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In response to Mazaheri et al.'s critique, we revisited our study (Valentini et al., 2022) on the relationship between peak alpha frequency (PAF) and pain. Their commentary prompted us to reassess our data to address the independence between slow and slowing alpha brain oscillations, as well as the predictivity of slow alpha oscillations in pain perception. Bayesian correlation analyses revealed mixed support for independence. Investigating predictivity, we found inconsistent associations between pre-PAF and unpleasantness ratings. We critically reflected on methodological and theoretical issues on the path to PAF validation as a pain biomarker. We emphasized the need for diversified methodology and analytical approaches as well as robust findings across research groups., Competing Interests: Declaration of competing interest The authors report no conflict of interest., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Purpose: Contrast-enhanced mammography (CEM) is an innovative imaging tool for breast cancer detection, involving intravenous injection of a contrast medium and the assessment of lesion enhancement in two phases: early and delayed. The aim of the study was to analyze the topographic concordance of lesions detected in the early- versus delayed phase acquisitions., Materials and Methods: Approved by the Ethics Committee (No. 118/20), this prospective study included 100 women with histopathological confirmed breast neoplasia (B6) at the Radiodiagnostics Department of the Maggiore della Carità Hospital of Novara, Italy from May 1, 2021, to October 17, 2022. Participants underwent CEM examinations using a complete protocol, encompassing both early- and delayed image acquisitions. Three experienced radiologists blindly analyzed the CEM images for contrast enhancement to determine the topographic concordance of the identified lesions. Two readers assessed the complete study (protocol A), while one reader assessed the protocol without the delayed phase (protocol B). The average glandular dose (AGD) of the entire procedure was also evaluated., Results: The analysis demonstrated high concordance among the three readers in the topographical identification of lesions within individual quadrants of both breasts, with a Cohen's κ > 0.75, except for the lower inner quadrant of the right breast and the retro-areolar region of the left breast. The mean whole AGD was 29.2 mGy. The mean AGD due to CEM amounted to 73% of the whole AGD (21.2 mGy). The AGD attributable to the delayed phase of CEM contributed to 36% of the whole AGD (10.5 mGy)., Conclusions: As we found no significant discrepancy between the readings of the two protocols, we conclude that delayed-phase image acquisition in CEM does not provide essential diagnostic benefits for effective disease management. Instead, it contributes to unnecessary radiation exposure., (© 2024. The Author(s).)

Introduction: Bcl-2 and Bcl-xL are the most studied anti-apoptotic members of Bcl-2 family proteins. We previously characterized both of them, not only for their role in regulating apoptosis and resistance to therapy in cancer cells, but also for their non-canonical functions, mainly including promotion of cancer progression, metastatization, angiogenesis, and involvement in the crosstalk among cancer cells and components of the tumor microenvironment. Our goal was to identify transcriptional signature and novel cellular pathways specifically modulated by Bcl-2., Methods: We performed RNAseq analysis of siRNA-mediated transient knockdown of Bcl-2 or Bcl-xL in human melanoma cells and gene ontology analysis to identify a specific Bcl-2 transcriptional signature. Expression of genes modulated by Bcl-2 and associated to Hippo pathway were validated in human melanoma, breast adenocarcinoma and non-small cell lung cancer cell lines by qRT-PCR. Western blotting analysis were performed to analyse protein expression of upstream regulators of YAP and in relation to different level of Bcl-2 protein. The effects of YAP silencing in Bcl-2 overexpressing cancer cells were evaluated in migration and cell viability assays in relation to different stiffness conditions. In vitro wound healing assays and co-cultures were used to evaluate cancer-specific Bcl-2 ability to activate fibroblasts., Results: We demonstrated the Bcl-2-dependent modulation of Hippo Pathway in cancer cell lines from different tumor types by acting on upstream YAP regulators. YAP inhibition abolished the ability of Bcl-2 to increase tumor cell migration and proliferation on high stiffness condition of culture, to stimulate in vitro fibroblasts migration and to induce fibroblasts activation., Conclusions: We discovered that Bcl-2 regulates the Hippo pathway in different tumor types, promoting cell migration, adaptation to higher stiffness culture condition and fibroblast activation. Our data indicate that Bcl-2 inhibitors should be further investigated to counteract cancer-promoting mechanisms., (© 2024. The Author(s).)

Soils and dusts can act as sinks for semivolatile lipophilic organic compounds and children ingest relatively large amounts of both soils and dusts. Following intake, sorbed chemicals may desorb (mobilize) and become available for intestinal absorption (bioaccessible). When chemicals are not degraded in the digestive tract, mobilization can approximate bioaccessibility. Alternatively, when gastrointestinal degradation of mobilized chemicals does occur, it can be useful to separate mobilization from bioaccessibility. In this study we used synthetic digestive fluids in a sequential, three-compartment (saliva, gastric, and intestinal) in vitro assay to construct mobilization and bioaccessibility models for 16 pesticides (log K ow 2.5-6.8) sorbed to 32 characterized soils and house dusts. To address the potential loss of mobilized pesticides due to absorption, the assays were repeated using a solid phase sorbent (tenax) added to the digestive fluid immediately after addition of the intestinal fluid components. We found that pesticide mobilization was predicted by pesticide log K ow and the carbon content of the soils and dusts. Pesticide loss measurably reduced the bioaccessibility of most pesticides, and bioaccessibility was largely predicted by log K ow and pesticide loss rate constants. Introduction of the sink increased mobilization by x̄ = 4 ± 6% (soil) and x̄ = 9 ± 7% (dust) while bioaccessibility increases were x̄ = 41 ± 21% (soil) and x̄ = 24 ± 12% (dust). The physicochemical properties of the soils, dusts, and pesticides used in this study successfully predicted the in vitro mobilization and bioaccessibility of the pesticides. This suggests that modeling of pesticide mobilization and bioaccessibility could reduce uncertainty in exposure and risk assessments., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Published by Elsevier Ltd.)

Per- and polyfluoroalkyl substances (PFAS) are regularly found in soils and dusts, both of which can be consumed by children at relatively high amounts. However, there is little data available to model the bioaccessibility of PFAS in soils and dusts when consumed or to describe how the physiochemical properties of PFAS and soils/dusts might affect bioaccessibility of these chemicals. Because bioaccessibility is an important consideration in estimating absorbed dose for exposure and risk assessments, in the current study, in vitro assays were used to determine bioaccessibility of 14 PFAS in 33 sets of soils and dusts. Bioaccessibility assays were conducted with and without a sink, which was used to account for the removal of PFAS due to their movement across the human intestine. Multiple linear regression with backward elimination showed that a segmented model using PFAS chain length, number of branches, and percent total organic carbon explained 78.0%-88.9% of the variability in PFAS bioaccessibility. In general, PFAS had significantly greater bioaccessibility in soils relative to dusts and the addition of a sink increased bioaccessibility in the test system by as much as 10.8% for soils and 20.3% for dusts. The results from this study indicate that PFAS bioaccessibility in soils and dusts can be predicted using a limited set of physical chemical characteristics and could be used to inform risk assessment models., (Published by Oxford University Press on behalf of the Society of Toxicology 2023.)

Low-grade chronic inflammation is a hallmark of ageing, associated with impaired tissue function and disease development. However, how cell-intrinsic and -extrinsic factors collectively establish this phenotype, termed inflammaging, remains poorly understood. We addressed this question in the mouse intestinal epithelium, using mouse organoid cultures to dissect stem cell-intrinsic and -extrinsic sources of inflammaging. At the single-cell level, we found that inflammaging is established differently along the crypt-villus axis, with aged intestinal stem cells (ISCs) strongly upregulating major histocompatibility complex class II (MHC-II) genes. Importantly, the inflammaging phenotype was stably propagated by aged ISCs in organoid cultures and associated with increased chromatin accessibility at inflammation-associated loci in vivo and ex vivo, indicating cell-intrinsic inflammatory memory. Mechanistically, we show that the expression of inflammatory genes is dependent on STAT1 signaling. Together, our data identify that intestinal inflammaging in mice is promoted by a cell-intrinsic mechanism, stably propagated by ISCs, and associated with a disbalance in immune homeostasis., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)

Breast cancer stands as the most frequent malignancy and leading cause of death among women. Early and accurate detection of this pathology represents a crucial factor in enhancing both incidence and mortality rates. Ultrasound (US) examination has been extensively adopted in clinical practice due to its non-invasiveness, affordability, ease of implementation, and wide accessibility, thus representing a valuable first-line diagnostic tool for the study of the mammary gland. In this scenario, recent developments in nanomedicine are paving the way for new interpretations and applications of US diagnostics, which are becoming increasingly personalized based on the molecular phenotype of each tumor, allowing for more precise and accurate evaluations. This review highlights the current state-of-the-art of US diagnosis of breast cancer, as well as the recent advancements related to the application of US contrast agents to the field of molecular diagnostics, still under preclinical study., (© 2023. The Author(s).)

