I diritti fondamentali nel sistema di compensazione italiano e nel contesto europeo

El presente trabajo de investigación pretende analizar aquellos elementos perturbadores que se interponen en el camino de la protección de la persona, en el caso de que la misma sufra daños no patrimoniales contra su integridad física y psíquica o contra su propia vida, daños por los cuales pretenda obtener una compensación. En primer lugar, se han analizado de forma preliminar aquellas cuestiones relacionadas con la evaluación efectiva del daño inmaterial. Los problemas, rectius las categorías de problemas, cuya investigación se hace necesaria, con el fin de un estudio más completo del daño biológico y de sus hermanos menores, son esencialmente dos: identificar el número de derechos realmente compensables y los límites que esta protección encuentra en la relación con agentes externos, en particular, los límites que se derivan del uso (o abuso) que los titulares de estos derechos hacen de ellos. A continuación, se ha procedido con la investigación de algunas de las diferentes categorías jurisprudenciales de indemnización por "daño a la persona": la ratio radica en el deseo de determinar los problemas críticos detrás de la aplicación práctica de las categorías de indemnización por daños biológicos, morales, existenciales, parentales etc… en caso de indemnización por daños que sean necesarios debido a una lesión de carácter no pecuniario. El propósito, de hecho, es verificar si, en estos casos particulares, las fórmulas de compensación mencionadas y los criterios para la liquidación del daño adoptado en los procedimientos civiles son válidos para garantizar a la víctima la compensación total y efectiva del daño sufrido, o si, en cambio, el principio de reparación integral da paso a un intento de reductio ad unum de los posibles elementos de daño. A partir de la investigación del contexto antes mencionado, ha sido luego posible identificar los fallos del sistema de compensación y de los criterios adoptados en sede civil para liquidar el daño que, de hecho, no permiten la protección total de la víctima del daño. Las operaciones descritas anteriormente, en realidad, han sido todas preliminares a un segundo paso de investigación, que ha consistido en la formulación de hipótesis de soluciones jurisprudenciales o legislativas que se aplicarán a nivel supranacional, a fin de proteger mejor a la parte perjudicada en el presente caso. De hecho, la perspectiva adoptada en el análisis plantea la necesidad de cruzar las fronteras nacionales para alcanzar la identificación de posibles perfiles de indemnización civil en el contexto europeo, dado que la protección de lesiones personales en ámbito civil requeriría un nivel de cumplimiento e implementación que basa sus raíces en una dimensión supranaciona

Intelligenza artificiale e responsabilitá civile: le nuove sfide in ambito sanitario

Ad ogni generazione di giuristi tocca il compito di misurare l’impatto della tecnologia sulle categorie ordinanti del diritto privato. Il giurista del primo ‘900 si è trovato a dover registrare i mutamenti che la motorizzazione del traffico stradale ha provocato sulla responsabilità civile, quello del secondo ‘900 ha dovuto fare i conti con le conseguenze che la diffusione di modelli riproduttivi di massa ha generato sul contratto; alla generazione di giuristi dell’ultima porzione del secolo scorso è toccato indagare il ruolo della digitalizzazione telematica sul contratto e sul diritto d’autore. Al giurista odierno spetta forse l’onere maggiore, quello di neutralizzare l’impatto che l’intelligenza artificiale sembra in grado di determinare su ogni ramo del diritto, con un grado di pervasività senza precedenti. In particolare, le strutture della responsabilità civile appaiono, a confronto con gli strumenti dell’intelligenza artificiale, spesso inadatte a rispondere efficacemente ai problemi che l’uso della robotica pone, specie se volgiamo lo sguardo all’ambito sanitario. Siamo di fronte alla nascita di nuovi sistemi di responsabilità, la cd. responsabilità civile algoritmica, o responsabilità robotica.; Each generation of lawyers has the task of measuring the impact of technology on the ordering categories of private law. The jurist of the early twentieth century had to record the changes that the motorization of road traffic caused on civil liability, that of the second twentieth century had to deal with the consequences that the spread of mass reproductive models generated on the contract; it was up to the generation of jurists of the last part of the last century to investigate the role of telematic digitization on contracts and copyright. Today’s jurist perhaps has the greatest burden, that of neutralizing the impact that artificial intelligence seems able to determine on every branch of law, with an unprecedented degree of pervasiveness. In particular, the structures of civil liability appear, in comparison with the tools of artificial intelligence, often unsuitable for responding effectively to the problems that the use of robotics raises, especially if we look at the health sector. We are facing the birth of new systems of responsibility, the so-called algorithmic civil liability, or robotic liability

A spatio-temporal dissection of the transcriptional activity of glucocorticoids in zebrafish: a new transgenic line as a living biosensor model.

