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IntroductionCPAP therapy treats various respiratory disorders. The overall performance of therapy delivery can be affected by the adopted circuit configuration. Recently, parallel to the canonical open configuration (OC), closed configurations (CC) have been proposed with potential advantages in terms of oxygen consumption, noise, airway dryness and contamination. However, the mechanical performance of CPAP devices in CC has been marginally investigated. The aim of this study is to clarify whether CPAP therapy delivered in CC configuration retains mechanical performances equivalent to that achieved in the canonical OC stipulated by the manufacturers.MethodsOCs and CCs implemented on seven different ventilation devices, classified as flowmeter, obstructive sleep apnoea device, and mechanical ventilator, were tested at different set CPAP levels. Mask and helmet interfaces were tested, and healthy, post-surgery and ARDS respiratory conditions were simulated. The mechanical performance was compared in terms of mean static pressure (Pmean), pressure oscillations, areas between pressure curve and Pmean during inspiration (Ai) and expiration (Ae), and the time in which the pressure curve remains above the Pmean along the expiration phase (T%).ResultsThe mechanical performances of CCs with helmet interface were comparable to canonical OCs used with mask interface. Globally, a CC supplied a reduced Pmean (on average, −1.3 cmH2O for the mask and −0.3 cmH2O for the helmet) and an increased ΔP, Ae and Ai (on average +0.5, +2.5, +2 times, respectively).ConclusionThe closed configuration proved its capability to effectively deliver CPAP therapy, thus making its intrinsic advantages available for future clinical use.

BACKGROUND: Coronary artery ectasias and aneurysms (CAE/CAAs) are among the less common forms of coronary artery disease, with undefined long-term outcomes and treatment strategies. AIMS: To assess the clinical characteristics, angiographic patterns, and long-term outcomes in patients with CAE, CAA, or both. METHODS: This 15-year (2006–2021) retrospective single-centre registry included 281 patients diagnosed with CAE/CAA via invasive coronary angiography. Major adverse cardiovascular events included all-cause death, non-fatal myocardial infarction, unplanned ischaemia-driven revascularisation, hospitalisation for heart failure, cerebrovascular events, and clinically overt bleeding. Time-dependent event risks for the CAE and CAA groups were assessed using Cox regression models and Kaplan-Meier curves. RESULTS: CAEs (n = 161, 57.3%) often had a multi-district distribution (45.8%), while CAAs (78, 27.8%) exhibited a single-vessel pattern (80%). The co-existence of CAAs and CAE was observed in 42 cases (14.9%), and multi-vessel obstructive coronary artery disease was prevalent (55.9% overall). Rates of major adverse cardiovascular events were 14.3% in-hospital and 38.1% at a median follow-up of 18.9 (interquartile range [IQR] 6.0–39.9) months. The presence of CAAs was associated with increased major adverse cardiovascular events risk in comparison to CAE (hazard ratio [HR] = 2.26, 95% confidence interval [CI] 1.38–3.69, p = 0.001), driven by a higher hazard ratio of non-fatal myocardial infarctions (HR = 5.00, 95% CI 1.66–15.0, p = 0.004) and unplanned ischaemia-driven revascularisation in both dilated (HR = 3.23, 95% CI 1.40–7.45, p = 0.006) and non-dilated coronary artery segments (HR 3.83, 95% CI 2.08–7.07, p = 0.001). CONCLUSIONS: Overlap between obstructive and dilated coronary artery disease is frequent. Among the spectrum of dilated coronary artery disease, the presence of a CAA was associated with worse long-term outcomes.

