Immunohistochemical evaluation of aquaporin-4 and its correlation with CD68, IBA-1, HIF-1α, GFAP, and CD15 expressions in fatal traumatic brain injury

Traumatic brain injury (TBI) is one of the leading causes of death and disability worldwide. Our understanding of its pathobiology has substantially increased. Following TBI, the following occur, edema formation, brain swelling, increased intracranial pressure, changes in cerebral blood flow, hypoxia, neuroinflammation, oxidative stress, excitotoxicity, and apoptosis. Experimental animal models have been developed. However, the difficulty in mimicking human TBI explains why few neuroprotective strategies, drawn up on the basis of experimental studies, have translated into improved therapeutic strategies for TBI patients. In this study, we retrospectively examined brain samples in 145 cases of death after different survival times following TBI, to investigate aquaporin-4 (AQP4) expression and correlation with hypoxia, and neuroinflammation in human TBI. Antibodies anti-glial fibrillary acid protein (GFAP), aquaporin-4 (AQP4), hypoxia induced factor-1α (HIF-1α), macrophage/phagocytic activation (CD68), ionized calcium-binding adapter molecule-1 (IBA-1), and neutrophils (CD15) were used. AQP4 showed a significant, progressive increase between the control group and groups 2 (one-day survival) and 3 (three-day survival). There were further increases in AQP4 immunopositivity in groups 4 (seven-day survival), 5 (14-dayssurvival), and 6 (30-day survival), suggesting an upregulation of AQP4 at 7 to 30 days compared to group 1. GFAP showed its highest expression in non-acute cases at the astrocytic level compared with the acute TBI group. Data emerging from the HIF-1α reaction showed a progressive, significant increase. Immunohistochemistry with IBA-1 revealed activated microglia starting three days after trauma and progressively increasing in the next 15 to 20 days after the initial trauma. CD68 expression demonstrated basal macrophage and phagocytic activation mostly around blood vessels. Starting from one to three days of survival after TBI, an increase in the number of CD68 cells was progressively observed; at 15 and 30 days of survival, CD68 showed the most abundant immunopositivity inside or around the areas of necrosis. These findings need to be developed further to gain insight into the mechanisms through which brain AQP4 is upregulated. This could be of the utmost clinicopathological importance

Controlling the nitrogen environment for optimal Rhodomonas salina production

The microalga Rhodomonas salina is a widely used species for rearing live feed organisms in the aquaculture feed market. A species-specific medium is an essential step towards enhancing productivity and decreasing production costs for microalgae cultivation. However, relevant aspects of medium composition such as nitrogen source and elemental ratio have not yet been characterized for this alga. This study aimed to optimize the following three aspects of culture media: 1) optimal ratio between nitrogen and phosphorus (N:P ratio); 2) preferred source of nitrogen; and 3) tolerance of R. salina towards free ammonia. To investigate this, we conducted a series of controlled laboratory experiments in shake flasks. Our experiments revealed a 45% increase in growth rate when an N:P ratio of 15:1 was used compared to the standard ratio of 25:1. Ammonium and nitrate were equally well accepted as a nitrogen source, however, a mix of ammonium and nitrate resulted in significant growth reduction. Free ammonia did not affect growth of the alga at the tested concentrations of up to 5 mg ammonia–nitrogen L−1. We conclude that for optimal R. salina cultivation, an N:P ratio of 15:1 is strongly preferred, as it leads to a significant increase in growth rate. Further, media with a single source of nitrogen promote faster growth over media with mixed sources, and ammonium may safely be used as a nitrogen source, since R. salina tolerates certain levels of free ammonia. Overall, this work provides insights into the optimal cultivation conditions for R. salina, allowing for more efficient and reliable production of this relevant species

Regulating pediatric off-label uses of medicines in the EU and USA: challenges and potential solutions

Background Off-label uses of medicines are common in pediatrics. The literature reports that at least one-third of children in hospital and up to 90% of newborns in neonatal intensive care units receive off-label prescriptions. Moreover, the lack of data on safety and efficacy in the pediatric population may sometimes increase the risk of adverse drug reactions. Objective This paper aims to (a) map the main gaps in the existing EU and US regulatory frameworks for pediatric drug development and off-label use and (b) propose potential solutions for further discussion. Conclusion The selected jurisdictions aim to limit off-label prescribing, but implementation levels generally seem low, including in pediatric settings. Subject to further research, we propose potential concerted actions and initiatives of international cooperation to fill this gap. In particular, regulators and pediatric societies could strengthen manufacturers' incentives to develop pediatric medicines, pediatricians' information about off-label uses, and patients' and parents' awareness

Autopsy findings in COVID-19-related deaths: a literature review

Although many clinical reports have been published, little is known about the pathological post-mortem findings from people who have died of the novel coronavirus disease. The need for postmortem information is urgent to improve patient management of mild and severe illness, and treatment strategies. The present systematic review was carried out according to the Preferred Reporting Items for Systematic Review (PRISMA) standards. A systematic literature search and a critical review of the collected studies were conducted. An electronic search of PubMed, Science Direct Scopus, Google Scholar, and Excerpta Medica Database (EMBASE) from database inception to June 2020 was performed. We found 28 scientific papers; the total amount of cases is 341. The major histological feature in the lung is diffuse alveolar damage with hyaline membrane formation, alongside microthrombi in small pulmonary vessels. It appears that there is a high incidence of deep vein thrombosis and pulmonary embolism among COVID-19 decedents, suggesting endothelial involvement, but more studies are needed. A uniform COVID-19 post-mortem diagnostic protocol has not yet been developed. In a time in which international collaboration is essential, standardized diagnostic criteria are fundamental requirements

Systemic immune-inflammation index predicts the clinical outcome in patients with metastatic renal cell cancer treated with sunitinib.

