Omission of axillary clearance in patients with early breast cancer and positive sentinel node biopsy treated with conservative surgery: application of the ACOSOG Z011 trial in clinical practice

Background: Axillary lymph node dissection (ALND) in presence of sentinel lymph node (SLN) metastases has been the standard in breast cancer (BC) patients for many years. Today, after the publication of the ACOSOG Z0011 trial, ALND is a procedure restricted to a dwindling group of patients with a clearly metastatic axilla. Material and methods: This was a prospective observational trial involving two Italian Breast Units: Policlinico di Sant’Orsola and San Raffaele hospital. Objective was to evaluate that the omission of ALND in patients with cT1-2 cN0 BC undergoing breast conserving surgery (BCS) and histological finding of metastases in 1 or 2 SLN is not associated with a worse prognostic outcome. Primary endpoint was overall survival (OS). Secondary endpoints were disease free survival (DFS) and locoregional recurrence. All BC patients treated between the 1st of November 2020 and 31st of July 2023 with cT1-2 cN0 BC, preoperative negative axillary ultrasound and 1 or 2 metastatic SLN treated with sentinel node biopsy (SLNB) alone entered the study. Results: 795 cT1-2 cN0 BC patients underwent BCS and SLNB. Ninety patients were included. Median age was 60 (52-68) years. Seventy-five patients (83%) had T1 tumor and 15 (17%) T2. Median tumor size was 16 mm (11-19). The median SLN removed was 2 (1-3). Eighty-one patients had 1 positive SLN (90%), while 9 had 2 SLN metastasis (10%). 39 (43%) micrometastases were identified and 51 macrometastasis (57%). All patients underwent radiotherapy. Seventeen (19%) performed adjuvant chemotherapy. Two received immunotherapy with trastuzumab and pertuzumab. Endocrine therapy was given to 84 (93%). At a median follow-up of 19 months (IQR 13-23) OS and DFS were 100%. No loco-regional recurrence was seen. Conclusion: The preliminary results of our study confirm that omitting ALND in patients meeting Z011 criteria is oncologically safe and should be the standard of care.

Development of strategies for vascular damage repair in Pulmonary Arterial Hypertension

Pulmonary arterial hypertension (PAH) is a progressive and rare disease with so far unclear pathogenesis, limited treatment options and poor prognosis. Unbalance of proliferation and migration in pulmonary arterial smooth muscle cells (PASMCs) is an important hallmark of PAH. In this research Sodium butyrate (BU) has been evaluated in vitro and in vivo models of PAH. This histone deacetylase inhibitor (HDACi) counteracted platelet-derived growth factor (PDGF)-induced ki67 expression in PASMCs, and arrested cell cycle mainly at G0/G1 phases. Furthermore, BU reduced the transcription of PDGFRbeta, and that of Ednra and Ednrb, two major receptors in PAH progression. Wound healing and pulmonary artery ring assays indicated that BU inhibited PDGF-induced PASMC migration. BU strongly inhibited PDGF-induced Akt phosphorylation, an effect reversed by the phosphatase inhibitor calyculinA. In vivo, BU showed efficacy in monocrotaline-induced PAH in rats. Indeed, the HDACi reduced both thickness of distal pulmonary arteries and right ventricular hypertrophy. Besides these studies, Serial Analysis of Gene Expression (SAGE) has be used to obtain complete transcriptional profiles of peripheral blood mononuclear cells (PBMCs) isolated from PAH and Healthy subjects. SAGE allows quantitative analysis of thousands transcripts, relying on the principle that a short oligonucleotide (tag) can uniquely identify mRNA transcripts. Tag frequency reflects transcript abundance. We enrolled patients naïve for a specific PAH therapy (4 IPAH non-responder, 3 IPAH responder, 6 HeritablePAH), and 8 healthy subjects. Comparative analysis revealed that significant differential expression was only restricted to a hundred of down- or up-regulated genes. Interestingly, these genes can be clustered into functional networks, sharing a number of crucial features in cellular homeostasis and signaling. SAGE can provide affordable analysis of genes amenable for molecular dissection of PAH using PBMCs as a sentinel, surrogate tissue. Altogether, these findings may disclose novel perspectives in the use of HDACi in PAH and potential biomarkers.

