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Alessandro Paolini,1 Giulia Battafarano,1 Valentina D’Oria,1 Francesco Mura,2 Simona Sennato,3 Valentina Mussi,4 Roberta Risoluti,5 Stefano Materazzi,5 Andrea Del Fattore,1 Andrea Masotti1 1Bambino Gesù Children’s Hospital, IRCCS, Research Laboratories, Rome 00146, Italy; 2Center for Nanotechnology for Engineering (CNIS), Sapienza University of Rome, Rome 00185, Italy; 3CNR-ISC UOS Sapienza and Physics Department, Sapienza University of Rome, Rome 00185, Italy; 4National Research Council, Institute for Microelectronics and Microsystems IMM-CNR, Roma 00133, Italy; 5Department of Chemistry, Sapienza University of Rome, Rome 00185, ItalyCorrespondence: Andrea MasottiBambino Gesù Children’s Hospital, IRCCS, Research Laboratories, Gene Expression-Microarrays Laboratory, Viale di San Paolo 15, Rome 00146, ItalyTel +39-06-68592650Fax +39-0668592904Email andrea.masotti@opbg.netBackground: The advent of 3D printing technology allowed the realization of custom devices that can be used not only in the everyday life but also in the nanotechnology and biomedical fields. In nanotechnology, the use of bi-dimensional nanostructures based on carbon nanotubes, generally referred as buckypapers, have received considerable attention for their versatility and potential application in many biomedical fields. Unfortunately, buckypapers are extremely hydrophobic and cannot be used in aqueous media to culture cells.Methods: A polymeric device able to accommodate buckypapers and facilitate cell growth was fabricated by using 3D printing technology. We imparted hydrophilicity to buckypapers by coating them with polyamidoamine (PAMAM) dendrimers.Results: We found that by using novel techniques such as polymer coating the buckypaper hydrophilicity increased, whereas the use of 3D printing technology allowed us to obtain custom devices that have been used to culture cells on buckypapers for many days. We characterized in details the morphology of these structures and studied for the first time the kinetic of cell proliferation. We found that these scaffolds, if properly functionalized, are suitable materials to grow cells for long time and potentially employable in the biomedical field.Conclusion: Although these materials are cytotoxic under certain circumstances, we have found a suitable coating and specific experimental conditions that encourage using buckypapers as novel scaffolds for cell growth and for potential applications in tissue repair and regeneration.Keywords: 3D printing, buckypaper, PAMAM dendrimer, cell proliferation, tissue regeneration, transfection

Antonella Celluzzi,1,* Alessandro Paolini,1,* Valentina D’Oria,1 Roberta Risoluti,2 Stefano Materazzi,2 Marco Pezzullo,1 Stefano Casciardi,3 Simona Sennato,4 Federico Bordi,4 Andrea Masotti1 1Bambino Gesù Children’s Hospital, IRCCS, Research Laboratories, 2Department of Chemistry, Sapienza University of Rome, 3Department of Occupational and Environmental Medicine, Epidemiology and Hygiene, National Institution for Insurance Against Accidents at Work (INAIL Research), Monte Porzio Catone, 4CNR-ISC UOS Roma, Department of Physics, Sapienza University of Rome, Roma, Italy *These authors contributed equally to this work Abstract: Recent findings in nanomedicine have revealed that carbon nanotubes (CNTs) can be used as potential drug carriers, therapeutic agents and diagnostics tools. Moreover, due to their ability to cross cellular membranes, their nanosize dimension, high surface area and relatively good biocompatibility, CNTs have also been employed as a novel gene delivery vector system. In our previous work, we functionalized CNTs with two polyamine polymers, polyethyleneimine (PEI) and polyamidoamine dendrimer (PAMAM). These compounds have low cytotoxicity, ability to conjugate microRNAs (such as miR-503) and, at the same time, transfect efficiently endothelial cells. The parameters contributing to the good efficiency of transfection that we observed were not investigated in detail. In fact, the diameter and length of CNTs are important parameters to be taken into account when evaluating the effects on drug delivery efficiency. In order to investigate the biophysical and biological contributions of polymer-coated CNTs in delivery of miRNAs to human cells, we decided to investigate three different preparations, characterized by different dimensions and aspect ratios. In particular, we took into account very small CNTs, a suspension of CNTs starting from the commercial product and a 2D material based on CNTs (ie, buckypapers [BPs]) to examine the transfection efficiency of a rigid scaffold. In conclusion, we extensively investigated the biophysical and biological contributions of polyamine-coated CNTs and bidimensional BPs in the delivery of miRNAs to human cells, in order to optimize the transfection efficiency of these compounds to be employed as efficient drug delivery vectors in biomedical applications. Keywords: carbon nanotubes, buckypapers, polyamines, cytotoxicity, cell transfection