Background: In this retrospective analysis, we explored the prognostic and predictive value of the systemic immune-inflammation index (SII), based on lymphocyte, neutrophil, and platelet counts, at baseline and changes at week 6 during first-line sunitinib in patients with metastatic renal cell cancer (RCC).Results: Patients were stratified into high SII (? 730) and low SII (< 730) groups. SII was associated with objective response, p < 0.0001. The median PFS was 6.3 months (95% CI 5.5–8.9) in patients with SII ? 730 and 18.7 months (95% CI 14.7–22.8) in those with SII < 730, p < 0.0001. The median OS was 43.6 months (95% CI 35.3–52.1) in patients with SII < 730, and 13.5 months (95% CI 9.8–18.5) in those with SII ? 730, p < 0.0001. In multivariate analysis, performance status, IMDC score and SII were able to predict OS (HR = 3.29, HR = 1.71 and HR = 1.79, respectively).Materials and Methods: We included 335 consecutive RCC patients treated with first-line sunitinib. The X-tile 3.6.1 software (Yale University, New Haven, CT) was used for bioinformatic analysis of the data to determine the cutoff value of SII. Progression-free survival (PFS), overall survival (OS) and their 95% confidence interval (95% CI) were estimated by Kaplan-Meier method and compared with logrank test. The impact of SII conversion at week 6 of treatment on PFS and OS was evaluated by Cox regression analyses.Conclusions: The SII and its changes during treatment represent a powerful prognostic indicator of clinical outcome in patients with metastatic RCC

Study of alteration and degradation products of a Roman marble sarcophagus located in the medieval cloister of the old st Cosimato’s Convent, now the new "Regina Margherita Hospital” (Rome)

Optical and SEM-EDS microscopy, FT-IR and mFT-IR spectroscopy investigations have been applied to the study of alteration and degradation products of a Roman sarcophagus located in the medieval cloister of St Cosimato’s Convent, now the new “Regina Margherita Hospital” (Rome) before restoration work. This scientific analysis was performed in order to compare it with a diagnosis report based on the restorer’s expertise. The sarcophagus is small and consists of a single block of fine-grained marble covered by a thick pink patina. The results obtained from this diagnostic investigation, allow us to draw important conclusions about the state of conservation of this archaeological artefact. The main components found in the samples analyzed are: black crust, calcium oxalate, organic matter due to recent conservation treatment, calcite, iron-oxides and coloured earths. In particular, the combined used of petrographical and mFT-IR analysis provided useful insights into the polishing and consolidation processes of archaeological artefacts

Reconsidering dependence on life-sustaining treatment as a criterion for assisted suicide: the Italian legal unicum in comparative perspective

The legalization of Medical Assisted Voluntary Death, including assisted suicide is spreading worldwide, alongside the recognition of the centrality of the patient’s right to self-determination even in case of therapeutic desistance. In Italy Law-no. 219/2017 has granted patients the option of refusing therapy including life-sustaining treatments even without justification. The present paper offers a critical analysis of the legal-normative aspects and ethical-clinical implications of constraining assisted suicide to dependence on life-sustaining treatments. Reviewing some of the key bioethical-legal pronouncements, we discuss the current Italian system on assisted suicide in which dependence on life-sustaining treatment, even after the recent Constitutional sentences, is still one of the mandatory requirements, despite several critical profiles. Through a literature overview on medical life-sustaining treatments notion, the dependence on them is analyzed and assessed in clinical, bioethical and validity terms as a requirement for access to assisted suicide. From this it appears how dependence on life-sustaining treatment constraint shows overly ambiguous definitional boundaries, with the risk of inhomogeneous interpretations especially in the Italian framework. Interestingly, our comparative analysis reveals that Italy is a global legal unicum among the main international systems regulating Medical Assisted Voluntary Death; which, conversely, tend to target the issue on terminally or irreversibly suffering patients, independently of dependence on life-sustaining treatment

Blood-brain barrier breakdown, central nervous system cell damage, and infiltrated T cells as major adverse effects in CAR-T-related deaths: a literature review

