Neo-linfogenesi nelle lesioni aterosclerotiche della biforcazione carotidea

INTRODUCTION: The indications for surgical or pharmacologic treatment of the patients with carotid stenosis are based on the macroscopic characteristics of the atherosclerotic carotid plaque (obtained by diagnostic investigations: ECD/CT). The stability of the plaque depends on inflammatory cells, inflammatory cytokines and on the neoangiogenesis. In the literature there are many studies about atherosclerotic plaques but the mechanism of development, progression and instability of the plaque are not known. A lot of studies show how the progression and instability of the plaques would depend on the formation of new blood vessels that are responsible for the migration of inflammatory plaques and intraplaque haemorrhage. The new \u201cactors\u201d in the atherosclerotic plaques are lymphatic vessels but their role is unknown. AIM: The aim of this project is to evaluate the inflammatory cells in atherosclerotic carotid plaque, the lymphatic and blood vessels, their location and their relationship with inflammatory cells in the plaque. MATERIAL AND METHOD: In this project, 31 patients with atherosclerotic carotid plaques were enrolled (10 plaques of patients were used to develop the method) and undergone to carotid endarterectomy (CEA) following ESVS guidelines at the Vascular Surgery Unit of the Ospedale di Circolo in Varese. For each clinical and biological data were collected. The carotid plaques were classified independently of clinical symptoms by their morphology (stenosis and plaque composition) by computed-tomography angiography into fatty/soft (< 60 Hounsfield units (HU), mixed 60>HU<129) and calcified (> 130 HU). The calcified plaques were excluded from this study. Carotid endarterectomy tissue samples, during CEA, were removed with minimal trauma and immediately placed in paraformaldehyde for histological evaluations researching inflammatory cells, lymphatic and blood vessels in the plaques. The Immunohistochemestry slides were processed with MIAquant that is a novel system for automatic segmentation, measurement and localization comparison of different biomarkers from serialized histological slices. This software would allow to analyze, with objective criteria, a large number of images but, especially, the entire surface of the section. RESULTS: In soft plaques there are a lot of inflammatory cells and vessels (lymphatic and blood) compared to mix plaques. Working on serial plaque sections, I focused on the localization of neovascularization, using Ulex to detect blood vessels and Podoplanin (D2-40) as marker of lymphatic vessels. In the soft plaques, the expression of D2-40 (lymphatic vessels) is near the lipidic core, unlike the blood vessels are localized in soft and mixed plaques, predominantly near the cap of the plaques and there isn\u2019t any relationship with the lipid core. For a better understanding of the origin and function of lymphatic and blood vessels inside atherosclerotic carotid plaques, some components of the inflammatory infiltrate were examined, particularly, I have analysed T lymphocytes (CD-3) and two different macrophage populations (CD-68, CD-163). In addition to this, their position was also evaluated against lymphatic and blood vessels. In atherosclerotic carotid plaques Lymphocytes (CD-3) are near blood vessels and are spread homogenously in the plaques. T lymphocytes and hematic vessels are not localized in the same territory of lymphatic vessels marked with D2-40. The macrophages cells (CD-68, CD-163) are located near the lipid core, in the same layer of lymphatic vessels. The colocation of lymphocytes with blood vessels and macrophages with lymphatic vessels were confirmed by double coloration with confocal microscopy. To achieve comparable and reproducible data, all soft plaque sections have been examined automatically, by MIAquant software, to obtain a localization image for each segmented marker (map), which shows the locations of all markers inside the atherosclerotic carotid plaques. Furthermore, the software allows to create an histogram with united density value of markers of all plaques. This software allow to confirm that macrophages cells and lymphatic vessels in all plaques are located near the lipid core, in particular at a distance of 0-400 pixels. Instead blood vessels and lymphocytes are spread homogenously. The co-location analysis of lymphatic vessels and macrophage