Narges Aberuyi,1 Soheila Rahgozar,1 Zohreh Khosravi Dehaghi,1 Alireza Moafi,2 Andrea Masotti,3,* Alessandro Paolini3,* 1Department of Biology, Faculty of Science, University of Isfahan, 2Department of Pediatric-Hematology-Oncology, Sayed-ol-Shohada Hospital, Isfahan University of Medical Sciences, Isfahan, Iran; 3Gene Expression – Microarrays Laboratory, Bambino Gesù Children’s Hospital-Istituto di Ricovero e Cura a Carattere Scientifico (IRCCS), Rome, Italy *These authors contributed equally to the manuscript Purpose: The aim of this work was to study the correlation between the expressions of the ABCA2 and ABCA3 genes at the mRNA and protein levels in children with acute lymphoblastic leukemia (ALL) and the effects of this association on multidrug resistance (MDR).Materials and methods: Sixty-nine children with de novo ALL and 25 controls were enrolled in the study. Mononuclear cells were isolated from the bone marrow. The mRNA levels of ABCA2 and ABCA3 were measured by real-time polymerase chain reaction (PCR). Samples with high mRNA levels were assessed for respective protein levels by Western blotting. Following the first year of treatment, persistent monoclonality of T-cell gamma receptors or immunoglobulin H (IgH) gene rearrangement was assessed and considered as the MDR. The tertiary structure of ABCA2 was predicted using Phyre2 and I-TASSER web systems and compared to that of ABCA3, which has been previously reported. Molecular docking was performed using DOCK 6.7.Results: Real-time quantitative PCR (qRT-PCR) showed high levels of ABCA2 and ABCA3 mRNAs in 13 and 17 samples, respectively. Among them, five and eight individuals demonstrated high levels of ABCA2 and ABCA3, respectively. Response to chemotherapy was significantly decreased (P=0.001) when the mRNA and protein of both genes were overexpressed compared to individuals with high transcriptional levels of either ABCA2 or ABCA3 alone. Close similarity between ABCA2 and ABCA3 structures was revealed by protein tertiary structure prediction, whereas molecular docking analysis suggested similar binding of chemotherapy drugs and therefore a potentially similar role in determining the MDR.Conclusion: Our findings suggested, for the first time, that quantification of the protein level of ABCA2 and ABCA3 transporters had a prognostic impact on pediatric ALL MDR. Furthermore, the tertiary structure of ABCA2 was predicted for the first time, and docking analysis revealed a possible compensatory effect between ABCA2 and ABCA3 transporters, which may contribute to the efflux of cytotoxic drugs and, ultimately, to chemoresistance. Keywords: ABCA2 transporter, ABCA3 transporter, multidrug resistance, childhood acute lymphoblastic leukemia, molecular docking, tertiary structure

Celluzzi A, Paolini A, D’Oria V, et al. Int J Nanomedicine. 2017;13:1–18.On page 13, an Acknowledgment section was missing. This should have read as:AcknowledgmentThis work received funding from the Italian Ministry of Health (Progetto Ricerca Finalizzata PE-2011–02347026).Read the original article.

Background and Aims: Hypoalbuminemia, as defined by serum albumin (SA) levels ≤35 g/L, is associated to venous and arterial thrombosis in general population and in patients at risk of cardiovascular disease. It is unknown if SA ≤35 g/L is also associated to portal vein thrombosis (PVT) in cirrhosis. Methods: Cirrhotic patients enrolled in the Portal vein thrombosis Relevance On Liver cirrhosis: Italian Venous thrombotic Events Registry (PRO-LIVER) study (n = 753), were followed-up for 2 years to assess the risk of PVT, that was diagnosed by Doppler ultrasonography. Child-Pugh classes, Model for End-Stage Liver Disease score, presence of hepatocellular carcinoma and laboratory variables including SA, D-dimer, and high-sensitivity C-reactive protein (hs-CRP) were measured at baseline. Results: SA ≤35 g/L was detected in 52% of patients. A logistic multivariate regression analysis showed that higher Child-Pugh class, hepatocellular carcinoma and thrombocytopenia were significantly associated to SA ≤35 g/L. In a subgroup of patients where data regarding hs-CRP and D-dimer were available, SA ≤35 g/L was inversely associated with hs-CRP and D-dimer. During the follow-up, a total of 61 patients experienced PVT. A Kaplan Meier survival analysis showed SA ≤35 g/L was associated to increased risk of PVT compared to SA >35 g/L (P = .005). A multivariate Cox proportional hazards regression analysis showed that male sex, lower platelet count, and SA ≤35 g/L remained associated to PVT after adjusting for confounding factors. Conclusion: Cirrhotic patients with SA ≤35 g/L are at higher risk of experiencing PVT compared to those with SA >35 g/L and could be considered as potential candidates to anticoagulant prophylaxis for PVT prevention.