Controllo dei parassiti nel rettilario del Bioparco di Roma

Poiché le infestazioni parassitarie dei rettili in cattività possono incidere sulla loro salute, è stato condotto uno studio sui parassiti intestinali e gli ectoparassiti su 213 rettili del Bioparco di Roma. In cattività, dove gli animali sono confinati in piccoli spazi, la concentrazione di parassiti può essere più alta. Alcuni di essi sono innocui, ma altri, specialmente in associazione allo stress, possono essere pericolosi, causare malattie e portare anche alla morte se non trattati. Inoltre, i rettili parassitati hanno una vita più breve, sono più suscettibili ad altre malattie, hanno scarsa fertilità, ridotta crescita ed alta mortalità. Tra Gennaio 2012 e Dicembre 2014 sono stati raccolti campioni da 213 rettili che non mostravano alcun segno clinico e che includevano 23 specie di sauri, 20 specie di cheloni, 20 specie di ofidi e 4 specie di alligatori. I campioni sono stati esaminati per la presenza di parassiti intestinali mediante copromicroscopia di tipo qualitativo previa sedimentazione e successiva flottazione, mentre la tecnica di Zihel-Neelsen è stata usata per la ricerca di oocisti di Cryptosporidium . I campioni di feci sottoposti ad esame copromicroscopico qualitativo sono risultati positivi per almeno una tipologia di endoparassiti nel 49% dei casi. Il 35% dei campioni è risultato positivo per ossiuri, il 22% per coccidi, l’1% per strongili ed il 5% per ascaridi. Degli strisci fecali colorati con la tecnica di Zihel-Neelsen, il 25% sono risultati positivi per Cryptosporidium spp. Ossiuri e coccidi sono stati più prevalenti nei sauri rispetto ai cheloni, agli ofidi e agli alligatori e nel 14% è stata osservata la presenza di entrambi. Inoltre, per quanto riguarda gli ectoparassiti, sono state rilevate uova di Myocoptes musculinus nel 24% dei rettili esaminati. Tutti gli strisci risultati positivi per Cryptosporidium spp. sono stati inviati all’Università di Santiago de Compostela per essere sottoposti alla tecnica PCR.

Controlled reproduction in Anguilla anguilla (L.): advanced studies on broodstock management, spawning techniques and system design for artificial seed production.

The thesis aims to expose the advances achieved in the practices of captive breeding of the European eel (Anguilla anguilla). Aspects investigated concern both approaches livestock (breeding selection, response to hormonal stimulation, reproductive performance, incubation of eggs) and physiological aspects (endocrine plasma profiles of players), as well as engineering aspects. Studies conducted on various populations of wild eel have shown that the main determining factor in the selection of wild females destined to captive breeding must be the Silver Index which may determine the stage of pubertal development. The hormonal induction protocol adopted, with increasing doses of carp pituitary extract, it has proven useful to ovarian development, with a synchronization effect that is positively reflected on egg production. The studies on the effects of photoperiod show how the condition of total darkness can positively influence practices of reproductions in captivity. The effects of photoperiod were also investigated at the physiological level, observing the plasma levels of steroids ( E2, T) and thyroid hormones (T3 and T4) and the expression in the liver of vitellogenin (vtg1 and vtg2) and estradiol membrane receptor (ESR1). From the comparison between spontaneous deposition and insemination techniques through the stripping is inferred as the first ports to a better qualitative and quantitative yield in the production of eggs capable of being fertilized, also the presence of a percentage of oocytes completely transparent can be used to obtain eggs at a good rate of fertility. Finally, the design and implementation of a system for recirculating aquaculture suited to meet the needs of species-specific eel showed how to improve the reproductive results, it would be preferable to adopt low-flow and low density incubation.