Over the last years computer modelling and simulation has emerged as an effective tool to support the total product life cycle of cardiovascular devices, particularly in the device preclinical evaluation and post-market assessment. Computational modelling is particularly relevant for heart valve prostheses, which require an extensive assessment of their hydrodynamic performance and of risks of hemolysis and thromboembolic complications associated with mechanically-induced blood damage. These biomechanical aspects are typically evaluated through a fluid-structure interaction (FSI) approach, which enables valve fluid dynamics evaluation accounting for leaflets movement. In this context, the present narrative review focuses on the computational modelling of bileaflet mechanical aortic valves through FSI approach, aiming to foster and guide the use of simulations in device total product life cycle. The state of the art of FSI simulation of heart valve prostheses is reviewed to highlight the variety of modelling strategies adopted in the literature. Furthermore, the integration of FSI simulations in the total product life cycle of bileaflet aortic valves is discussed, with particular emphasis on the role of simulations in complementing and potentially replacing the experimental tests suggested by international standards. Simulations credibility assessment is also discussed in the light of recently published guidelines, thus paving the way for a broader inclusion of in silico evidence in regulatory submissions. The present narrative review highlights that FSI simulations can be successfully framed within the total product life cycle of bileaflet mechanical aortic valves, emphasizing that credible in silico models evaluating the performance of implantable devices can (at least) partially replace preclinical in vitro experimentation and support post-market biomechanical evaluation, leading to a reduction in both time and cost required for device development.

Abstract Intrinsic permeability describes the ability of a porous medium to be penetrated by a fluid. Considering porous scaffolds for tissue engineering (TE) applications, this macroscopic variable can strongly influence the transport of oxygen and nutrients, the cell seeding process, and the transmission of fluid forces to the cells, playing a crucial role in determining scaffold efficacy. Thus, accurately measuring the permeability of porous scaffolds could represent an essential step in their optimization process. In literature, several methods have been proposed to characterize scaffold permeability. Most of the currently adopted approaches to assess permeability limit their applicability to specific scaffold structures, hampering protocols standardization, and ultimately leading to incomparable results among different laboratories. The content of novelty of this study is in the proposal of an adaptable test bench and in defining a specific testing protocol, compliant with the ASTM International F2952-22 guidelines, for reliable and repeatable measurements of the intrinsic permeability of TE porous scaffolds. The developed permeability test bench (PTB) exploits the pump-based method, and it is composed of a modular permeability chamber integrated within a closed-loop hydraulic circuit, which includes a peristaltic pump and pressure sensors, recirculating demineralized water. A specific testing protocol was defined for characterizing the pressure drop associated with the scaffold under test, while minimizing the effects of uncertainty sources. To assess the operational capabilities and performance of the proposed test bench, permeability measurements were conducted on PLA scaffolds with regular (PS) and random (RS) micro-architecture and on commercial bovine bone matrix-derived scaffolds (CS) for bone TE. To validate the proposed approach, the scaffolds were as well characterized using an alternative test bench (ATB) based on acoustic measurements, implementing a blind randomized testing procedure. The consistency of the permeability values measured using both the test benches demonstrated the reliability of the proposed approach. A further validation of the PTB’s measurement reliability was provided by the agreement between the measured permeability values of the PS scaffolds and the theory-based predicted permeability value. Once validated the proposed PTB, the performed measurements allowed the investigation of the scaffolds’ transport properties. Samples with the same structure (guaranteed by the fused-deposition modeling technique) were characterized by similar permeability values, and CS and RS scaffolds showed permeability values in agreement with the values reported in the literature for bovine trabecular bone. In conclusion, the developed PTB and the proposed testing protocol allow the characterization of the intrinsic permeability of porous scaffolds of different types and dimensions under controlled flow regimes, representing a powerful tool in view of providing a reliable and repeatable framework for characterizing and optimizing scaffolds for TE applications.

BackgroundComputational fluid dynamics (CFD) is emerging as an effective technology able to improve procedural outcomes and enhance clinical decision-making in patients with coronary artery disease (CAD). The present study aims to assess the state of knowledge, use and clinical acceptability of CFD in the diagnosis and treatment of CAD.MethodsWe realized a 20-questions international, anonymous, cross-sectional survey to cardiologists to test their knowledge and confidence on CFD as a technology applied to patients suffering from CAD. Responses were recorded between May 18, 2022, and June 12, 2022.ResultsA total of 466 interventional cardiologists (mean age 48.4 ± 8.3 years, males 362), from 42 different countries completed the survey, for a response rate of 45.9%. Of these, 66.6% declared to be familiar with the term CFD, especially for optimization of existing interventional techniques (16.1%) and assessment of hemodynamic quantities related with CAD (13.7%). About 30% of respondents correctly answered to the questions exploring their knowledge on the pathophysiological role of some CFD-derived quantities such as wall shear stress and helical flow in coronary arteries. Among respondents, 85.9% would consider patient-specific CFD-based analysis in daily interventional practice while 94.2% declared to be interested in receiving a brief foundation course on the basic CFD principles. Finally, 87.7% of respondents declared to be interested in a cath-lab software able to conduct affordable CFD-based analyses at the point-of-care.ConclusionsInterventional cardiologists reported to be profoundly interested in adopting CFD simulations as a technology supporting decision making in the treatment of CAD in daily practice.

