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IntroductionBrain metastases (BrM), which commonly arise in patients with melanoma, breast cancer and lung cancer, are associated with a poor clinical prognosis. In this context, the tumor microenvironment (TME) plays an important role since it either promotes or inhibits tumor progression. Our previous studies have characterized the immunosuppressive microenvironment of glioblastoma (GBM). The aim of this study is to compare the immune profiles of BrM and GBM in order to identify potential differences that may be exploited in their differential treatment.MethodsTumor and/or blood samples were taken from 20 BrM patients and 19 GBM patients. Multi-parametric flow cytometry was used to evaluate myeloid and lymphoid cells, as well as the expression of immune checkpoints in the TME and blood. In selected cases, the immunosuppressive ability of sorted myeloid cells was tested, and the ex vivo proliferation of myeloid, lymphoid and tumor cell populations was analyzed.ResultsHigh frequencies of myeloid cells dominated both the BrM and GBM landscapes, but a higher presence of tumor-associated macrophages was observed in GBM, while BrM were characterized by a significant presence of tumor-infiltrating lymphocytes. Exhaustion markers were highly expressed in all T cells from both primary and metastatic brain tumors. Ex vivo analysis of the cell cycle of a single sample of a BrM and of a GBM revealed subsets of proliferating tumor cells and blood-derived macrophages, but quiescent resident microglial cells and few proliferating lymphocytes. Macrophages sorted from a single lung BrM exhibited a strong immunosuppressive activity, as previously shown for primary GBM. Finally, a significant expansion of some myeloid cell subsets was observed in the blood of both GBM and BrM patients.DiscussionOur results define the main characteristics of the immune profile of BrM and GBM, which are distinguished by different levels of immunosuppressive myeloid cells and lymphocytes devoid of effector function. Understanding the role of the different cells in establishing the metastatic setting is critical for improving the therapeutic efficacy of new targeted immunotherapy strategies.

Assessing the cognitive abilities of students in academic contexts can provide valuable insights for teachers to identify their cognitive profile and create personalized teaching strategies. While numerous studies have demonstrated promising outcomes in clustering students based on their cognitive profiles, effective comparisons between various clustering methods are lacking in the current literature.In this study, we aim to compare the effectiveness of two clustering techniques to group students based on their cognitive abilities including general intelligence, attention, visual perception, working memory, and phonological awareness. 292 students, aged 11–15 years, participated in the study.A two-level approach based on the joint use of Kohonen's Self-Organizing Map (SOMs) and k-means clustering algorithm was compared with an approach based on the k-means clustering algorithm only. The resulting profiles were then predicted via AdaBoost and ANN supervised algorithms.The results showed that the two-level approach provides the best solution for this problem while the ANN algorithm was the winner in the classification problem.These results laying the foundations for developing a useful instrument for predicting the students’ cognitive profile.

Abstract Myeloid-derived suppressor cells (MDSCs) are a heterogeneous population of immunosuppressive cells developing from myeloid progenitors, which are enriched in pathological conditions such as cancer, and are known to inhibit the functions of effector T cells. During aging, several changes occur both at the adaptive and innate immune system level, in a process defined as immunoscenescence. In particular, the low-grade inflammation state observed in the elderly appears to affect hematopoiesis. We previously demonstrated that the combination of GM-CSF and G-CSF drives the in vitro generation of bone marrow-derived MDSCs (BM-MDSCs) from precursors present in human bone marrow aspirates of healthy donors, and that these cells are endowed with a strong immune suppressive ability, resembling that of cancer-associated MDSCs. In the present work we investigated BM-MDSCs induction and functional ability in a cohort of pediatric versus elderly donors. To this aim, we analyzed the differences in maturation stages and ability to suppress T cell proliferation. We found that the ex vivo distribution of myeloid progenitors is similar between pediatric and elderly individuals, whereas after cytokine treatment a significant reduction in the more immature compartment is observed in the elderly. Despite the decreased frequency, BM-MDSCs maintain their suppressive capacity in aged donors. Taken together, these results indicate that in vitro induction of MDSCs from the BM is reduced with aging and opens new hypotheses on the role of age-related processes in myelopoiesis.