Managing invasive populations of Anoplophora chinensis and A. glabripennis in Lombardy

Two Asian longhorned beetles (Coleoptera: Cerambycidae), commonly known as Citrus Longhorned Beetle (CLB), Anoplophora chinensis (Forster), and Asian Longhorned Beetle (ALB), A. glabripennis (Motschulsky), are considered the most destructive wood borers introduced in Lombardy (northern Italy). This research aimed at (1) improving laboratory rearing methods for the biological control agent Aprostocetus anoplophorae (Hym.: Eulophidae), an egg parasitoid specific to CLB, and defining release techniques allowing its establishment; (2) test the efficacy of the sentinel tree technique for the early detection of CLB; and (3) evaluating the efficacy of traps baited with artificial lures in attracting adults of ALB and possibly CLB. Several problems were faced while rearing the egg parasitoid in laboratory. It appeared that the rate of parasitism of the hosts could depend on the age of the host eggs and/or age of the laying parasitoid females. Data results from the field experiments about A. anoplophorae release-capture showed that the percentage of slits containing a CLB egg was particularly low on most sentinel trees and the percentage of CLB eggs that were killed, because of natural predators, was high. Only one egg amongst those exposed was attacked by the released parasitoid. These negative results were anyway very useful, since they provided evidence and information on the type of host plants to be used, the time necessary for the exposure of the plants to the egg-laying CLB females, the number of laying parasitoid females to be inserted per cage. The sentinel trees technique revealed to be not successful; signs and symptoms of CLB presence were not recorded during the two seasons of field observations (2012-2013). Extremely positive was instead the trial with artificial lures carried out during summer 2013. A total of 32 beetles were captured (4 ALB and 28 CLB) deploying 50 baited traps.

Multi-scale analysis of active landslides using two-pass differential interferometry

Landslides are common features of the landscape of the north-central Apennine mountain range and cause frequent damage to human facilities and infrastructure. Most of these landslides move periodically with moderate velocities and, only after particular rainfall events, some accelerate abruptly. Synthetic aperture radar interferometry (InSAR) provides a particularly convenient method for studying deforming slopes. We use standard two-pass interferometry, taking advantage of the short revisit time of the Sentinel-1 satellites. In this paper we present the results of the InSAR analysis developed on several study areas in central and Northern Italian Apennines. The aims of the work described within the articles contained in this paper, concern: i) the potential of the standard two-pass interferometric technique for the recognition of active landslides; ii) the exploration of the potential related to the displacement time series resulting from a two-pass multiple time-scale InSAR analysis; iii) the evaluation of the possibility of making comparisons with climate forcing for cognitive and risk assessment purposes. Our analysis successfully identified more than 400 InSAR deformation signals (IDS) in the different study areas corresponding to active slope movements. The comparison between IDSs and thematic maps allowed us to identify the main characteristics of the slopes most prone to landslides. The analysis of displacement time series derived from monthly interferometric stacks or single 6-day interferograms allowed the establishment of landslide activity thresholds. This information, combined with the displacement time series, allowed the relationship between ground deformation and climate forcing to be successfully investigated. The InSAR data also gave access to the possibility of validating geographical warning systems and comparing the activity state of landslides with triggering probability thresholds.

Cross-sensors analysis of multispectral Earth observation data for environmental monitoring

Time Series Analysis of multispectral satellite data offers an innovative way to extract valuable information of our changing planet. This is now a real option for scientists thanks to data availability as well as innovative cloud-computing platforms, such as Google Earth Engine. The integration of different missions would mitigate known issues in multispectral time series construction, such as gaps due to clouds or other atmospheric effects. With this purpose, harmonization among Landsat-like missions is possible through statistical analysis. This research offers an overview of the different instruments from Landsat and Sentinel missions (TM, ETM, OLI, OLI-2 and MSI sensors) and products levels (Collection-2 Level-1 and Surface Reflectance for Landsat and Level-1C and Level-2A for Sentinel-2). Moreover, a cross-sensors comparison was performed to assess the interoperability of the sensors on-board Landsat and Sentinel-2 constellations, having in mind a possible combined use for time series analysis. Firstly, more than 20,000 pairs of images almost simultaneously acquired all over Europe were selected over a period of several years. The study performed a cross-comparison analysis on these data, and provided an assessment of the calibration coefficients that can be used to minimize differences in the combined use. Four of the most popular vegetation indexes were selected for the study: NDVI, EVI, SAVI and NDMI. As a result, it is possible to reconstruct a longer and denser harmonized time series since 1984, useful for vegetation monitoring purposes. Secondly, the spectral characteristics of the recent Landsat-9 mission were assessed for a combined use with Landsat-8 and Sentinel-2. A cross-sensor analysis of common bands of more than 3,000 almost simultaneous acquisitions verified a high consistency between datasets. The most relevant discrepancy has been observed in the blue and SWIRS bands, often used in vegetation and water related studies. This analysis was supported with spectroradiometer ground measurements.

