Tecnologie fotolitografiche e soft-litografiche per la realizzazione di patterns biomolecolari su superfici antifouling di perfluoropolieteri

LAUREA MAGISTRALEQuesto lavoro di ricerca si propone di realizzare patterns biomolecolari per l’immobilizzazione di globuli rossi infettati dal Plasmodium della malaria su un substrato anti-fouling e fouling-release. Un array di eparina, che risulta altamente selettivo e consente la disposizione ordinata di cellule target sul substrato, è stato depositato su una superficie di perfluoropolietere-uretano-dimetacrilato (PFPE-DMA). Due tipi di materiali sono stati esplorati e le loro performance messe a confronto, decidendo infine di utilizzare un blend composto, 50:50, dai due PFPE-DMAs di diverso peso molecolare che permettesse di mediare i loro vantaggi e svantaggi. Diverse strategie sono state adottate per la realizzazione di tali patterns sfruttando la capacità di biomolecole con gruppi fotoreattivi di legarsi al substrato, contenente gruppi metacrilici reattivi, in seguito all'esposizione alla luce UV: (i) legare, tramite micro-contanct printing, eparina biotinilata ad un pattern sottostante di avidina, a sua volta autoassemblata su un pattern di fotobiotina legato chimicamente al substrato, sfruttando la nota affinità tra avidina e biotina; (ii) eparina funzionalizzata con gruppi metacrilici è stata legata chimicamente, tramite fotolitografia, al substrato. Dispositivi microfluidici sono stati realizzati con due metodologie, la fotolitografia e la replica molding. Sono state inoltre realizzate superfici in PFPE-DMAs con microstrutture, le cui estremità sono state funzionalizzate con patterns biomolecolari di eparina, tramite micro-contact printing. Dispositivi microfluidici possono integrare superfici 2D e 3D funzionalizzate con l’array di eparina permettendo un'immobilizzazione maggiormente selettiva delle cellule target in condizioni dinamiche, nelle quali è possibile sfruttare al massimo le potenzialità fouling-release dei PFPE-DMAs utilizzati. Test cellulari statici effettuati sugli arrays di eparina biotinilata, incubati con un campione di sangue infetto, hanno confermato che le cellule formano agglomerati in corrispondenza degli spots, mentre le proprietà antifouling dei substrati non favoriscono l'adesione aspecifica sul resto della superficie non coperta dal pattern.The aim of this work, is realize heparin array by exploiting photolithografic process and micro-contact-printing in order to capture red blood cells infected by Plasmodium Falciparum (pRBCs), the malaria parasite, onto anti-fouling/fouling release UV-curable perfluoropolyether (PFPE-DMA) substrate. Two PFPE-DMAs materials were evaluated and their performance compared. Finally, it was decided to use a blend composed of two different molecular weight PFPE-DMAs in equal parts. Heparin was chosen because of its role as competitors towards pRBCs, which during the blood phase bind healthy red blood cells, with serious consequences including death. Heparin arrays are highly selective thus allowing an orderly arrangement of target cells on the substrate. Two different strategies were employed for heparin patterning: (i) biotinylated-heparin was linked to avidin patterned on an underlying photobiotin array, chemically linked to the substrate and obtained by μ-contact printing, exploiting the high affinity between biotin and avidin; (ii) methacrylated-heparin was directly bound to the unreacted double bonds of a partially cross-linked PFPE surface through a photo-lithographic process. The first method is characterized by high versatility, indeed any biotinylated molecule may bind the avidin array, and this method offers the possibility of isolating different rare cells. The photolithographic method allows the pattern realization in a single step, because it provides a direct grafting of the biomolecule onto the substrate. The patterned surfaces could be integrated in microfluidic devices. These devices were produced by two methods, photolithography and replica molding. Also microstructured PFPE-DMA surfaces were created, and the microstructure ends were functionalized by micro-contact printing with heparin pattern. Microfluidic devices with integrated surface functionalized with biomolecular baits allow more selective immobilization of target cells under dynamic conditions, thus better exploiting the antifouling and fouling-release PFPE-DMAs properties. Furthermore, the use of microstructured microchannels allows a better interaction biomolecule bait-target cell

