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Gastro-entero-pancreatic neuroendocrine tumors (GEP-NETs) are rare diseases encompassing pancreatic (PanNETs) and ileal NETs (SINETs), characterized by heterogeneous somatostatin receptors (SSTRs) expression. Treatments for inoperable GEP-NETs are limited, and SSTR-targeted Peptide Receptor Radionuclide Therapy (PRRT) achieves variable responses. Prognostic biomarkers for the management of GEP-NET patients are required. 18F-FDG uptake is a prognostic indicator of aggressiveness in GEP-NETs. This study aims to identify circulating and measurable prognostic miRNAs associated with 18F-FDG-PET/CT status, higher risk and lower response to PRRT.MethodsWhole miRNOme NGS profiling was conducted on plasma samples obtained from well-differentiated advanced, metastatic, inoperable G1, G2 and G3 GEP-NET patients enrolled in the non-randomized LUX (NCT02736500) and LUNET (NCT02489604) clinical trials prior to PRRT (screening set, n= 24). Differential expression analysis was performed between 18F-FDG positive (n=12) and negative (n=12) patients. Validation was conducted by Real Time quantitative PCR in two distinct well-differentiated GEP-NET validation cohorts, considering the primary site of origin (PanNETs n=38 and SINETs n=30). The Cox regression was applied to assess independent clinical parameters and imaging for progression-free survival (PFS) in PanNETs. In situ RNA hybridization combined with immunohistochemistry was performed to simultaneously detect miR and protein expression in the same tissue specimens. This novel semi-automated miR-protein protocol was applied in PanNET FFPE specimens (n=9). In vitro functional experiments were performed in PanNET models.ResultsWhile no miRNAs emerged to be deregulated in SINETs, hsa-miR-5096, hsa-let-7i-3p and hsa-miR-4311 were found to correlate with 18F-FDG-PET/CT in PanNETs (p-value:18F-FDG-PET/CT positive PanNETs with worse prognosis after PRRT (p-value:68Gallium-DOTATOC captation values (p-value:SSTR2 when ectopically expressed in PanNET cells (p-value:Conclusionshsa-miR-5096 well performs as a biomarker for 18F-FDG-PET/CT and as independent predictor of PFS. Moreover, exosome-mediated delivery of hsa-miR-5096 may promote SSTR2 heterogeneity and thus resistance to PRRT.

Planet formation imprints signatures on the physical structures of disks. In this paper, we present high-resolution ($\sim$50 mas, 8 au) Atacama Large Millimeter/submillimeter Array (ALMA) observations of 1.3 mm dust continuum and CO line emission toward the disk around the M3.5 star 2MASS J04124068+2438157. The dust disk consists only of two narrow rings at radial distances of 0.47 and 0.78 arcsec ($\sim$70 and 116 au), with Gaussian $\sigma$ widths of 5.6 and 8.5 au, respectively. The width of the outer ring is smaller than the estimated pressure scale height by $\sim25\%$, suggesting dust trapping in a radial pressure bump. The dust disk size, set by the location of the outermost ring, is significantly larger (by $3\sigma$) than other disks with similar millimeter luminosity, which can be explained by an early formation of local pressure bump to stop radial drift of millimeter dust grains. After considering the disk's physical structure and accretion properties, we prefer planet--disk interaction over dead zone or photoevaporation models to explain the observed dust disk morphology. We carry out high-contrast imaging at $L'$ band using Keck/NIRC2 to search for potential young planets, but do not identify any source above $5\sigma$. Within the dust gap between the two rings, we reach a contrast level of $\sim$7 mag, constraining the possible planet below $\sim$2--4 $M_{\rm Jup}$. Analyses of the gap/ring properties suggest a $\sim$Saturn mass planet at $\sim$90 au is likely responsible for the formation of the outer ring, which can be potentially revealed with JWST., Comment: 15 pages, 5 figures. Accepted for publication in ApJ

IntroductionExtracellular vesicles (EVs) and particles (EPs) represent reliable biomarkers for disease detection. Their role in the inflammatory microenvironment of severe COVID-19 patients is not well determined. Here, we characterized the immunophenotype, the lipidomic cargo and the functional activity of circulating EPs from severe COVID-19 patients (Co-19-EPs) and healthy controls (HC-EPs) correlating the data with the clinical parameters including the partial pressure of oxygen to fraction of inspired oxygen ratio (PaO2/FiO2) and the sequential organ failure assessment (SOFA) score.MethodsPeripheral blood (PB) was collected from COVID-19 patients (n=10) and HC (n=10). EPs were purified from platelet-poor plasma by size exclusion chromatography (SEC) and ultrafiltration. Plasma cytokines and EPs were characterized by multiplex bead-based assay. Quantitative lipidomic profiling of EPs was performed by liquid chromatography/mass spectrometry combined with quadrupole time-of-flight (LC/MS Q-TOF). Innate lymphoid cells (ILC) were characterized by flow cytometry after co-cultures with HC-EPs or Co-19-EPs.ResultsWe observed that EPs from severe COVID-19 patients: 1) display an altered surface signature as assessed by multiplex protein analysis; 2) are characterized by distinct lipidomic profiling; 3) show correlations between lipidomic profiling and disease aggressiveness scores; 4) fail to dampen type 2 innate lymphoid cells (ILC2) cytokine secretion. As a consequence, ILC2 from severe COVID-19 patients show a more activated phenotype due to the presence of Co-19-EPs.DiscussionIn summary, these data highlight that abnormal circulating EPs promote ILC2-driven inflammatory signals in severe COVID-19 patients and support further exploration to unravel the role of EPs (and EVs) in COVID-19 pathogenesis.