Musculoskeletal disorders, research quality, and pelvic floor health in sports: exploring current evidence and new perspectives

Background. Pelvic floor dysfunction (PFD) is an umbrella term that includes a myriad of conditions such as urinary (UI) and anal incontinence, pelvic organ prolapse, pelvic pain, and sexual dysfunction. Literature showed high prevalence rates of PFD among athletes, especially UI, with high-impact sports have been linked with an increased risk of developing symptoms. However, comprehensive research summarising PFD prevalence across sexes, exploring treatment options, and the absence of a standardised referral screening tool are notable gaps. Misinformation is also prevalent in the sports medicine field. Methods. This doctoral project comprises four studies addressing different aspects of pelvic health in athletes. The first two studies were scoping reviews of epidemiological PFD data in male and female athletes, as well as available interventions. Study 3 concerned the development of a new screening tool for PFD in female athletes, aiming to guide sports medicine clinicians in referring patients to PFD specialists through a worldwide Delphi consensus. Study 4 summarised all previous findings, integrating data into an infographic. Results and conclusions. In Study 1, the findings of 100 articles on PFD in both sexes have been collected, highlighting a higher prevalence of studies on female athletes evaluating UI across multiple sports. Other conditions remain rarely investigated. Study 2 found a diverse range of interventions for female PFD, with a notable emphasis on conservative approaches. Recommendations for clinical practice often relied on the transferability of results from the nonathlete population or expert opinions. In Study 3, 41 international experts took part in the consensus development of the Pelvic Floor Dysfunction-ScrEeNing Tool IN fEmale athLetes (PFD-SENTINEL). It incorporates a cluster of PFD symptoms, items (risk factors, clinical, and sports-related characteristics), and a clinical algorithm. Lastly, Study 4 included ten evidence-based information with a relative description concerning pelvic floor health in athletes.

Epidemiological studies on toxoplasma gondii infection in a one health perspective

The objective of the present Ph.D. thesis was to investigate with a One Health approach the epidemiological patterns of T. gondii infection in Italy, to better understand the transmission dynamics of the parasite, following different research lines. The results of a retrospective analysis in animals and human showed the widespread distribution of T. gondii in the study area, with specific antibodies found in various animal species and human populations, indicating its constant presence across diverse environments. The environment plays a significant role in T. gondii's epidemiology. Migratory aquatic birds, rodents, wolves, and wild boars were investigated as sentinels of their spread, highlighting the potential transmission across geographic areas and infection risks for wildlife in natural settings. The study also provided insights into seroprevalence in wolves. Dogs, subjected to serological investigations exhibited risk factors for T. gondii infection, such as cohabitation with cats, coprophagy behaviours, and continuous outdoor. Correlation between serological evidence of exposure to T. gondii and pathological anxiety in large-size dogs was observed, and the consumption of raw meat was associated with a higher risk of infection in these animals. Results of the investigations conducted in this thesis, demonstrate the dynamic nature of T. gondii infection in cattle, characterized by new infections and declining antibody levels over the production cycle. The study also describes a co-infection between T. gondii and Sarcocystis hominis in bovine eosinophilic myositis. In the final part of the Thesis, a comprehensive genotyping of T. gondii in Italy reveals the predominance of Type II strains, particularly in cases of ovine abortion and fatal toxoplasmosis among captive Lemur catta. This approach enhances our understanding of the parasite's genetic diversity and transmission patterns, vital for effective management of its impact on human and animal health in Italy.