cells was further scrutinized by way of immunofluorescence in confocal microscopy. Taking advantage of the resolution power of confocal microscopy, it observed that only rarely we can find tubular structures attributable to one or collapsed lymphatic vessels, this is frequently seen in structures that could also be isolated cells near macrophage cells CD68. In some cases, cells seem that they express both markers. To characterise macrophage cells and to better understand how they could influence new lymph genesis, an analysis of macrophage cells CD-68 was done on the confocal microscope with doublemarkings and I observed the presence in the plaques of cells that show positive labelling for CD-68 and they don\u2019t express HLA-DR. Such features are characteristic for myeloid-derived suppressor cells (MDSC) of the monocyte-macrophage line. DISCUSSION: These data show that the lymphatic and blood vessels are located in different place in the carotid atherosclerotic plaque. The blood vessels co-locate with lymphocyte CD-3 cells, these cells are important to determinate the development of the lesions, in fact they can cause apoptosys and they produce cytokines that promote the endothelial proliferation and develop of neo-vessels. Furthermore they reduce macrophages proliferation and so they cause plaque instability. The lymphatic vessels, near the lipid core in atherosclerotic plaque, could be the attempt to absorb and metabolize lipids and cholesterol. In fact in physiological conditions they have a key role in the reverse transport of lipid and cholesterol in peripheral blood. These data are further support by the co-location of lymphatic vessels and macrophage cells, in fact it is known that the macrophage cells in atherosclerosis are important in the process of phagocytosis of lipid and cholesterol performing the role of \u201cscavenger\u201d. Macrophages localize in the proximity of the lymphatic vessels and co-localize with themselves, probably because they represent inflammatory cells mainly involved in the lymphogenesis process. They determine lymphogenesis via two methods: the production of paracrine signals represents the first one on behalf of macrophages; the second one is symbolized by the differentiation of macrophages in lymphatic endothelial cells. The first mechanism of macrophages that supports lymphogenesis, is determined by the up-regulation of VEGF-C during the inflammatory process inside the atherosclerotic plaque. In fact, macrophages produce TNF-\u3b1 that activates the TNFR1 receptor facilitating the creating of the VEGF-C, which activates VEGFR-3 promoting the development and the activation of lymphatic endothelial cells. In addition to the paracrine mechanism, macrophages contribute to lymphogenesis using transdifferentiation in lymphatic endothelial cells. Through this investigation, I have also recognized the presence of myeloid-derived suppressor cells of the monocyte-macrophage line inside the atherosclerotic carotid plaques. These cells inside neoplasms have a dual function: trying to inhibit T lymphocytes activity from avoiding self-inflicted damage, although, at the same time, they facilitate neo-angiogenesis and neo-lymphogenesis, and thus tumor\u2019s growth and its dissemination. In atherosclerotic carotid plaques, these cells will probably inhibit T cells\u2019 activity, as they do not localize near such cells, and they foster lymphogenesis more extensively that angiogenesis, given their localization close to lymphatic vessels and not near blood vessels. CONCLUSIONS: Lymphatic vessels are new actors in atherosclerotic carotid plaques but it is unknown if they determine the progression, stability or instability of the plaques. This project might be an optimal start point to continue the study trying to identify the functions of myeloid-derived suppressor cells in particular, by pinpointing the cytokines that they produce, to understand that type of activity they possess inside the atherosclerotic plaque. Therefore, this investigation, regardless of the riveting and not yet identified data in the literature, it represents the preliminary phase of other possible examinations of atherosclerotic carotid plaques. The inspection of new plaques and additional markers will allow us to acquire significant results for an overall assessment of the plaque, and hopefully, to the identification of areas at a higher atheroembolic risk inside the atherosclerotic carotid plaque

