Paleopathological Approach to the Study of a Christian Relic: The Case of the Blessed Maria Lorenza Longo

The Blessed Maria Lorenza Longo, founder of the hospital of Santa Maria del Popolo degli Incurabili and the Order of the Capuchin Poor Clares in Naples, Italy, died on 21 October 1539 and was recently beatified on 9 October 2021. The relic, a fully skeletonized cranium, underwent visual and radiological inspection. The biological profile supports the at¬tribution of female sex of the relic, whereas the age at death is estimated to be younger than that reported by historians. A paleopathological survey was conducted to evaluate the historical reports of poisoning or rheumatoid arthritis affecting Maria Longo. Given the limited skeletal data, it was not possible to confirm the presence of these claims. No obvious in¬dicators of dietary deficiencies were observed, and the tertiary syphilis hypothesized by textual sources was excluded. Postmortem alterations of the relic were clearly visible on the superior aspect of the cranium and testified to the worship of the relic

Thyroid Cancer and Fibroblasts

Thyroid cancer is the most common type of endocrine cancer, and its prevalence continue to rise. Non-metastatic thyroid cancer patients are successfully treated. However, looking for new therapeutic strategies is of great importance for metastatic thyroid cancers that still lead to death. With respect to this, the tumor microenvironment (TME), which plays a key role in tumor progression, should be considered as a new promising therapeutic target to hamper thyroid cancer progression. Indeed, thyroid tumors consist of cancer cells and a heterogeneous and ever-changing niche, represented by the TME, which contributes to establishing most of the features of cancer cells. The TME consists of extracellular matrix (ECM) molecules, soluble factors, metabolites, blood and lymphatic tumor vessels and several stromal cell types that, by interacting with each other and with tumor cells, affect TME remodeling, cancer growth and progression. Among the thyroid TME components, cancer-associated fibroblasts (CAFs) have gained more attention in the last years. Indeed, recent important evidence showed that thyroid CAFs strongly sustain thyroid cancer growth and progression by producing soluble factors and ECM proteins, which, in turn, deeply affect thyroid cancer cell behavior and aggressiveness. Hence, in this article, we describe the thyroid TME, focusing on the desmoplastic stromal reaction, which is a powerful indicator of thyroid cancer progression and an invasive growth pattern. In addition, we discuss the origins and features of the thyroid CAFs, their influence on thyroid cancer growth and progression, their role in remodeling the ECM and their immune-modulating functions. We finally debate therapeutic perspectives targeting CAFs

Impairment of T cell development and acute inflammatory response in HIV-1 Tat transgenic mice

Immune activation and chronic inflammation are hallmark features of HIV infection causing T-cell depletion and cellular immune dysfunction in AIDS. Here, we addressed the issue whether HIV-1 Tat could affect T cell development and acute inflammatory response by generating a transgenic mouse expressing Tat in lymphoid tissue. Tat-Tg mice showed thymus atrophy and the maturation block from DN4 to DP thymic subpopulations, resulting in CD4(+) and CD8(+) T cells depletion in peripheral blood. In Tat-positive thymus, we observed the increased p65/NF-κB activity and deregulated expression of cytokines/chemokines and microRNA-181a-1, which are involved in T-lymphopoiesis. Upon LPS intraperitoneal injection, Tat-Tg mice developed an abnormal acute inflammatory response, which was characterized by enhanced lethality and production of inflammatory cytokines. Based on these findings, Tat-Tg mouse could represent an animal model for testing adjunctive therapies of HIV-1-associated inflammation and immune deregulation

RNA-Based Assay for Next-Generation Sequencing of Clinically Relevant Gene Fusions in Non-Small Cell Lung Cancer

Gene fusions represent novel predictive biomarkers for advanced non-small cell lung cancer (NSCLC). In this study, we validated a narrow NGS gene panel able to cover therapeutically-relevant gene fusions and splicing events in advanced-stage NSCLC patients. To this aim, we first assessed minimal complementary DNA (cDNA) input and the limit of detection (LoD) in different cell lines. Then, to evaluate the feasibility of applying our panel to routine clinical samples, we retrospectively selected archived lung adenocarcinoma histological and cytological (cell blocks) samples. Overall, our SiRe RNA fusion panel was able to detect all fusions and a splicing event harbored in a RNA pool diluted up to 2 ng/µL. It also successfully analyzed 46 (95.8%) out of 48 samples. Among these, 43 (93.5%) out of 46 samples reproduced the same results as those obtained with conventional techniques. Intriguingly, the three discordant results were confirmed by a CE-IVD automated real-time polymerase chain reaction (RT-PCR) analysis (Easy PGX platform, Diatech Pharmacogenetics, Jesi, Italy). Based on these findings, we conclude that our new SiRe RNA fusion panel is a valid and robust tool for the detection of clinically relevant gene fusions and splicing events in advanced NSCLC

La rilevazione dello stato mutazionale di EGFR su campione citologico di neoplasia polmonare a cellule non piccole: il ruolo dell' anatomo patologo nella validazione analitica e clinica.