Abstract In bone tissue engineering research, bioreactors designed for replicating the main features of the complex native environment represent powerful investigation tools. Moreover, when equipped with automation, their use allows reducing user intervention and dependence, increasing reproducibility and the overall quality of the culture process. In this study, an automated uni-/bi-directional perfusion bioreactor combinable with pulsed electromagnetic field (PEMF) stimulation for culturing 3D bone tissue models is proposed. A user-friendly control unit automates the perfusion, minimizing the user dependency. Computational fluid dynamics simulations supported the culture chamber design and allowed the estimation of the shear stress values within the construct. Electromagnetic field simulations demonstrated that, in case of combination with a PEMF stimulator, the construct can be exposed to uniform magnetic fields. Preliminary biological tests on 3D bone tissue models showed that perfusion promotes the release of the early differentiation marker alkaline phosphatase. The histological analysis confirmed that perfusion favors cells to deposit more extracellular matrix (ECM) with respect to the static culture and revealed that bi-directional perfusion better promotes ECM deposition across the construct with respect to uni-directional perfusion. Lastly, the Real-time PCR results of 3D bone tissue models cultured under bi-directional perfusion without and with PEMF stimulation revealed that the only perfusion induced a ~ 40-fold up-regulation of the expression of the osteogenic gene collagen type I with respect to the static control, while a ~ 80-fold up-regulation was measured when perfusion was combined with PEMF stimulation, indicating a positive synergic pro-osteogenic effect of combined physical stimulations.

An experimental set-up is presented for the in vitro characterization of the fluid dynamics in personalized phantoms of healthy and stenosed coronary arteries. The proposed set-up was fine-tuned with the aim of obtaining a compact, flexible, low-cost test-bench for biomedical applications. Technically, velocity vector fields were measured adopting a so-called smart-PIV approach, consisting of a smartphone camera and a low-power continuous laser (30 mW). Experiments were conducted in realistic healthy and stenosed 3D-printed phantoms of left anterior descending coronary artery reconstructed from angiographic images. Time resolved image acquisition was made possible by the combination of the image acquisition frame rate of last generation commercial smartphones and the flow regimes characterizing coronary hemodynamics (velocities in the order of 10 cm/s). Different flow regimes (Reynolds numbers ranging from 20 to 200) were analyzed. The smart-PIV approach was able to provide both qualitative flow visualizations and quantitative results. A comparison between smart-PIV and conventional PIV (i.e., the gold-standard experimental technique for bioflows characterization) measurements showed a good agreement in the measured velocity vector fields for both the healthy and the stenosed coronary phantoms. Displacement errors and uncertainties, estimated by applying the particle disparity method, confirmed the soundness of the proposed smart-PIV approach, as their values fell within the same range for both smart and conventional PIV measured data (≈5% for the normalized estimated displacement error and below 1.2 pixels for displacement uncertainty). In conclusion, smart-PIV represents an easy-to-implement, low-cost methodology for obtaining an adequately robust experimental characterization of cardiovascular flows. The proposed approach, to be intended as a proof of concept, candidates to become an easy-to-handle test bench suitable for use also outside of research labs, e.g., for educational or industrial purposes, or as first-line investigation to direct and guide subsequent conventional PIV measurements.

