Archivio Istituzionale della Ricerca - Università degli Studi della Campania "Luigi Vanvitelli"
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Production of bio-oil and biochar from digestate via pyrolysis in a mechanical fluidized bed reactor
No full textThe rising pressure to reduce greenhouse gas emissions calls for effective strategies to valorize organic waste streams. This study investigates the potential of slow pyrolysis to convert digestate, a byproduct of anaerobic digestion, into biochar and bio-oil using a mechanically stirred fluidized bed reactor. Experiments were done at 400, 450, 500, and 550 degrees C, with an additional run at 500 degrees C which employed a catalyst bed composed of processderived biochar. As the temperature increased from 400 to 500 degrees C, the biochar yield decreased from 68 % to 50 %, while the bio-oil yield increased from 21 % to 30 %. The presence of the biochar bed further reduced biochar formation by approximately 5 %, enhancing vapor production. GC-MS analysis revealed that the bio-oil was primarily composed of carbonyl compounds, sterols, alcohols, and phenolic derivatives. These results demonstrate the influence of temperature and a biochar catalyst on product distribution and composition. Overall, the study supports pyrolysis as a viable pathway for digestate valorization and sustainable carbon recovery, contributing to emissions mitigation and improved resource management
Antitumor activity and mechanism of action of S-adenosylmethionine in prostate cancer cells
No full textProstate cancer (PCa) is the most common male malignancy in Western countries, with
approximately 1.6 million new cases annually. The etiology of PCa is not yet fully understood,
but the main causes are attributable to environmental and genetic factors. Because of the
fundamental role in tumor progression of the androgen receptor, hormonal therapy or androgen
deprivation therapy induces tumor regression and apoptosis in hormone-sensitive cells.
However, metastatic castration-resistant prostate cancer (mCRPC), an advanced stage of
prostate cancer, does not respond to conventional therapy due to androgen receptor variants
that promote aberrant activation and neoplastic progression.
In recent years, scientific research has focused on the development and identification
of natural compound–based drugs. Many natural compounds appear to be effective in
enhancing the therapeutic effect of common chemotherapeutic drugs and could be used in
combination therapy, reducing side effects and overcoming drug resistance mechanisms.
Among natural molecules, S-adenosyl-L-methionine (AdoMet or SAM) has been extensively
studied as a therapeutic compound, and its application, alone or in combination with other
molecules, is emerging as a potential effective strategy for cancer treatment and
chemoprevention. AdoMet is a sulfur-containing nucleoside ubiquitous in all living organisms
that plays an indispensable role in cellular metabolism, participating in fundamental
biochemical reactions, the most relevant being transmethylation, transsulfuration, and
polyamine biosynthesis.
The research carried out during my PhD program is part of a broader project aimed at
developing new and alternative therapeutic approaches for the treatment of prostate cancer,
with the goal of addressing the increased mortality of patients affected by this aggressive tumor.
During the three years of my PhD, I evaluated the antiproliferative effects of AdoMet on
mCRPC cells and the potential of this sulfonium compound to overcome multidrug resistance
mechanisms induced by treatment with cabazitaxel (CBZ), one of the most commonly used
chemotherapeutic drugs in clinical practice for prostate cancer treatment. Recent findings have
highlighted the role of AdoMet as a modulator of a wide variety of signaling pathways involved
in carcinogenesis, including cell proliferation, apoptosis, and autophagy. It is important to note
that AdoMet is an FDA-approved dietary supplement, which makes it suitable for therapeutic
purposes without the common contraindications of chemotherapeutic drugs.
To assess the antiproliferative effects of AdoMet, cell viability assays were first
performed after treatment with increasing doses of AdoMet (from 62 μM to 1 mM) for different
time points (24, 48, and 72 hr) using two PCa cell lines, PC-3 and DU 145. The results showed
that AdoMet exerts an inhibitory effect on cell proliferation in a dose- and time-dependent
manner, inducing a 50% reduction in cell viability after 72 hr at a concentration of 400 μM.
Subsequently, to evaluate the chemosensitizing effects of AdoMet on the antiproliferative
activity of CBZ, cell viability assays were performed after 72 hr of treatment with increasing
doses of CBZ, alone or in combination with 400 μM AdoMet. The results suggest that mCRPC
cells are more sensitive to CBZ treatment when combined with AdoMet, confirming that
AdoMet can enhance the cytotoxic effect of CBZ.
Glutathione (GSH) is a key component of the cellular antioxidant system. In cancer
cells, elevated GSH levels neutralize reactive oxygen species (ROS) and detoxify xenobiotics.