Single-cell RNA sequencing has become an important method to identify cell types, delineate the trajectories of cell differentiation in whole organisms, and understand the heterogeneity in cellular responses. Nevertheless, sample collection and processing remain a severe bottleneck for single-cell RNA sequencing experiments. Cell isolation protocols often lead to significant changes in the transcriptomes of cells, requiring novel methods to preserve cell states. Here, we developed and benchmarked protocols using glyoxal as a fixative for single-cell RNA sequencing applications. Using Drop-seq methodology, we detected a large number of transcripts and genes from glyoxal-fixed Drosophila cells after single-cell RNA sequencing. The effective glyoxal fixation of transcriptomes in Drosophila and human cells was further supported by a high correlation of gene expression data between glyoxal-fixed and unfixed samples. Accordingly, we also found highly expressed genes overlapping to a large extent between experimental conditions. These results indicated that our fixation protocol did not induce considerable changes in gene expression and conserved the transcriptome for subsequent single-cell isolation procedures. In conclusion, we present glyoxal as a suitable fixative for Drosophila cells and potentially cells of other species that allow high-quality single-cell RNA sequencing applications., Competing Interests: Conflicts of interest The author(s) declare no conflict of interest., (© The Author(s) 2023. Published by Oxford University Press on behalf of The Genetics Society of America.)

BH3 mimetics, targeting the Bcl-2 family anti-apoptotic proteins, represent a promising therapeutic opportunity in cancers. ABT-199, the first specific Bcl-2 inhibitor, was approved by FDA for the treatment of several hematological malignancies. We have recently discovered IS21, a novel pan BH3 mimetic with preclinical antitumor activity in several tumor types. Here, we evaluated the efficacy of IS21 and other BH3 mimetics, both as single agents and combined with the currently used antineoplastic agents in T-cell acute lymphoblastic leukemia, ovarian cancer, and melanoma. IS21 was found to be active in T-cell acute lymphoblastic leukemia, melanoma, lung, pancreatic, and ovarian cancer cell lines. Bcl-xL and Mcl-1 protein levels predicted IS21 sensitivity in melanoma and ovarian cancer, respectively. Exploring IS21 mechanism of action, we found that IS21 activity depends on the presence of BAX and BAK proteins: complexes between Bcl-2 and Bcl-xL proteins and their main binding partners were reduced after IS21 treatment. In combination experiments, BH3 mimetics sensitized leukemia cells to chemotherapy, ovarian cancer cells and melanoma models to PARP and MAPK inhibitors, respectively. We showed that this enhancing effect was related to the potentiation of the apoptotic pathway, both in hematologic and solid tumors. In conclusion, our data suggest the use of inhibitors of anti-apoptotic proteins as a therapeutic strategy to enhance the efficacy of anticancer treatment., (© 2023. The Author(s).)

The Terror Management Theory (TMT) offered a great deal of generative hypotheses that have been tested in a plethora of studies. However, there is a still substantive lack of clarity about the interpretation of TMT-driven effects and their underlying neurological mechanisms. Here, we aimed to expand upon previous research by introducing two novel methodological manipulations aimed to enhance the effects of mortality salience (MS). We presented participants with the idea of the participants' romantic partner's death as well as increased the perceived threat of somatosensory stimuli. Linear mixed modelling disclosed the greater effects of MS directed at one's romantic partner on pain perception (as opposed to the participant's own mortality). The theta event-related oscillatory activity measured at the vertex of the scalp was significantly lower compared to the control condition. We suggest that MS aimed at one's romantic partner can result in increased effects on perceptual experience; however, the underlying neural activities are not reflected by a classical measure of cortical arousal.

Cross-individual pain assessment models based on electroencephalography (EEG) allow pain assessment in individuals who cannot report pain (e.g., unresponsive patients). The main obstacle to the generalisation of pain assessment models is the individual variation of brain responses to pain. Hence, we took the individual variation into account in cross-individual model development. We developed two convolutional neural networks (CNN) sharing an encoder architecture. One CNN predicts pain, while the other predicts the identity of an individual. We performed a leave-one-out (LOO) test with the exclusion of each subject and applied evidence accumulation to it for validating the pain prediction model's performance, where the binary classifier involves the states of pain (Hot) and resting state (Eyes-open). The mean accuracy produced by the LOO tests was 57.81% (max: 73.33%), and the mean accuracy of evidence accumulation achieved 69.75% (max: 100.00%). The individual recognition model achieved an accuracy of 99.63%. Nevertheless, when we acquired the most similar subject to a novel subject using the individual recognition model, where the most similar subject was used to train a subject-wise pain prediction model. The accuracy of predicting the pain-related conditions of the novel subject by the subject-wise model was only 53.73% (max: 79.50%). Therefore, the approach to utilising the features related to individual variation extracted by the CNN model needs more investigation for improving cross-individual pain assessment.Clinical relevance- This model can be applied to assess pain from EEG signals at the bedside with future improvement, which can help caretakers of unresponsive patients.

Brief painful laser and innocuous tactile stimuli have been associated with an increase of neuronal oscillations in the gamma range. Although it is indicated that event-related gamma oscillations may be highly variable across individuals, to date no study has systematically investigated interindividual variability and individual stability of induced gamma synchronization. Here, we addressed this question using two EEG datasets. The first dataset contains two repeated sessions of tactile and painful stimulation from 22 participants. The second dataset contains a single session of painful stimulation from 48 participants. In the first dataset, we observed gamma responses in the majority of the included participants. We found a broad interindividual variety of gamma magnitudes, time-frequency (TF) response patterns, and scalp topographies. Some participants showed a gamma response with individually unique time-frequency patterns, others did not exhibit any gamma response. This was reproducible and therefore stable; subjects with a large gamma magnitude in the first session showed a large gamma magnitude and a similar response pattern in the follow-up session. The second dataset confirmed the large between-subject variability, but only a fraction of the included participants exhibited laser-induced gamma synchronization. Our results indicate that current EEG measures do not reflect the complex reality of the diverse individual response patterns to brief pain and touch experiences. The present findings question whether a similar phenomenon would be observed in other neuroscience domains. Group results may be replicable, but could be driven by a subgroup of the sample. NEW & NOTEWORTHY The interpretation of gamma activity in response to noxious and innocuous somatosensory stimuli has sparked controversy. Here, we show that participants' gamma oscillations measured through electroencephalography vary. Although some participants do not show a distinct gamma response, others exhibit stable and reliable response patterns in terms of time, frequency, and magnitude.

Suicide ranks among the 10 leading causes of death worldwide; in Italy, almost 4000 persons per year die by suicide. Studies focusing on suicidality in Italian inpatient samples before the COVID-19 pandemic are scant. We, thus, aimed to define sociodemographic and clinical variables associated with suicidal ideation (SI) and deliberate self-harm (DSH) in a sample of inpatients admitted to a Psychiatric Inpatient Unit. This retrospective study was conducted in the Psychiatric Inpatient Unit of the Perugia Hospital, from January 2018 to December 2019. Sociodemographic and clinical characteristics, including diagnostic and treatment features, were collected from the medical records of subjects admitted for suicidality-related phenomena, namely DSH and SI. The prevalence of suicidality-related phenomena in the sample ( n = 850) was 14.12% (n=120) and was mainly due to DSH ( n = 84; 70%). Subjects hospitalized due to these conditions were more frequently females, separated, and displayed a higher prevalence of personality disorders, especially borderline personality disorder. People in the suicidality-related phenomena subgroup were more often committed involuntarily and reported multiple hospitalizations less frequently than other inpatients. They were receiving community treatment in a higher percentage of cases, and lithium was prescribed more frequently than among inpatients who were hospitalized for reasons other than suicidality. Our study provides a further characterization of psychiatric inpatients who experience SI or perform DSH. Targeted treatment strategies should be considered for subjects suffering from personality disorders who experience suicidality-related phenomena., (Copyright © 2023 Wolters Kluwer Health, Inc. All rights reserved.)