Demographic and clinical characteristics of patients involuntarily hospitalized in an Italian psychiatric ward: a 1-year retrospective analysis

Background: In Italy, psychiatric compulsory treatments are regulated by Law 180 of 13-5-1978 that establishes three criteria: 1) acute psychiatric conditions requiring urgent treatment, 2) patient’s refusal of treatment, 3) inpatient treatment is necessary and cannot be postponed. Aim: To highlight demographic and clinical risk factors for involuntary treatments. Methods: We retrospectively collected all hospitalizations in the Service of Psychiatric Diagnosis and Treatment of a northern Italian town from 1-1-2015 to 31-12-2015. We statistically compared demographic and clinical variables related to voluntarily and involuntarily admitted patients and their hospitalizations. Results: We divided our sample into patients voluntarily hospitalized (PVH=236) and involuntarily (PIH=160) according to their voluntary (VH= 304) and involuntary (IH=197) hospitalizations. PIH were older than PVH and, more frequently, lived alone and were unemployed (p<0.001). “Acute worsening of psychopathology” for IH and “Suicidality” for VH were the prevalent reasons (p<0.001). IH was longer than VH (p<0.001). Among PIH, the most frequent diagnoses were “Schizophrenia and Other Psychosis” (ICD-9-CM) and “Ineffective Impulse Control + Disturbed Personal Identity” (NANDA-I) (p<0.001). During hospitalizations, PIH more often than PVH presented aggressive behavior (p<0.001). At discharge, PIH were more frequently sent to another psychiatric ward or protected facility with long-acting injectable antipsychotics (p<0.001). Conclusions: Our involuntarily admitted patients were affected by severe psychiatric disorders with social maladjustment and required complex therapeutic and rehabilitative programs to counteract aggressive behaviour, poor therapeutic compliance and prolonged hospitalizations. The assessment of patients’ characteristics can help clinicians recognize who are at risk for compulsory treatment and prevent it

Minority stress and mental health among LGBT populations: an update on the evidence

INTRODUCTION: In the past five years, researchers have increasingly turned to the study of mental health outcomes in LGBT populations. The present paper summarizes recent literature on the relationship between minority stress experienced by sexual minorities and mental health. EVIDENCE ACQUISITION: PsycINFO, PubMed, and the EBSCO Psychology and Behavioral Science Collection were searched for papers concerning minority stress and mental health disparities in LGBT populations, published between 1 January 2014 and 30 June 2018. All collected papers were screened using the following criteria: study involving >50 individuals; written in English; focusing on clinical outcomes of depression, suicidality, and substance use in relation to experienced minority stress. EVIDENCE SYNTHESIS: Sixty-two papers were included in this review. Findings are reported under three main headings: studies primarily focused on depression, studies concerning suicidality and suicide attempts, and papers analyzing the correlation between substance use and minority stress in LGBT populations. The included studies supported the minority stress model as a framework to better explain disparities in mental health outcomes in sexual minority populations. Higher rates of depression, suicidality, and substance use are reported in LGBT populations, as are the related minority stressors analyzed. CONCLUSIONS: Sexual minorities still face numerous mental health disparities. Research indicates that the levels of minority stressors positively predict mental health outcomes. Specific policies designed to support the civil rights of sexual minorities may help to overcome such inequalities

Early death in Munchausen Syndrome: a case report

This case contributes to raising awareness and understanding of the complex clinical presentations of Munchausen syndrome (MS). Education of staff to the seriousness and genuineness of this disorder should be implemented, especially in hospital units, in order to detect such complex clinical situations quickly and accurately, preventing iatrogenic risks

The efficacy of interprofessional simulation in improving collaborative attitude between nursing students and residents in medicine. A study protocol for a randomised controlled trial