Investigating multisensory integration in human early visual and auditory areas with intracranial electrophysiological recordings: insights and perspectives

Cross-modal processing and multisensory integration (MSI) can be observed at early stages of sensory processing in the cortex. However, the neurophysiological mechanisms underlying these processes and how they vary across sensory systems remain elusive. The aim of this study was to investigate how cross-modal processing and MSI are reflected in power and phase of oscillatory neuronal activity at different temporal scales in different sensory cortices. To this goal, we recorded stereo-electroencephalographic (SEEG) responses from early visual (calcarine and pericalcarine) and auditory (Heschl’s gyrus and planum temporale) regions in patients with drug-resistant epilepsy while performing an audio-visual oddball task. To Investigate crossmodal processing and MSI in the power domain of oscillatory activity, we explored a wide range of frequency bands (theta/alpha band: 5-13Hz; beta band: 13-30 Hz; gamma band: 30-80 Hz; high-gamma band: 80-200 Hz) during the first 150 ms post-stimulus onset. Differently, to investigate crossmodal processing and MSI in the phase domain of oscillatory activity, we explored a narrow range of frequency bands (theta/alpha band: 5-13Hz; beta band: 13-30 Hz; gamma band: 30-80 Hz) during the first 300 ms post-stimulus onset. In the power domain, we showed that cross-modal processing occurs mainly in the high-gamma band (80-200Hz) in both cortices. However, we evidenced that the way MSI is expressed across modalities differs considerably: in the visual cortex, MSI relies mainly on the beta band, however it is also evident, to a lesser extent, in the gamma and high-gamma band, while the auditory cortex reveals widespread MSI in the high-gamma band and, to a lesser extent, across the gamma band and the other investigated frequency bands. In the phase domain, we showed that cross-modal processing is differently expressed across modalities: in the auditory cortex it induces an increased phase concentration index (PCI) in ongoing oscillatory activity across all the investigated frequency bands, while, in the visual cortex, it induces an increased PCI particularly evident in the theta/alpha band with few or no effect respectively in the gamma and beta band. Importantly in both cortices, the most part of the COIs showing increased PCI, were not accompanied by a concomitant increase in power. These results indicate that in both auditory and visual cortex, cross-modal processing induces a pure phase resetting of the oscillatory activity. During MSI processing we observed, in both cortices, a stronger increase in PCI, in comparison to the intramodal processing, in the theta/alpha band and in the gamma band. Our results confirm the presence of cross-modal information representations at neuronal populations level and conform to a model where the cross-modal input induces phase-locked modulation of the ongoing oscillations. Importantly, our data showed that the way MSI is expressed in power modulations differs between the investigated sensory cortices suggesting the presence of different types of neurophysiological interactions during this process. These results are discussed in the framework of the current literature

I professionisti del sociale: crisi del welfare state, crisi economica

This article analyzes the present condition of the professional social workers through their practice activities and their biographies. The context of the ethnographic research is Naples. The first part of the paper describes the experience of social operators that work in the cooperatives of the third sector and in the private social organizations. The second one presents a focus about the condition of the social workers that take care of the homeless. The theoretical perspective of this paper is the critical approach to the “selective welfare policy” produced by the liberal governmentality. The authors focus on the distortions created by the system of governance and its impact on social workers and “users”. From the point of view of professional social workers, this article reflects on how “work, health and family” become an “individual risk” as a result of the transformation of welfare policies

Effects of mutational loss of nucleoside kinases on deoxyadenosine 5'-phosphate/deoxyadenosine substrate cycle in cultured CEM and V79 cells.

