322590 research outputs found
Sort by
VIRAL SAFETY EVALUATION OF BIOTECHNOLOGY PRODUCTS: DEVELOPMENT AND OPTIMIZATIONOF RELIABLE VIRAL CLEARANCE STRATEGIES
No full textThis manuscript brings together the research activities carried out throughout my doctoral program and
reflects the dual nature of the project, which developed at the interface between academic research and
industrial application. The thesis is part of a broader framework dedicated to the study, optimization, and
validation of viral clearance strategies within downstream processing for monoclonal antibody (mAb)
production, with the overarching goal of establishing procedures that are effective, reproducible, and fully
transferable to the biopharmaceutical industry.
During the first year, thanks to a structured training period at the Department of Pharmaceutical Sciences
(DISFARM), University of Milan, I consolidated essential skills in preparative chromatography and conducted
an extensive literature analysis on viral clearance approaches. This effort culminated in a review article,
entitled ‘Viral Clearance in Biopharmaceutical Manufacturing: Current Strategies, Challenges, and Future
Directions’, prepared in collaboration with Prof. Altomare and recently submitted, and positively revised, for
publication in Biotechnology Advances (Elsevier). Presented here as the first part of the thesis, the review
provides the reader with an up-to-date overview of the principles, technologies, and challenges associated
with viral removal within DSP, offering the conceptual foundation necessary to contextualize the
experimental core of the project.
The second part of the thesis is dedicated to the experimental work conducted jointly at DISFARM and
Eurofins Biolab Srl., where I established, optimized, and subsequently transferred to an industrial setting a
purification protocol based on Protein A chromatography and Ion Exchange Chromatography (IEX) for mAb
manufacturing. This workflow was integrated with viral removal and inactivation studies performed at the
Virology Unit of Eurofins, employing model viruses such as X-MuLV and MVM and combining
chromatographic systems, filtration technologies, and advanced biological assays. This section provides an
in-depth description of the methodologies adopted, the rationale behind the operational strategies, and a
critical discussion of the results, with particular focus on process scalability, robustness, and industrial
transferability.
Overall, the thesis aims to guide the reader through a coherent path that begins with the current state of viral
clearance technologies, progresses through the optimization of a laboratory-scale purification process, and
culminates in its validation under industrial conditions. The overarching objective is to provide a
comprehensive, structured, and forward-looking perspective on viral removal strategies for therapeutic
antibody production, highlighting the scientific and technical contributions developed over the course of the
PhD program
INVESTIGATION OF RESILIENCE AND ITS GENETICSTHROUGH THE USE OF LONGITUDINAL DATA
No full textThis thesis investigates resilience in dairy cattle using an integrated approach that combines phenotypic characterization, simulation of perturbations, and genomic analysis. The research was motivated by the need to breed animals capable of maintaining productivity amid the environmental and economic challenges facing modern agriculture. Thanks to close collaboration with a medium-large dairy farm in Northern Italy, we had access to a comprehensive dataset of daily milk records and genotype information, which formed the
basis for the analysis.
Over the past twenty years, resilience assessment in livestock has become feasible thanks to new technologies that enable the collection of high-frequency longitudinal data. Resilience indicators are recently defined proxies that combine routinely collected data, such as milk yield records, to capture an animal’s response to environmental challenges. Although it has been demonstrated that resilience indicators possess some genetic merit to some extent, their true significance and methodological foundations require further investigation to enhance the understanding of these phenotypes and their application in breeding programs.
Lactation curve models aiming to represent the unperturbed state of each individual cow are essential for deriving meaningful resilience indicators; however, their impact on these indicators has not been systematically evaluated.
This thesis addresses these gaps through three main studies: (1) characterization of resilience indicators derived from various lactation curve models; (2) a simulation study evaluating the ability of resilience indicators to capture true genetic resilience; (3) a genome-wide association study (GWAS) identifying genomic regions associated with resilience indicators. Additionally, one further contribution is included: (4) construction of a copy
number variation map for the population studied. The characterization study demonstrated that methodological choices in lactation curve modelling significantly influence phenotypic resilience indicator values and cow rankings.