Depletion of intracellular GSH makes neoplastic cells more vulnerable to oxidative stress and
chemotherapeutics. Analysis of GSH levels after treatment with 400 μM AdoMet and 0.7 nM
or 1.5 nM CBZ, administered individually or in combination for 48 and 72 hr in DU 145 and
PC-3 cell lines, revealed a significant reduction of GSH in cells treated with the combination
therapy compared to controls and single-agent treatments. This effect was more pronounced
after 72 hr, especially in DU 145 cells. The substantial decrease in GSH suggests that the
combined treatment effectively compromises the antioxidant defenses of mCRPC cells,
increasing their sensitivity to oxidative damage. Supporting this, Western blot analysis showed
that AdoMet, CBZ, and especially their combination reduced the expression levels of key
antioxidant enzymes involved in maintaining redox homeostasis and protecting against ROS-
induced damage, such as GPX4 and catalase. Compounds that increase ROS represent a
promising therapeutic strategy, as intensifying oxidative stress in cancer cells, combined with
GSH depletion, enhances their susceptibility to chemotherapy. Intracellular ROS levels were
also measured by flow cytometry using a CellROX assay, a weakly fluorescent dye that, upon
ROS oxidation, emits a strong and stable green fluorescence. In DU 145 cells, single treatments
did not alter ROS levels, while the combination of AdoMet and CBZ led to a 1.6-fold increase.
In PC-3 cells, AdoMet induced a slight increase and CBZ a more marked rise, but the combined
therapy produced the most significant effect, with a 2.4-fold increase compared to control.
Furthermore, to evaluate whether high ROS levels caused DNA damage in mCRPC cells, the
expression of γH2AX, the phosphorylated form of histone H2AX and a sensitive marker of
genomic damage, was analyzed. Immunofluorescence and Western blot showed increased
γH2AX signal intensity in treated nuclei, particularly in the combination treatment,
highlighting the synergistic interaction between the drug and the sulfonium compound in
causing DNA damage.
The results obtained led me to investigate morphological alterations in mCRPC cells
after treatment with AdoMet and/or CBZ, to study the effects of the sulfonium compound on
cell death by mitotic catastrophe, a mechanism considered tumor-suppressive as it directs cells
unable to complete mitosis toward death. Mitotic catastrophe is an aberrant form of mitosis
characterized mainly by misassembled spindles and abnormal chromosome arrangements.
These morphological features were used to study this phenomenon. Specifically,
immunofluorescence was performed using an α-tubulin antibody to visualize the microtubule
network and mitotic spindle, while chromosomes were stained with Hoechst 33258. Images
revealed well-organized mitotic spindles and properly aligned chromosomes on the metaphase
plate in control cells, whereas AdoMet- and CBZ-treated cells showed disorganized
chromosomes and absence of a mitotic spindle. This spindle disorganization was even more
evident when AdoMet and CBZ were administered in combination. To further investigate the
antiproliferative effects of AdoMet, CBZ, and their combination in DU 145 and PC-3 cell lines,
the main mechanism of cell death, apoptosis, was assessed by FACS analysis. After 72 hr, a
significant increase in apoptosis was observed in cells treated with the combination compared
to single treatments. These FACS results were confirmed by Western blot analysis of cleaved
poly ADP-ribose polymerase (PARP), which was particularly evident in the combination
treatment, confirming the synergistic activation of apoptosis by AdoMet and CBZ.
In conclusion, these results confirm that AdoMet exerts pleiotropic effects in promoting
tumor cell death by modulating ROS and spindle integrity and suggest the possible use of this
important and versatile physiological compound, in combination with CBZ, for the
development of low-cost therapeutic strategies targeting prostate cancer patients.
AdoMet is one of the most studied epigenetic regulators and a known modulator of
miRNAs in many biological processes, including cancer progression. To gain new insights into
the molecular mechanisms underlying AdoMet’s antitumor activity and to evaluate whether
AdoMet can act as an epigenetic regulator of miRNAs in mCRPC cells, we analyzed the
expression profile of miRNAs involved in PCa pathogenesis. To this end, quantitative Real-
Time PCR (qRT-PCR) experiments were performed after treatment with 400 μM AdoMet for
72 hr in DU 145 cell line. The analysis showed that AdoMet treatment was able to modulate
the expression levels of miR-34a and miR-21. MiR-34a is a tumor suppressor involved in
regulating pro-apoptotic genes and cell cycle control. Conversely, miR-21, often upregulated
in prostate cancer, promotes tumor progression by inhibiting tumor suppressor genes such as
PTEN, contributing to increased cellular aggressiveness and treatment resistance.