The majority of patients with Follicular Lymphoma (FL) experience subsequent phases of remission and relapse, making the disease "virtually" incurable. To predict the outcome of FL patients at diagnosis, various clinical-based prognostic scores have been proposed; nonetheless, they continue to fail for a subset of patients. Gene expression profiling has highlighted the pivotal role of the tumor microenvironment (TME) in the FL prognosis; nevertheless, there is still a need to standardize the assessment of immune-infiltrating cells for the prognostic classification of patients with early or late progressing disease. We studied a retrospective cohort of 49 FL lymph node biopsies at the time of the initial diagnosis using pathologist-guided analysis on whole slide images, and we characterized the immune repertoire for both quantity and distribution (intrafollicular, IF and extrafollicular, EF) of cell subsets in relation to clinical outcome. We looked for the natural killer (CD56), T lymphocyte (CD8, CD4, PD1) and macrophage (CD68, CD163, MA4A4A)-associated markers. High CD163/CD8 EF ratios and high CD56/MS4A4A EF ratios, according to Kaplan-Meier estimates were linked with shorter EFS (event-free survival), with the former being the only one associated with POD24. In contrast to IF CD68+ cells, which represent a more homogeneous population, higher in non-progressing patients, EF CD68+ macrophages did not stratify according to survival. We also identify distinctive MS4A4A+CD163-macrophage populations with different prognostic weights. Enlarging the macrophage characterization and combining it with a lymphoid marker in the rituximab era, in our opinion, may enable prognostic stratification for low-/high-grade FL patients beyond POD24. These findings warrant validation across larger FL cohorts.

Graft versus host disease (GVHD) is a major complication of allogeneic hematopoietic stem cell transplantation (HSCT). Rabbit anti-T lymphocyte globulin (ATLG) in addition to calcineurin inhibitors and antimetabolites is a suitable strategy to prevent GVHD in several transplant settings. Randomized studies already demonstrated its efficacy in terms of GVHD prevention, although the effect on relapse remains the major concern for a wider use. Tailoring of ATLG dose on host characteristics is expected to minimize its side effects (immunological reconstitution, relapse, and infections). Here, day -6 to day +15 pharmacokinetics of active ATLG serum level was first assayed in an explorative cohort of 23 patients by testing the ability of the polyclonal serum to bind antigens on human leukocytes. Significantly lower levels of serum active ATLG were found in the patients who developed GVHD (ATLG_AUCCD45: 241.52 ± 152.16 vs. 766.63 +/- 283.52 (μg*day)/ml, p = 1.46e-5). Consistent results were obtained when the ATLG binding capacity was assessed on CD3+ and CD3+/CD4+ T lymphocytes (ATLG_AUCCD3: 335.83 ± 208.15 vs. 903.54 ± 378.78 (μg*day)/ml, p = 1.92e-4; ATLG_AUCCD4: 317.75 ± 170.70 vs. 910.54 ± 353.35 (μg*day)/ml, p = 3.78e-5. Concomitantly, at pre-infusion time points, increased concentrations of CD69+ extracellular vesicles (EVs) were found in patients who developed GVHD (mean fold 9.01 ± 1.33; p = 2.12e-5). Consistent results were obtained in a validation cohort of 12 additional ATLG-treated HSCT patients. Serum CD69+ EVs were mainly represented in the nano (i.e. 100 nm in diameter) EV compartment and expressed the leukocyte marker CD45, the EV markers CD9 and CD63, and CD103, a marker of tissue-resident memory T cells. The latter are expected to set up a host pro-inflammatory cell compartment that can survive in the recipient for years after conditioning regimen and contribute to GVHD pathogenesis. In summary, high levels of CD69+ EVs are significantly correlated with an increased risk of GVHD, and they may be proposed as a tool to tailor ATLG dose for personalized GVHD prevention.

Introduction: Three-dimensional (3D) multicellular spheroids are fundamental in vitro tools for studying in vivo tissues. Volume is the main feature used for evaluating the drug/treatment effects, but several other features can be estimated even from a simple 2D image. For high-content screening analysis, the bottleneck is the segmentation stage, which is essential for detecting the spheroids in the images and then proceeding to the feature extraction stage for performing morphotypic analysis. Problem: Today, several tools are available for extracting morphological features from spheroid images, but all of them have pros and cons and there is no general validated solution. Thanks to new deep learning models, it is possible to standardize the process and adapt the analysis to big data. Novelty: Starting from the first version of AnaSP, an open-source software suitable for estimating several morphological features of 3D spheroids, we implemented a new module for automatically segmenting 2D brightfield images of spheroids by exploiting convolutional neural networks. Results: Several deep learning segmentation models (i.e., VVG16, VGG19, ResNet18, ResNet50) have been trained and compared. All of them obtained very interesting results and ResNet18 ranked as the best-performing. Conclusions: A network based on an 18-layer deep residual architecture (ResNet-18) has been integrated into AnaSP, releasing AnaSP 2.0, a version of the tool optimized for high-content screening analysis. The source code, standalone versions, user manual, sample images, video tutorial, and further documentation are freely available at: https://sourceforge.net/p/anasp .

The imaging gallery demonstrates that the number of EZH2 immunolabelled nuclei parallels those decorated by Ki-67 in both FS-DFSP (left) and DFSP (right) cases.

Top 50 upregulated and downregulated genes in FS-DFSP vs DFSP and pathway analyses.

Principal Component Analysis of DFSP and FS-DFSP gene expression profiles.

Clinical/pathologic characteristics of the patients from whom the tissue samples molecularly profiled were derived; bold/underlined type indicates the tumor tissue from which the sample for the molecular analysis was derived.