The simulation study revealed that current resilience indicators capture only a fraction of true genetic resilience, with the best model-indicator combinations achieving correlations of 0.15 to 0.25 with simulated resilience breeding values. The genome-wide association study identified genomic regions associated with resilience indicators, confirming the polygenic nature of the trait and highlighting genes involved in immune response, energy metabolism, and tissue integrity. Additionally, the copy number variation map provided a valuable genomic resource for future integrated analyses.
These findings advance our understanding of resilience as a complex, multifaceted trait while providing practical tools for its genetic improvement. The practical implications are clear: breeding programs should incorporate resilience indicators derived from meaningfully selected lactation curve models, set realistic expectations for genetic progress given the modest correlations with true resilience, and balance selection for resilience alongside production and other traits through appropriately weighted selection indices. Looking ahead, continued refinement of resilience phenotypes, functional characterization of candidate genes and economic analyses will advance breeding for resilience
When do prediction markets return average beliefs? Experimental evidence
No full textIn prediction markets, prices can be interpreted as the average belief of the traders under restrictive theoretical assumptions, i.e. specific risk preferences and the Prior Information Equilibrium. The validity of these assumptions depends on the specific market institution and on the composition of the market in terms of risk preferences. In this paper we test in a laboratory experiment the main elements that should affect the distance between prices and average beliefs, manipulating the market institution and the market composition. We do not find that risk preferences significantly affect prices. We find instead that in the double auction– where at least partial information aggregation is expected – prices are closer to the average belief than in the call auction – where, instead, belief aggregation is expected. We show that traders update beliefs in the direction of observed prices, rather than of the true state
LDL-C target achievement after adding evinacumab in 2 patients with autosomal recessive hypercholesterolemia
No full textBACKGROUND: Autosomal recessive hypercholesterolemia (ARH) is a rare form of genetic hypercholesterolemia
consequent to pathogenic variants in the low-density lipoprotein receptor adaptor protein 1
(LDLRAP1) gene, coding for a protein responsible for moving LDL-receptor (LDL-R) to its site of activity.
ARH is characterized by very high levels of LDL cholesterol (LDL-C), leading to aggressive and frequently
premature atherosclerotic cardiovascular disease (ASCVD). Lowering of LDL-C is the main target of
treatment; however, classical lipid-lowering agents, for example, statins, frequently have a modest response,
in view of their selective LDL-R–raising activity.
OBJECTIVE: Among newer agents with an LDL-R–independent mechanism, evinacumab has been
shown to be effective in homozygous familial hypercholesterolemia, but few data are available
in LDLRAP1 variant carriers.
METHODS: We here report 2 cases of this extremely rare form of familial hypercholesterolemia with
a surprising response to evinacumab. Evinacumab was added to maximally tolerated background
therapy.
RESULTS: In the first patient, who had severe ASCVD and a prior inadequate response to statins,
proprotein convertase subtilisin/kexin type 9 (PCSK9) inhibitors, and lomitapide, evinacumab reduced
time-averaged LDL-C by 82% (from 549 to 62 mg/dL). In the second patient, evinacumab achieved a
sustained 73.8% LDL-C reduction, maintaining levels < 55 mg/dL and allowing discontinuation of
the PCSK9 inhibitor.
CONCLUSION: These cases demonstrate a marked and clinically meaningful LDL-C–lowering effect
of evinacumab in ARH, supporting its use as an effective LDL-R–independent therapeutic option
Modelling the effect of gravitational instabilities on the subduction zone initiation at passive margins
No full textSubduction is a widely studied process, but the mechanisms leading to the initiation of a new subduction zone are poorly understood. Two main types of subduction zone initiation (SZI) are currently recognised: induced, when tectonic convergence is dominant, and spontaneous, when SZI is mainly driven by local forces, i.e., the negative buoyancy due to the gravitational instability of the plate (Stern 2004; Stern and Gerya 2018; Crameri et al. 2020). The passive margins are potential sites for spontaneous SZI due to the natural gravitational instabilities that characterize these geodynamic settings, caused by density, composition, strength and temperature lateral contrasts, topographic discontinuity and sedimentary loading. These instabilities should favor the initiation of a new subduction zone (Stern and Gerya 2018; Lallemand and Arcay 2021; Arcay et al. 2020), but previous research suggests that these local forces are not strong enough to develop a self-sustained subduction and that the collapse of a passive margin without horizontal tectonic forcing would require an unlikely coincidence of multiple weakening mechanisms, casting doubt on the feasibility of spontaneous subduction under present-day tectonic conditions (Lallemand and Arcay 2021; Arcay et al. 2020).