Glucocorticoids (GCs) are steroid hormones exerting several essential functions during embryo development and physiological processes in adult life. They bind to their specific receptor, which upon ligand binding migrates to the nucleus, dimerizes and binds to specific glucocorticoid responsive elements (GREs) on DNA, which regulate in a positive or negative (nGREs) way the transcription of target genes. Besides, glucocorticoid receptor can indirectly regulate gene transcription, by means of protein-protein interactions with other transcription factors. This thesis is focused on the transcriptional analysis of GCs activity in the zebrafish model organism. In the first part of my PhD, we set and validated a glucocorticoid responsive zebrafish transgenic line (ia20), in which the green fluorescent protein EGFP (Enhanced-GFP) is located downstream of nine tandem GRE repeats. Thus, this line enables us to follow in vivo GCs transcriptional activity, by means of fluorescence observation. Endogenous fluorescence is first ubiquitously detectable during somitogenesis, becoming then more localized in discrete sites during the first days of development, where it persists until adulthood. EGFP signal increases in a dose-dependent way in response to different DEX concentrations, while gr inactivation through antisense morpholino causes a strong fluorescence decrease and the loss of DEX responsiveness. To validate the ia20 line specificity, we exposed embryos to different steroids at the same concentration: we observed a signal increase only in response to GCs treatments, while fluorescence was not modified by mineralocorticoids, androgens and estrogens treatments. Besides, we collected larvae every 2 hours for 28 hours and demonstrated that the ia20 line is sensitive to the daily endogenous GCs concentration fluctuations due to the circadian rhythm. For all these features, we can consider the ia20 line a potent biosensor to analyze GCs activity from a spatio-temporal point of view. In the last part of this thesis, we put the ia20 line in grs357 mutant background. In these mutants, the transcriptional activity of the glucocorticoid receptor mediated by a direct DNA binding is missing because of a single amino acid substitution in the DNA binding domain. The receptor function not requiring a direct DNA binding is maintained in mutants. Indeed, the mutation is not lethal in homozygosis. This line was generated in order to characterize the possible cross-reactivity among different steroid hormones, since glucocorticoid, mineralocorticoid, androgen and progestin receptors share the ability to bind GREs (that for this reason are also called “Hormone Responsive Elements”, HREs). In the ia20 line in grs357 background we observed a clear fluorescence decrease and a complete loss of GCs responsiveness at 3 dpf (days post fertilization), even at high GCs concentrations, suggesting that, at least during early development, the ia20 line is able to respond only to GCs. We also observed a decrease of endogenous fluorescence intensity at later stages, at 21 dpf and in some tissues in adults (but the latter result derives from a preliminary experiment). Besides, preliminary experiments on adult fish suggest that liver and gut only respond to glucocorticoid receptor in the ia20 line, because these tissues do not show endogenous fluorescence and do not respond to DEX in mutant background. In gonads, instead, and although to lesser extent in muscle, we hypothesize that androgens and progestins receptors may contribute to GREs-mediated transcriptional activity

Sujidades leves em alimentos extrusados para cães comercializados a granel e em embalagem fechada na região da Grande Florianópolis

Foi avaliada a presença de sujidades leves (insetos, ácaros e pelos de roedores) em alimentos extrusados para cães comercializados a granel e em embalagem fechada, bem como as condições ambientais de comercialização. Foram analisadas 25 amostras a granel e 25 embalagens fechadas. Quanto a sujidades leves, 34% tiveram alguma sujidade detectada, sendo 30% delas em alimentos comercializados a granel. A principal sujidade encontrada foram os fragmentos de insetos, 40% nas amostras a granel e 8% em embalagem fechada. Em relação à condição de comercialização, dos 16 comércios visitados, 4 tinham condições higiênicas sanitárias baixas com alimentos expostos ao ambiente, acesso a outros animais e local empoeirado. Condições ambientais inadequadas, como as observadas nos alimentos a granel, associadas à falta de boas práticas de comercialização, contribuem para o aumento da presença de vetores, como roedores e insetos

Clinical Features, Cardiovascular Risk Profile, and Therapeutic Trajectories of Patients with Type 2 Diabetes Candidate for Oral Semaglutide Therapy in the Italian Specialist Care