Additional fusion genes identified in FS-DFSP.

Supplemental methodological information

Expression analysis (qRT-PCR) of H19, HOTAIR and EZH2 in DFSP and FS-DFSP samples. Statistical p values (unpaired Student's t test) are indicated.

Purpose: To report on imatinib mesylate (IM) in patients with metastatic dermatofibrosarcoma protuberans (DFSP)/fibrosarcomatous (FS)-DFSP and on the impact of the treatment on tumor biology.Experimental design: Ten consecutive patients treated with IM from 2007 to 2015 for a metastatic relapse from DFSP/FS-DFSP were identified. FISH analysis for COL1A1-PDGFB was performed. Two IM-treated and 4 naïve FS-DFSP were transcriptionally profiled by RNAseq on HiScanSQ platform. Differential gene expression was analyzed with edgeR (Bioconductor), followed by hierarchical clustering and Principal Component Analysis.Results: All cases featured fibrosarcomatous in the metastasis and retained the COL1A1-PDGFB. Best RECIST response was: 8 partial response, 1 stable disease, and 1 progressive disease. Median progression-free survival was 11 months. Five patients received surgery after IM and all relapsed. IM was restored in 4 patients with a new response. After IM, the most upregulated genes included those encoding for immunoglobulins and those affecting functions and differentiation of endothelial cells. Pathway enrichment analysis revealed upregulation in genes involved in antigen processing and presentation, natural killer–mediated cytotoxicity, and drug and xenobiotics metabolism. Conversely, a significant down-regulation of kinase signaling pathways was detected.Conclusions: All metastatic cases were fibrosarcomatous. Most patients responded to IM, but PFS was shorter than reported in published series which included both DFSP and FS-DFSP. All patients operated after IM had a relapse, suggesting that IM cannot eradicate metastatic cases and that the role of surgery is limited. Transcriptional profile of naïve and posttreatment samples pointed the contribution of immune infiltrates in sustaining the response to IM. Clin Cancer Res; 22(4); 837–46. ©2015 AACR.

Supplementary Table 1. Supervised analysis highlighted the differential expression of 250 genes, with a p value

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Supplementary Figure 1 from Synergistic Proapoptotic Activity of Recombinant TRAIL Plus the Akt Inhibitor Perifosine in Acute Myelogenous Leukemia Cells

Supplementary Figure 1 from Activity of the Novel Dual Phosphatidylinositol 3-Kinase/Mammalian Target of Rapamycin Inhibitor NVP-BEZ235 against T-Cell Acute Lymphoblastic Leukemia

Supplementary Figure 2 from Activity of the Novel Dual Phosphatidylinositol 3-Kinase/Mammalian Target of Rapamycin Inhibitor NVP-BEZ235 against T-Cell Acute Lymphoblastic Leukemia

Recent investigations have documented that constitutively activated phosphatidylinositol 3-kinase (PI3K)/Akt/mammalian target of rapamycin (mTOR) signaling is a common feature of T-cell acute lymphoblastic leukemia (T-ALL), where it strongly influences growth and survival. These findings lend compelling weight for the application of PI3K/Akt/mTOR inhibitors in T-ALL. However, our knowledge of PI3K/Akt/mTOR signaling in T-ALL is limited and it is not clear whether it could be an effective target for innovative therapeutic strategies. Here, we have analyzed the therapeutic potential of the dual PI3K/mTOR inhibitor PI-103, a small synthetic molecule of the pyridofuropyrimidine class, on both T-ALL cell lines and patient samples, which displayed constitutive activation of PI3K/Akt/mTOR signaling. PI-103 inhibited the growth of T-ALL cells, including 170-kDa P-glycoprotein overexpressing cells. PI-103 cytotoxicity was independent of p53 gene status. PI-103 was more potent than inhibitors that are selective only for PI3K (Wortmannin, LY294002) or for mTOR (rapamycin). PI-103 induced G0-G1 phase cell cycle arrest and apoptosis, which was characterized by activation of caspase-3 and caspase-9. PI-103 caused Akt dephosphorylation, accompanied by dephosphorylation of the Akt downstream target, glycogen synthase kinase-3β. Also, mTOR downstream targets were dephosphorylated in response to PI-103, including p70S6 kinase, ribosomal S6 protein, and 4E-BP1. PI-103 strongly synergized with vincristine. These findings indicate that multitargeted therapy toward PI3K and mTOR alone or with existing drugs may serve as an efficient treatment toward T-ALL cells, which require up-regulation of PI3K/Akt/mTOR signaling for their survival and growth. [Cancer Res 2009;69(8):3520–28]

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Supplementary Figure 1 from Dual Inhibition of Class IA Phosphatidylinositol 3-Kinase and Mammalian Target of Rapamycin as a New Therapeutic Option for T-Cell Acute Lymphoblastic Leukemia

Supplementary Figure 2 from Dual Inhibition of Class IA Phosphatidylinositol 3-Kinase and Mammalian Target of Rapamycin as a New Therapeutic Option for T-Cell Acute Lymphoblastic Leukemia

Supplementary Figure 3 from Dual Inhibition of Class IA Phosphatidylinositol 3-Kinase and Mammalian Target of Rapamycin as a New Therapeutic Option for T-Cell Acute Lymphoblastic Leukemia

Supplementary Figure 2 from Synergistic Proapoptotic Activity of Recombinant TRAIL Plus the Akt Inhibitor Perifosine in Acute Myelogenous Leukemia Cells