Background: Laser-Doppler Vibrometry (LDV) is a laser-based technique that allows measuring the motion of moving targets with high spatial and temporal resolution. To demonstrate its use for the measurement of carotid-femoral pulse wave velocity, a prototype system was employed in a clinical feasibility study. Data were acquired for analysis without prior quality control. Real-time application, however, will require a real-time assessment of signal quality. In this study, we (1) use template matching and matrix profile for assessing the quality of these previously acquired signals; (2) analyze the nature and achievable quality of acquired signals at the carotid and femoral measuring site; (3) explore models for automated classification of signal quality.Methods: Laser-Doppler Vibrometry data were acquired in 100 subjects (50M/50F) and consisted of 4–5 sequences of 20-s recordings of skin displacement, differentiated two times to yield acceleration. Each recording consisted of data from 12 laser beams, yielding 410 carotid-femoral and 407 carotid-carotid recordings. Data quality was visually assessed on a 1–5 scale, and a subset of best quality data was used to construct an acceleration template for both measuring sites. The time-varying cross-correlation of the acceleration signals with the template was computed. A quality metric constructed on several features of this template matching was derived. Next, the matrix-profile technique was applied to identify recurring features in the measured time series and derived a similar quality metric. The statistical distribution of the metrics, and their correlates with basic clinical data were assessed. Finally, logistic-regression-based classifiers were developed and their ability to automatically classify LDV-signal quality was assessed.Results: Automated quality metrics correlated well with visual scores. Signal quality was negatively correlated with BMI for femoral recordings but not for carotid recordings. Logistic regression models based on both methods yielded an accuracy of minimally 80% for our carotid and femoral recording data, reaching 87% for the femoral data.Conclusion: Both template matching and matrix profile were found suitable methods for automated grading of LDV signal quality and were able to generate a quality metric that was on par with the signal quality assessment of the expert. The classifiers, developed with both quality metrics, showed their potential for future real-time implementation.

Hemodynamics interacts with the cellular components of human vessels, influencing function and healthy status. Locally acting hemodynamic forces have been associated—by a steadily increasing amount of scientific evidence—with nucleation and evolution of atherosclerotic plaques in several vascular regions, resulting in the formulation of the ‘hemodynamic risk hypothesis’ of the atherogenesis. At the level of coronary arteries, however, the complexity of both anatomy and physiology made the study of this vascular region particularly difficult for researchers. Developments in computational fluid dynamics (CFD) have recently allowed an accurate modelling of the intracoronary hemodynamics, thus offering physicians a unique tool for the investigation of this crucial human system by means of advanced mathematical simulations. The present review of CFD applications in coronary artery disease was set to concisely offer the medical reader the theoretical foundations of quantitative intravascular hemodynamics—reasoned schematically in the text in its basic (i.e., pressure and velocity) and derived quantities (e.g., fractional flow reserve, wall shear stress and helicity)—along with its current implications in clinical research. Moreover, attention was paid in classifying computational modelling derived from invasive and non-invasive imaging modalities with unbiased remarks on the advantages and limitations of each procedure. Finally, an extensive description—aided by explanatory figures and cross references to recent clinical findings—was presented on the role of near-wall hemodynamics, in terms of shear stress, and of intravascular flow complexity, in terms of helical flow.

Coronary atherosclerosis is a leading cause of illness and death in Western World and its mechanisms are still non completely understood. Several animal models have been used to 1) study coronary atherosclerosis natural history and 2) propose predictive tools for this disease, that is asymptomatic for a long time, aiming for a direct translation of their findings to human coronary arteries. Among them, swine models are largely used due to the observed anatomical and pathophysiological similarities to humans. However, a direct comparison between swine and human models in terms of coronary hemodynamics, known to influence atherosclerotic onset/development, is still lacking. In this context, we performed a detailed comparative analysis between swine- and human-specific computational hemodynamic models of coronary arteries. The analysis involved several near-wall and intravascular flow descriptors, previously emerged as markers of coronary atherosclerosis initiation/progression, as well as anatomical features. To do that, non-culprit coronary arteries (18 right–RCA, 18 left anterior descending–LAD, 13 left circumflex–LCX coronary artery) from patients presenting with acute coronary syndrome were imaged by intravascular ultrasound and coronary computed tomography angiography. Similarly, the three main coronary arteries of ten adult mini-pigs were also imaged (10 RCA, 10 LAD, 10 LCX). The geometries of the imaged coronary arteries were reconstructed (49 human, 30 swine), and computational fluid dynamic simulations were performed by imposing individualized boundary conditions. Overall, no relevant differences in 1) wall shear stress-based quantities, 2) intravascular hemodynamics (in terms of helical flow features), and 3) anatomical features emerged between human- and swine-specific models. The findings of this study strongly support the use of swine-specific computational models to study and characterize the hemodynamic features linked to coronary atherosclerosis, sustaining the reliability of their translation to human vascular disease.