Introduction: This study aimed to address therapeutic inertia in the management of type 2 diabetes (T2D) by investigating the potential of early treatment with oral semaglutide. Methods: A cross-sectional survey was conducted between October 2021 and April 2022 among specialists treating individuals with T2D. A scientific committee designed a data collection form covering demographics, cardiovascular risk, glucose control metrics, ongoing therapies, and physician judgments on treatment appropriateness. Participants completed anonymous patient questionnaires reflecting routine clinical encounters. The preferred therapeutic regimen for each patient was also identified. Results: The analysis was conducted on 4449 patients initiating oral semaglutide. The population had a relatively short disease duration (42%  60% of patients, and more often than sitagliptin or empagliflozin. Conclusion: The study supports the potential of early implementation of oral semaglutide as a strategy to overcome therapeutic inertia and enhance T2D management

Association of Variants in the SPTLC1 Gene With Juvenile Amyotrophic Lateral Sclerosis

Importance: Juvenile amyotrophic lateral sclerosis (ALS) is a rare form of ALS characterized by age of symptom onset less than 25 years and a variable presentation.Objective: To identify the genetic variants associated with juvenile ALS.Design, Setting, and Participants: In this multicenter family-based genetic study, trio whole-exome sequencing was performed to identify the disease-associated gene in a case series of unrelated patients diagnosed with juvenile ALS and severe growth retardation. The patients and their family members were enrolled at academic hospitals and a government research facility between March 1, 2016, and March 13, 2020, and were observed until October 1, 2020. Whole-exome sequencing was also performed in a series of patients with juvenile ALS. A total of 66 patients with juvenile ALS and 6258 adult patients with ALS participated in the study. Patients were selected for the study based on their diagnosis, and all eligible participants were enrolled in the study. None of the participants had a family history of neurological disorders, suggesting de novo variants as the underlying genetic mechanism.Main Outcomes and Measures: De novo variants present only in the index case and not in unaffected family members.Results: Trio whole-exome sequencing was performed in 3 patients diagnosed with juvenile ALS and their parents. An additional 63 patients with juvenile ALS and 6258 adult patients with ALS were subsequently screened for variants in the SPTLC1 gene. De novo variants in SPTLC1 (p.Ala20Ser in 2 patients and p.Ser331Tyr in 1 patient) were identified in 3 unrelated patients diagnosed with juvenile ALS and failure to thrive. A fourth variant (p.Leu39del) was identified in a patient with juvenile ALS where parental DNA was unavailable. Variants in this gene have been previously shown to be associated with autosomal-dominant hereditary sensory autonomic neuropathy, type 1A, by disrupting an essential enzyme complex in the sphingolipid synthesis pathway.Conclusions and Relevance: These data broaden the phenotype associated with SPTLC1 and suggest that patients presenting with juvenile ALS should be screened for variants in this gene.</p

Mortality and pulmonary complications in patients undergoing surgery with perioperative SARS-CoV-2 infection: an international cohort study

Background: The impact of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) on postoperative recovery needs to be understood to inform clinical decision making during and after the COVID-19 pandemic. This study reports 30-day mortality and pulmonary complication rates in patients with perioperative SARS-CoV-2 infection. Methods: This international, multicentre, cohort study at 235 hospitals in 24 countries included all patients undergoing surgery who had SARS-CoV-2 infection confirmed within 7 days before or 30 days after surgery. The primary outcome measure was 30-day postoperative mortality and was assessed in all enrolled patients. The main secondary outcome measure was pulmonary complications, defined as pneumonia, acute respiratory distress syndrome, or unexpected postoperative ventilation. Findings: This analysis includes 1128 patients who had surgery between Jan 1 and March 31, 2020, of whom 835 (74·0%) had emergency surgery and 280 (24·8%) had elective surgery. SARS-CoV-2 infection was confirmed preoperatively in 294 (26·1%) patients. 30-day mortality was 23·8% (268 of 1128). Pulmonary complications occurred in 577 (51·2%) of 1128 patients; 30-day mortality in these patients was 38·0% (219 of 577), accounting for 81·7% (219 of 268) of all deaths. In adjusted analyses, 30-day mortality was associated with male sex (odds ratio 1·75 [95% CI 1·28–2·40], p\textless0·0001), age 70 years or older versus younger than 70 years (2·30 [1·65–3·22], p\textless0·0001), American Society of Anesthesiologists grades 3–5 versus grades 1–2 (2·35 [1·57–3·53], p\textless0·0001), malignant versus benign or obstetric diagnosis (1·55 [1·01–2·39], p=0·046), emergency versus elective surgery (1·67 [1·06–2·63], p=0·026), and major versus minor surgery (1·52 [1·01–2·31], p=0·047). Interpretation: Postoperative pulmonary complications occur in half of patients with perioperative SARS-CoV-2 infection and are associated with high mortality. Thresholds for surgery during the COVID-19 pandemic should be higher than during normal practice, particularly in men aged 70 years and older. Consideration should be given for postponing non-urgent procedures and promoting non-operative treatment to delay or avoid the need for surgery. Funding: National Institute for Health Research (NIHR), Association of Coloproctology of Great Britain and Ireland, Bowel and Cancer Research, Bowel Disease Research Foundation, Association of Upper Gastrointestinal Surgeons, British Association of Surgical Oncology, British Gynaecological Cancer Society, European Society of Coloproctology, NIHR Academy, Sarcoma UK, Vascular Society for Great Britain and Ireland, and Yorkshire Cancer Research