Abstract Background Myeloid derived suppressor cells (MDSCs) and tumor-associated macrophages (TAMs) are two of the major players involved in the inhibition of anti-tumor immune response in cancer patients, leading to poor prognosis. Selective targeting of myeloid cells has therefore become an attractive therapeutic strategy to relieve immunosuppression and, in this frame, we previously demonstrated that lipid nanocapsules (LNCs) loaded with lauroyl-modified gemcitabine efficiently target monocytic MDSCs in melanoma patients. In this study, we investigated the impact of the physico-chemical characteristics of LNCs, namely size and surface potential, towards immunosuppressive cell targeting. We exploited myeloid cells isolated from glioblastoma patients, which play a relevant role in the immunosuppression, to demonstrate that tailored nanosystems can target not only tumor cells but also tumor-promoting cells, thus constituting an efficient system that could be used to inhibit their function. Results The incorporation of different LNC formulations with a size of 100 nm, carrying overall positive, neutral or negative charge, was evaluated on leukocytes and tumor-infiltrating cells freshly isolated from glioblastoma patients. We observed that the maximum LNC uptake was obtained in monocytes with neutral 100 nm LNCs, while positively charged 100 nm LNCs were more effective on macrophages and tumor cells, maintaining at low level the incorporation by T cells. The mechanism of uptake was elucidated, demonstrating that LNCs are incorporated mainly by caveolae-mediated endocytosis. Conclusions We demonstrated that LNCs can be directed towards immunosuppressive cells by simply modulating their size and charge thus providing a novel approach to exploit nanosystems for anticancer treatment in the frame of immunotherapy.

BackgroundGlioblastoma (GBM) is the most commonly occurring primary malignant brain tumor, and it carries a dismal prognosis. Focusing on the tumor microenvironment may provide new insights into pathogenesis, but no clinical tools are available to do this. We hypothesized that the infiltration of different leukocyte populations in the tumoral and peritumoral brain tissues may be measured by magnetic resonance imaging (MRI).MethodsPre-operative MRI was combined with immune phenotyping of intraoperative tumor tissue based on flow cytometry of myeloid cell populations that are associated with immune suppression, namely, microglia and bone marrow-derived macrophages (BMDM). These cell populations were measured from the central and marginal areas of the lesion identified intraoperatively with 5-aminolevulinic acid-guided surgery. MRI features (volume, mean and standard deviation of signal intensity, and fractality) were derived from all MR sequences (T1w, Gd+ T1w, T2w, FLAIR) and ADC MR maps and from different tumor areas (contrast- and non-contrast-enhancing tumor, necrosis, and edema). The principal components of MRI features were correlated with different myeloid cell populations by Pearson’s correlation.ResultsWe analyzed 126 samples from 62 GBM patients. The ratio between BMDM and microglia decreases significantly from the central core to the periphery. Several MRI-derived principal components were significantly correlated (p

BackgroundAlthough gliomas are confined to the central nervous system, their negative influence over the immune system extends to peripheral circulation. The immune suppression exerted by myeloid cells can affect both response to therapy and disease outcome. We analyzed the expansion of several myeloid parameters in the blood of low- and high-grade gliomas and assessed their relevance as biomarkers of disease and clinical outcome.MethodsPeripheral blood was obtained from 134 low- and high-grade glioma patients. CD14+, CD14+/p-STAT3+, CD14+/PD-L1+, CD15+ cells and four myeloid-derived suppressor cell (MDSC) subsets, were evaluated by flow cytometry. Arginase-1 (ARG1) quantity and activity was determined in the plasma. Multivariable logistic regression model was used to obtain a diagnostic score to discriminate glioma patients from healthy controls and between each glioma grade. A glioblastoma prognostic model was determined by multiple Cox regression using clinical and myeloid parameters.ResultsChanges in myeloid parameters associated with immune suppression allowed to define a diagnostic score calculating the risk of being a glioma patient. The same parameters, together with age, permit to calculate the risk score in differentiating each glioma grade. A prognostic model for glioblastoma patients stemmed out from a Cox multiple analysis, highlighting the role of MDSC, p-STAT3, and ARG1 activity together with clinical parameters in predicting patient’s outcome.ConclusionsThis work emphasizes the role of systemic immune suppression carried out by myeloid cells in gliomas. The identification of biomarkers associated with immune landscape, diagnosis, and outcome of glioblastoma patients lays the ground for their clinical use.