Past research indicates that patients' reports of pain are often met with skepticism and that observers tend to underestimate patients' pain. The mechanisms behind these biases are not yet fully understood. One relevant domain of inquiry is the interaction between the emotional valence of a stranger's expression and the onlooker's trustworthiness judgment. The emotion overgeneralization hypothesis posits that when facial cues of valence are clear, individuals displaying negative expressions (e.g., disgust) are perceived as less trustworthy than those showing positive facial expressions (e.g., happiness). Accordingly, we hypothesized that facial expressions of pain (like disgust) would be judged more untrustworthy than facial expressions of happiness. In two separate studies, we measured trustworthiness judgments of four different facial expressions (i.e., neutral, happiness, pain, and disgust), displayed by both computer-generated and real faces, via both explicit self-reported ratings (Study 1) and implicit motor trajectories in a trustworthiness categorization task (Study 2). Ratings and categorization findings partly support our hypotheses. Our results reveal for the first time that when judging strangers' facial expressions, both negative expressions were perceived as more untrustworthy than happy expressions. They also indicate that facial expressions of pain are perceived as untrustworthy as disgust expressions, at least for computer-generated faces. These findings are relevant to the clinical setting because they highlight how overgeneralization of emotional facial expressions may subtend an early perceptual bias exerted by the patient's emotional facial cues onto the clinician's cognitive appraisal process., Competing Interests: Declaration of competing interest The authors declare that there is no conflict of interest., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

Microbial biofilm is of paramount importance in the development of mucositis or peri-implantitis in patients with dental implants. This study was designed to investigate whether an electromagnetic field at high frequency waves directly applied on 33 titanium implants could remove experimentally-induced Enterococcus faecalis bacterial biofilm. A specially designed device (X-IMPLANT) was used to generate the electromagnetic field, with output power of 8 W, supply frequency (action/pause) 3/2s, and an output frequency of 625±5% kHz in plastic devices containing the biofilm-covered implants immersed in sterile saline. The bacterial biofilm on both treated and untreated control implants was quantitatively measured by phenol red-based Bio-Timer-Assay reagent. The kinetic analysis of the curves showed that the electrical treatment generated by the X-IMPLANT device completely removed the bacterial biofilm after 30 minutes of treatment (p<0.01). Elimination of the biofilm was also confirmed by chromatic observation in the macro-method. Our data seem to indicate that the procedure could be considered for clinical application in peri-implantitis to counteract bacterial biofilm on dental implants.

This work correlates the impact of robotization on employment and households' income at the regional scale with the level of investment in R&D and education policies. This kind of policy, by raising the qualitative and quantitative levels of human capital, contributes to improving the complementarity effect between humans and robots, thus mitigating the substitution effect. To this end, we compute the Adjusted Penetration of Robots (APR) (a metric used to measure the extent to which robots are being used in a particular industry or sector) at the sectoral level, combining the International Federation of Robotics database for the stock of robots, EUROSTAT Regional database, and the STructural ANalysis database on 150 NUTS-2 regions of the Euro area. We then perform a spatial stacked-panel analysis on the investment in R&D and education level. Results supports the idea that regions that invest more in R&D and have higher levels of human capital can turn the risk of robotization into an increase in both income and "quantity of work," by enhancing complementarity between robots and the labor force. On the contrary, regions investing less in R&D and having lower levels of human capital may suffer a reduction in households' disposable income., (© The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature 2023, Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.)

Aim: According to the Italian national statistical institute, severe bacterial infections (SBI) in Italy are responsible for 1.7% of mortality under 5 years of age and their recognition is often challenging, especially in the first stages of the disease. We tried to estimate the prevalence of SBI in our target population and to identify signs and symptoms that could guide in the initial evaluation of a child with a possible SBI., Methods: We designed a prospective, multicentre study and enrolled patients aged 0-14 years at the first evaluation to the emergency department with an acute illness lasting a maximum of 5 days. The presence of variables suggestive of SBI was collected for every enrolled patient. One week after the enrolment, every patient was followed up by telephone., Results: SBI is more likely to be detected with the 'gut feeling' in both univariate and multivariate models (univariate OR: 7.16, 95% CI: 4.08-12.56; multivariate OR: 5.34, 95% CI: 2.78-10.25), while abnormal breathing pattern resulted significative only in univariate model (OR 3.83, 95% CI: 1.98-7.40). Nevertheless, their associated sensitivity is low., Conclusion: SBI is uncommon in the absence of paediatricians' gut feeling and abnormal respiratory pattern., (© 2022 The Authors. Acta Paediatrica published by John Wiley & Sons Ltd on behalf of Foundation Acta Paediatrica.)

Chemotherapy regimens and radiotherapy are common strategies to fight cancer. In women, these therapies may cause side effects such as premature ovarian insufficiency (POI) and infertility. Clinical strategies to protect the ovarian reserve from the lethal effect of cancer therapies needs better understanding of the mechanisms underlying iatrogenic loss of follicle reserve. Recent reports demonstrate a critical role for p53 and CHK2 in the oocyte response to different DNA stressors, which are commonly used to treat cancer. Here we review the molecular mechanisms underlying the DNA damage stress response (DDR) and discuss crosstalk between DDR and signaling pathways implicated in primordial follicle activation.

Our current economic and political structures have an increasingly devastating impact on the Earth's climate and ecosystems: we are facing a biospheric emergency, with catastrophic consequences for both humans and the natural world on which we depend. Life scientists - including biologists, medical scientists, psychologists and public health experts - have had a crucial role in documenting the impacts of this emergency, but they have failed to drive governments to take action in order to prevent the situation from getting worse. Here we, as members of the movement Scientist Rebellion, call on life scientists to re-embrace advocacy and activism - which were once hallmarks of academia - in order to highlight the urgency and necessity of systemic change across our societies. We particularly emphasise the need for scientists to engage in nonviolent civil resistance, a form of public engagement which has proven to be highly effective in social struggles throughout history., Competing Interests: FR, EV, GR, TS, AN, LA, MM, SH, FO, CG, JH No competing interests declared, (© 2022, Racimo et al.)

Monitoring semi-natural grasslands is difficult and costly because they occur in highly dynamic and extremely complex habitat mosaics. We combined the use of a low-cost unmanned aerial vehicle (UAV) and Structure from Motion (SfM) photogrammetry to acquire high spatial resolution (∼1.5 cm pixel) RGB imagery. After image classification through Object-Based Image Analysis (OBIA), we accurately were able to distinguish three semi-natural grassland types, one of which is a habitat of conservation concern. The use of orthomosaics, digital elevation models (DEMs), and canopy height models (CHMs) yielded excellent overall classification accuracies (>89%) assessed through both remotely validated and ground-truthed points. We identified two layers of woody vegetation with a user's (UA) and producer's (PA) accuracies >73% and three grassland types: closed grassland (UA = 94%; PA = 97%), open grassland habitat (UA = 97%; PA = 93%) and open grasslands with soil erosion (UA = 96%; PA = 98%). The grassland types differed substantially in the cover of vegetation, rocks, stones, and bare soil measured in the field, as well as in the number and relative cover of the habitat diagnostic species. The proposed methodology is highly promising for mapping and monitoring semi-natural grassland of conservation concern in support of tailored management actions., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022. Published by Elsevier Ltd.)