Supplementary Table 1 from Dual Inhibition of Class IA Phosphatidylinositol 3-Kinase and Mammalian Target of Rapamycin as a New Therapeutic Option for T-Cell Acute Lymphoblastic Leukemia

In the last few years, steps forward in the knowledge of the biology of soft tissue sarcomas (STS) have led to the development of new therapeutic strategies, including immunotherapy.This review outlines the recent findings on immunological features and provides a synopsis of the results of clinical trials with different immunotherapy approaches in STS, discussing criticisms, and how the efficacy of immunotherapy could be improved.The heterogeneity of STS has limited generalized approaches of immunotherapy in the disease. Clinical decisions should encompass a comprehensive characterization of the tumor microenvironment (TME), marked by intra-histotype diversity. Profiling of immune cells, checkpoint molecules, and antigen target/HLA expression is deemed to reshape the classical histotype classification for a selection of the most appropriate immune-based treatment. In a synergistic view, tumor-directed treatments, designed on the genetic and epigenetic histotype make-up, should be monitored for their immunomodulant effect and applied to ensure or amplify immunotherapy response. In light of the dynamic nature of the TME, this immunomonitoring should be conducted at baseline and during treatment, for improved therapeutic decisions and rational sequence of treatment combination, pursuing an immunological marker approach by histotype guidance.

Peripheral T-cell lymphomas not otherwise specified (PTCL/NOS) is the commonest subtype of PTCL. NF–kB related molecules have been found to be variably expressed in PTCL/NOS, suggesting a potential involvement of the NF–kB system in their pathogenesis. However, the actual contribution of NF–kB molecular programs to the PTCL/NOS landscape has not been investigated yet. In this study, we assessed in a large series of PTCL/NOS, the activation status of NF–kB programs and investigated the prognostic impact of such NF–kB expression. Moreover, we explored the possible role of NF–kB inhibitors. We studied the gene expression profiles of 180 PTCL cases and tested two different drugs, the IKK inhibitor BMS-345541 and the proteasome inhibitor Bortezomib, in four PTCL cell lines. We found that most cases (84%) presented with some degree of NF–kB activation, based on the expression of REL and RELA. Functionally, the latter was strictly related with TCR signaling activation, while REL was at least partially TCR independent. We also identified genes related with NF–kB activation in this setting that were mainly involved in cell proliferation and apoptosis inhibition. Further, by reverse engineering we defined the transcriptional network of both REL and RELA in PTCLs that only partially overlapped. On the clinical ground, we found that RELA expression was related to a significantly poorer overall survival, with similar trends for REL. However, most remarkably, when all the three genes were considered together, cases with at least one gene over-expressed, showed a dramatically inferior overall survival (28.67 vs. 56.018 months; p = 0.004). Finally, we showed that NF–kB pharmacological inhibition was associated with cell cycle arrest and cell death in NF–kB positive PTCL cells. In conclusion, we extensively explored NF–kB activation in PTCL/NOS, documenting its negative prognostic role. Further, we showed that NF–kB inhibition might represent a rational therapeutic approach in selected cases.

AimsTo assess survival predictivity of baseline blood cell differential count (BCDC) discretization methods in acutely ill adults visiting the emergency room over one-year.MethodsRetrospective cohort study on one-year survival of adults reporting to the emergency room of the A. Manzoni Hospital (Italy) during 2020. Automated BCDC analysis performed at baseline, assessed hemoglobin, red cell mean volume and distribution width (RDW), platelet distribution width (PDW), platelet-hematocrit, absolute red blood cells, white blood cells, neutrophils, lymphocytes, monocytes, eosinophils, basophils, and platelets. Discretization cutoffs were defined by: Benchmark laboratory reference values and Tailored (maximally selected rank statistics for linear and sigmoid-shaped distributed variables; optimal-equal hazard ratio (HR) method for U-shaped distributed variables. Explanatory variables (age, gender, inward admission) were analyzed using Cox multivariable regression. Receiver operating characteristic curves used the sum of Cox-significant variables in each method.ResultsOf 11052 patients (median age 67 years, interquartile range (IQR) 51–81, 48% female), 59% (n=6489) were discharged and 41% (n=4563) were admitted. After a 306-day median follow up (IQR 208–417 days), 9455 (86%) patients were alive and 1597 (14%) deceased. Increased HRs were associated with age > 73 years (HR=4.29 CI 3.78–4.87), and hospital admission (HR=2.05, CI 1.83–2.29). Age, sex, hemoglobin, mean corpuscular volume, RDW, PDW, neutrophils, lymphocytes and eosinophils were significant in overall. Benchmark included basophils and platelet count (area under the ROC curve (AUROC) 0.78). Tailored included monocyte counts and PCT (AUROC of 0.82).ConclusionsTailored discretization of BCDC provided meaningful insight regarding acute patient survival.Key messagesWhat is already known on this topicInformation on survival predictivity of BCDC is scarce, particularly in acutely ill patients considering that reference values are based on the general population.What this study addsLaboratory reference interval values predicting survival were hemoglobin, RDW, MCV, neutrophil, lymphocyte, eosinophil, and basophils counts, PLT, and PDW, independently of sex, age, and acute inward admission. Survival predictivity was improved by discretization of hemoglobin, RDW, MCV, neutrophil, lymphocyte, eosinophil, and monocyte counts, and PDW, according to the maximally selected rank statistics and optimal-equal HR method.How this study might affect research, practice, or policyBaseline BCDC discretized by tailored methods may be a useful biomarker for hazard warning in acute illness.