The functions of a deoxynucleoside kinase and a deoxynucleotidase can give rise to substrate cycles in which the two enzymes catalyze in opposite directions the irreversible interconversion of a deoxynucleoside 5'-monophosphate (dNMP) and its deoxynucleoside. Earlier evidence showed that pyrimidine dNMP cycles occur in cultured cells and participate in the regulation of the size of dNMP pools there by affecting the transport of deoxyribonucleosides across the cell membrane. Here, we apply an isotope flow method using labeled adenine as precursor of dAMP and DNA to quantify deoxyadenosine excretion as a measure of the catabolic activity of a putative dAMP/deoxyadenosine cycle. A comparison of human CEM lymphoblasts and hamster V79 fibroblasts, including mutant cells lacking kinases for the phosphorylation of deoxyadenosine, shows a much lower deoxyadenosine excretion in CEM cells (0.05% of dATP synthesized by reduction of ADP) as compared with V79 cells (4% of dATP). Mutational loss of deoxycytidine kinase increases these values to 0.3% in CEM cells and to 10% in V79 cells. This strongly suggests the presence of a dAMP/deoxyadenosine cycle in both CEM and V79 cells. Additional loss of adenosine kinase only marginally affects deoxyadenosine excretion in CEM cells. The small excretion of deoxyadenosine (also in the absence of both kinases) demonstrates that in CEM cells the in situ activity of the deoxynucleotidase affecting the dAMP/deoxyadenosine substrate cycle is very low and that the cycle has mainly an anabolic function there

Predictive Analysis of Healthcare-Associated Blood Stream Infections in the Neonatal Intensive Care Unit Using Artificial Intelligence: A Single Center Study

Background: Neonatal infections represent one of the six main types of healthcare-associated infections and have resulted in increasing mortality rates in recent years due to preterm births or problems arising from childbirth. Although advances in obstetrics and technologies have minimized the number of deaths related to birth, different challenges have emerged in identifying the main factors affecting mortality and morbidity. Dataset characterization: We investigated healthcare-associated infections in a cohort of 1203 patients at the level III Neonatal Intensive Care Unit (ICU) of the “Federico II” University Hospital in Naples from 2016 to 2020 (60 months). Methods: The present paper used statistical analyses and logistic regression to identify an association between healthcare-associated blood stream infection (HABSIs) and the available risk factors in neonates and prevent their spread. We designed a supervised approach to predict whether a patient suffered from HABSI using seven different artificial intelligence models. Results: We analyzed a cohort of 1203 patients and found that birthweight and central line catheterization days were the most important predictors of suffering from HABSI. Conclusions: Our statistical analyses showed that birthweight and central line catheterization days were significant predictors of suffering from HABSI. Patients suffering from HABSI had lower gestational age and birthweight, which led to longer hospitalization and umbilical and central line catheterization days than non-HABSI neonates. The predictive analysis achieved the highest Area Under Curve (AUC), accuracy and F1-macro score in the prediction of HABSIs using Logistic Regression (LR) and Multi-layer Perceptron (MLP) models, which better resolved the imbalanced dataset (65 infected and 1038 healthy)

The Impacts of Modeling Choices on the Inference of the Circumgalactic Medium Properties from Sunyaev-Zeldovich Observations

As the signal-to-noise of Sunyaev-Zeldovich (SZ) cross-correlation measurements of galaxies improves our ability to infer properties about the circumgalactic medium (CGM), we will transition from being limited by statistical uncertainties to systematic uncertainties. Using thermodynamic profiles of the CGM created from the IllustrisTNG (The Next Generation) simulations we investigate the importance of specific choices in modeling the galaxy sample. These choices include different sample selections in the simulation (stellar vs. halo mass, color selections) and different fitting models (matching by the shape of the mass distribution, inclusion of a two-halo term). We forward model a mock galaxy sample into projected SZ observable profiles and fit these profiles to a generalized Navarro-Frenk-White profile using forecasted errors of the upcoming Simons Observatory experiment. We test the number of free parameters in the fits and show that this is another modeling choice that yields different results. Finally, we show how different fitting models can reproduce parameters of a fiducial profile, and show that the addition of a two-halo term and matching by the mass distribution of the sample are extremely important modeling choices to consider.Comment: 22 pages, 9 figures, comments welcom

Improvement of skeletal muscle performance in ageing by the metabolic modulator Trimetazidine