In the past two decades, relevant advances have been made in the generation of engineered cardiac constructs to be used as functional in vitro models for cardiac research or drug testing, and with the ultimate but still challenging goal of repairing the damaged myocardium. To support cardiac tissue generation and maturation in vitro, the application of biomimetic physical stimuli within dedicated bioreactors is crucial. In particular, cardiac-like mechanical stimulation has been demonstrated to promote development and maturation of cardiac tissue models. Here, we developed an automated bioreactor platform for tunable cyclic stretch and in situ monitoring of the mechanical response of in vitro engineered cardiac tissues. To demonstrate the bioreactor platform performance and to investigate the effects of cyclic stretch on construct maturation and contractility, we developed 3D annular cardiac tissue models based on neonatal rat cardiac cells embedded in fibrin hydrogel. The constructs were statically pre-cultured for 5 days and then exposed to 4 days of uniaxial cyclic stretch (sinusoidal waveform, 10% strain, 1 Hz) within the bioreactor. Explanatory biological tests showed that cyclic stretch promoted cardiomyocyte alignment, maintenance, and maturation, with enhanced expression of typical mature cardiac markers compared to static controls. Moreover, in situ monitoring showed increasing passive force of the constructs along the dynamic culture. Finally, only the stretched constructs were responsive to external electrical pacing with synchronous and regular contractile activity, further confirming that cyclic stretching was instrumental for their functional maturation. This study shows that the proposed bioreactor platform is a reliable device for cyclic stretch culture and in situ monitoring of the passive mechanical response of the cultured constructs. The innovative feature of acquiring passive force measurements in situ and along the culture allows monitoring the construct maturation trend without interrupting the culture, making the proposed device a powerful tool for in vitro investigation and ultimately production of functional engineered cardiac constructs.

During embryonic morphogenesis, the heart undergoes a complex series of cellular phenotypic maturations (e.g., transition of myocytes from proliferative to quiescent or maturation of the contractile apparatus), and this involves stiffening of the extracellular matrix (ECM) acting in concert with morphogenetic signals. The maladaptive remodeling of the myocardium, one of the processes involved in determination of heart failure, also involves mechanical cues, with a progressive stiffening of the tissue that produces cellular mechanical damage, inflammation, and ultimately myocardial fibrosis. The assessment of the biomechanical dependence of the molecular machinery (in myocardial and non-myocardial cells) is therefore essential to contextualize the maturation of the cardiac tissue at early stages and understand its pathologic evolution in aging. Because systems to perform multiscale modeling of cellular and tissue mechanics have been developed, it appears particularly novel to design integrated mechano-molecular models of heart development and disease to be tested in ex vivo reconstituted cells/tissue-mimicking conditions. In the present contribution, we will discuss the latest implication of mechanosensing in heart development and pathology, describe the most recent models of cell/tissue mechanics, and delineate novel strategies to target the consequences of heart failure with personalized approaches based on tissue engineering and induced pluripotent stem cell (iPSC) technologies.

