Biochemical Signatures of Doppel Protein in Human Astrocytomas to Support Prediction in Tumor Malignancy

Doppel (Dpl) is a membrane-bound glycoprotein mainly expressed in the testis of adult healthy people. It is generally absent in the central nervous system, but its coding gene sequence is ectopically expressed in astrocytoma specimens and in derived cell lines. In this paper, we investigated the expression and the biochemical features of Dpl in a panel of 49 astrocytoma specimens of different WHO malignancy grades. As a result, Dpl was expressed in the majority of the investigated specimens (86%), also including low grade samples. Importantly, Dpl exhibited different cellular localizations and altered glycan moieties composition, depending on the tumor grade. Most low-grade astrocytomas (83%) showed a membrane-bound Dpl, like human healthy testis tissue, whereas the majority of high-grade astrocytomas (75%) displayed a cytosolic Dpl. Deglycosylation studies with N-glycosidase F and/or neuraminidase highlighted defective glycan moieties and an unexpected loss of sialic acid. To find associations between glial tumor progression and Dpl biochemical features, predictive bioinformatics approaches were produced. In particular, Decision tree and Nomogram analysis showed well-defined Dpl-based criteria that separately clustered low-and high-grade astrocytomas. Taken together, these findings show that in astrocytomas, Dpl undergoes different molecular processes that might constitute additional helpful tools to characterize the glial tumor progression

A novel approach for the purification and proteomic analysis of pathogenic immunglobulin free light chains from serum

An excess of circulating monoclonal free immunoglobulin light chains (FLC) is common in plasma cell disorders. A subset of FLC, as amyloidogenic ones, possess intrinsic pathogenicity. Because of their complex purification, little is known on the biochemical features of serum FLC, possibly related to their pathogenic spectrum. We developed an immunopurification approach to isolate serum FLC from patients with monoclonal gammopathies, followed by proteomic characterization. Serum monoclonal FLC were detected and quantified by immunofixation and immunonephelometry. Immunoprecipitation was performed by serum incubation with agarose beads covalently linked to polyclonal anti-κ or λ FLC antibodies. Isolated FLC were analyzed by SDS-PAGE, 2D-PAGE, immunoblotting, mass spectrometry (MS). Serum FLC were immunoprecipitated from 15 patients with ALλ amyloidosis (serum λ FLC range: 98-2350mg/L), 5 with ALκ amyloidosis and 1 with κ light chain (LC) myeloma (κ FLC range: 266-2660mg/L), and 3 controls. Monoclonal FLC were the prevalent eluted species in patients. On 2D-PAGE, both λ and κ FLC originated discrete spots with multiple pI isoforms. The nature of eluted FLC and coincidence with the LC sequence from the bone marrow clone was confirmed by MS, which also detected post-translational modifications, including truncation, tryptophan oxidation, cysteinylation, peptide dimerization. Serum FLC were purified in soluble form and adequate amounts for proteomics, which allowed studying primary sequence and detecting post-translational modifications. This method is a novel instrument for studying the molecular bases of FLC pathogenicity, allowing for the first time the punctual biochemical description of the circulating forms

Biblioteche accademiche in rete per la terza missione? Valorizzazione, inclusione e nuove opportunità strategiche

In recent years, Italian academic libraries have undertaken third mission activities linked to the enhancement of their heritage: library science scholars have already been studying this kind of initiatives, and today the Italian national agency for the evaluation of universities and research institutes (Anvur) considers such actions as a result of the institutional commitment of a university library. A workshop, organized by the Library System of the University of Ferrara together with the Italian library association during the Stelline congress 2022 (Milan, 11 march), analyzed the case study of the bibliotour Storie di libri e palazzi, set up by the University of Ferrara; the workshop also explored new strategic opportunities for the enhancement of academic libraries’ heritage, firstly the inclusion of all audiences, with specific reference to Goal 10 (“Reduce inequalities”) of the United Nations 2030 Agenda. Furthermore, this perspective embraces potentially the entire corpus of recommendations provided by the Agenda. The paper is intended not only to report the main ideas that emerged during the workshop held in Milan but, starting from them, also to suggest building a network of academic libraries, in order to facilitate the communication of third mission initiatives and their public outreach. Therefore, full adherence to the inclusive dimension and the commitment to sustainable development are two new strategic opportunities for academic library activities related to heritage enhancement. AIB has been supporting both strategies for several years, by means of concrete actions, such as the “Maria A. Abenante” Award, as well as organizational structures, like the Observatory on library and sustainable development (Obiss).In anni recenti le biblioteche accademiche italiane hanno intrapreso attività di terza missione legate alla valorizzazione del proprio patrimonio, entro una linea d’azione già in parte esplorata dalla letteratura biblioteconomica, ed oggi anche oggetto d’indagine dell’Agenzia nazionale di valutazione del sistema universitario e della ricerca (Anvur) quale espressione delle funzioni istituzionali di una biblioteca di ateneo. Un workshop, organizzato dal Sistema bibliotecario dell’Università degli studi di Ferrara e dall’Associazione italiana biblioteche in occasione del convegno delle Stelline 2022 (Milano, 11 marzo), ha analizzato il caso-studio dell’itinerario biblioturistico “Storie di libri e palazzi” attivo presso l’ateneo estense, e ha consentito di approfondire la riflessione su nuove opportunità strategiche entro le quali contestualizzare l’azione bibliotecaria di valorizzazione del patrimonio culturale universitario. Si tratta, in primis, dell’impegno per l’inclusione di tutti i pubblici, con specifico riferimento all’Obiettivo 10 (“Ridurre le diseguaglianze”) dell’Agenda 2030 delle Nazioni Unite, ma potenzialmente la prospettiva abbraccia l’intero corpus di raccomandazioni contenute nel documento. Il presente contributo intende restituire i principali spunti emersi durante il laboratorio milanese e, a partire da essi, aprirsi alla concreta proposta di una rete fra le biblioteche accademiche, utile ad agevolare la comunicazione delle iniziative di terza missione e a favorire il raggiungimento dei pubblici. A questo scopo, nuove opportunità strategiche per le biblioteche di ateneo che operano sul fronte della valorizzazione sono individuate nella piena adesione alla dimensione inclusiva e nell’impegno per lo sviluppo sostenibile, istanze che da tempo AIB supporta attraverso iniziative concrete, come l’istituzione del Premio Maria A. Abenante, e specifici assetti organizzativi, quale l’Osservatorio biblioteche e sviluppo sostenibile (OBISS)