Abstract Background To test whether known prognosticators of COVID-19 maintained their stratification ability across age groups. Methods We performed a retrospective study. We included all patients (n = 2225), who presented to the Emergency Department of the Careggi University Hospital for COVID-19 in the period February 2020—May 2021, and were admitted to the hospital. The following parameters were analyzed as dichotomized: 1) SpO2/FiO2 ≤ or > 214; 2) creatinine 82). The primary end-point was in-hospital mortality. Results By the univariate analysis, the aforementioned dichotomized variables demonstrated a significant association with in-hospital mortality in all subgroups. We introduced them in a multivariate model: in G1 SpO2/FiO2 ≤ 214 (Relative Risk, RR 15.66; 95%CI 3.98–61,74), in G2 creatinine ≥ 1.1 mg/L (RR 2.87, 95%CI 1.30–6.32) and LDH ≥ 250 UI/L (RR 8.71, 95%CI 1,15–65,70), in G3 creatinine ≥ 1.1 mg/L (RR 1.98, 95%CI 1,17–3.36) and CRP ≥ 60 ng/L (RR 2.14, 95%CI 1.23–3.71), in G4 SpO2/FiO2 ≤ 214 (RR 5.15, 95%CI 2.35–11.29), creatinine ≥ 1.1 mg/L (RR 1.75, 95%CI 1.09–2.80) and CRP ≥ 60 ng/L (RR 1.82, 95%CI 1.11–2.98) were independently associated with an increased in-hospital mortality. Conclusions A mild to moderate respiratory failure showed an independent association with an increased mortality rate only in youngest and oldest patients, while kidney disease maintained a prognostic role regardless of age.

Sustainable food production implements circular economic system, valuing side streams and minimizing waste. This study was aimed to develop a new food by fermenting a blend of dehulled sunflower seed protein powder (SSPP) and reconstituted bovine sweet whey powder (RSWP). Blends were inoculated with Lactococcus lactis B12 alone or in association with Saccharomyces cerevisiae L12, and fermentation proceeded until reaching pH 4.8. After in vitro static gastrointestinal digestion, RSWP and SSPP proteins were highly proteolyzed and the soluble nitrogen content was 69–71% of total nitrogen. In digests, 42–75 unique peptides were identified, and most of them weighed 500–1000 Da. Free amino acids accounted for 202–228 mg/g protein in digests. Few bioactive peptides derived from RSWP were identified. These findings demonstrated strong degradability of RSWP and SSPP proteins during digestion and shed light on nutritional properties exploitable for food applications of the developed fermented blend.

Abstract Background Paediatric-type diffuse High-Grade Gliomas (PDHGG) are highly heterogeneous tumours which include distinct cell sub-populations co-existing within the same tumour mass. We have previously shown that primary patient-derived and optical barcoded single-cell-derived clones function as interconnected networks. Here, we investigated the role of exosomes as a route for inter-clonal communication mediating PDHGG migration and invasion. Results A comprehensive characterisation of seven optical barcoded single-cell-derived clones obtained from two patient-derived cell lines was performed. These analyses highlighted extensive intra-tumour heterogeneity in terms of genetic and transcriptional profiles between clones as well as marked phenotypic differences including distinctive motility patterns. Live single-cell tracking analysis of 3D migration and invasion assays showed that the single-cell-derived clones display a higher speed and longer travelled distance when in co-culture compared to mono-culture conditions. To determine the role of exosomes in PDHGG inter-clonal cross-talks, we isolated exosomes released by different clones and characterised them in terms of marker expression, size and concentration. We demonstrated that exosomes are actively internalized by the cells and that the inhibition of their biogenesis, using the phospholipase inhibitor GW4689, significantly reduced the cell motility in mono-culture and more prominently when the cells from the clones were in co-culture. Analysis of the exosomal miRNAs, performed with a miRNome PCR panel, identified clone-specific miRNAs and a set of miRNA target genes involved in the regulation of cell motility/invasion/migration. These genes were found differentially expressed in co-culture versus mono-culture conditions and their expression levels were significantly modulated upon inhibition of exosome biogenesis. Conclusions In conclusion, our study highlights for the first time a key role for exosomes in the inter-clonal communication in PDHGG and suggests that interfering with the exosome biogenesis pathway may be a valuable strategy to inhibit cell motility and dissemination for these specific diseases.