La terapia mirata del carcinoma del polmone non a piccole cellule (NSCLC) con inibitori del dominio tirosin-chinasico (anti-TKI) del recettore del fattore di crescita epidermico (EGFR) è efficace solo nei pazienti con mutazioni di tale gene. Le mutazioni più frequenti sono rappresentate da delezioni nell'esone 19 e dalla mutazione puntiforme (L858R) nell'esone 21. A causa della scoperta in fase avanzara di tali neoplasie, spesso la diagnosi di carcinoma polmonare è esclusivamente citologica; il DNA estratto dai campioni citologici viene poi utilizzato per determinare lo stato mutazionale di EGFR. Tuttavia, la possibilità che le mutazioni siano distribuite nel tessuto polmonare eterogeneamente è tutt'ora discussa. E' possibile che i campioni citologici non siano del tutto rappresentativi dello stato mutazionale dell'intera neoplasia e quindi, il valore predittivo dell' analisi mutazionale effettuata esclusivamente su citologico è ancora dubbia. Questa Tesi Sperimentale esamina il ruolo del patologo nella gestione del materiale citologico da NSCLC, dalla valutazione pre-analitica del vetrino, alla supervisione delle fasi analitiche dei test molecolari e, infine, alla sintesi finale nel report diagnostico. È stata inoltre studiata la validità del campione citologico come substrato biologico adeguato nella predizione della risposta alla terapia anti-TKI

Biopsic Sampling (cancer) comprehensive sampling and sample preparation

Virtually all forms of organ injury involve structural alterations in cells and tissues. The practice of pathology is devoted to investigating and diagnosing diseases by studying these morphologic changes.1 This can be achieved by two different but complementary approaches. Histopathology involves both the macroscopic and light microscopic examination of tissues, whose architecture is retained; cytopathology specimens are composed of cells that are dissociated from their surrounding tissues without preserving the histologic architecture. Histopathology is usually the primary mode for tumor diagnosis, which is supplemented with prognostic information by tumor grading and staging.1 This section examines the sampling issues related to istopathology, and the following section focuses on cytopathology

Thyroid Carcinoma: Molecular Signature by Histotype-Specific Mutations and Gene Expression Patterns Diagnostic, Prognostic and Therapeutic Value of Gene Signatures

Although the most important application of novel molecular markers is that on cytology, translation from the bench to the FNA is complex . To make DNA- and/or RNA-based testing cost-effective on cytology a close integration with morphology is needed . Thus, the informativeness of the sample for cytology needs to be preserved to keep the accuracy of microscopy high. To this end, each of the steps of traditional cytology, such as preparation of FNA material, search for morphological criteria, assignment to the correct diagnostic class, and suggestion of the appropriate post FNA options should not be altered by molecular analysis. Then, this latter can refine cytology. The final result is to effectively stratify into high-risk and low-risk categories, and the indeterminate cytology classes identified by the BSRTC . Thus, cytological specimens should be properly handled to provide both morphological and molecular information . Our method of preparation of FNA to harvest material sufficient for both tests was recently validated on a series of 128 routinely performed FNA . The rationale behind our sample collection method was to ensure first an adequate cytological diagnosis and, then, to exploit part of the diagnostic material for molecular testing . Thus, two passes from different areas of the lesion are performed. A representative air-dried Diff-Quik stained smear is prepared within few minutes and reviewed on site ] . In 44 cases, the cytological evidences were sufficient for morphological assessment and the third pass was directly collected in RNA or DNA buffer extraction. Conversely, in 84 cases the specimen was either deemed inadequate by the onsite evaluation or required an additional ethanol-fixed Papanicolaou-stained smear to better evaluate nuclear morphology. Thus, a third pass was dedicated to the preparation of an additional smears and only needle rinsing was collected for BRAF testing. Higher average of extracted DNA concentration was observed in the dedicated pass group (25.9 vs. 7.95 ng/ m l). However, the rate of successful exon 15 BRAF amplification was similar with (43/44; 97.7%) or without (79/84; 94%) the dedicated pass. Thus, our protocol is suitable for both tests. When necessary, BRAF testing may also be performed on the residual samples of thyroid nodules, without interfering with routine cytology. Similarly, as far as mRNA markers are concerned, we have shown that in most samples, qRT-PCR analysis does not interfere with cytology . In fact, in a recent study on UbcH10 expression, including 84 cases with a cytological diagnosis of either follicular neoplasm ( n = 57) or suspicious for malignancy ( n = 27), we found that most (73.8%) cases were adequate for both test