A spatio-temporal dissection of the transcriptional activity of glucocorticoids in zebrafish: a new transgenic line as a living biosensor model.

Glucocorticoids (GCs) are steroid hormones exerting several essential functions during embryo development and physiological processes in adult life. They bind to their specific receptor, which upon ligand binding migrates to the nucleus, dimerizes and binds to specific glucocorticoid responsive elements (GREs) on DNA, which regulate in a positive or negative (nGREs) way the transcription of target genes. Besides, glucocorticoid receptor can indirectly regulate gene transcription, by means of protein-protein interactions with other transcription factors. This thesis is focused on the transcriptional analysis of GCs activity in the zebrafish model organism. In the first part of my PhD, we set and validated a glucocorticoid responsive zebrafish transgenic line (ia20), in which the green fluorescent protein EGFP (Enhanced-GFP) is located downstream of nine tandem GRE repeats. Thus, this line enables us to follow in vivo GCs transcriptional activity, by means of fluorescence observation. Endogenous fluorescence is first ubiquitously detectable during somitogenesis, becoming then more localized in discrete sites during the first days of development, where it persists until adulthood. EGFP signal increases in a dose-dependent way in response to different DEX concentrations, while gr inactivation through antisense morpholino causes a strong fluorescence decrease and the loss of DEX responsiveness. To validate the ia20 line specificity, we exposed embryos to different steroids at the same concentration: we observed a signal increase only in response to GCs treatments, while fluorescence was not modified by mineralocorticoids, androgens and estrogens treatments. Besides, we collected larvae every 2 hours for 28 hours and demonstrated that the ia20 line is sensitive to the daily endogenous GCs concentration fluctuations due to the circadian rhythm. For all these features, we can consider the ia20 line a potent biosensor to analyze GCs activity from a spatio-temporal point of view. In the last part of this thesis, we put the ia20 line in grs357 mutant background. In these mutants, the transcriptional activity of the glucocorticoid receptor mediated by a direct DNA binding is missing because of a single amino acid substitution in the DNA binding domain. The receptor function not requiring a direct DNA binding is maintained in mutants. Indeed, the mutation is not lethal in homozygosis. This line was generated in order to characterize the possible cross-reactivity among different steroid hormones, since glucocorticoid, mineralocorticoid, androgen and progestin receptors share the ability to bind GREs (that for this reason are also called “Hormone Responsive Elements”, HREs). In the ia20 line in grs357 background we observed a clear fluorescence decrease and a complete loss of GCs responsiveness at 3 dpf (days post fertilization), even at high GCs concentrations, suggesting that, at least during early development, the ia20 line is able to respond only to GCs. We also observed a decrease of endogenous fluorescence intensity at later stages, at 21 dpf and in some tissues in adults (but the latter result derives from a preliminary experiment). Besides, preliminary experiments on adult fish suggest that liver and gut only respond to glucocorticoid receptor in the ia20 line, because these tissues do not show endogenous fluorescence and do not respond to DEX in mutant background. In gonads, instead, and although to lesser extent in muscle, we hypothesize that androgens and progestins receptors may contribute to GREs-mediated transcriptional activity.I glucocorticoidi sono ormoni steroidei con un ruolo di fondamentale importanza durante lo sviluppo embrionale e in molti processi fisiologici durante tutta la vita. Svolgono la loro funzione legandosi al loro recettore che in risposta al legame migra nel nucleo, dimerizza e si lega a specifiche sequenze di DNA responsive ai glucocorticoidi (glucocorticoid responsive elements, GREs). In questo modo regolano positivamente o negativamente (nGREs) la trascrizione di geni bersaglio. Il recettore per i glucocorticoidi può inoltre regolare la trascrizione genica in modo indiretto, mediante interazioni proteina-proteina con altri fattori di trascrizione. Questo lavoro di tesi si è focalizzato sull’analisi trascrizionale dell’attività dei glucocorticoidi nell’organismo modello zebrafish. Durante la prima parte del percorso di dottorato è stata messa a punto e validata una linea transgenica di zebrafish (ia20) responsiva ai glucocorticoidi, nella quale la