In this work we explore if and how an initial gravitational phase, even if it doesn’t lead to a passive margin collapse resulting in a spontaneous subduction initiation, induces weakening and deformation in the margin, and how this influences the induced SZI and the eventual subduction style once convergence begins to affect the margin. induced SZI and the eventual subduction style once convergence begins to affect the margin.
We performed 225 2D simulations using the finite-element code FALCON (Regorda et al. 2023) on a domain 3000 Km wide and 700 Km deep, representing a passive margin composed by a 20 Myr old oceanic lithosphere and a 90 Km thick continental lithosphere.
The models undergo a first gravitational phase, simulated with free-slip lateral boundaries, that can last for 0, 10, 20 or 30 Myr. After the gravitational phase, the convergence begins, simulated by imposing a convergence velocity of 0.01, 0.05, 0.1, 0.25, 0.5 or 1 cm/yr at the lateral boundaries, along all the lithospheric thicknesses. To investigate the response of the margin to the gravitational instability, we set up 3 viscous weakening intervals and 3 plastic laws, for a total of 9 weakening combinations.
The failure and the evolution of the passive margin have been explored in post processing, by evaluating the area within a certain distance from the trench in which the strain rate is higher than 2 10-14 1/s, and the mean strain rate within this area.
During the gravitational phase, the models with a strong enough weakening combination show a strain rate localization, controlled by the plastic weakening law in the surface and by the viscous weakening interval in depth, that slowly fades over time. Once the convergence begins, this damaged zone is reactivated, and if the convergence velocity is high enough it evolves in a subduction plane. The subduction zone initiation starts before in the models that experienced strain rate localization during the gravitational phase, leading to the conclusion that the gravitational instabilities can trigger weakening processes capable of affecting the passive margin stability and the impact of the tectonic convergence on it.
Acknowledgments
This research has been supported by the ASI - Agenzia Spaziale Italiana - project ”NGGM-MAGIC - A breakthrough in understanding the dynamics of the Earth”. Contract number n. 2023-22.HH.0 ASI-UNIMI
References
Arcay, Diane, Serge Lallemand, Sarah Abecassis, and Fanny Garel (2020). “Can subduction initiation at a transform fault be spontaneous?” In: Solid Earth 11. DOI: 10.5194/se-11-37-2020.
Crameri, Fabio et al. (2020). “A transdisciplinary and community-driven database to unravel subduction zone initiation”. In: Nature Communications. DOI: 10.1038/s41467-020-17522-9.
Lallemand, Serge and Diane Arcay (2021). “Subduction initiation from the earliest stages to self-sustained subduction: Insights from the analysis of 70 Cenozoic sites”. In: Earth-Science Reviews 221. DOI: 10.1016/j.earscirev.2021.103779.
Regorda, Alessandro, Cedric Thieulot, Iris van Zelst, Zoltán Erdős, Julia Maia, and Susanne Buiter (2023). “Rifting Venus: Insights From Numerical Modeling”. In: Journal of Geophysical Research: Planets 128. DOI: 10.1029/2022JE007588.
Stern, Robert J. (2004). “Subduction initiation: Spontaneous and induced”. In: Earth and Planetary Science Letters 226. DOI: 10.1016/j.epsl.2004.08.007.
Stern, Robert J. and Taras Gerya (2018). “Subduction initiation in nature and models: A review”. In: Tectonophysics 746. DOI: 10.1016/j.tecto.2017.10.014
Carriage of rare APOB variants predisposes to severe steatotic liver disease and hepatocellular carcinoma
No full textBackground: MASLD has a substantial inherited component. Rare variants in Apolipoprotein B gene (APOB) have been implicated in susceptibility to liver steatosis, but their role in disease progression and outcomes is unclear.
