available at 10.1186/s40658-022-00469-9. Polyethylene (30 µm), polyvinylidene chloride (11 µm), and polyvinyl chloride (8 µm) and laminated films of polypropylene, ethylene vinyl alcohol copolymer , and polyethylene (104 µm) were used as plastic films. The numbers in parentheses are the film thicknesses. The first three plastic films are commercially available and used for wrapping perishable food materials to maintain freshness. The laminated film was developed to pack dried fish flakes so that they are not damaged by oxygen and high humidity. As rubber sheets, pieces of latex rubber gloves  and nitrile rubber gloves  were used. These gloves are currently used as PPE against COVID-19 AcNO3 solutio125I]NaI solution was dropped on the materials, strong radioactivity was detected in pieces of filter paper under polyethylene film, polyvinyl chloride film, and latex rubber, in that order. No significant radioactivity was detected in pieces of filter paper under the polyvinylidene chloride film, the laminated film, or the nitrile rubber sheet (Fig. 211At]NaAt solution.When the [eet Fig. . These r111In]InCl3, [201Tl]TlCl, and [99mTc]NaTcO4 solutions were dropped on the materials, no significant radioactivity was detected in each piece of filter paper even after 15 h of exposure (Additional file When 11InInCl3The radioactivity of pieces of filter paper is shown in Fig. 211At, a promising alpha-particle-emitting radionuclide for TAT, is popular in Japan because of its relatively wide availability. Many researchers in Japan have been engaged in studies using radionuclides. However, this element is regarded as an enigmatic element [211At can easily volatilize and contaminate the environment [211At]NaAt solution is dependent on the type of shielding material. 225Ac is another popular alpha-particle-emitting radionuclide, and the [225Ac]AcCl3 solution penetrated films and rubber only minimally. Because the numbers of alpha particles emitted during the single decay of 211At and 225Ac atoms are 1 and 4, respectively, 225Ac is a stronger alpha-particle-emitting radionuclide than 211At. Considering these findings, we believe that the penetration of the [211At]NaAt solution of films and rubber was not induced by the direct destruction of materials by emitted alpha particles. The penetration depends on the chemical properties of the shielding materials. 225Ac, which is cationic in a solution, and other cationic radionuclides 111In and 201Tl also failed to penetrate the shielding materials. In contrast, 125I, which is a halogen and is anionic in [125I]NaI solution in the same manner as 211At, showed a similar trend to that of 211At. However, another popular radioactive anion, [99mTc]TcO4−, did not penetrate the material. These results suggest that halogenic radionuclides under anionic conditions are likely to penetrate plastic films and rubber. element , and mosironment . Therefo211At and were considered ineffective in protecting against the contamination due to 211At. However, the polyvinylidene chloride film, the unique laminated film, and the nitrile rubber glove were resistant to the penetration by the [211At]AtNa solution. Although the details of the differences in chemical properties of materials between these two groups, easily permeable and non-permeable for 211At, were unrevealed, the permeability of the [211At]AtNa solution was correlated with that of gas or water barrier properties according to open data [211At for plastic films and rubber gloves (data are shown in Additional file 211At, polyethylene is inexpensive, and bags made of this material are sold for radioactive waste disposal and are actually used in experiments with 211At [211At can easily penetrate them (data are shown in Additional file 211At solution. Hence, we must wear nitrile gloves and wrap or cover specimens with polyvinylidene chloride film or a laminated film while working with compounds including 211At to avoid unnecessary internal radiation exposure.Among the materials examined in this study, two types of plastic film, polyethylene and polyvinyl chloride, and the latex rubber glove were penetrated by pen data . The resth 211At . Latex r211At anion can easily penetrate at least two types of plastic films, polyethylene and polyvinyl chloride, and latex rubber. The permeability of 211At may depend on its chemical properties as a halogen that becomes an anion in a water solution. When we deal with the 211At anion, we must wear nitrile gloves and wrap or cover specimens using a polyvinylidene chloride film or a laminated film.Our preliminary experiments indicated that the Additional file 1. Supplementary figures and tables.