Abstract Background Systemic and local immune suppression plays a significant role in glioma progression. Glioma microenvironment contains both brain-resident microglial cells (MG) and bone marrow-derived macrophages (BMDM), but the study of their functional and immune regulatory activity has been hampered until now by the lack of markers allowing a proper identification and isolation to collect pure populations. Methods Myeloid and lymphoid infiltrate were characterized in grade II, III and IV gliomas by multicolor flow cytometry, along with the composition of the cell subsets of circulating myeloid cells. Macrophages were sorted and tested for their immunosuppressive ability. Moreover, following preoperative administration of 5-aminolevulinic acid to patients, distinct areas of tumor lesion were surgically removed and analyzed, based on protoporphyrin IX fluorescence emission. Results The immune microenvironment of grade II to grade IV gliomas contains a large proportion of myeloid cells and a small proportion of lymphocytes expressing markers of dysfunctional activity. BMDM and resident MG cells were characterized through a combination of markers, thus permitting their geographical identification in the lesions, their sorting and subsequent analysis of the functional characteristics. The infiltration by BMDM reached the highest percentages in grade IV gliomas, and it increased from the periphery to the center of the lesion, where it exerted a strong immunosuppression that was, instead, absent in the marginal area. By contrast, MG showed little or no suppression. Functional differences, such as iron metabolism and phagocytosis, characterized resident versus blood-derived macrophages. Significant alterations in circulating monocytes were present in grade IV patients, correlating with accumulation of tumor macrophages. Conclusions Grade IV gliomas have an alteration in both circulating and tumor-associated myeloid cells and, differently from grade II and III gliomas, show a significant presence of blood-derived, immune suppressive macrophages. BMDM and MG have different functional properties.

Although several screening tests for recognizing early signs of reading and spelling difficulties have been developed, brief and methodologically grounded tools for teachers are very limited. The present study aimed to lay the foundation for a new screening tool for teachers: the Checklist for early Indicators of risk Factors in Reading Ability (CI-FRA). The proposed checklist consists of 20 items, based on a 7-point Likert scale, and it investigates five domains: reading, writing, attention, and motor skills. Six hundred sixty-seven children were evaluated by 40 teachers during the first year of primary school and, longitudinally, in the second year. Exploratory factor analysis and confirmatory factor analysis (CFA) were applied to verify structural validity. Concurrent validity was assessed by Spearman correlation to analyze the link between CI-FRA and reading and spelling standardized tests and cognitive tests. Reliability was assessed by Cronbach α and interclass correlation coefficient. The CFA reported a three-factor structure as the optimal solution, including language (reading and writing), visuospatial attention, and fine motor skills subscales. Good reliability, good internal consistency, and acceptable test–retest indices were found. Concurrent validity was confirmed by significant correlations between CI-FRA total score and standardized reading and spelling test, as well as by correlations between CI-FRA subscales and neuropsychological standardized test scores. Preliminary evaluation of sensitivity by receiver operating characteristic curves showed that the CI-FRA score has particularly high sensitivity and specificity for word reading speed deficit. In conclusion, the results confirm that CI-FRA is a theoretically grounded and statistically valid tool that could help the teachers to screen for early signs of reading and spelling difficulties.