Protoplanetary disks emit radiation across a broad range of wavelengths, requiring a multiwavelength approach to fully understand their physical mechanisms and how they form planets. Observations at sub-millimeter to centimeter wavelengths can provide insights into the thermal emission from dust, free-free emission from ionized gas, and possible gyro-synchrotron emission from the stellar magnetosphere. This Letter focuses on CX Tau, a ${\sim}0.4\,M_\odot$ star with an extended gas emission and a compact and apparently structureless dust disk, with an average millimeter flux when compared to Class II sources in Taurus. We present Karl G. Jansky Very Large Array (VLA) observations in 4 bands (between 9.0 mm and 6.0 cm) and combine them with archival data from the Atacama Large Millimeter/submillimeter Array (ALMA), the Submillimeter Array (SMA) and the Plateau de Bure Interferometer (PdBI). Such a multiwavelength approach allows to separate the dust continuum from other emissions. After isolating the dust thermal emission, we derived an upper limit of the dust disk extent at 1.3 cm which is consistent with theoretical predictions of a radial drift-dominated disk. Centimeter data show a peculiar behavior: deep observations at 6.0 cm did not detect the source, while at 1.3 cm the flux density is anomalously higher than adjacent bands. Intraband spectral indices suggest a dominant contribution from free-free emission, whereas gyro-synchrotron emission is excluded. To explain these observations, we propose strong variability of the free-free emission with timescales shorter than a month. Another possible interpretation is the presence of anomalous microwave emission from spinning dust grains., Comment: Accepted for publication in A&A Letters; 13 pages, 13 figures, 1 table

Aims In patients undergoing cardiac implantable electronic device (CIED) intervention, routine pre-procedure antibiotic prophylaxis is recommended. A more powerful antibiotic protocol has been suggested in patients at high risk of infection. Stratification of individual infective risk could guide the prophylaxis before CIED procedure. Methods and results Patients undergoing CIED surgery were stratified according to the Shariff score in low and high infective risk. Patients in the ‘low-risk’ group were treated with only two antibiotic administrations while patients in the ‘high-risk’ group were treated with a prolonged 9-day protocol, according to renal function and allergies. We followed-up patients for 250 days with clinical outpatient visit and electronic control of the CIED. As primary endpoint, we evaluated CIED-related infections. A total of 937 consecutive patients were enrolled, of whom 735 were stratified in the ‘low-risk’ group and 202 in the ‘high-risk’ group. Despite different risk profiles, CIED-related infection rate at 250 days was similar in the two groups (8/735 in ‘low risk’ vs. 4/202 in ‘high risk’, P = 0.32). At multivariate analysis, active neoplasia, haematoma, and reintervention were independently associated with CIED-related infection (HR 5.54, 10.77, and 12.15, respectively). Conclusion In a large cohort of patients undergoing CIED procedure, an antibiotic prophylaxis based on individual stratification of infective risk resulted in similar rate of infection between groups at high and low risk of CIED-related infection.

The present work is aimed at presenting the data, methods and criteria that are being used for the assessment of ecosystem collapse risk in Italy.

Hemolytic uremic syndrome (HUS), the leading cause of acute renal failure in children (< 3 years), is mainly related to Shiga toxins (Stx)-producing Escherichia coli (STEC) infections. STEC are confined to the gut resulting in hemorrhagic colitis, whereas Stx are delivered in blood to target kidney and brain, with unclear mechanisms, triggering HUS in 5 to 15% of infected children. Stx were found on circulating cells, free in sera (soluble Stx) or in blood cell-derived microvesicles (particulate Stx), whereby the relationship between these forms of circulating toxins is unclear. Here, we have examined 2,846 children with bloody diarrhea and found evidence of STEC infection in 5%. Twenty patients were enrolled to study the natural course of STEC infections before the onset of HUS. In patients, Stx were found to be associated to circulating cells and/or free and functionally active in sera. In most children, Stx were bound to neutrophils when high amounts of toxins were found in feces. Time-course analysis showed that Stx increased transiently in patients' sera while the decrease of toxin amount on leukocytes was observed. Notably, patients who recovered (85%) displayed different settings than those who developed HUS (15%). The distinctive feature of the latter group was the presence in blood of particulate Stx2 (Stx2 sedimented at g-forces corresponding to 1 μm microvesicles) the day before diagnosis of HUS, during the release phase of toxins from circulating cells. This observation strongly suggests the involvement of blood cell-derived particulate Stx2 in the transition from hemorrhagic colitis to HUS.

Diet is a major driver of gut microbiota variation and plays a role in metabolic disorders, including metabolic syndrome (MS). Mycorrhized foods from symbiotic agriculture (SA) exhibit improved nutritional properties, but potential benefits have never been investigated in humans. We conducted a pilot interventional study on 60 adults with ≥ 1 risk factors for MS, of whom 33 consumed SA-derived fresh foods and 27 received probiotics over 30 days, with a 15-day follow-up. Stool, urine and blood were collected over time to explore changes in gut microbiota, metabolome, and biochemical, inflammatory and immunologic parameters, previous dietary habits were investigated through a validated food-frequency questionnaire. The baseline microbiota showed alterations typical of metabolic disorders, mainly an increase in Coriobacteriaceae and a decrease in health-associated taxa, which were partly reversed after the SA-based diet. Improvements were observed in metabolome, MS presence (two out of six subjects no longer had MS) or components. Changes were more pronounced with less healthy baseline diets. Probiotics had a marginal, not entirely favorable, effect, although one out of three subjects no longer suffered from MS. These findings suggest that improved dietary patterns can modulate the host microbiota and metabolome, counteracting the risk of developing MS.