Background: A few months after the COVID-19 pandemic onset, knowledge of SARS-CoV-2 infection and outcomes and treatments blew up. This paper aimed to evaluate the features of a Tuscany COVID-19 hospitalized cohort and to identify risk factors for COVID-19 severity. Methods: This retrospective observational COVID-19 cohort study (1 March 2020–1 March 2021) was conducted on patients ≥ 18 years old, admitted to Tuscany Hospital, and subjected to follow-up within 12 months after discharge. Patients were enrolled at Pisana, Senese and Careggi University Hospitals, and South East, North West, and Center Local Hospitals. Results: 2888 patients (M = 58.5%, mean age = 66.2 years) were enrolled, of whom 14.3% (N = 413) were admitted to an intensive care unit. Smokers were 25%, and overweight and obese 65%. The most used drugs were corticosteroids, antacids, antibiotics, and antithrombotics, all antiviral drugs, with slight differences between 2020 and 2021. A strong association was found between outcomes of evolution towards critical COVID-19 (non-invasive mechanical ventilation (NIV) and/or admission to intensive care) and smoking (RR = 4.91), ex-smoking (RR = 3.48), overweight (RR = 1.30), obese subjects (RR = 1.62), comorbidities (aRR = 1.38). The alteration of liver enzymes (aspartate aminotransferase, alanine aminotransferase, or gamma-glutamyl transpeptidase) was associated with NIV (aOR = 2.28). Conclusions: Our cohort, characterized by patients with a mean age of 66.2 years, showed 65% of patients were overweight and obese. Smoking/ex-smoking, overweight/obesity, and other comorbidities were associated with COVID-19 adverse outcomes. The findings also demonstrated that alterations in liver enzymes were associated with worse outcomes.

Background: Body image perception can significantly influence various aspects of adolescent lives. The study analyzed the relationship between body image satisfaction and sports participation in adolescents, examining various factors that contribute to body image concerns and their implications for sports engagement. Methods: A total of 237 schoolchildren were recruited from lower secondary Italian schools. Anthropometric characteristics were measured directly. Assessment of body image perception was performed using Body Silhouette Charts for preadolescent children. The Italian version of the International Physical Activity Questionnaire for Adolescents questionnaire was administered to assess physical activity (PA) levels at school and during leisure time in the last 7 days. Analysis of variance was used to test differences in PA levels, while multiple regression models were carried out to assess possible predictors of body dissatisfaction. Results: In total, 42.6% of children were not satisfied with their figure and 23.2% were very dissatisfied; among them, the vast majority would have liked to be thinner. The dissatisfaction and satisfaction were similar in boys and girls. The frequency of satisfaction with one’s body image was higher in children who practiced extracurricular sports compared to those who did not. The percentage of dissatisfaction with one’s body image was similar in the two groups, but the frequency of children being very dissatisfied was double in the group that does not practice extracurricular sport (31.2% vs. 17.7%). Body dissatisfaction increases with increasing body mass index in both genders but decreases in children involved in extracurricular sports. Conclusions: Encouraging teenagers to engage in regular physical activity should be a key component of therapies supporting positive body image. This study found a relationship between extracurricular sports and body image satisfaction, suggesting that physical activity protects teenagers’ body image satisfaction.