BACKGROUND: The loss of muscle mass (sarcopenia) and the associated reduced muscle strength are key limiting factors for elderly people's quality of life. Improving muscle performance does not necessarily correlate with increasing muscle mass. In fact, particularly in the elderly, the main explanation for muscle weakness is a reduction of muscle quality rather than a loss of muscle mass, and the main goal to be achieved is to increase muscle strength. The effectiveness of Trimetazidine (TMZ) in preventing muscle functional impairment during ageing was assessed in our laboratory. METHODS: Aged mice received TMZ or vehicle for 12 consecutive days. Muscle function was evaluated at the end of the treatment by a grip test as well as by an inverted screen test at 0, 5, 7 and 12 days of TMZ treatment. After sacrifice, muscles were stored for myofiber cross‐sectional area assessment and myosin heavy chain expression evaluation by western blotting. RESULTS: Chronic TMZ treatment does not affect the mass of both gastrocnemius and tibialis anterior muscles, while it significantly increases muscle strength. Indeed, both latency to fall and grip force are markedly enhanced in TMZ‐treated versus untreated mice. In addition, TMZ administration results in higher expression of slow myosin heavy chain isoform and increased number of small‐sized myofibers. CONCLUSIONS: We report here some data showing that the modulation of skeletal muscle metabolism by TMZ increases muscle strength in aged mice. Reprogramming metabolism might therefore be a strategy worth to be further investigated in view of improving muscle performance in the elderly

Generation of induced pluripotent stem cells (iPSCs) from patient with Cri du Chat Syndrome

Abstract The Cri du Chat Syndrome (CdCS) is a genetic disease resulting from variable size deletion occurring on the short arm of chromosome 5. The main clinical features are a high-pitched monochromatic cry, microcephaly, severe psychomotor and mental retardation with characteristics of autism spectrum disorders such as hand flapping, obsessive attachments to objects, twirling objects, repetitive movements, and rocking. We reprogrammed to pluripotency peripheral blood mononuclear cells derived from a patient carrying large deletion on the short arm of chromosome 5, using a commercially available non-integrating expression system. The iPSCs expressed pluripotency markers and differentiated in the three embryonic germ layers

Hypoallergenic fragment of Par j 2 increases functional expression of Toll-like receptors in atopic children

Parietaria judaica (Par j) is one of the main causes of allergy in the Mediterranean countries. The activation of Toll-like receptor 4 (TLR4) by lipopolysaccharide (LPS) inhibits nasal inflammation of atopic children

Prognostic Value of Mid-Regional Proadrenomedullin Sampled at Presentation and after 72 Hours in Septic Patients Presenting to the Emergency Department: An Observational Two-Center Study

The prognostic value of mid-regional proADM (MR-proADM) in septic patients presenting to the emergency department (ED) is not well established. In this prospective observational study enrolling septic patients evaluated in two EDs, MR-proADM was measured at arrival (t0) and after 72 h (t72). MR-proADM(%change) was calculated as follows: (MR-proADM(t72h) − MR-proADM(t0))/MR-proADM(t0). In total, 147 patients were included in the study, including 109 with a final diagnosis of sepsis and 38 with septic shock, according to the Sepsis-3 criteria. The overall 28-day mortality (outcome) rate was 12.9%. The AUC for outcome prognostication was 0.66 (95% CI 0.51–0.80) for MR-proADM(t0), 0.77 (95% CI 0.63–0.92) for MR-proADM(t72) and 0.74 (95% CI 0.64–0.84) for MR-proADM(%change). MR-proADM(t0) ≥ 2.78 nmol/L, MR-proADM(t72) ≥ 2.7 nmol/L and MR-proADM(%change) ≥ −15.2% showed statistically significant log-rank test results and sensitivity/specificity of 81/65%, 69/80% and 75/70% respectively. In regression analysis, MR-proADM(%change) was a significant outcome predictor both in univariate and multivariate analysis, after adjustment for age, SOFA and APACHEII scores, providing up to 80% of added prognostic value. In conclusion, time trends of MR-proADM may provide additional insights for patient risk stratification over single sampling. MR-proADM levels sampled both at presentation and after 72 h predicted 28-day survival in septic patients presenting to the ED