Abstract Within the circular economy framework, our study aims to assess the rhamnolipid production from winery and olive oil residues as low-cost carbon sources by nonpathogenic strains. After evaluating various agricultural residues from those two sectors, Burkholderia thailandensis E264 was found to use the raw soluble fraction of nonfermented (white) grape marcs (NF), as the sole carbon and energy source, and simultaneously, reducing the surface tension to around 35 mN/m. Interestingly, this strain showed a rhamnolipid production up to 1070 mg/L (13.37 mg/g of NF), with a higher purity, on those grape marcs, predominately Rha-Rha C14-C14, in MSM medium. On olive oil residues, the rhamnolipid yield of using olive mill pomace (OMP) at 2% (w/v) was around 300 mg/L (15 mg/g of OMP) with a similar CMC of 500 mg/L. To the best of our knowledge, our study indicated for the first time that a nonpathogenic bacterium is able to produce long-chain rhamnolipids in MSM medium supplemented with winery residues, as sole carbon and energy source. Key points • Winery and olive oil residues are used for producing long-chain rhamnolipids (RLs). • Both higher RL yields and purity were obtained on nonfermented grape marcs as substrates. • Long-chain RLs revealed stabilities over a wide range of pH, temperatures, and salinities

Vineyard and olive farming are deeply rooted in the economy, culture, and even the welfare of Mediterranean countries. Although numerous approaches and methods were developed to valorize their agricultural residues, several challenges remain to overcome moving towards the Circular Economy (CE). Conventional treatments of winery and olive mill wastes are becoming increasingly expensive, demanding significant amounts of effort, resources, and energy for safe waste discharge. Therefore, the need to recycle, reuse, and recover energy and valuable biocompounds from the wastes of these two sectors become apparent. In the context of increasing waste production, climate change, coupled with dramatic rises in demand from a much larger global population and customers, CE offers the prospect of a better future. Microbes are able to produce diverse biosurfactants (BS) containing both hydrophilic and hydrophobic moieties that could interact with surfaces, lower surface and interfacial tensions, form micelles, and emulsify immiscible substances. Among several BS classes, rhamnolipids are glycolipids produced by some gram-negative bacteria with various potential applications in numerous fields, including bioremediation of contaminated soils. In contrast to several biosurfactants, this particular class of glycolipids is among the economically competitive biomolecules with its synthetic equivalents. For that, we highlighted the main opportunities, the challenges, and the future aspects towards the production of this particular type of glycolipids using winery and olive oil wastes. Therefore, we proposed the critical points that should be assumed through the path of successful strategies for rhamnolipid production using these two potential agricultural residues. This review might help at boosting the current attempts to find novel CE models. Graphic Abstract: [Figure not available: see fulltext.]

Polycythemia Vera (PV) is a myeloproliferative neoplasm with increased risk of thrombosis and progression to myelofibrosis. Chronic inflammation is commonly observed in myeloproliferative neoplasms including PV. The inflammatory network includes the extracellular vesicles (EVs), which play a role in cell-cell communication. Recent evidence points to circulating microbial components/microbes as potential players in hemopoiesis regulation. To address the role of EVs in PV, here we investigated phenotype and microbial DNA cargo of circulating EVs through multidimensional analysis. Peripheral blood and feces were collected from PV patients (n=38) and healthy donors (n=30). Circulating megakaryocyte (MK)- and platelet (PLT)-derived EVs were analyzed by flow cytometry. After microbial DNA extraction from feces and isolated EVs, the 16S rDNA V3-V4 region was sequenced. We found that the proportion of circulating MK-derived EVs was significantly decreased in PV patients as compared with the healthy donors. By contrast, the proportion of the PLT-derived EVs was increased. Interestingly, PV was also associated with a microbial DNA signature of the isolated EVs with higher diversity and distinct microbial composition than the healthy counterparts. Of note, increased proportion of isolated lipopolysaccharide-associated EVs has been demonstrated in PV patients. Conversely, the gut microbiome profile failed to identify a distinct layout between PV patients and healthy donors. In conclusion, PV is associated with circulating EVs harbouring abnormal phenotype and dysbiosis signature with a potential role in the (inflammatory) pathogenesis of the disease.

Class III stars are those in star forming regions without large non-photospheric infrared emission, suggesting recent dispersal of their protoplanetary disks. We observed 30 class III stars in the 1-3 Myr Lupus region with ALMA at ${\sim}856��$m, resulting in 4 detections that we attribute to circumstellar dust. Inferred dust masses are $0.036{-}0.093M_\oplus$, ${\sim}1$ order of magnitude lower than any previous measurements; one disk is resolved with radius ${\sim}80$ au. Two class II sources in the field of view were also detected, and 11 other sources, consistent with sub-mm galaxy number counts. Stacking non-detections yields a marginal detection with mean dust mass ${\sim}0.0048M_\oplus$. We searched for gas emission from the CO J=3-2 line, and present its detection to NO Lup inferring a gas mass ($4.9 {\pm} 1.1$) ${\times}10^{-5} M_\oplus$ and gas-to-dust ratio $1.0{\pm}0.4$. Combining our survey with class II sources shows a gap in the disk mass distribution from $0.09{-}2M_\oplus$ for ${>}0.7M_\odot$ Lupus stars, evidence of rapid dispersal of mm-sized dust from protoplanetary disks. The class III disk mass distribution is consistent with a population model of planetesimal belts that go on to replenish the debris disks seen around main sequence stars. This suggests that planetesimal belt formation does not require long-lived protoplanetary disks, i.e., planetesimals form within ${\sim}$2 Myr. While all 4 class III disks are consistent with collisional replenishment, for two the gas and/or mid-IR emission could indicate primordial circumstellar material in the final stages of protoplanetary disk dispersal. Two class III stars without sub-mm detections exhibit hot emission that could arise from ongoing planet formation processes inside ${\sim}1$ au., 30 pages, 20 figures. This is a pre-copyedited, author-produced PDF of an article accepted for publication in MNRAS following peer review