Although the relationship between developmental dyslexia (DD) and the risk of occurrence of internalizing symptomatology has been widely investigated in the extant literature, different findings have been reported. In this study, two experiments with two general purposes are presented. The first study investigates whether the differences in the severity of internalizing symptoms between DD and controls are greater in students attending secondary school than in those attending primary school. Sixty-five DD and 169 controls attending primary and secondary school took part in the first study. The diagnosis of dyslexia was obtained from standardized reading tests; internalizing symptom severity was assessed with the Self Administrated Psychiatric Scales for Children and Adolescents questionnaire. The results showed that adolescents with dyslexia had an increased level of self-perceived anxiety, depression and somatic symptoms, whereas no significant differences between DD and controls emerged in childhood. In the second study, a cohort of adolescents attending secondary school (DD = 44; controls = 51) was closely analyzed to clarify whether contextual and subjective factors could contribute toward exacerbating the risk of internalizing symptomatology at that age. Internalizing symptom severity was assessed with the Child Behavior Checklist, Youth Self Report questionnaire, decision-making factors were measured with the Melbourne Decision Making Questionnaire, and student’s quality of life was gaged using the Clipper test. The results showed that high levels of internalizing symptoms in DD were associated with a low level of self-esteem and the tendency to react to problematic situations with hyperactivation. By contrast, positive relationships with peers were associated with low symptom severity. In conclusion, the intensified internalizing symptoms that could emerge in adolescents in association with the presence of dyslexia are predicted by social protective and risk factors that are associated with symptom severity. Accordingly, the results suggest that remediation programs for dyslexia should include implementing motivation strategies, self-esteem enhancement activities and building peers networks that, starting in childhood, can prevent the appearance of internalizing symptoms.

We studied the vascular flora of the isthmus of Feniglia, a Nature Reserve in southern Tuscany stretching between the Italian Peninsula and Mt. Argentario. Since the nineteenth century, the area has undergone significant environmental modifications due to intensive grazing and deforestation. Later, rehabilitation interventions were carried out, including reforestation and planting of dune species, making it an interesting protected area to study for its botanical aspects. Therefore, we aim to compile the floristic inventory of this distinctive Mediterranean area by integrating old and new data. The checklist comprises 502 specific and subspecific taxa of vascular plants. The life-form spectrum shows a predominance of therophytes, followed by hemicryptophytes. The chorological spectrum highlights the dominance of Mediterranean species, followed by Euromediterranean and Eurasian species. The presence of 15 species of regional importance, 3 species of the Italian red list and 3 Italian endemics (Ornithogalum exscapum Ten., Limonium multiforme Pignatti, Linaria purpurea (L.) Mill.) is noteworthy. Despite the environmental changes undergone in this area, our study reports the presence of remarkable species, including rare ones such as Atriplex littoralis L., Ruppia spiralis L. ex Dumort., and species at their distribution margin in the Italian Peninsula, such as Staphisagria macrosperma Spach, Juncus sorrentinoi Parl. and Maresia nana (DC.) Batt. Additionally the presence of 26 alien species, of which 15 are invasive, 9 naturalized and 2 casual, indicates a relatively low presence of alien species. Notwithstanding the significant environmental changes that the Feniglia isthmus has experienced in the last century and the high tourist pressure during the summer, the presence of the protected area suggested the effectiveness and benefits of such a form of protection for plant diversity.

Orchidaceae is one of the largest and most diverse families of flowering plants in the world but also one of the most threatened. Climate change is a global driver of plant distribution and may be the cause of their disappearance in some regions. Forest orchids are associated with specific biotic and abiotic environmental factors, that influence their local presence/absence. Changes in these conditions can lead to significant differences in species distribution. We studied three forest orchids belonging to different genera (Cephalanthera, Epipactis and Limodorum) for their potential current and future distribution in a protected area (PA) of the Northern Apennines. A Habitat Suitability Model was constructed for each species based on presence-only data and the Maximum Entropy algorithm (MaxEnt) was used for the modelling. Climatic, edaphic, topographic, anthropogenic and land cover variables were used as environmental predictors and processed in the model. The aim is to identify the environmental factors that most influence the current species distribution and the areas that are likely to contain habitats suitable for providing refuge for forest orchids and ensuring their survival under future scenarios. This will allow PA authorities to decide whether to invest more resources in conserving areas that are potential refuges for threatened species.

In this contribution, new data concerning the distribution of vascular flora alien to Italy are presented. It includes new records, confirmations, exclusions, and status changes for Italy or for Italian administrative regions. Nomenclatural and distribution updates published elsewhere are provided as Suppl. material 1.

In this contribution, new data concerning the distribution of vascular flora alien to Italy are presented. It includes new records, confirmations, exclusions, and status changes for Italy or for Italian administrative regions. Nomenclatural and distribution updates published elsewhere are provided as Suppl. material 1.