Background CAR-T-related deaths observed worldwide are rare. The underlying pathogenetic mechanisms are the subject of study, as are the findings that enable diagnosis. A systematic literature search of the PubMed database and a critical review of the collected studies were conducted from the inception of this database until January 2023. The aim of the study is to determine when death is related to CAR-T cell therapy and to develop a shareable diagnostic algorithm.Methods The database was searched by combining and meshing the terms ("CAR-t" OR "CART") AND ("Pathology" OR "Histology" OR "Histological" OR "Autopsy") AND ("Heart" OR "Cardiac" OR "Nervous System" OR "Kidney" OR "Liver") with 34 results and also the terms: [(Lethal effect) OR (Death)] AND (CAR-T therapy) with 52 results in titles, abstracts, and keywords [all fields]. One hundred scientific articles were examined, 14 of which were additional records identified through other sources. Fifteen records were included in the review.Results Neuronal death, neuronal edema, perivascular edema, perivascular and intraparenchymal hemorrhagic extravasation, as well as perivascular plasmatodendrosis, have been observed in cases with fatal cerebral edema. A cross-reactivity of CAR-T cells in cases of fatal encephalopathy can be hypothesized when, in addition to the increased vascular permeability, there is also a perivascular lymphocyte infiltrate, which appears to be a common factor among most authors.Conclusion Most CAR-T-related deaths are associated with blood-brain barrier breakdown, central nervous system cell damage, and infiltrated T cells. Further autopsies and microscopic investigations would shed more light on the lethal toxicity related to CAR-T cells. A differential diagnosis of CAR-T-related death is crucial to identifying adverse events. In this article, we propose an algorithm that could facilitate the comparison of findings through a systematic approach. Despite toxicity cases, CAR-T therapy continues to stand out as the most innovative treatment within the field of oncology, and emerging strategies hold the promise of delivering safer therapies in future

Regulation of miRNAs as new tool for cutaneous vitality lesions demonstration in ligature marks in deaths by hanging

This study aims to demonstrate that the application of miRNA expression in forensic pathology, in cases of hanging, applying the method on skin samples. The proposed investigative protocol allowed us to highlight a different miRNA expression in the skin ligature marks of subjects who died by hanging compared to healthy skin control samples. The results obtained showed an increase in the expression of miRNAs recognized as regulators of the inflammatory response in skin lesions such as miR125a-5p and miR125b-5p. Furthermore, overexpression of additional miRNAs – miR214a-3p, miR128-3p, miR130a-3p, and miR92a-3p – with anti-inflammatory activity was highlighted. It was possible to document a statistical significance to control skin samples only for miR103a-3p (p < 0.05), miR214-3p and miR92a-3p (p < 0.01) The upregulation of miR222-3p and miR150-5p, respectively related to mast-cell activation and neutrophils after the application of traumatic stimuli supports the immunohistochemical data showed in literature. The diagnostic accuracy of miRNAs could expand the range of diagnostic tools available in the assessment of the vitality of a lesion

A Sustainable Approach for the Management and Valorization of Underwater Cultural Heritage: New Perspectives from the TECTONIC Project

Documentation and conservation of underwater cultural heritage (UCH) are crucial to preserving humankind’s history and traditions, safeguarding tangible testimonies of past human life while ensuring its accessibility to future generations. The TECTONIC (Technological Consortium TO develop sustainability of underwater Cultural Heritage) project is promoting an intersectoral collaboration between academic and non-academic professionals (i.e., archaeologists, conservators, geologists, engineers, etc.) working on different topics related to UCHs, to find solutions to the issues still existing in the field. The overall aim is the exchange of skills for the improvement and assessment of innovative materials and techniques to develop solutions and marketable products for the conservation and management of the UCH, sustainably. To achieve its overall aim, TECTONIC is undertaking activities driven by the following objectives: (a) the study, documentation, and mapping of selected UCHs; (b) the creation of decision-support tools for UCH risk assessment in a changing environment; (c) the initiation of conservation studies and protocols for conservation activities; (d) the development of open and low-cost robotic solutions for the inspection of UCH; and (e) the raising of public awareness and knowledge about UCH. All the objectives are devoted to stimulating new sustainable ideas that would bring the growth of cultural tourism and the development of new marketable products by capitalizing on the research results.Fil: Ricca, Michela. Università della Calabria; ItaliaFil: Alexandrakis, George. Foundation For Research And Technology ? Hellas.; GreciaFil: Bonazza, Alessandra. Consiglio Nazionale Delle Ricerche. Istituto Di Scienze Dell Atmosfera E del Clima.; ItaliaFil: Bruno, Fabio. Università della Calabria; ItaliaFil: Petriaggi, Barbara Davidde. Instituto Superiore per la Conservazione ed il Restauro; ItaliaFil: Elkin, Dolores Carolina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Secretaría de Cultura de la Nación. Dirección Nacional de Cultura y Museos. Instituto Nacional de Antropología y Pensamiento Latinoamericano; ArgentinaFil: Lagudi, Antonio. Università della Calabria; ItaliaFil: Nicolas, Stephane. Centre d'Activité des Playes ZE Jean Monnet; FranciaFil: Novák, Michal. Synpo; República ChecaFil: Papatheodorou, George. University Of Patras; GreciaFil: Prieto, Javier. Universidad de Salamanca; EspañaFil: Ricci, Marco. Università della Calabria; ItaliaFil: Vasilijevic, Antonio. H2O Robotics; CroaciaFil: La Russa, Mauro Francesco. Università della Calabria; Italia. Foundation For Research And Technology ? Hellas.; Greci