Recent research proposed that the slowing of individual alpha frequency (IAF) could be an objective marker of pain. However, it is unclear whether this research can fully address the requirements of specificity and sensitivity of IAF to the pain experience. Here, we sought to develop a robust methodology for assessing the specificity of the relationship between alpha oscillations and acute tonic pain in healthy individuals. We recorded electroencephalography (EEG) of 36 volunteers during consecutive 5-minute sessions of painful hot water immersion, innocuous warm water immersion and aversive, non-painful auditory stimulus, matched by unpleasantness to the painful condition. Participants rated stimulus unpleasantness throughout each condition. We isolated two regions of the scalp displaying peak alpha activity across participants: centro-parietal (CP) and parieto-occipital (PO) ROI. In line with previous research our findings revealed decreased IAF during hot compared with warm stimulation, however the effect was not specific for pain as we found no difference between hot and sound in the CP ROI (compared to baseline). In contrast, the PO ROI reported the same pattern of differences, but their direction was opposite to the CP in that this ROI revealed faster frequency during hot condition than controls. Finally, we show that IAF in both ROIs did not mediate the relationship between the experimental manipulation and the affective experience. Altogether, these findings emphasize the importance of a robust methodological and analytical design to disclose the functional role of alpha oscillations during affective processing. Likewise, they suggest the absence of a causal role of IAF in the generation of acute pain experience in healthy individuals., Competing Interests: Declaration of Competing Interest The authors report no conflict of interest., (Copyright © 2022. Published by Elsevier Inc.)

The complexity of brain activity involved in the generation of the experience of pain makes it hard to identify neural markers able to predict pain states. The within and between subjects variability of pain hinders the predictive potential of machine learning models trained across participants. This challenge can be tackled by implementing deep learning classifiers based on convolutional neural networks (CNNs). We targeted phase-based connectivity in the alpha band recorded with electroencephalography (EEG) during resting states and sensory conditions (eyes open [O] and closed [C] as resting states, and warm [W] and hot [H] water as sensory conditions). Connectivity features were extracted and re-organized as square matrices, because CNNs are effective in detecting the patterns from 2D data. To assess the classifier performance we implemented two complementary approaches: we 1) trained and tested the classifier with data from all participants, and 2) using a leave-one-out approach, that is excluding one participant at a time during training while using their data as a test set. The accuracy of binary classification between pain condition (H) and eyes open resting state (O) was 94.16% with the first approach, and 61.01 % with the leave-one-out approach. Clinical relevance-Further validation of the CNN classifier may help caregivers track the rehabilitation of chronic pain patients and dynamically modify the therapy. Further refinement of the model may allow its application in critical care setting with unresponsive patients to identify pain-like states otherwise incommunicable to medical personnel.

Patient-derived organoids resemble the biology of tissues and tumors, enabling ex vivo modeling of human diseases. They have heterogeneous morphologies with unclear biological causes and relationship to treatment response. Here, we use high-throughput, image-based profiling to quantify phenotypes of over 5 million individual colorectal cancer organoids after treatment with >500 small molecules. Integration of data using multi-omics modeling identifies axes of morphological variation across organoids: Organoid size is linked to IGF1 receptor signaling, and cystic vs. solid organoid architecture is associated with LGR5 + stemness. Treatment-induced organoid morphology reflects organoid viability, drug mechanism of action, and is biologically interpretable. Inhibition of MEK leads to cystic reorganization of organoids and increases expression of LGR5, while inhibition of mTOR induces IGF1 receptor signaling. In conclusion, we identify shared axes of variation for colorectal cancer organoid morphology, their underlying biological mechanisms, and pharmacological interventions with the ability to move organoids along them., (© 2022. The Author(s).)

The concern about the offspring's health is one of the reasons for a reduced family size of women with rheumatic diseases (RD). Increased risk of autoimmune diseases (AD) and neurodevelopmental disorders (ND) has been reported in children born to patients with RD. Within a nationwide survey about reproductive issues of women with RD, we aimed at exploring the long-term outcome of their children. By surveying 398 patients who received their diagnosis of RD during childbearing age (before the age of 45), information about the offspring were obtained from 230 women who declared to have had children. A total of 148 (64.3%) patients were affected by connective tissue diseases (CTD) and 82 (35.7%) by chronic arthritis. Data on 299 children (156 males, 52.1%; mean age at the time of interview 17.1 ± 9.7 years) were collected. Twelve children (4.0%), who were born to patients with CTD in 75% of the cases, were affected by AD (8 cases of celiac disease). Eleven children had a certified diagnosis of ND (3.6%; 6 cases of learning disabilities); 9 of them were born to mothers with CTD (5 after maternal diagnosis). No association was found between ND and prenatal exposure to either maternal autoantibodies or anti-rheumatic drugs. Absolute numbers of offspring affected by AD and ND were low in a multicentre cohort of Italian women with RD. This information can be helpful for the counselling about reproductive issues, as the health outcomes of the offspring might not be an issue which discourage women with RD from having children., (© 2021. The Author(s).)

Bcl-2 family anti-apoptotic proteins are overexpressed in several hematological and solid tumors, and contribute to tumor formation, progression, and resistance to therapy. They represent a promising therapeutic avenue to explore for cancer treatment. Venetoclax, a Bcl-2 inhibitor is currently used for hematological malignancies or is undergoing clinical trials for either hematological or solid tumors. Despite these progresses, ongoing efforts are focusing on the identification and development of new molecules targeting Bcl-2 protein and/or other family members. Methods: Machine learning guided virtual screening followed by surface plasmon resonance, molecular docking and pharmacokinetic analyses were performed to identify new inhibitors of anti-apoptotic members of Bcl-2 family and their pharmacokinetic profile. The sensitivity of cancer cells from different origin to the identified compounds was evaluated both in in vitro (cell survival, apoptosis, autophagy) and in vivo (tumor growth in nude mice) preclinical models. Results: IS20 and IS21 were identified as potential new lead compounds able to bind Bcl-2, Bcl-xL and Mcl-1 recombinant proteins. Molecular docking investigation indicated IS20 and IS21 could bind into the Beclin-1 BH3 binding site of wild type Bcl-2, Bcl-xL and Mcl-1 proteins. In particular, although the IS21 docked conformation did not show a unique binding mode, it clearly showed its ability in flexibly adapting to either BH3 binding sites. Moreover, both IS20 and IS21 reduced cell viability, clonogenic ability and tumor sphere formation, and induced apoptosis in leukemic, melanoma and lung cancer cells. Autophagosome formation and maturation assays demonstrated induction of autophagic flux after treatment with IS20 or IS21. Experiments with z-VAD-fmk, a pan-caspase inhibitor, and chloroquine, a late-stage autophagy inhibitor, demonstrated the ability of the two compounds to promote apoptosis by autophagy. IS21 also reduced in vivo tumor growth of both human leukemia and melanoma models. Conclusion: Virtual screening coupled with in vitro and in vivo experimental data led to the identification of two new promising inhibitors of anti-apoptotic proteins with good efficacy in the binding to recombinant Bcl-2, Bcl-xL and Mcl-1 proteins, and against different tumor histotypes., Competing Interests: Competing Interests: The authors have declared that no competing interest exists., (© The author(s).)

Abstract: Infective uvulitis is a rare condition in children. In this report, we describe the case of a 4-year old-patient who presented a group A Streptococcus pharyngitis with uvulitis. No signs of epiglottitis were detected at nasal fibroscopy. She recovered rapidly with intravenous antibiotic therapy and 2 days of corticosteroid. Uvulitis is usually caused by group A Streptococcus or Haemophilus influentiae, but also other bacteria can be detected. Uvulitis can be isolated, or it can occur with epiglottitis and become an emergency., Competing Interests: Disclosure: The authors declare no conflict of interest., (Copyright © 2020 Wolters Kluwer Health, Inc. All rights reserved.)

Aim: B-cell lymphoma-2 (Bcl-2)-like protein-10 (Bcl2L10) is the less studied member of Bcl-2 family proteins, with the controversial role in different cancer histotypes. Very recently, Bcl2L10 expression in melanoma tumor specimens and its role in melanoma response to therapy have been demonstrated. Here, the involvement of Bcl2L10 on the in vitro and in vivo properties associated with melanoma aggressive features has been investigated., Methods: Endogenous Bcl2L10 protein expression was detected by western blotting analysis in a panel of patient-derived and commercially available human melanoma cells. In vitro assays to evaluate clonogenicity, cell proliferation, cell migration, cell invasion, and in vitro capillary-like structure formation [vasculogenic mimicry (VM)] have been performed by using human melanoma cells stably overexpressing Bcl2L10 or transiently transfected for loss/gain function of Bcl2L10, grown under two- or three-dimensional (3D) conditions Xenograft melanoma model was employed to evaluate in vivo tumor growth and angiogenesis., Results: Results demonstrated that Bcl2L10 acts as an inducer of in vitro cell migration, invasion, and VM, while in vitro cell proliferation, in vivo tumor growth, as well as colony formation properties were not affected. Dissecting different signaling pathways, it was found that Bcl2L10 positively affects the phosphorylation of extracellular-signal-regulated kinase (ERK) and the expression of markers of cell invasion, such as urokinase plasminogen activator receptor (uPAR) and matrix metalloproteinases (MMPs). Of note, Bcl2L10-dependent in vitro migration, invasion, and VM are linked to uPAR. Bcl2L10 also negatively regulates the intracellular calcium level. Finally, reduced invasion capability in 3D spheroid invasion assay of melanoma cells transiently overexpressing Bcl2L10 was observed after treatment with inhibitors of MMPs and uPAR., Conclusions: Overall, data reported in this paper provide evidence supporting a positive role of Bcl2L10 in melanoma aggressive features., Competing Interests: The authors declare that there are no conflicts of interest., (© The Author(s) 2022.)