Background: Effective collaboration among health professionals is an essential component to ensure quality of care. Many adverse events experienced by patients are attributed to misunderstanding or poor communication among members of the interprofessional team. Interprofessional simulation is a learning strategy used to improve collaboration and facilitate communication between medical and nursing students. Aim of the work: To determine the efficacy of educational program based on high-fidelity interprofessional simulation aimed at improving collaborative attitude. Method: For this purpose, a protocol for a planned single-center, non-blinded and Randomized Controlled Trial (RCT) was chosen. The present has been approved by the Ethics Committee of Area Vasta Emilia Nord (Italy) (n° 479/2018). All students attending the second and third year of nursing and all resident physicians in anesthesia, reanimation, intensive care and pain management of University of Modena and Reggio Emilia, will be recruited and randomly assigned to two groups. The Experimental Group (EG) will receive an educational intervention based on high-fidelity simulation and the Control Group (CG) will attend a traditional classroom lesson. Jefferson Scale of Attitudes toward Physician-Nurse Collaboration (JSAPNC) and Readiness for Interprofessional Learning Scale (RIPLS) will be administered before and after the educational program in both the EG and CG. Conclusion: Expected outcomes is that, at the end of the study, nursing students and resident physicians who participated in the interprofessional simulation show significantly higher levels of interprofessional collaboration compared to the CG, evaluated through the JSAPNC

An RNA-seq study in Friedreich ataxia patients identified hsa-miR-148a-3p as a putative prognostic biomarker of the disease

friedreich ataxia (FRDA) is a life-threatening hereditary ataxia; its incidence is 1:50,000 individuals in the caucasian population. a unique therapeutic drug for FRDA, the antioxidant omaveloxolone, has been recently approved by the US food and drug administration (FDA). FRDA is a multi-systemic neurodegenerative disease; in addition to a progressive neurodegeneration, FRDA is characterized by hypertrophic cardiomyopathy, diabetes mellitus and musculoskeletal deformities. cardiomyopathy is the predominant cause of premature death. the onset of FRDA typically occurs between the ages of 5 and 15. given the complexity and heterogeneity of clinical features and the variability of their onset, the identification of biomarkers capable of assessing disease progression and monitoring the efficacy of treatments is essential to facilitate decision making in clinical practice. we conducted an RNA-seq analysis in peripheral blood mononuclear cells from FRDA patients and healthy donors, identifying a signature of small non-coding RNAs (sncRNAs) capable of distinguishing healthy individuals from the majority of FRDA patients. among the differentially expressed sncRNAs, microRNAs are a class of small non-coding endogenous RNAs that regulate posttranscriptional silencing of target genes. In FRDA plasma samples, hsa-miR-148a-3p resulted significantly upregulated. the analysis of the receiver operating characteristic (ROC) curve, combining the circulating expression levels of hsa-miR-148a-3p and hsa-miR-223-3p (previously identified by our group), revealed an area under the curve (AUC) of 0.86 (95%, confidence Interval 0.77-0.95; p-value &lt; 0.0001). an in silico prediction analysis indicated that the IL6ST gene, an interesting marker of neuroinflammation in FRDA, is a common target gene of both miRNAs. our findings support the evaluation of combined expression levels of different circulating miRNAs as potent epi-biomarkers in FRDA. moreover, we found hsa-miR-148a-3p significantly over-expressed in Intermediate and late-onset friedreich ataxia patients' group (IOG and LOG, respectively) compared to healthy individuals, indicating it as a putative prognostic biomarker in this pathology

2022-RA-767-ESGO Machine learning to implement the accuracy of magnetic resonance imaging (MRI) in the detection of lymph node metastasis in patients with locally advance cervical cancer treated with neoadjuvant chemotherapy

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The rapid spread of SARS-COV-2 Omicron variant in Italy reflected early through wastewater surveillance