Nuclear mRNA retention and aberrant doppel protein expression in human astrocytic tumor cells

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Amyloidogenic light chains impair plasma cell survival

Systemic light chain amyloidosis (AL) is a clonal plasma cell disorder characterized by the deposition of misfolded immunoglobulin light chains (LC) as insoluble fibrils in organs. The lack of suitable models has hindered the investigation of the disease mechanisms. Our aim was to establish AL LC-producing plasma cell lines and use them to investigate the biology of the amyloidogenic clone. We used lentiviral vectors to generate cell lines expressing LC from patients suffering from AL amyloidosis. The AL LC-producing cell lines showed a significant decrease in proliferation, cell cycle arrest, and an increase in apoptosis and autophagy as compared with the multiple myeloma LC-producing cells. According to the results of RNA sequencing the AL LC-producing lines showed higher mitochondrial oxidative stress, and decreased activity of the Myc and cholesterol pathways. The neoplastic behavior of plasma cells is altered by the constitutive expression of amyloidogenic LC causing intracellular toxicity. This observation may explain the disparity in the malignant behavior of the amyloid clone compared to the myeloma clone. These findings should enable future in vitro studies and help delineate the unique cellular pathways of AL, thus expediting the development of specific treatments for patients with this disorder

Clinical outcome with different doses of low-molecular-weight heparin in patients hospitalized for COVID-19

A pro-thrombotic milieu and a higher risk of thrombotic events were observed in patients with CoronaVirus disease-19 (COVID-19). Accordingly, recent data suggested a beneficial role of low molecular weight heparin (LMWH), but the optimal dosage of this treatment is unknown. We evaluated the association between prophylactic vs. intermediate-to-fully anticoagulant doses of enoxaparin and in-hospital adverse events in patients with COVID-19. We retrospectively included 436 consecutive patients admitted in three Italian hospitals. Outcome according to the use of prophylactic (4000IU) vs. higher (>4000IU) daily dosage of enoxaparin was evaluated. The primary end-point was in-hospital death. Secondary outcome measures were in-hospital cardiovascular death, venous thromboembolism, new-onset acute respiratory distress syndrome (ARDS) and mechanical ventilation. A total of 287 patients (65.8%) were treated with the prophylactic enoxaparin regimen and 149 (34.2%) with a higher dosing regimen. The use of prophylactic enoxaparin dose was associated with a similar incidence of all-cause mortality (25.4% vs. 26.9% with the higher dose; OR at multivariable analysis, including the propensity score: 0.847, 95% CI 0.400-0.1.792; p=0.664). In the prophylactic dose group, a significantly lower incidence of cardiovascular death (OR 0.165), venous thromboembolism (OR 0.067), new-onset ARDS (OR 0.454) and mechanical intubation (OR 0.150) was observed. In patients hospitalized for COVID-19, the use of a prophylactic dosage of enoxaparin appears to be associated with similar in-hospital overall mortality compared to higher doses. These findings require confirmation in a randomized, controlled study

Trends in Net Survival from Vulvar Squamous Cell Carcinoma in Italy (1990–2015)

Objective: In many Western countries, survival from vulvar squamous cell carcinoma (VSCC) has been stagnating for decades or has increased insufficiently from a clinical perspective. In Italy, previous studies on cancer survival have not taken vulvar cancer into consideration or have pooled patients with vulvar and vaginal cancer. To bridge this knowledge gap, we report the trend in survival from vulvar cancer between 1990 and 2015. (2) Methods: Thirty-eight local cancer registries covering 49% of the national female population contributed the records of 6274 patients. Study endpoints included 1- and 2-year net survival (NS) calculated using the Pohar-Perme estimator and 5-year NS conditional on having survived two years (5|2-year CNS). The significance of survival trends was assessed with the Wald test on the coefficient of the period of diagnosis, entered as a continuous regressor in a Poisson regression model. (3) Results: The median patient age was stable at 76 years. One-year NS decreased from 83.9% in 1990–2001 to 81.9% in 2009–2015 and 2-year NS from 72.2% to 70.5%. Five|2-year CNS increased from 85.7% to 86.7%. These trends were not significant. In the age stratum 70–79 years, a weakly significant decrease in 2-year NS from 71.4% to 65.7% occurred. Multivariate analysis adjusting for age group at diagnosis and geographic area showed an excess risk of death at 5|2-years, of borderline significance, in 2003–2015 versus 1990–2002. (4) Conclusions: One- and 2-year NS and 5|2-year CNS showed no improvements. Current strategies for VSCC control need to be revised both in Italy and at the global level