The trimeric CCAAT-binding NF-Y is a "pioneer" Transcription Factor -TF- known to cooperate with neighboring TFs to regulate gene expression. Genome-wide analyses detected a precise stereo-alignment -10/12 bp- of CCAAT with E-box elements and corresponding colocalization of NF-Y with basic-Helix-Loop-Helix (bHLH) TFs. We dissected here NF-Y interactions with USF1 and MAX. USF1, but not MAX, cooperates in DNA binding with NF-Y. NF-Y and USF1 synergize to activate target promoters. Reconstruction of complexes by structural means shows independent DNA binding of MAX, whereas USF1 has extended contacts with NF-Y, involving the USR, a USF-specific amino acid sequence stretch required for trans-activation. The USR is an intrinsically disordered domain and adopts different conformations based on E-box-CCAAT distances. Deletion of the USR abolishes cooperative DNA binding with NF-Y. Our data indicate that the functionality of certain unstructured domains involves adapting to small variation in stereo-alignments of the multimeric TFs sites., (Copyright © 2021. Published by Elsevier B.V.)

The peripersonal space (PPS) is a special portion of space immediately surrounding the body, where the integration between tactile stimuli delivered on the body and auditory or visual events emanating from the environment occurs. Interestingly, PPS can widen if a tool is employed to interact with objects in the far space. However, electrophysiological evidence of such tool-use dependent plasticity in the human brain is scarce. Here, in a series of three experiments, participants were asked to respond to tactile stimuli, delivered to their right hand, either in isolation (unimodal condition) or combined with auditory stimulation, which could occur near (bimodal-near) or far from the stimulated hand (bimodal-far). According to multisensory integration spatial rule, when bimodal stimuli are presented at the same location, we expected a response enhancement (response time - RT - facilitation and event-related potential - ERP - super-additivity). In Experiment 1, we verified that RT facilitation was driven by bimodal input spatial congruency, independently from auditory stimulus intensity. In Experiment 2, we showed that our bimodal task was effective in eliciting the magnification of ERPs in bimodal conditions, with significantly larger responses in the near as compared to far condition. In Experiment 3 (main experiment), we explored tool-use driven PPS plasticity. Our audio-tactile task was performed either following tool-use (a 20-min reaching task, performed using a 145 cm-long rake) or after a control cognitive training (a 20-min visual discrimination task) performed in the far space. Following the control training, faster RTs and greater super-additive ERPs were found in bimodal-near as compared to bimodal-far condition (replicating Experiment 2 results). Crucially, this far-near differential response was significantly reduced after tool-use. Altogether our results indicate a selective effect of tool-use remapping in extending the boundaries of PPS. The present finding might be considered as an electrophysiological evidence of tool-use dependent plasticity in the human brain., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Background: The present study investigates the impact of the Coronavirus diseases 2019 (Covid-19) pandemic on the subjective experience of pregnant women, as well as the impact of the pandemic on this population in terms of psychopathological correlates., Subjects and Methods: Pregnant women referring to the Section of Obstetrics and Gynecology of the General Hospital of Perugia, Italy, were recruited from 1 st May, 2021 to 15 th June, 2021. Socio-demographic and clinical data was collected, as well as information regarding the Covid-19 pandemic impact on the subjective experience of pregnancy. Psychopathology was evaluated by means of the State-Trait Anxiety Inventory Form Y (STAI-Y), the Symptom Checklist-90 (SCL-90) and the Prenatal Distress Measure (Pre-DM). Descriptive analyses were performed. Significant associations between distress symptoms and the collected sociodemographic and clinical variables were assessed by using the Pearson correlation (p<0.05)., Results: 25 women were included in the study. Among these, 18 (72%) reported that the Covid-19 pandemic negatively impacted their experience of pregnancy. Were detected an average Pre-DM total score of 7.28±4.33 and an average state anxiety scale value of 35.56±9.21 and an average trait anxiety scale value of 34.04±7.44 at the STAI-Y. A global severity index > 1 at SCL-90 was detected in 8.3% of the sample., Conclusions: The identification of antepartum distress and the early treatment of perinatal psychopathology represent a priority during the Covid-19 pandemic era.

Background: Candidates for bariatric surgery undergo a multidisciplinary evaluation in the pre-operative phase, including a psychiatric visit aimed at the screening for psychiatric comorbidities, including feeding and eating disorders (FEDs), which are shortcomings to the intervention or predictors of worse prognosis. The presence of FEDs, such as Binge Eating Disorder (BED) and Bulimia Nervosa (BN), is associated with higher rates of other psychiatric disorders. Furthermore, there is evidence of the association between obesity and Depressive Disorders, as well as B and C Cluster Personality Disorders. The aim of this study was to evaluate the presence of psychiatric comorbidities among a population of candidates for bariatric surgery., Subjects and Methods: Subjects were recruited at the outpatient service of the Section of Psychiatry, Clinical Psychology and Rehabilitation of the General Hospital/University of Perugia after being referred by surgeons. Psychiatric comorbidities were investigated by means of the Structured Clinical Interview for DSM-5 Disorders. Subjects underwent specific assessment with scales for the evaluation of FEDs, namely Binge Eating Scale, Obesity Questionnaire, Bulimia Test-Revised and Body Shape Questionnaire., Results: The sample consisted of 101 subjects: 43 (42.6%) were diagnosed with at least one psychiatric disorder, including FEDs. In particular, 30 subjects (29.7%) presented at least one FED, among which the most frequent were FED not otherwise specified (24.1%) and BED (6.8%). Moreover, 26 subjects (25.7%) were diagnosed with at least one psychiatric disorder other than FEDs, such as Personality Disorders (17.1%), with a higher prevalence of B and C Cluster Disorders. Depressive Disorders were detected in 5% of the sample., Conclusions: Subjects undergoing bariatric surgery often display psychiatric comorbidities, more frequently one or more FEDs. The systematic screening of these conditions should be implemented in the clinical practice in order to provide early intervention strategies and adequate monitoring.

Background: Suicidality is a widespread phenomenon with a dramatic burden worldwide. The Coronavirus disease 2019 (Covid-19) pandemic determined a relevant impact on mental health, due to the infection itself and its socio-economic consequences. The present study is aimed at analyzing the prevalence of suicidality during the Covid-19 pandemic among subjects requiring a psychiatric consultation in an emergency setting., Subjects and Methods: Socio-demographic and clinical information was collected at the emergency department of the General Hospital of Perugia from June 1 st , 2020 to January 31 st , 2021. Data was entered into an electronic datasheet and retrospectively analysed. Pearson's bivariate correlation was performed in order to assess significant associations between suicide-related variables and specific socio-demographic and clinical features (p<0.05)., Results: Among 447 subjects included in the analysis, 109 (24.4%) showed suicidality-related phenomena, particularly suicide attempts (SA) (n=44, 9.8%), suicidal ideation (SI) (n=41, 9.2%), non suicidal self-injury (NSSI) (n=31, 6.9%), that in some cases co-occurred. A statistically significant association was detected between NSSI and living with marital family (p=0.024) and between suicidality-related phenomena and adjustment disorders (p=0.018). None of the examined subjects reported a previous positivity for Covid-19 and neither did their relatives., Conclusions: The present study confirms the impact of the Covid-19 pandemic on suicide-related phenomena. Consultation psychiatry fulfills a key role in the early detection and clinical management of these conditions, that require targeted intervention strategies.