To broaden the study of miRNA regulation after AdoMet treatment, an exploratory
bioinformatic analysis was performed. In particular, edgeR, an R-based software for RNA-seq
data analysis that applies negative binomial distribution models to estimate variability and
detect differential gene expression, was used. Enrichment analyses through GO (Gene
Ontology) and KEGG (Kyoto Encyclopedia of Genes and Genomes) were then conducted to
interpret the biological pathways involved. These preliminary analyses form the basis for a
more in-depth interpretation of the data, allowing hypotheses on the potential pathways and
biological processes regulated by AdoMet. Further studies will be required to validate and
clarify the functional significance of these findings
The Potential of Material and Product Passports for the Circular Management of Heritage Buildings
No full textInterventions on Heritage Buildings (HBs) involve significant challenges due to their tangi-
ble (embodied in the material, architectural, physical and technical integrity of the cultural
asset), and intangible values (linked to socio-historical–cultural and collective identity,
memory, customs and symbols meanings), which must be preserved while also adapting
to current sustainability and circular economy goals. However, current conservation and
management practices often lack systematic tools to trace, assess, and organise material
and component information, hindering the implementation of circular strategies. In line
with the European Union’s objectives for climate neutrality and resource efficiency and suf-
ficiency, Material and Product Passports (MPPs) have emerged as digital tools that enhance
data traceability, interoperability and transparency throughout a building’s lifecycle. This
paper examines the potential of MPPs to support circular management of HBs by analysing
the structure of MPPs and outlining the information flows generated by rehabilitation,
maintenance and adaptive reuse strategies. A mixed methods approach, combining litera-
ture review and data structure analysis, is adopted to identify how the different categories
of data produced during maintenance, rehabilitation and adaptive reuse processes can be
integrated into MPP modules. The research highlights the conceptual opportunities of
MPPs to document and interlink historical, cultural, and technical data, thereby improving
decision-making and transparency across intervention stages. The analysis suggests that
adapting MPPs to the specificities of historic contexts, such as authenticity preservation,
reversibility, and contextual sensitivity, can foster innovative, sustainable, and circular
practices in the conservation and management of HBs
Pre-feasibility analysis of a cold ironing system for cruise ships. An engineering, economic and environmental analysis
No full textThe present paper presents a pre-feasibility study for the development of a cold ironing system for cruise ships in the Italian context. The aim of the study is to understand if the development of a cold ironing system is more sustainable from the energy, environmental, and economic point of view with respect to onboard generation. To this end, a simple model is developed for the estimation of energy consumption, pollutant, and carbon emissions of a cruise ship while hotelling in port. The model allows to assess the level of consumption and emissions for both onboard generation and cold ironing. Since the parameters of the model are affected by uncertainty, a Monte Carlo analysis is also developed to understand the overall range of variability of the results. Then, a financial model is developed to estimate an appropriate tariff for the cold ironing system and to compare with the fuel cost. Results show that savings of 3ktoe/year of energy, 200 t/day of CO2, 9 M€2021 in operating cost, 12 M€2021 in externality costs can be achieved if a cold ironing system is used. Monte Carlo analysis shows that there is a probability of 95% to save between 10.3 and 8.1 M€ in operating costs
DIGITAL CHILD LABOUR. EMERGING CHALLENGES FOR ITALIAN LABOUR LAW
No full textThe evolution of the digital economy has redefined the boundaries of child
labour, giving rise to new forms of exploitation and vulnerability alongside traditional ones.
From content production on social media to employment in the competitive video game
industry, children today are exposed to risks that are often unrecognised and inadequately
regulated. This paper examines the new frontiers of child labour in the digital age, with a
particular focus on the Italian context, highlighting the main regulatory gaps and emerging
legal challenges. In particular, it analyses the capacity of the current Italian legislative
framework to prevent and combat the most insidious forms of online child exploitation. The
analysis reveals the inadequacy of the Italian legal system to respond to the complexities of
the new digital ecosystem. In conclusion, some guidelines are outlined for a reform aimed at
strengthening the protection of minors, promoting an integrated and cross-sectoral approach
Edizione di papiri paraletterari e documentari dalla Collezione Milano Vogliano
No full textEdition of 32 texts from greek papyri of the roman, byzantine and arab period of the Collection of the University of Milan. The first 15 texts are from the so-called ‘Archive of Petaus‘, while the following 17 are miscellaneous texts dating from the IV to the VIII century
A line-search based SGD algorithm with Adaptive Importance Sampling
No full textStochastic Gradient methods are widely used in the field of supervised learning associated with big data. In this context, importance sampling-based algorithms have been proposed to minimize the variance of the stochastic gradient by introducing practical strategies to approximate the optimal sampling distribution, which is otherwise only theoretically accessible. In this paper, we propose a scheme that combines stochastic gradient descent with adaptive importance sampling with automatic step-size selection based on a stochastic Armijo-type line-search. This approach makes the method robust to the choice of the initial step-size, which would otherwise require a tuning phase that is computationally expensive or even impractical in certain big data scenarios. Moreover, we introduce different mini-batch variants to foster the practical acceleration of the original scheme. Finally, numerical experiments are presented on real datasets to validate the proposed method in the context of supervised classification problems