Context. Observations at millimeter wavelengths of bright protoplanetary disks have shown the ubiquitous presence of structures such as rings and spirals in the continuum emission. The derivation of the underlying properties of the emitting material is nontrivial because of the complex radiative processes involved. Aims. In this paper we analyze new observations from the Atacama Large Millimeter/submillimeter Array (ALMA) and the Karl G. Jansky Very Large Array (VLA) at high angular resolution corresponding to 5 – 8 au to determine the dust spatial distribution and grain properties in the ringed disk of HD 163296. Methods. We fit the spectral energy distribution as a function of the radius at five wavelengths from 0.9 to 9 mm, using a simple power law and a physical model based on an analytic description of radiative transfer that includes isothermal scattering. We considered eight dust populations and compared the models' performance using Bayesian evidence. Results. Our analysis shows that the moderately high optical depth (τ>1) at λ ≤ 1.3 mm in the dust rings artificially lower the millimeter spectral index, which should therefore not be considered as a reliable direct proxy of the dust properties and especially the grain size. We find that the outer disk is composed of small grains on the order of 200 µm with no significant difference between rings at 66 and 100 au and the adjacent gaps, while in the innermost ~30 au, larger grains (≥mm) could be present. We show that the assumptions on the dust composition have a strong impact on the derived surface densities and grain size. In particular, increasing the porosity of the grains to 80% results in a total dust mass about five times higher with respect to grains with 25% porosity. Finally, we find that the derived opacities as a function of frequency deviate from a simple power law and that grains with a lower porosity seem to better reproduce the observations of HD 163296. Conclusions. While we do not find evidence of differential trapping in the rings of HD 163296, our overall results are consistent with the postulated presence of giant planets affecting the dust temperature structure and surface density, and possibly originating a second-generation dust population of small grains., Astronomy & Astrophysics, 664, ISSN:0004-6361, ISSN:1432-0746

The 1.33 mm survey of protoplanetary discs in the Taurus molecular cloud found annular gaps and rings to be common in extended sources (>~55 au), when their 1D visibility distributions were fit parametrically. We first demonstrate the advantages and limitations of nonparametric visibility fits for data at the survey's 0.12" resolution. Then we use the nonparametric model in Frankenstein ('frank') to identify new substructure in three compact and seven extended sources. Among the new features we identify three trends: a higher occurrence rate of substructure in the survey's compact discs than previously seen, underresolved (potentially azimuthally asymmetric) substructure in the innermost disc of extended sources, and a 'shoulder' on the trailing edge of a ring in discs with strong depletion at small radii. Noting the shoulder morphology is present in multiple discs observed at higher resolution, we postulate it is tracing a common physical mechanism. We further demonstrate how a super-resolution frank brightness profile is useful in motivating an accurate parametric model, using the highly structured source DL Tau in which frank finds two new rings. Finally we show that sparse (u, v) plane sampling may be masking the presence of substructure in several additional compact survey sources., Accepted to MNRAS

(Abridged) Exoplanetary research has provided us with exciting discoveries of planets around very low-mass (VLM) stars (e.g., TRAPPIST-1 and Proxima Centauri). However, current theoretical models strive to explain planet formation in these conditions and do not predict the development of giant planets. Recent high-resolution observations from ALMA of the disk around CIDA 1, a VLM star in Taurus, show substructures hinting at the presence of a massive planet. We aim to reproduce the dust ring of CIDA 1, observed in the dust continuum emission in ALMA Band 7 (0.9 mm) and Band 4 (2.1 mm), along with its $^{12}$CO (J=3-2) and $^{13}$CO (J=3-2) channel maps, assuming the structures are shaped by the interaction of the disk with a massive planet. We seek to retrieve the mass and position of the putative planet. We model the protoplanetary disk with a set of hydrodynamical simulations, varying the mass and locations of the embedded planet. We compute the dust and gas emission using radiative transfer simulations, and, finally, we obtain the synthetic observations treating the images as the actual ALMA observations. Our models indicate that a planet with a minimum mass of $\sim1.4\,\text{M}_\text{Jup}$ orbiting at a distance of $\sim 9-10$ au can explain the morphology and location of the observed dust ring at Band 7 and Band 4. We can reproduce the low spectral index ($\sim 2$) observed where the dust ring is detected. Our synthetic images reproduce the morphology of the $^{12}$CO and $^{13}$CO observed channel maps where the cloud absorption allowed a detection. Applying an empirical relation between planet mass and gap width in the dust, we predict a maximum planet mass of $\sim4 - 8\,\text{M}_\text{Jup}$. Our results suggest the presence of a massive planet orbiting CIDA 1, thus challenging our understanding of planet formation around VLM stars., Accepted for publication in A&A, 22 pages, 21 figures, 3 tables

Recent observations have revealed that most proto-planetary discs show a pattern of bright rings and dark gaps. However, most of the high-resolution observations have focused only on the continuum emission. In this Paper we present high-resolution ALMA band 7 (0.89mm) observations of the disc around the star CI Tau in the $^{12}$CO & $^{13}$CO $J=3$-2 and CS $J=7$-6 emission lines. Our recent work demonstrated that the disc around CI Tau contains three gaps and rings in continuum emission, and we look for their counterparts in the gas emission. While we find no counterpart of the third gap and ring in $^{13}$CO, the disc has a gap in emission at the location of the second continuum ring (rather than gap). We demonstrate that this is mostly an artefact of the continuum subtraction, although a residual gap still remains after accounting for this effect. Through radiative transfer modelling we propose this is due to the inner disc shadowing the outer parts of the disc and making them colder. This raises a note of caution in mapping high-resolution gas emission lines observations to the gas surface density - while possible, this needs to be done carefully. In contrast to $^{13}$CO, CS emission shows instead a ring morphology, most likely due to chemical effects. Finally, we note that $^{12}$CO is heavily absorbed by the foreground preventing any morphological study using this line.