The proteomic analysis of plasma holds great promise to advance precision medicine and identify biomarkers of disease. However, it is likely that many potential biomarkers circulating in plasma originate from other tissues and are only present in low abundances in the plasma. Accurate detection and quantification of low abundance proteins by standard mass spectrometry approaches remain challenging. In addition, it is difficult to link low abundance plasma proteins back to their specific tissues or organs of origin with confidence. To address these challenges, we developed a mass spectrometry approach based on the use of tandem mass tags (TMT) and a tissue reference sample. By applying this approach to nonhuman primate plasma samples, we were able to identify and quantify 820 proteins by using a kidney tissue homogenate as reference. On average, 643 ± 16 proteins were identified per plasma sample. About 58% of proteins identified in replicate experiments were identified both times. A ratio of 50 μg kidney protein to 10 μg plasma protein, and the use of the TMT label with the highest molecular weight (131) for the kidney reference yielded the largest number of proteins in the analysis, and identified low abundance proteins in plasma that are prominently found in the kidney. Overall, this methodology promises efficient quantification of plasma proteins potentially released from specific tissues, thereby increasing the number of putative disease biomarkers for future study.  The promise of precision medicine for improved patient care and disease management critically depends on early disease biomarker signatures that predict health outcomes, facilitate early diagnosis, and inform treatment . Ideallym/z) for each ion that is detected in the sample. Subsequently, individual peptide ions are fragmented and a fragment MS spectrum (MS2) is collected that aids in the determination of the amino acid sequence of the initial peptide [Over the past two decades, liquid chromatography-coupled tandem mass spectrometry (MS) has become the primary methodology for untargeted proteomics analysis, including biomarker discovery. In a standard workflow, peptides go through two levels of MS analysis ,14,15. T peptide . Low abu peptide . Therefo peptide . These dTandem mass tag (TMT) labeling is a widely used technique in proteomics for the identification and quantification of proteins ,19. TMTsIn this study, we describe an adaptation of this approach for the identification of low-abundance tissue leakage proteins in plasma. With this approach, any peptide identified during MS analysis in a tissue reference sample can subsequently be quantified in individual plasma samples using the TMT reporter ions, and would be detected even if present in low amounts. This mass spectrometry plasma proteomics methodology to discover novel disease markers will expand our ability to detect proteins from specific tissues in plasma samples, and expand the list of putative tissue-derived biomarker proteins for many diseases and tissues.Papio hamadrayas) that were part of a pedigreed baboon colony from the Southwest National Primate Research Center at the Texas Biomedical Research Institute, San Antonio, Texas [All samples used in this study were collected from female olive baboons  for removal of the 14 highly abundant proteins. Both plasma and homogenized tissues were precipitated using acetone. For unlabeled proteomics , 100 μg of the resulting tissue proteins were dissolved in 100 mM ammonium bicarbonate (ABC)  buffer and followed by three major steps: 25 mM dithiothreitol (DTT)  for reduction, 13.875 mM iodoacetamide (IAA)  for acetylation, and 2.5 μg trypsin for overnight digestion. The digested samples were further fractionated using the Thermo pH fractionation kit (Pierce High pH Reversed-Phase Peptide Fractionation). Mass Spectrometry: For TMT proteomics, 100 μg of total precipitated proteins from kidney cortex, medulla, and plasma in 100 mM TEAB were processed using 200 mM TCEP, 375 mM IAA, and 2.5 μg trypsin digestion. Cortex, medulla, and plasma digests were labeled with the designated TMT10plex channel  reagents (see m/z = 400–1600) at a resolution of 120,000 with automatic gain control (AGC) of 1 × 106 and maximum injection time of 50 ms. MS/MS data acquisition was done using higher-energy C-trap dissociation (HCD) mode in Orbitrap detection at a resolution of 50,000 with an AGC target of 1.25 × 105 and maximum injection time of 86 ms. All data acquisition was carried out using Thermo Scientific Xcalibur software  and data analysis was completed in MaxQuant  against P. anubis proteome database  using the default parameters for TMT or LFQ analysis. Modifications were set to the default variable modifications (oxidation (M) and acetyl (protein N-term)) and fixed modifications (carbamidomethyl). Individual peptide mass tolerance was selected to filter peptides according to individual peptide mass tolerances. False discovery rates (FDR) were set to 0.01 at both the PSM and the protein level. MaxQuant output files (proteingroups) were manually curated, and contaminant matches or protein identifications with no measured reporter ion intensities across all samples were removed. All results files can be found in the After fractionation, eight fractions were pooled to four  and 1 μg of each sample was loaded on a PepMap RSLC C18 easy-spray column  using Easy-nLC 1200 coupled to an Orbitrap Lumos Tribrid Mass Spectrometer . Peptides were separated using a 2-h gradient of Mobile phase A (0.1% formic acid in 95:5 water: acetonitrile) and Mobile Phase B (0.1% formic acid in 80:20 acetonitrile: water). The following gradient program was used for peptide elution: 2–30% B in 85 minutes, 30–95% B in 30 minutes, and 95% B in 5 minutes. Data were acquired in MS1 scan mode  samples for development and optimization of our method. Seven plasma samples were collected as part of a recent study  and kidnWe implemented an isotopic labeling strategy using tandem mass tags (TMT) to identify and quantify tissue-derived proteins. In this approach, a tissue homogenate sample is used as a reference sample and labeled with one TMT, and then combined with plasma samples labeled with complementary TMTs. We used the Thermo Scientific TMT10plex Isobaric Mass Labeling Reagent Set for the experiments described here. To optimize our methodology, we focused on the putative use of kidney homogenate samples (cortex and medulla) to identify additional proteins in plasma samples. Proteins from tissue homogenate and plasma proteins were mixed in different ratios to assess the optimal protocol to maximize protein identification in plasma samples, and maximize reproducibility between replicate