Methods: We investigated APOB rare variants in a case-control cohort of people with advanced MASLD vs. healthy controls (n = 510/261), a family-based study (n = 43 and literature meta-analysis), the Million Veteran Program cohort (MVP, n = 94,885) and the UK Biobank (UKBB, n = 417,657).
Results: In the clinical cohort, APOB variants were enriched in people with advanced MASLD (OR 13.8, 95% c.i. 2.7-70.7, P = 0.002) and associated with lower circulating lipids, but higher MASLD activity and fibrosis (P < 0.05). In the family study, APOB variants segregated with hepatic steatosis and fibrosis (P < 0.05). Cross-ancestry meta-analysis of the study cohorts yielded pooled ORs for cirrhosis and hepatocellular carcinoma of 1.82, 95% c.i. 1.33-2.49 and 3.53, 95% c.i. 2.09-5.98, respectively. Variants affecting specifically ApoB100 had a three-fold greater impact on hepatic lipid metabolism compared to those impairing also ApoB48 and were specifically protective against coronary artery disease (P < 0.05). Variants affected cirrhosis risk similarly, but ApoB48/100 had a larger impact on hepatocellular carcinoma (P < 0.05).
Conclusions: Rare APOB variants predispose to advanced MASLD and HCC, with distinct contributions from disrupted VLDL and chylomicrons secretion. These findings highlight the interplay between hepatic and intestinal lipid handling, suggesting that APOB genotyping may enhance MASLD risk stratification and case identification.
Funding: European Union, Italian Ministry of Health, Swedish Research Council, Veteran health administration, NIH
Promozione delle pari opportunità e contrasto alle diseguaglianze di genere nella disciplina del fenomeno sportivo
No full textIl contributo analizza il fenomeno sportivo sia nei termini delle diseguaglianze di genere che lo affliggono sia per quel che riguarda il tessuto normativo che, a vario livello, promuove la parità di genere, tanto a livello di pratica sportiva quanto per ciò che riguarda le cariche tecniche e dirigenziali. Dopo aver ricostruito, dal punto di vista storico, alcune importanti tappe dell'emancipazione femminile attraverso lo sport, l'analisi presta particolare attenzione, nella ricostruzione del quadro normativo, alle potenzialità dell'art. 33, comma 7, Cost
Use of the Lüscher Color Test in Pediatric Dentistry: A Prospective Study in Behaviorally Challenging Pediatric Dental Patients Undergoing Conscious Sedation
No full textBackground: Dental anxiety is common in pediatric dentistry and may hinder care, particularly in behaviorally challenging children. Most anxiety measures rely on verbal report,
which can be unreliable in young patients. This study explored whether the Lüscher Color
Test, a non-verbal psychological instrument, shows associations with established anxiety
proxies in a pediatric dental sedation setting. Methods: In this single-center prospective observational study, 100 children aged 4–12 years referred for dental treatment in a conscious
sedation unit were recruited; 80 completed the protocol (exclusion rate 20%). N2O/O2
inhalation sedation was not randomized and was selected by the clinician based on clinical
judgement. Anxiety was assessed pre- and post-operative using the Lüscher Color Test,
heart rate (HR) monitoring, and the Visual Facial Anxiety Scale (VFAS). The primary outcome was the pre–post change in the Lüscher anxiety index calculated as the pre-operative
score minus the post-operative score (∆ = pre − post). Associations between changes in
anxiety measures and demographic/clinical variables were examined. Results: Anxiety
scores decreased after treatment for both the Lüscher Color Test and VFAS (both p < 0.001).
Change in Lüscher scores was positively associated with HR reduction (Spearman r = 0.68;
p < 0.01), whereas VFAS change showed a weaker association (r = 0.28; p < 0.05). In
regression analyses, treatment-related variables were explored; however, given the observational design and subgroup imbalance, these findings should be interpreted cautiously.
Conclusions: Although pre–post scores suggested a reduction in anxiety, the Lüscher Color
Test should be considered an exploratory, complementary non-verbal measure rather than
a validated diagnostic instrument. In the multivariable logistic regression, nitrous oxide
sedation showed only a non-significant trend toward greater anxiety reduction (p = 0.07).
Further studies with appropriate validation frameworks and stronger designs are needed
before clinical implementation can be recommended