In this contribution, new data concerning the distribution of vascular flora alien to Italy are presented. It includes new records, confirmations, exclusions, and status changes for Italy or for Italian administrative regions. Nomenclatural and distribution updates published elsewhere are provided as Suppl. material 1.

In this contribution, new data concerning the distribution of vascular flora alien to Italy are presented. It includes new records, confirmations, exclusions, and status changes for Italy or for Italian administrative regions. Furthermore, three new combinations are proposed. Nomenclatural and distribution updates published elsewhere are provided as Suppl. material 1.

Abstract The split between conservation science and real‐world application is an ongoing issue despite several calls for unification. Researchers are empowered to partially bridge the research‐implementation gap by making their findings more accessible. Cypripedium calceolus is the most recognizable orchid of the European flora, and is currently facing habitat change and fragmentation, in addition to threats from collectors and illegal traders. Although several studies have focused on the ecological and genetic features of the species, a comprehensive account of how such aspects can be translated into concrete conservation recommendations is still missing. In this study, we describe microsatellite genetic variation in 188 individuals from different Italian populations of C. calceolus. Our results indicate the need for immediate conservation action for the most isolated populations in the Central Apennines and north‐western Italy. Although our genetic findings are specific to the Italian populations, our aim is to review ecological and population genetic aspects in C. calceolus and their implications for conservation against the existing threats. Therefore, our detailed guidelines for translocation, habitat management and post‐translocation monitoring can be used to inform conservation strategies in threatened populations of C. calceolus across its range.

In this contribution, new data concerning the distribution of vascular flora alien to Italy are presented. It includes new records, confirmations, exclusions, and status changes for Italy or for Italian administrative regions of taxa in the genera Bunias, Calocedrus, Calycanthus, Celosia, Clerodendrum, Convolvulus, Crassula, Cyclamen, Datura, Dicliptera, Eragrostis, Erigeron, Gamochaeta, Gazania, Impatiens, Kolkwitzia, Leucaena, Ludwigia, Medicago, Muscari, Nigella, Oenothera, Opuntia, Paulownia, Petroselinum, Phyllostachys, Physalis, Pseudosasa, Quercus, Reynoutria, Roldana, Saccharum, Sedum, Semiarundinaria, Senecio, Sisyrinchium, Solanum, Sporobolus, Tulipa, Vachellia, Verbena, and Youngia. Nomenclatural and distribution updates published elsewhere are provided as Suppl. material 1.

In this contribution, new data concerning the distribution of native vascular flora in Italy are presented. It includes new records, confirmations, exclusions, and status changes to the Italian administrative regions for taxa in the genera Ajuga, Chamaemelum, Clematis, Convolvulus, Cytisus, Deschampsia, Eleocharis, Epipactis, Euphorbia, Groenlandia, Hedera, Hieracium, Hydrocharis, Jacobaea, Juncus, Klasea, Lagurus, Leersia, Linum, Nerium, Onopordum, Persicaria, Phlomis, Polypogon, Potamogeton, Securigera, Sedum, Soleirolia, Stachys, Umbilicus, Valerianella, and Vinca. Nomenclatural and distribution updates, published elsewhere, and corrigenda are provided as Suppl. material 1.

In this contribution, new data concerning the distribution of vascular flora alien to Italy are presented. It includes new records, confirmations, and status changes for Italy or for Italian administrative regions of taxa in the genera Abies, Actinidia, Aloe, Amaryllis, Anredera, Arctotheca, Bidens, Cardiospermum, Celosia, Commelina, Cotoneaster, Cyclamen, Eclipta, Euphorbia, Grevillea, Hedera, Hibiscus, Impatiens, Juglans, Kalanchoe, Koelreuteria, Lindernia, Melinis, Myriophyllum, Nandina, Nicotiana, Oenothera, Oxalis, Parthenocissus, Phoenix, Phyllanthus, Physalis, Plumbago, Pteris, Quercus, Setaria, Symphytum, Tagetes, and Washingtonia. Nomenclatural and distribution updates, published elsewhere are provided as Suppl. material 1.