Background: The impact of intrauterine growth restriction (IUGR) on lung function in very preterm children is largely unknown as current evidence is mainly based on studies in children born small for gestational age but not necessarily with IUGR., Methods: Spirometry, transfer factor of the lung for carbon monoxide (TLco), and lung clearance index (LCI) were cross-sectionally evaluated at 8.0-15.0 years of age in children born <32 weeks of gestation with IUGR (n = 28) and without IUGR (n = 67). Controls born at term (n = 67) were also included., Results: Very preterm children with IUGR had lower mean forced expired volume in the first second (FEV 1 ) z-score than those with normal fetal growth (∆ -0.66, 95% confidence interval (CI) -1.12, -0.19), but not significant differences in LCI (∆ +0.24, 95% CI -0.09, 0.56) and TLco z-score (∆ -0.11, 95% CI -0.44, 0.23). The frequency of bronchopulmonary dysplasia (BPD) in the two groups was, respectively, 43% and 10% (P = 0.003). IUGR was negatively associated with FEV 1 (B = -0.66; P = 0.004), but the association lost significance (P = 0.05) when adjusting for BPD., Conclusions: IUGR has an impact on conducting airways function of very preterm children at school age, with part of this effect being mediated by BPD. Ventilation inhomogeneity and diffusing capacity, instead, were not affected., Impact: IUGR does not necessarily imply a low birthweight for gestational age (and vice versa). While a low birthweight is associated with worse respiratory outcomes, the impact of IUGR on lung function in premature children is largely unknown. IUGR affects conducting airways function in school-age children born <32 weeks with IUGR, but not ventilation inhomogeneity and diffusing capacity. The impact of IUGR on FEV 1 seems mainly related to the higher risk of BPD in this group., (© 2020. The Author(s), under exclusive licence to the International Pediatric Research Foundation, Inc.)

Tissue homeostasis is achieved by balancing stem cell maintenance, cell proliferation and differentiation, as well as the purging of damaged cells. Elimination of unfit cells maintains tissue health; however, the underlying mechanisms driving competitive growth when homeostasis fails, for example, during tumorigenesis, remain largely unresolved. Here, using a Drosophila intestinal model, we find that tumor cells outcompete nearby enterocytes (ECs) by influencing cell adhesion and contractility. This process relies on activating the immune-responsive Relish/NF-κB pathway to induce EC delamination and requires a JNK-dependent transcriptional upregulation of the peptidoglycan recognition protein PGRP-LA. Consequently, in organisms with impaired PGRP-LA function, tumor growth is delayed and lifespan extended. Our study identifies a non-cell-autonomous role for a JNK/PGRP-LA/Relish signaling axis in mediating death of neighboring normal cells to facilitate tumor growth. We propose that intestinal tumors "hijack" innate immune signaling to eliminate enterocytes in order to support their own growth., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Background: The past two decades have seen a particular focus towards high-frequency neural activity in the gamma band (>30 Hz). However, gamma band activity shares frequency range with unwanted artefacts from muscular activity., New Method: We developed a novel approach to remove muscle artefacts from neurophysiological data. We re-analysed existing EEG data that were decomposed by a blind source separation method (independent component analysis, ICA), which helped to better spatially and temporally separate single muscle spikes. We then applied an adapting algorithm that detects these singled-out muscle spikes., Results: We obtained data almost free from muscle artefacts; we needed to remove significantly fewer artefact components from the ICA and we included more trials for the statistical analysis compared to standard ICA artefact removal. All pain-related cortical effects in the gamma band have been preserved, which underlines the high efficacy and precision of this algorithm., Conclusions: Our results show a significant improvement of data quality by preserving task-relevant gamma oscillations of presumed cortical origin. We were able to precisely detect, gauge, and carve out single muscle spikes from the time course of neurophysiological measures without perturbing cortical gamma. We advocate the application of the tool for studies investigating gamma activity that contain a rather low number of trials, as well as for data that are highly contaminated with muscle artefacts. This validation of our tool allows for the application on event-free continuous EEG, for which the artefact removal is more challenging., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Essential oils (EOs) have been recently emerging for their promising biological activities in preventing tumorigenesis or progression of different tumor histotypes, including melanoma. In this study, we investigated the antitumor activity of a panel of EOs in different tumor models. The ability of Melaleuca alternifolia (tea tree oil) and its main component, terpinen-4-ol, to sensitize the target therapy currently used for melanoma treatment was also assessed. Our results demonstrated that EOs differently affect the viability of human cancer cells and led us to select six EOs effective in melanoma and lung cancer cells, without toxic effects in human fibroblasts. When combined with dabrafenib and/or trametinib, Melaleuca alternifolia synergistically reduced the viability of melanoma cells by activating apoptosis. Through machine learning classification modeling, α-terpineol, tepinolene, and terpinen-4-ol, three components of Melaleuca alternifolia, were identified as the most likely relevant components responsible for the EO's antitumor effect. Among them, terpinen-4-ol was recognized as the Melaleuca alternifolia component responsible for its antitumor and proapoptotic activity. Overall, our study holds promise for further analysis of EOs as new anticancer agents and supports the rationale for their use to improve target therapy response in melanoma.

Hypoxia, a condition of oxygen deprivation, is considered a hallmark of tumor microenvironment regulating several pathways and promoting cancer progression and resistance to therapy. Semaphorins, a family of about 20 secreted, transmembrane and GPI-linked glycoproteins, and their cognate receptors (plexins and neuropilins) play a pivotal role in the crosstalk between cancer and stromal cells present in the tumor microenvironment. Many studies reported that some semaphorins are involved in the development of a permissive tumor niche, guiding cell-cell communication and, consequently, the development and progression, as well as the response to therapy, of different cancer histotypes, including melanoma.In this review we will summarize the state of art of semaphorins regulation by hypoxic condition in cancer with different origin. We will also describe evidence about the ability of semaphorins to affect the expression and activity of transcription factors activated by hypoxia, such as hypoxia-inducible factor-1. Finally, we will focus our attention on findings reporting the role of semaphorins in melanocytes transformation, melanoma progression and response to therapy. Further studies are necessary to understand the mechanisms through which semaphorins induce their effect and to shed light on the possibility to use semaphorins or their cognate receptors as prognostic markers and/or therapeutic targets in melanoma or other malignancies.

Background: Colorectal cancer (CRC) remains a serious health concern worldwide. Despite advances in diagnosis and treatment, about 15 to 30% of stage II CRC patients subjected to tumor resection with curative intent, develop disease relapse. Moreover, the therapeutic strategy adopted after surgery is not consensual for these patients. This supports the imperative need to find new prognostic and predictive biomarkers for stage II CRC., Methods: For this purpose, we used a one-hospital series of 227 stage II CRC patient samples to assess the biomarker potential of the immunohistochemical expression of MUC2 mucin and CDX2 and SOX2 transcription factors. The Kaplan-Meier method was used to generate disease-free survival curves that were compared using the log-rank test, in order to determine prognosis of cases with different expression of these proteins, different mismatch repair (MMR) status and administration or not of adjuvant chemotherapy., Results: In this stage II CRC series, none of the studied biomarkers showed prognostic value for patient outcome. However low expression of MUC2, in cases with high expression of CDX2, absence of SOX2 or MMR-proficiency, conferred a significantly worst prognosis. Moreover, cases with low expression of MUC2 showed a significantly clear benefit from treatment with adjuvant chemotherapy., Conclusion: In conclusion, we observe that patients with stage II CRC with low expression of MUC2 in the tumor respond better when treated with adjuvant chemotherapy. This observation supports that MUC2 is involved in resistance to fluorouracil-based adjuvant chemotherapy and might be a promising future predictive biomarker in stage II CRC patients.

