Prospects of microwave spectrometry for vascular stent monitoring. Towards a non-invasive and non-ionizing follow-up alternative

[eng] Throughout this thesis we have assessed the prospects of microwave spectrometry (MWS) as a non-ionizing non-invasive monitoring alternative for stented patients in a very early proof-of-concept stage. In Chapter 1 we have provided a generalist retrospective medical background along with a state-of-the-art summary of existing microwave-based stent monitoring approaches. First, we have introduced cardiovascular diseases in general, and ischemic heart disease in particular. Next we have reviewed how percutaneous coronary interventions addressed the medical problem represented by atherosclerosis, giving a special emphasis to balloon angioplasty, bare-metal stenting and drug-eluting stenting. We have further exposed how the outcomes of such revolutionary strategies were compromised by the high rates of post-procedural complications, making unavoidable the invasive and ionizing follow-up of stented patients. Finally, we have summarized existing non-invasive and non-ionizing stent monitoring alternatives based in microwave techniques. In Chapter 2 we have introduced the working principle of our MWS setup. We have first presented how this arrangement can obtain the absorbance of a stent as a function of the frequency and the incidence angle of the microwave fields. We have also shown how these data are combined in a single two-dimensional chart, and how we recognize therein the characteristic resonance frequencies of stents at a glance. As an example, we have presented a typical absorbance diagram to illustrate the general features of such resonances. In particular we have highlighted that these resonances are discrete and have multi-lobed angular patterns. In Chapter 3 we have characterized many stents having a wide variety of nominal sizes to better understand their characteristic resonances in terms of microwave scattering. First, we have found that the resonance frequency obeys a reciprocal dependence on the stent length. This has allowed us to obtain an empirical expression for such relationship just by adjusting two fitting parameters. However, we have not been able to find an analogous expression for the dependence on the stent diameter. In any case, while investigating the latter, we have unexpectedly uncovered how the particular stent architecture influences the corresponding resonance frequencies. By gathering all these individual results we have finally suggested a straightforward half-theoretical half-empirical model linking the resonance frequencies of stents with their structural integrity (through their length), with their particular architecture (through the scaling factor), as well as with their surrounding medium (through the dielectric permittivity and the magnetic permeability). We have also theoretically estimated the resonance frequencies of implanted stents from their corresponding values in free space conditions, showing that in vivo resonance frequencies should be around one order of magnitude smaller than their free space counterparts. Finally, in Chapters 4 and 5 we have explored the potential diagnostic capabilities of MWS in two possible scenarios: stent fracture (SF) and in-stent neoatherosclerosis (ISNA). We have started both chapters reviewing the incidence, the medical implications, and the mechanism of these two stent-related complications. SF has been evaluated in Chapter 4 by means of two “fracture tests” consisting in a successive series of strut cuts. We have shown that MWS provides qualitative indicators for single and multiple strut fractures (downshift of the fundamental resonance frequency), and also quantitative indicators for single or multiple complete transverse linear SFs (split and upshift of that frequency). ISNA has been evaluated in Chapter 6 by means of four ``cholesterol tests'' consisting in a gradual process of increasing cholesterol deposition. We have shown that MWS provides an indicator for a growing presence of cholesterol around a stent (downshift of the fundamental resonance frequency). We have concluded this chapter calculating the theoretical evolution of the resonance frequencies along a cholesterol deposition process, estimating the upper limit for the resonance frequency displacement. Taking together the results we have reported in Chapters 5 and 6, we have shown that MWS could potentially warn about SF and ISNA

Characterization of p38α signaling networks in cancer cells using quantitative proteomics and phosphoproteomics