Abstract Controlled large-scale production of human pluripotent stem cells (hPSCs) is indispensable for their envisioned clinical translation. Aiming at advanced process development in suspension culture, the sensitivity of hPSC media to continuous peristaltic pump-based circulation, a well-established technology extensively used in hydraulically-driven bioreactors, was investigated. Unexpectedly, conditioning of low protein media (i.e. E8 and TeSR-E8) in a peristaltic pump circuit induced severe viability loss of hPSCs cultured as aggregates in suspension. Optical, biochemical, and cytological analyses of the media revealed that the applied circulation mode resulted in the reduction of the growth hormone insulin by precipitation of micro-sized particles. Notably, in contrast to insulin depletion, individual withdrawal of other medium protein components (i.e. bFGF, TGFβ1 or transferrin) provoked minor reduction of hPSC viability, if any. Supplementation of the surfactant glycerol or the use of the insulin analogue Aspart did not overcome the issue of insulin precipitation. In contrast, the presence of bovine or human serum albumin (BSA or HSA, respectively) stabilized insulin rescuing its content, possibly by acting as molecular chaperone-like protein, ultimately supporting hPSC maintenance. This study highlights the potential and the requirement of media optimization for automated hPSC processing and has broad implications on media development and bioreactor-based technologies.

Alzheimer's Disease (AD) is related with the abnormal aggregation of amyloid β-peptides Aβ1−40 and Aβ1−42, the latter having a polymorphic character which gives rise to U- or S-shaped fibrils. Elucidating the role played by the nanoscale-material architecture on the amyloid fibril stability is a crucial breakthrough to better understand the pathological nature of amyloid structures and to support the rational design of bio-inspired materials. The computational study here presented highlights the superior mechanical behavior of the S-architecture, characterized by a Young's modulus markedly higher than the U-shaped architecture. The S-architecture showed a higher mechanical resistance to the enforced deformation along the fibril axis, consequence of a better interchain hydrogen bonds' distribution. In conclusion, this study, focusing the attention on the pivotal multiscale relationship between molecular phenomena and material properties, suggests the S-shaped Aβ1−42 species as a target of election in computational screen/design/optimization of effective aggregation modulators.

A marked interest has recently emerged regarding the analysis of the wall shear stress (WSS) vector field topological skeleton in cardiovascular flows. Based on dynamical system theory, the WSS topological skeleton is composed of fixed points, i.e., focal points where WSS locally vanishes, and unstable/stable manifolds, consisting of contraction/expansion regions linking fixed points. Such an interest arises from its ability to reflect the presence of near-wall hemodynamic features associated with the onset and progression of vascular diseases. Over the years, Lagrangian-based and Eulerian-based post-processing techniques have been proposed aiming at identifying the topological skeleton features of the WSS. Here, the theoretical and methodological bases supporting the Lagrangian- and Eulerian-based methods currently used in the literature are reported and discussed, highlighting their application to cardiovascular flows. The final aim is to promote the use of WSS topological skeleton analysis in hemodynamic applications and to encourage its application in future mechanobiology studies in order to increase the chance of elucidating the mechanistic links between blood flow disturbances, vascular disease, and clinical observations.

TP53 is the most mutated gene in all cancers. The mutant protein also accumulates in cells. The high frequency of p53 mutations makes the protein a promising target for anti-cancer therapy. Only a few molecules have been found, using in vitro screening, to reactivate the mutant protein. APR-246 is currently the most successful mutant p53 activator, which reactivates the transcriptional activity of p53 by covalently binding to C124 of the protein. We have recently created in silico models of G245S-mp53 in its apo and DNA-bound forms. In this paper we further report on our in silico screening for potential activators of G245S-mp53. We filtered the ZINC15 database (13 million compounds) to only include drug-like molecules with moderate to standard reactivity. Our filtered database of 130,000 compounds was screened using the DOCKTITE protocol in the Molecular Operating Environment software. We performed covalent docking at C124 of G245S-mp53 to identify potential activators of the mutant protein. The docked compounds were ranked using a consensus scoring approach. We also used ADMET Predictor™ to predict pharmacokinetics and the possible toxicities of the compounds. Our screening procedure has identified compounds, mostly thiosemicarbazones and halo-carbonyls, with the best potential as G245S-mp53 activators, which are described in this work. Based on its binding scores and ADMET risk score, compound 2 is likely to have the best potential as a G245S-mp53 activator compared to the other top hits.