proteina fluorescente EGFP (Enhanced-GFP) si trova a valle di nove ripetizioni in tandem della sequenza GRE. Mediante l’osservazione della fluorescenza è quindi possibile monitorare in vivo l’attività trascrizionale dei glucocorticoidi. La fluorescenza endogena inizia ad essere apprezzabile a partire dalla somitogenesi ed è inizialmente ubiquitaria, per poi localizzarsi in siti discreti nei primi giorni di sviluppo dove rimane rilevabile anche in età adulta. In risposta al trattamento con diverse concentrazioni del glucocorticoide sintetico desametasone (DEX), il segnale della EGFP aumenta in modo dose-dipendente, mentre l’inattivazione del trascritto per il gr, ottenuta mediante l’iniezione di un morpholino antisenso, provoca una considerevole diminuzione del segnale e la mancata risposta al DEX. La specificità della linea è stata validata trattando gli embrioni con diversi steroidi alla stessa concentrazione: il segnale aumenta solo in risposta ai glucocorticoidi mentre rimane invariato nei trattamenti con mineralocorticoidi, androgeni ed estrogeni. Inoltre, un campionamento ogni 2 ore nell’arco di un periodo di tempo di 28 ore ha dimostrato che la linea transgenica ia20 è sensibile alle variazioni di concentrazione dei glucocorticoidi endogeni dovute al ritmo circadiano. Queste caratteristiche rendono la linea ia20 un efficace biosensore per analizzare l’attività dei glucocorticoidi in vivo dal punto di vista spaziale e temporale. Durante l’ultimo periodo di dottorato, la linea ia20 è stata messa in background mutante grs357. In questi mutanti, l’attività trascrizionale del recettore dei glucocorticoidi legata ad un’interazione diretta con il DNA viene persa a causa di una singola sostituzione amminoacidica a livello del dominio di legame al DNA. Vengono invece mantenute le altre funzioni di controllo della trascrizione del recettore. La linea è stata generata con lo scopo di caratterizzare l’eventuale presenza di cross-reattività da parte dei diversi ormoni steroidei, in quanto i recettori per i glucocorticoidi, mineralocorticoidi, androgeni e progestinici condividono la capacità di legarsi ai siti GRE (definiti per questo motivo anche “Hormone Responsive Elements”, HREs). Nella linea ia20 in background mutante abbiamo osservato una notevole riduzione nella fluorescenza endogena ed una totale perdita di responsività ai glucocorticoidi anche a concentrazioni elevate su embrioni di tre giorni. Questo risultato suggerisce la mancanza di cross-reattività fra i diversi ormoni steroidei nei primi giorni di sviluppo, e questo a causa della mancata espressione di alti livelli dei recettori degli altri ormoni steroidei. Per quanto riguarda gli stadi più avanzati, la diminuzione nella fluorescenza endogena nei mutanti è stata osservata anche a tre settimane e in alcuni tessuti dell’adulto, in un esperimento preliminare che dovrà essere in seguito approfondito. Esperimenti preliminari su pesci adulti suggeriscono inoltre che la fluorescenza osservata nella linea ia20 su fegato ed intestino derivi solo dall’attivazione del recettore dei glucocorticoidi, in quanto nel mutante questi tessuti non presentano alcun segnale di fluorescenza e non rispondono al trattamento con il DEX. Per quanto riguarda le gonadi, e in misura minore il tessuto muscolare, il segnale fluorescente presente negli esemplari mutanti e la mancata risposta al DEX, possono essere spiegati dall’attività trascrizionale determinata dai recettori per gli androgeni e i progestinici

A living biosensor model to dynamically trace glucocorticoid transcriptional activity during development and adult life in zebrafish

Glucocorticoids (GCs) modulate many cellular processes through the binding of the glucocorticoid receptor (GR) to specific responsive elements located upstream of the transcription starting site or within an intron of GC target genes. Here we describe a transgenic fish line harboring a construct with nine GC-responsive elements (GREs) upstream of a reporter (EGFP) coding sequence. Transgenic fish exhibit strong fluorescence in many known GC-responsive organs. Moreover, its enhanced sensitivity allowed the discovery of novel GC-responsive tissue compartments, such as fin, eyes, and otic vesicles. Long-term persistence of transgene expression is seen during adult stages in several organs. Pharmacological and genetic analysis demonstrates that the transgenic line is highly responsive to drug administration and molecular manipulation. Moreover, reporter expression is sensitively and dynamically modulated by the photoperiod, thus proving that these fish are an in vivo valuable platform to explore GC responsiveness to both endogenous and exogenous stimuli