The distributions and abundances of molecules in protoplanetary disks are powerful tracers of the physical and chemical disk structures. The abundance ratios of HCN and its isomer HNC are known to be sensitive to gas temperature. Their line ratios might therefore offer a unique opportunity to probe the properties of the emitting gas. Using the 2D thermochemical code DALI, we ran a set of disk models accounting for different stellar and disk properties, with an updated chemical network for the nitrogen chemistry. These modeling results were then compared with observations, including new ALMA observations of HNC $J=3-2$ for the TW Hya disk and HNC $J=1-0$ for 29 disks in Lupus. Similar to CN, HCN and HNC have brighter line emission in models with larger disk flaring angles and higher UV fluxes. HNC and HCN are predicted to be abundant in the warm surface layer and outer midplane region, which results in ring-shaped emission patterns. However, the precise emitting regions and emission morphology depend on the probed transition, as well as on other parameters such as C and O abundances. The modeled HNC-to-HCN line intensity ratio increases from $, accepted for publication in A&A

Recently, treatment of immune‐mediated thrombotic thrombocytopenic purpura (ITTP) has changed with the advent of caplacizumab in clinical practice. The International Working Group (IWG) has recently integrated the ADAMTS‐13 activity/autoantibody monitoring in consensus outcome definitions. We report three ITTP cases during the coronavirus disease 2019 pandemic, that received a systematic evaluation of ADAMTS‐13 activity and autoantibodies. We describe how the introduction of caplacizumab and ADAMTS‐13 monitoring could change the management of ITTP patients and discuss whether therapeutic choices should be based on the clinical response alone. ADAMTS‐13 activity/antibodies were assessed every 5 days. Responses were evaluated according to updated IWG outcome definitions. These kinetics, rather than clinical remission, guided the therapy, allowing early and safe caplacizumab discontinuation and sensible administration of rituximab. Caplacizumab was cautiously discontinued after achieving ADAMTS‐13 complete remission. These cases illustrate that prospective ADAMTS‐13 evaluation and use of updated IWG definitions may improve real‐life patients’ management in the caplacizumab era.

The DSHARP survey evidenced the ubiquity of substructure in the mm dust distribution of large, bright protoplanetary discs. Intriguingly, these datasets have yet higher resolution information that is not recovered in a CLEAN image. We first show that the intrinsic performance of the CLEAN algorithm is resolution-limited. Then analyzing all 20 DSHARP sources using the 1D, super-resolution code Frankenstein (frank), we accurately fit the 1D visibilities to a mean factor of 4.3 longer baseline than the Fourier transform of the CLEAN images and a factor of 3.0 longer baseline than the transform of the CLEAN component models. This yields a higher resolution brightness profile for each source, identifying new substructure interior to 30 au in multiple discs; resolving known gaps to be deeper, wider, and more structured; and known rings to be narrower and brighter. Across the survey, high contrast gaps are an average 14% wider and 44% deeper in the frank profiles relative to CLEAN, and high contrast rings are an average 26% narrower. Categorizing the frank brightness profiles into trends, we find that the relative scarcity of features interior to 30 au in the survey's CLEAN images is an artifact of resolving power, rather than an intrinsic rarity of inner disc (or compact disc) substructure. Finally the rings in the frank profiles are narrower than the previously inferred deconvolved widths, indicating smaller alpha / St ratios in the local gas disc., Comment: Revised version, accepted to MNRAS. Changes: added CLEAN .model profiles to Sec.3 and 4, quantified gaps/rings in frank profiles in Sec.5 and Table 3, expanded analysis of alpha/St in Sec.5.3.1. All frank fits are now available at https://zenodo.org/record/5587841

We analyse spatially resolved ALMA observations at 0.9, 1.3, and 3.1 mm for the 26 brightest protoplanetary discs in the Lupus star-forming region. We characterise the discs multi-wavelength brightness profiles by fitting the interferometric visibilities in a homogeneous way, obtaining effective disc sizes at the three wavelengths, spectral index profiles and optical depth estimates. We report three fundamental discoveries: first, the millimeter continuum size - luminosity relation already observed at 0.9 mm is also present at 1.3 mm with an identical slope, and at 3.1 mm with a steeper slope, confirming that emission at longer wavelengths becomes increasingly optically thin. Second, when observed at 3.1 mm the discs appear to be only 9% smaller than when observed at 0.9 mm, in tension with models of dust evolution which predict a starker difference. Third, by forward modelling the sample of measurements with a simple parametric disc model, we find that the presence of large grains ($a_\mathrm{max}>1 $mm) throughout the discs is the most favoured explanation for all discs as it reproduces simultaneously their spectral indices, optical depth, luminosity, and radial extent in the 0.9-1.3 mm wavelength range. We also find that the observations can be alternatively interpreted with the discs being dominated by optically thick, unresolved, substructures made of mm-sized grains with a high scattering albedo., MNRAS Accepted. 20 pages, 12 figures, 5 tables. Detailed fit results in Appendix E (27 pages, 78 figures), available as supplementary material. Machine-readable version of Table 1, Table 2, and Table 3 available at https://zenodo.org/record/4756381