experiments. For example, a ratio of 5:1 (kidney tissue to plasma) contains 50 μg peptides from the kidney tissue homogenate for medulla and cortex (labeled with 126 and 127N reporter ions), and is combined with 10 μg peptides from each plasma sample . Since ip = 7 × 10−11–2 × 10−23). The method TMT 5:1 identified on average 457 ± 19 proteins, the 10:1 ratio 499 ± 3 proteins, and TMT 5:2 401 ± 4 proteins. We also reversed the sample order in additional experiments where the cortex and medulla peptides were labeled with the highest molecular weight reporter ions (130C and 131). In this reversed experiment (method TMT 5:1 reverse), we identified on average 643 ± 16 proteins per plasma sample. As shown in When only proteins are considered that are identified in both replicate experiments, method TMT 5:1 reverse identified 477 proteins compared to 266, 288, and 261 for TMT 5:1. TMT 5:2, and TMT 10:1 experiments, respectively . Clearlyp = 0.00165), suggesting that this method enhances the ability to detect low abundance proteins in plasma that may be derived from the kidney. As shown in http://www.proteinatlas.org, accessed on 4 January 2023) identifies 452 genes that have at least four-fold higher mRNA expression levels in the kidney compared to any other tissues. We used this gene set to annotate the proteins identified by the different methods described above. About 50 of the proteins identified by the method TMT 5:1 reverse are annotated as kidney-enriched. These five proteins  are only detected by the method TMT 5:1 reverse. All of these proteins are expressed in the proximal tubules of the kidney. In contrast, only one protein identified by the LFQ analysis of the plasma samples (GPX3) is included in the list of kidney-enriched proteins. This annotation demonstrates that a high proportion of the additional proteins identified by the TMT labeling approach are kidney-enriched, and may serve as potential biomarkers for kidney function or dysfunction. For example, the gene solute carrier family 4 member 4 (SLC4A4) encodes a sodium bicarbonate cotransporter, and the gene has been associated with renal tubular acidosis [The Human Protein Atlas  results in the largest number of unique proteins identified, the highest number of proteins per plasma sample, and the results are most reproducible between replicate experiments, suggesting that this is the best experimental design to enhance the discovery of putative tissue leakage proteins in plasma samples using TMT-labeled proteomics mass spectrometry.Analysis of tissue leakage proteins in plasma samples using untargeted mass spectrometry approaches remains a challenge. Only a small fraction of proteins account for over 95% of the total protein content in plasma, and these highly abundant plasma proteins impede effective detection and quantification of the remaining, less abundant proteins (REF). However, low abundance proteins may oftentimes be the very proteins that are indicative of disease processes in particular tissues and organs. As tissue deteriorates, tissue-derived proteins are released into the blood stream in small quantities, particularly during early stages of disease development. Such proteins are of particular interest as potential disease biomarkers, especially if they are likely released as part of the disease process. However, they remain challenging to identify and quantify. While some recent and more sensitive  mass spectrometry approaches improve the detection and quantification of low abundance proteins, linking them to particular pathophysiological processes in tissues or organs remains difficult. Recent adaptations of tandem mass tag (TMT) labeling in proteomics have shown that a TMT labeling approach can be used to effectively identify proteins at very low abundance if the analysis also includes a reference sample. In single cell proteomics, the SCoPE-MS approach uses a reference sample of proteins extracted from several hundred cells for protein identification in MS1 and MS2, and then uses proteins labeled with other TMT reporters to quantify those proteins in individual cells . In our Our mass spectrometry data clearly demonstrate that the use of a kidney protein extract as a reference sample in a TMT experiment dramatically increases the number of proteins identified in plasma samples. The optimal methodology uses the heaviest TMT reporter ion for the reference sample. This likely minimizes reporter ion interference with other reporter ion channels, and leads to the identification and quantification of 820 proteins in these plasma samples. A substantial fraction of the proteins only identified by the optimal methodology (that we named TMT 5:1 reverse) is also identified in the kidney homogenate, but is much more abundant in the kidney, supporting our hypothesis that this approach will identify kidney proteins (leakage proteins) that are much less abundant in plasma. Obviously, not all proteins identified in our analysis are exclusively expressed in the kidney. In fact, analysis of GTEx data suggests that some proteins are significantly enriched in particular tissues, but very few are exclusively expressed in one or only few tissues . As we dOur analysis only included a small set of four animals from the initial study. Given our sample size and the short duration of the experiment, we would not expect to detect any proteins that are associated with sodium exposure or blood pressure variation at this point. Clearly, future studies will explore the usefulness of the approach described here to identify sets of protein biomarkers that are associated with particular kidney diseases. Given a set of over 800 proteins that can be monitored and quantified using our TMT labeling approach, this significantly increases the probability of finding kidney-derived biomarkers using this untargeted mass spectrometry approach.There are other technologies being promoted for plasma proteome analysis such as Somascan, an aptamer-based affinity analysis ,26, and Overall, our pilot work has allowed us to quantify proteins that are in low abundance and difficult to detect using standard label-free proteomics strategies and even more difficult to link back to an organ or tissue. The innovation of introducing TMT-labels with tissue reference samples in multiplexing enables us to explore the biological significance of specific tissue proteins in plasma, and extend the number of putative biomarkers for diseases. Clearly, this work can be performed with reference samples from other disease tissues as well , opening the door to future discoveries of early disease biomarker signatures that are tissue- or organ-specific and link to the underlying pathophysiology of a disease.