The SARS-CoV-2 Omicron variant emerged in South Africa in November 2021, and has later been identified worldwide, raising serious concerns. A real-time RT-PCR assay was designed for the rapid screening of the Omicron variant, targeting characteristic mutations of the spike gene. The assay was used to test 737 sewage samples collected throughout Italy (19/21 Regions) between 11 November and 25 December 2021, with the aim of assessing the spread of the Omicron variant in the country. Positive samples were also tested with a real-time RT-PCR developed by the European Commission, Joint Research Centre (JRC), and through nested RT-PCR followed by Sanger sequencing. Overall, 115 samples tested positive for Omicron SARS-CoV-2 variant. The first occurrence was detected on 7 December, in Veneto, North Italy. Later on, the variant spread extremely fast in three weeks, with prevalence of positive wastewater samples rising from 1.0% (1/104 samples) in the week 5–11 December, to 17.5% (25/143 samples) in the week 12–18, to 65.9% (89/135 samples) in the week 19–25, in line with the increase in cases of infection with the Omicron variant observed during December in Italy. Similarly, the number of Regions/Autonomous Provinces in which the variant was detected increased fromone in the first week, to 11 in the second, and to 17 in the last one. The presence of the Omicron variant was confirmed by the JRC real-time RT-PCR in 79.1% (91/115) of the positive samples, and by Sanger sequencing in 66% (64/97) of PCR amplicons

The rapid spread of SARS-COV-2 Omicron variant in Italy reflected early through wastewater surveillance

The SARS-CoV-2 Omicron variant emerged in South Africa in November 2021, and has later been identified worldwide, raising serious concerns. A real-time RT-PCR assay was designed for the rapid screening of the Omicron variant, targeting characteristic mutations of the spike gene. The assay was used to test 737 sewage samples collected throughout Italy (19/21 Regions) between 11 November and 25 December 2021, with the aim of assessing the spread of the Omicron variant in the country. Positive samples were also tested with a real-time RT-PCR developed by the European Commission, Joint Research Centre (JRC), and through nested RT-PCR followed by Sanger sequencing. Overall, 115 samples tested positive for Omicron SARS-CoV-2 variant. The first occurrence was detected on 7 December, in Veneto, North Italy. Later on, the variant spread extremely fast in three weeks, with prevalence of positive wastewater samples rising from 1.0% (1/104 samples) in the week 5-11 December, to 17.5% (25/143 samples) in the week 12-18, to 65.9% (89/135 samples) in the week 19-25, in line with the increase in cases of infection with the Omicron variant observed during December in Italy. Similarly, the number of Regions/Autonomous Provinces in which the variant was detected increased from one in the first week, to 11 in the second, and to 17 in the last one. The presence of the Omicron variant was confirmed by the JRC real-time RT-PCR in 79.1% (91/115) of the positive samples, and by Sanger sequencing in 66% (64/97) of PCR amplicons. In conclusion, we designed an RT-qPCR assay capable to detect the Omicron variant, which can be successfully used for the purpose of wastewater-based epidemiology. We also described the history of the introduction and diffusion of the Omicron variant in the Italian population and territory, confirming the effectiveness of sewage monitoring as a powerful surveillance tool

The clinical effectiveness of an integrated multidisciplinary evidence-based program to prevent intraoperative pressure injuries in high-risk children undergoing long-duration surgical procedures: a quality improvement study

The prevention of hospital-acquired pressure injuries (HAPIs) in children undergoing long-duration surgical procedures is of critical importance due to the potential for catastrophic sequelae of these generally preventable injuries for the child and their family. Long-duration surgical procedures in children have the potential to result in high rates of HAPI due to physiological factors and the difficulty or impossibility of repositioning these patients intraoperatively. We developed and implemented a multi-modal, multi-disciplinary translational HAPI prevention quality improvement program at a large European Paediatric University Teaching Hospital. The intervention comprised the establishment of wound prevention teams, modified HAPI risk assessment tools, specific education, and the use of prophylactic dressings and fluidized positioners during long-duration surgical procedures. As part of the evaluation of the effectiveness of the program in reducing intraoperative HAPI, we conducted a prospective cohort study of 200 children undergoing long-duration surgical procedures and compared their outcomes with a matched historical cohort of 200 children who had undergone similar surgery the previous year. The findings demonstrated a reduction in HAPI in the intervention cohort of 80% (p &lt; 0.01) compared to the comparator group when controlling for age, pathology, comorbidity, and surgical duration. We believe that the findings demonstrate that it is possible to significantly decrease HAPI incidence in these highly vulnerable children by using an evidence-based, multi-modal, multidisciplinary HAPI prevention strategy