Objective: This study aimed to evaluate the efficacy of laser-activated irrigation using photon-induced photoacoustic streaming (PIPS ® ) and photoactivated disinfection (PAD) techniques and their combination to improve penetration and activation of toluidine blue in the endodontic space of teeth experimentally infected with Enterococcus faecalis . Materials and methods: Twenty-seven extracted single-root teeth were instrumented, sterilized, and infected with E. faecalis and divided into seven groups of three teeth each: Group A [sodium hypochlorite (NaClO) 5% hand irrigation], Group B [NaClO 5% hand irrigation+ethylenediaminetetraacetic acid (EDTA)+NaClO 5% activated by PIPS], Group C (EDTA+NaClO 5% activated by PIPS), Group D (toluidine blue activated by PAD), Group E (toluidine blue activated by PIPS and PAD), Group F (NaClO 5% hand irrigation+toluidine blue activated by PAD), and Group G (NaClO 5% hand irrigation+toluidine blue activated by PIPS and PAD). Finally, positive and negative group controls were prepared. The presence of biofilms after the treatments was assessed by the BioTimer assay. PIPS was performed with an Er:YAG laser (2940 nm, LightWalker, Fotona ® d.o.o., Slovenia) at 20 mJ, 15 Hz, 0.3 W, and 50-μs pulse duration. PAD was performed with a 635 nm diode laser (Smart M, Lasotronix ® , Poland) at 400 mW in continuous wave (CW). Results: When NaClO was used, significant decontamination ( p  ≤ 0.05) was obtained in all experimental groups with respect to the positive control, other than Group G. Irrigation with EDTA+NaClO activated by PIPS produced a higher level of decontamination than Group A ( p  ≤ 0.05). Significant results in reducing biofilm load compared with the control and Group A were observed when NaClO was coupled with toluidine blue activated by PAD ( p  ≤ 0.05). Conclusions: Disinfection of root canals can be obtained using a combination of different irrigants, photosensitizers, and activation protocols. EDTA+NaClO using the PIPS protocol and toluidine blue activated by PAD (both preceded by NaClO irrigation) can be considered effective tools. The possibility of replacing NaClO with toluidine blue, whatever the method of activation, should be further investigated.

Current theories of object perception emphasize the automatic nature of perceptual inference. Repetition suppression (RS), the successive decrease of brain responses to repeated stimuli, is thought to reflect the optimization of perceptual inference through neural plasticity. While functional imaging studies revealed brain regions that show suppressed responses to the repeated presentation of an object, little is known about the intra-trial time course of repetition effects to everyday objects. Here, we used event-related potentials (ERPs) to task-irrelevant line-drawn objects, while participants engaged in a distractor task. We quantified changes in ERPs over repetitions using three general linear models that modeled RS by an exponential, linear, or categorical "change detection" function in each subject. Our aim was to select the model with highest evidence and determine the within-trial time-course and scalp distribution of repetition effects using that model. Model comparison revealed the superiority of the exponential model indicating that repetition effects are observable for trials beyond the first repetition. Model parameter estimates revealed a sequence of RS effects in three time windows (86-140, 322-360, and 400-446 ms) and with occipital, temporoparietal, and frontotemporal distribution, respectively. An interval of repetition enhancement (RE) was also observed (320-340 ms) over occipitotemporal sensors. Our results show that automatic processing of task-irrelevant objects involves multiple intervals of RS with distinct scalp topographies. These sequential intervals of RS and RE might reflect the short-term plasticity required for optimization of perceptual inference and the associated changes in prediction errors and predictions, respectively, over stimulus repetitions during automatic object processing., (© 2018 The Authors. European Journal of Neuroscience published by Federation of European Neuroscience Societies and John Wiley & Sons Ltd.)

Background: Therapeutic approaches to fibromyalgia (FM) are shifting towards a combined multi-treatment approach to tackle the variety of symptoms experienced in FM. Importantly, little is known about FM patients' attitude towards the available treatments., Methods: A cross-sectional web survey obtained responses from 464 individuals who satisfied diagnostic criteria for FM. Respondents were asked to report which treatments they adopted in their past, present and intend to adopt in the future. They also rated their level of well-being, depression, anxiety and pain catastrophizing., Results: Data revealed a predominantly combined multi-treatment approach in a sample characterized by middle-aged, Caucasian women. Respondents reported pervasive use of pharmacological therapy but had also adopted non-pharmacological treatment in the past. Future intentions clustered on alternative treatment or no treatment. Regression analyses revealed that pharmacological treatment in the past was predictive of both pharmacological and non-pharmacological treatments in the present. However, use of non-pharmacological treatment in the past was uniquely predictive of its reuse in the present and future. This pattern was also accounted for by individual differences in pain magnification and well-being in the past., Conclusions: Treatment preferences of FM individuals reveal an ambivalent combination of heavy reliance on pharmacological treatment with lower but consistent reliance on non-pharmacological treatment for those individuals who used it in the past and present. This finding may inform longitudinal research into the relationship between pharmacological and non-pharmacological treatment preference in FM patients, which could in turn inform guidelines for individualized therapeutic plans for clinicians., Significance: Individuals with fibromyalgia reported the use of non-pharmacological and pharmacological treatments in the past but a predominant use of a pharmacological approach overall. Patterns of treatment experienced in the past were differentially related to future preferences. Pharmacological treatment in the past was likely to lead to both pharmacological and non-pharmacological choices in the present. However, non-pharmacological treatment in the past was more likely to be chosen again in the present and future, but unlikely to lead to a pharmacological choice., (© 2020 European Pain Federation - EFIC®.)

Introduction: Colorectal cancer arises in a multistep process of carcinogenesis from normal mucosa. The earliest precursor might be a morphologically inconspicuous precancerous field, harboring cancer-associated mutations., Methods: We systematically analyzed genetic alterations in 77 tissue samples from 30 patients with sporadic colorectal neoplasms (18 large adenomas and 12 adenocarcinomas) and matched adjacent normal mucosa (N = 30), as well as normal rectal tissue (N = 17). We profiled mutations associated with colorectal cancer by targeted sequencing of 46 genetic loci using 157 custom amplicons and a median depth of 42,655 reads per loci., Results: Multiple mutations were found in colorectal neoplasms, most frequently in APC, KRAS, and TP53. In a subgroup of 11 of 30 patients, alterations were also detected in non-neoplastic mucosa. These mutations were divergent from those in matched neoplasms. The total alteration count and the allele frequency of mutations were higher in neoplasms compared with those in adjacent tissues. We found that younger patients (≤70 years) are less likely affected by mutations in non-neoplastic mucosa than older patients (>70 years, P = 0.013), although no association was found for other variables, including type, location and differentiation of neoplasia, and previous history of polyps., Discussion: Our data show that cancer-associated mutations can be found in non-neoplastic tissues in a subgroup of patients with colorectal neoplasms. Further studies are needed to specify the risk of occurrence and recurrence of neoplasia in this patient population.

Purpose: Our goal was to evaluate the usefulness of apparent diffusion coefficient (ADC) ratios in discriminating true from false positives in multiparametric (mp) prostate MRI in clinical practice., Methods: We retrospectively evaluated 98 prostate lesions in a series of 73 patients who had undergone prostate mpMRI and standard 12-core prostatic biopsy in our institution from 2016 to 2018. Two experienced radiologists performed double blind ADC value quantifications of both MRI-identified lesions and apparently benign contralateral prostatic parenchyma in a circular region of interest (ROI) of ∼10 mm 2 . The ratios between the mean values of both measurements (i.e., ADC ratio mean) and between the minimum value of the lesion and the maximum value of the benign parenchyma (i.e., ADC ratio min-max) were automatically calculated. The malignancy of all lesions was determined through biopsy according to Gleason score (GS ≥ 6) and localization., Results: For Reader 1, the area under the ROC curve (AUC) of ADC ratio mean and ADC ratio min-max were 0.72 and 0.67, respectively, whereas for Reader 2 these values were 0.74 and 0.71, respectively. The best cut-off values for ADC ratio means were ≥ 0.5 (Reader 1) and ≥ 0.6 (Reader 2), with a sensitivity of 76.3 % and 84.2 % and a specificity of 51.7 % and 50 %, respectively. Moreover, based on a threshold of 0.6, no clinically significant prostate cancer (csPCa) was missed by Reader 1, while only one went unnoticed by Reader 2., Conclusion: The ADC ratio is a useful and moderately accurate complementary tool to diagnose prostate cancer in the mp-MRI., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Background: Understanding the signalling pathways involved in angiogenesis, and developing anti-angiogenic drugs are one of the major focuses on cancer research. Herein, we assessed the effect of CPTH6, a lysine acetyltransferase inhibitor and anti-tumoral compound, on angiogenesis-related properties of both endothelial and cancer cells., Methods: The in vitro effect of CPTH6 on protein acetylation and anti-angiogenic properties on endothelial and lung cancer cells was evaluated via wound healing, trans-well invasion and migration, tube formation, immunoblotting and immunofluorescence. Matrigel plug assay, zebrafish embryo and mouse xenograft models were used to evaluate in vivo anti-angiogenic effect of CPTH6., Results: CPTH6 impaired in vitro endothelial angiogenesis-related functions, and decreased the in vivo vascularization both in mice xenografts and zebrafish embryos. Mechanistically, CPTH6 reduced α-tubulin acetylation and induced accumulation of acetylated microtubules in the perinuclear region of endothelial cells. Interestingly, CPTH6 also affected the angiogenesis-related properties of lung cancer cells, and conditioned media derived from CPTH6-treated lung cancer cells impaired endothelial cells morphogenesis. CPTH6 also modulated the VEGF/VEGFR2 pathway, and reshaped cytoskeletal organization of lung cancer cells. Finally, anti-migratory effect of CPTH6, dependent on α-tubulin acetylation, was also demonstrated by genetic approaches in lung cancer cells., Conclusion: Overall, this study indicates that α-tubulin acetylation could play a role in the anti-angiogenic effect of CPTH6 and, more in general, it adds information to the role of histone acetyltransferases in tumor angiogenesis, and proposes the inhibition of these enzymes as an antiangiogenic therapy of cancer.