In this contribution, the conservation status assessment of four vascular plants according to IUCN categories and criteria are presented. It includes the assessments of Aurinia leucadea (Guss.) K.Koch, Chondrilla chondrilloides (Ard.) H.Karst., Daphne cneorum L., and Ophioglossum azoricum C.Presl at regional level (Italy).

In this contribution, new data concerning the distribution of native vascular flora in Italy are presented. It includes new records, confirmations and status changes to the Italian administrative regions for taxa in the genera Alchemilla, Arundo, Bupleurum, Clematis, Clinopodium, Cota, Crassula, Cytisus, Euphorbia, Hieracium, Isoëtes, Lamium, Leontodon, Linaria, Lychnis, Middendorfia, Ophrys, Philadelphus, Pinus, Sagina, Sedum, Taeniatherum, Tofieldia, Triticum, Veronica, and Vicia. Nomenclature and distribution updates, published elsewhere, and corrigenda are provided as supplementary material.

In this contribution, new data concerning the distribution of vascular flora alien to Italy are presented. It includes new records, confirmations, exclusions, and status changes for Italy or for Italian administrative regions of taxa in the genera Acalypha, Acer, Canna, Cardamine, Cedrus, Chlorophytum, Citrus, Cyperus, Epilobium, Eucalyptus, Euphorbia, Gamochaeta, Hesperocyparis, Heteranthera, Lemna, Ligustrum, Lycium, Nassella, Nothoscordum, Oenothera, Osteospermum, Paspalum, Pontederia, Romulea, Rudbeckia, Salvia, Sesbania, Setaria, Sicyos, Styphnolobium, Symphyotrichum, and Tradescantia. Nomenclature and distribution updates, published elsewhere, and corrigenda are provided as supplementary material.

In this contribution, new data concerning the distribution of vascular flora alien to Italy are presented. It includes new records, and confirmations for Italy or for Italian administrative regions of taxa in the genera Albizia, Anredera, Bougainvillea, Cardamine, Cenchrus, Cephalaria, Ceratochloa, Cytisus, Datura, Delosperma, Euonymus, Freesia, Hylotelephium, Lantana, Musa, Physalis, Rotala, Styphnolobium, Trachycarpus, and Tradescantia. Nomenclature and distribution updates, published elsewhere, and corrections are provided as supplementary material.

Isoëtes sabatina is an aquatic quillwort endemic to Italy. It is one of the rarest quillworts in Europe, and is critically endangered due to restricted range and to the continuous decline of both population and habitat quality. This study aims to develop an optimized protocol to reproduce and grow I. sabatina sporelings. Mature and immature megaspores were mixed with mature microspores to evaluate the influence of the developmental stage on germination and sporeling development. Two substrates, distilled water and water-agar medium, were tested for germination and sporeling emergence, and three substrates, sand, lake sediment and water-agar, were tested for transplants. A high percentage of megaspore germination (a total of 79.1%) was obtained in both substrates, higher for mature than immature spores. A total of 351 sporelings were produced in distilled water and water-agar cultures, with similar percentages (64.5% and 69.6%, respectively). The development stage of the megaspores affected both germination and sporeling development. Sporeling emergence showed significantly higher percentages in mature megaspores than immature ones (69.6% vs. 11.6%, respectively), with 85% of germinated spores developing sporelings. Only transplants over water-agar medium were successful. This protocol could be useful for the propagation of sporelings as the key step towards the planning of in situ actions to save this Mediterranean quillwort from extinction.

The increasing emphasis on terrestrial orchid conservation has led to conservation actions for a wide range of threatened Mediterranean species. Many terrestrial orchids are currently at great risk for extinction as a result of a multiplicity of threatening processes. We focus on orchid seed germination capabilities in vitro, specifically germination capability of a threatened species, Anacamptis palustris, compared to three other more common species (A. laxiflora, A. morio, and A. papilionacea), and also discuss its potential impact on orchid distribution and conservation. Asymbiotic germination tests were performed with mature seeds using BM-1 medium. In vitro seed germination and protocorm developmental stages were evaluated up to 20 weeks after sowing. Significant differences in seedling development were detected among the species, and a correlation was evident between the rarity of the species and their germinability. Thus, the presence of intrinsic, biological factors that affect and limit the distribution of A. palustris may exist.