The Josephin Domain (JD), i.e. the N-terminal domain of Ataxin 3 (At3) protein, is an interesting example of competition between physiological function and aggregation risk. In fact, the fibrillogenesis of Ataxin 3, responsible for the spinocerebbellar ataxia 3, is strictly related to the JD thermodynamic stability. Whereas recent NMR studies have demonstrated that different JD conformations exist, the likelihood of JD achievable conformational states in solution is still an open issue. Marked differences in the available NMR models are located in the hairpin region, supporting the idea that JD has a flexible hairpin in dynamic equilibrium between open and closed states. In this work we have carried out an investigation on the JD conformational arrangement by means of both classical molecular dynamics (MD) and Metadynamics employing essential coordinates as collective variables. We provide a representation of the free energy landscape characterizing the transition pathway from a JD open-like structure to a closed-like conformation. Findings of our in silico study strongly point to the closed-like conformation as the most likely for a Josephin Domain in water.

A versatile bioreactor suitable for dynamic suspension cell culture under tunable shear stress conditions has been developed and preliminarily tested culturing cancer cell spheroids. By adopting simple technological solutions and avoiding rotating components, the bioreactor exploits the laminar hydrodynamics establishing within the culture chamber enabling dynamic cell suspension in an environment favourable to mass transport, under a wide range of tunable shear stress conditions. The design phase of the device has been supported by multiphysics modelling and has provided a comprehensive analysis of the operating principles of the bioreactor. Moreover, an explanatory example is herein presented with multiphysics simulations used to set the proper bioreactor operating conditions for preliminary in vitro biological tests on a human lung carcinoma cell line. The biological results demonstrate that the ultralow shear dynamic suspension provided by the device is beneficial for culturing cancer cell spheroids. In comparison to the static suspension control, dynamic cell suspension preserves morphological features, promotes intercellular connection, increases spheroid size (2.4-fold increase) and number of cycling cells (1.58-fold increase), and reduces double strand DNA damage (1.5-fold reduction). It is envisioned that the versatility of this bioreactor could allow investigation and expansion of different cell types in the future.

The transcription factor p53 is a potent tumor suppressor dubbed as the “guardian of the genome” because of its ability to orchestrate protective biological outputs in response to a variety of oncogenic stresses. Mutation and thus inactivation of p53 can be found in 50% of human tumors. The majority are missense mutations located in the DNA binding region. Among them, G245S is known to be a structural hotspot mutation. To understand the behaviors and differences between the wild-type and mutant, both a dimer of the wild type p53 (wt-p53) and its G245S mutant (G245S-mp53), complexed with DNA, were simulated using molecular dynamics for more than 1 μs. wt-p53 and G245S-mp53 apo monomers were simulated for 1 μs as well. Conformational analyses and binding energy evaluations performed underline important differences and therefore provide insights to understand the G245S-mp53 loss of function. Our results indicate that the G245S mutation destabilizes several structural regions in the protein that are crucial for DNA binding when found in its apo form and highlight differences in the mutant-DNA complex structure compared to the wt protein. These findings not only provide means that can be applied to other p53 mutants but also serve as structural basis for further studies aimed at the development of cancer therapies based on restoring the function of p53.

Spinocerebellar ataxia (SCA) 3, the most common form of SCA, is a neurodegenerative rare disease characterized by polyglutamine tract expansion and self-assembly of Ataxin3 (At3) misfolded proteins into highly organized fibrillar aggregates. The At3 N-terminal Josephin Domain (JD) has been suggested as being responsible for mediating the initial phase of the At3 double-step fibrillogenesis. Several issues concerning the residues involved in the JD's aggregation and, more generally, the JD clumping mechanism have not been clarified yet. In this paper we present an investigation focusing on the JD protein-protein interaction by means of molecular modeling. Our results suggest possible aminoacids involved in JD contact together with local and non-local effects following JD dimerization. Surprisingly, JD conformational changes following the binding may involve ubiquitin binding sites and hairpin region even though they do not pertain to the JD interaction surfaces. Moreover, the JD binding event has been found to alter the hairpin open-like conformation toward a closed-like arrangement over the simulated timescale. Finally, our results suggest that the JD aggregation might be a multi-step process, with an initial fast JD-JD binding mainly driven by Arg101, followed by slower structural global rearrangements involving the exposure to the solvent of Leu84-Trp87, which might play a role in a second step of JD aggregation.