Abstract Background Fibroblast growth factor-23 (FGF23) has been associated to left ventricular (LV) hypertrophy and heart failure (HF) severity. We aimed to investigate the clinical correlates and prognostic value of intact FGF23 (iFGF23) in HF patients. Methods Patients with stable HF and left ventricular ejection fraction (LVEF) 50.9 pg/mL. LVEF decreased from the first iFGF23 tertile to the third tertile (p = 0.014). N-terminal pro-B-type natriuretic peptide (NT-proBNP) increased from the first to the third tertile (p = 0.001), while peak oxygen consumption decreased (p

Abstract Bed bug has become a major public health pest worldwide. Infestation may result in numerous negative health effects. Homeless shelters are one of the most habitats that can be infested with bed bugs, a few studies have focused on bed bug infestations in these settings. We conducted a survey of infestations of bed bugs in a homeless shelter in southern France, using an innovative seven-level scale (0–6) to assess the degree of infestation, MALDI TOF-MS to identify bed bugs, and a biomolecular tool to detect bacteria. Bed bug infestations were documented in 13% (9/68) of investigated rooms. A total of 184 bed bugs were collected and morphologically identified as Cimex lectularius. MALDI TOF–MS analysis allowed us to obtain high-quality MS spectra for all 184 specimens, to correctly identify all specimens, and included 178/184 (97%) Log Score Values higher than 1.8. Among the bacteria tested, Wolbachia sp. DNA was found in 149/184 (81%) of the bed bugs, and one sample was positive for Coxiella burnetii, the agent of Q fever. Our study is the first of its kind that offers new perspectives for increasing public awareness of the conditions in homeless shelters.

The measurement of partial pressures of oxygen (O2) and carbon dioxide (CO2) is fundamental for evaluating a patient’s conditions in clinical practice. There are many ways to retrieve O2/CO2 partial pressures and concentrations. Arterial blood gas (ABG) analysis is the gold standard technique for such a purpose, but it is invasive, intermittent, and potentially painful. Among all the alternative methods for gas monitoring, non-invasive transcutaneous O2 and CO2 monitoring has been emerging since the 1970s, being able to overcome the main drawbacks of ABG analysis. Clark and Severinghaus electrodes enabled the breakthrough for transcutaneous O2 and CO2 monitoring, respectively, and in the last twenty years, many innovations have been introduced as alternatives to overcome their limitations. This review reports the most recent solutions for transcutaneous O2 and CO2 monitoring, with a particular consideration for wearable measurement systems. Luminescence-based electronic paramagnetic resonance and photoacoustic sensors are investigated. Optical sensors appear to be the most promising, giving fast and accurate measurements without the need for frequent calibrations and being suitable for integration into wearable measurement systems.

Abstract Background It is important for non-Native persons to understand that the meaning of culture to Native American/Indigenous Peoples is not about esteem, taste or music but rather is described as a cognitive map on how to be. Native American/Indigenous culture can be thought of as all the things and ways in which Native/Indigenous people understand who they are, where they come from and how they are to interact with others. Hundreds of years across many generations have taught that culture-based activities and interventions improve Native/Indigenous health and wellbeing. We explore if increased Native American culture/cultural connectedness is associated with better mental health/well-being and physical health. Methods We analyzed data from a two-phased study (N = 259 and N = 102) of 361 urban Native Americans in California (2018–2021). The 29 items validated Cultural Connectedness Scale-California (CCS-CA) measured Native culture/cultural connectedness. Mental health/well-being and physical health were assessed using the: modified Herth Hope Index (mHHI), Satisfaction with Life (SWL), Center for Epidemiologic Studies Depression Scale-Revised (CESD-R-10), Substance Abuse (CAGE-AID), and Health Related Quality of Life (HRQOL). We conducted Pearson correlations and stepwise regression analyses with CCS-CA as the independent (predictor) variable to explore our main research questions: 1) Is increased Native American/Indigenous culture associated with: 1) better mental health/well-being; and 2) better physical health? Results Increased Native/Indigenous culture (CCS-CA scores) is significantly associated with better mental health/well-being (mHHI, p

Abstract In human cells BRAF oncogene is invariably expressed as a mix of two coding transcripts: BRAF-ref and BRAF-X1. These two mRNA isoforms, remarkably different in the sequence and length of their 3′UTRs, are potentially involved in distinct post-transcriptional regulatory circuits. Herein, we identify PARP1 among the mRNA Binding Proteins that specifically target the X1 3′UTR in melanoma cells. Mechanistically, PARP1 Zinc Finger domain down-regulates BRAF expression at the translational level. As a consequence, it exerts a negative impact on MAPK pathway, and sensitizes melanoma cells to BRAF and MEK inhibitors, both in vitro and in vivo. In summary, our study unveils PARP1 as a negative regulator of the highly oncogenic MAPK pathway in melanoma, through the modulation of BRAF-X1 expression.

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