Introduction: Survivors of extreme prematurity may have disrupted lung development. We hypothesized that the multiple breath washout (MBW) index Scond, which is intended to reflect ventilation inhomogeneity from the conducting airways, could be a sensitive marker of respiratory impairment in this group., Methods: Spirometry, TLco, and MBW were cross-sectionally evaluated at 8 to 14 years of age in children born at <28 weeks between 2004 and 2010 in Udine, Italy. Age-matched controls born at term were also included. Bronchopulmonary dysplasia (BPD) was defined as oxygen-dependence at 36 weeks postmenstrual age. The limits of normal were the 5th percentile of the reference population (Global Lung Initiative) for spirometry and TLco and the 95th percentile of controls for Lung Clearance Index, Scond, and Sacin from MBW., Results: Results were obtained in 47 extremely preterm children (53% boys, mean ± standard deviation age 11.3 ± 2.0 years, 40% with BPD) and 60 controls (50% boys, 11.6 ± 1.9 years). There were significant differences between preterm children and controls in all lung function outcomes, except for Sacin. Among children born <28 weeks, Scond tended to be frequently abnormal than FEV 1 z-score (29% vs 14%, P = .06). At multivariable linear regression, in the preterm group, current asthma was significantly associated with a higher Scond (B = 0.019, 95% confidence interval, 0.000-0.038), whereas BPD was not., Conclusion: Almost a third of extremely preterm children at school age showed Scond alterations that affected also children without BPD. Longitudinal studies should clarify the prognostic meaning of Scond abnormalities in this group., (© 2020 Wiley Periodicals, Inc.)

Adherence to antihypertensive medication is an important challenge that doctors often face in the treatment of hypertension. Good adherence is crucial to prevent cardiovascular complications. In consequence, the present study aimed at determining the prevalence of adherence to antihypertensive treatment and identifying associated clinical variables. A multicenter cross-sectional study was conducted in 12 cities of Argentina. A systematic sampling was performed in order to select patients with hypertension and under pharmacological treatment for at least 6 months. Physicians took three BP measurements, and the level of adherence was assessed using the self-administered Morisky questionnaire (MMAS-8). Participants were classified into three levels of adherence: high adherence-MMAS score of 8; medium adherence-MMAS scores of 6 to <8; and low adherence-MMAS scores of <6. A total of 1111 individuals (62 ± 12 years old, women 49.4%) were included in the present analysis; 159 (14.3%), 329 (29.6%) and 623 (56.1%) patients had low, medium, and high adherence, respectively. The prevalence of controlled hypertension increased only in high adherent patients: 42.8%, 42.2%, and 64.5% for low, medium, and high adherence groups, respectively. Similarly, systolic BP was lower only in the high adherence group. High educational level (OR 3.47, 95% CI 2.68-4.49) and diuretic treatment (OR 0.64, 95% CI 0.47-0.88) were independent predictors of high adherence. In conclusion, more than a half of treated hypertensive patients had a high level of adherence. These patients had lower BP values and higher control levels. A high educational level predicts high adherence., (© 2020 Wiley Periodicals, Inc.)

Context: Soil erosion is one of the main threats driving soil degradation across the globe with important impacts on crop yields, soil biota, biogeochemical cycles, and ultimately human nutrition., Objectives: Here, using an empirical model, we present a global and temporally explicit assessment of soil erosion risk according to recent (2001-2013) dynamics of rainfall and vegetation cover change to identify vulnerable areas for soils and soil biodiversity., Methods: We used an adaptation of the Universal Soil Loss Equation together with state of the art remote sensing models to create a spatially and temporally explicit global model of soil erosion and soil protection. Finally, we overlaid global maps of soil biodiversity to assess the potential vulnerability of these soil communities to soil erosion., Results: We show a consistent decline in soil erosion protection over time across terrestrial biomes, which resulted in a global increase of 11.7% in soil erosion rates. Notably, soil erosion risk systematically increased between 2006 and 2013 in relation to the baseline year (2001). Although vegetation cover is central to soil protection, this increase was mostly driven by changes in rainfall erosivity. Globally, soil erosion is expected not only to have an impact on the vulnerability of soil conditions but also on soil biodiversity with 6.4% (for soil macrofauna) and 7.6% (for soil fungi) of these vulnerable areas coinciding with regions with high soil biodiversity., Conclusions: Our results indicate that an increasing proportion of soils are degraded globally, affecting not only livelihoods but also potentially degrading local and regional landscapes. Similarly, many degraded regions coincide with and may have impacted high levels of soil biodiversity.

Background: A bidirectional crosstalk between tumor cells and the surrounding microenvironment contributes to tumor progression and response to therapy. Our previous studies have demonstrated that bcl-2 affects melanoma progression and regulates the tumor microenvironment. The aim of this study was to evaluate whether bcl-2 expression in melanoma cells could influence tumor-promoting functions of tumor-associated macrophages, a major constituent of the tumor microenvironment that affects anticancer immunity favoring tumor progression., Methods: THP-1 monocytic cells, monocyte-derived macrophages and melanoma cells expressing different levels of bcl-2 protein were used. ELISA, qRT-PCR and Western blot analyses were used to evaluate macrophage polarization markers and protein expression levels. Chromatin immunoprecipitation assay was performed to evaluate transcription factor recruitment at specific promoters. Boyden chamber was used for migration experiments. Cytofluorimetric and immunohistochemical analyses were carried out to evaluate infiltrating macrophages and T cells in melanoma specimens from patients or mice., Results: Higher production of tumor-promoting and chemotactic factors, and M2-polarized activation was observed when macrophages were exposed to culture media from melanoma cells overexpressing bcl-2, while bcl-2 silencing in melanoma cells inhibited the M2 macrophage polarization. In agreement, the number of melanoma-infiltrating macrophages in vivo was increased, in parallel with a greater expression of bcl-2 in tumor cells. Tumor-derived interleukin-1β has been identified as the effector cytokine of bcl-2-dependent macrophage reprogramming, according to reduced tumor growth, decreased number of M2-polarized tumor-associated macrophages and increased number of infiltrating CD4 + IFNγ + and CD8 + IFNγ + effector T lymphocytes, which we observed in response to in vivo treatment with the IL-1 receptor antagonist kineret. Finally, in tumor specimens from patients with melanoma, high bcl-2 expression correlated with increased infiltration of M2-polarized CD163 + macrophages, hence supporting the clinical relevance of the crosstalk between tumor cells and microenvironment., Conclusions: Taken together, our results show that melanoma-specific bcl-2 controls an IL-1β-driven axis of macrophage diversion that establishes tumor microenvironmental conditions favoring melanoma development. Interfering with this pathway might provide novel therapeutic strategies., Competing Interests: Competing interests: None declared., (© Author(s) (or their employer(s)) 2020. Re-use permitted under CC BY. Published by BMJ.)