p38α (encoded by MAPK14) is a protein kinase that regulates cellular responses to almost all types of environmental and intracellular stresses. Upon activation, p38α phosphorylates many substrates both in the cytoplasm and nucleus, allowing this pathway to regulate a wide variety of cellular processes. While the role of p38α in the stress response has been widely investigated, its implication in cell homeostasis is less understood. To investigate the signaling networks regulated by p38α in proliferating cancer cells, we performed quantitative proteomic and phosphoproteomic analyses in breast cancer cells in which this pathway had been either genetically targeted or chemically inhibited. Our study identified with high confidence 35 proteins and 82 phosphoproteins (114 phosphosites) that are modulated by p38α, and highlighted the implication of various protein kinases, including MK2 and mTOR, in the p38α-regulated signaling networks. Moreover, functional analyses revealed an important contribution of p38α to the regulation of cell adhesion, DNA replication and RNA metabolism. Indeed, we provide experimental evidence supporting that p38α facilitates cancer cell adhesion, and showed that this p38α function is likely mediated by the modulation of the adaptor protein ArgBP2. Collectively, our results illustrate the complexity of the p38α regulated signaling networks, provide valuable information on p38α-dependent phosphorylation events in cancer cells, and document a mechanism by which p38α can regulate cell adhesion.Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved

Ex vivo assessment and in vivo validation of non-invasive stent monitoring techniques based on microwave spectrometry

Some conditions are well known to be directly associated with stent failure, including in-stent re-occlusion and stent fracture. Currently, identification of these high-risk conditions requires invasive and complex procedures. This study aims to assess microwave spectrometry (MWS) for monitoring stents non-invasively. Preliminary ex vivo data are presented to move to in vivo validation. Fifteen mice were assigned to receive subcutaneous stent implantations (n¿=¿10) or sham operations (n¿=¿5). MWS measurements were carried out at 0, 2, 4, 7, 14, 22, and 29 days of follow-up. Additionally, 5 stented animals were summited to micro-CT analyses at the same time points. At 29 days, 3 animals were included into a stent fracture subgroup and underwent a last MWS and micro-CT analysis. MWS was able to identify stent position and in-stent stenosis over time, also discerning significant differences from baseline measures (P¿<¿0.001). Moreover, MWS identified fractured vs. non-fractured stents in vivo. Taken together, MWS emerges as a non-invasive, non-ionizing alternative for stent monitoring. MWS analysis clearly distinguished between in-stent stenosis and stent fracture phenomena.Peer ReviewedPostprint (published version

Ex vivo assessment and in vivo validation of non-invasive stent monitoring techniques based on microwave spectrometry

Some conditions are well known to be directly associated with stent failure, including in-stent re-occlusion and stent fracture. Currently, identification of these high-risk conditions requires invasive and complex procedures. This study aims to assess microwave spectrometry (MWS) for monitoring stents non-invasively. Preliminary ex vivo data are presented to move to in vivo validation. Fifteen mice were assigned to receive subcutaneous stent implantations (n = 10) or sham operations (n = 5). MWS measurements were carried out at 0, 2, 4, 7, 14, 22, and 29 days of follow-up. Additionally, 5 stented animals were summited to micro-CT analyses at the same time points. At 29 days, 3 animals were included into a stent fracture subgroup and underwent a last MWS and micro-CT analysis. MWS was able to identify stent position and in-stent stenosis over time, also discerning significant differences from baseline measures (P < 0.001). Moreover, MWS identified fractured vs. non-fractured stents in vivo. Taken together, MWS emerges as a non-invasive, non-ionizing alternative for stent monitoring. MWS analysis clearly distinguished between in-stent stenosis and stent fracture phenomena

Biomarker candidates for progression and clinical management of COVID-19 associated pneumonia at time of admission

COVID-19 pathophysiology is currently not fully understood, reliable prognostic factors remain elusive, and few specific therapeutic strategies have been proposed. In this scenario, availability of biomarkers is a priority. MS-based Proteomics techniques were used to profile the proteome of 81 plasma samples extracted in four consecutive days from 23 hospitalized COVID-19 associated pneumonia patients. Samples from 10 subjects that reached a critical condition during their hospital stay and 10 matched non-severe controls were drawn before the administration of any COVID-19 specific treatment and used to identify potential biomarkers of COVID-19 prognosis. Additionally, we compared the proteome of five patients before and after glucocorticoids and tocilizumab treatment, to assess the changes induced by the therapy on our selected candidates. Forty-two proteins were differentially expressed between patients' evolution groups at 10% FDR. Twelve proteins showed lower levels in critical patients (fold-changes 1.20-3.58), of which OAS3 and COG5 found their expression increased after COVID-19 specific therapy. Most of the 30 proteins over-expressed in critical patients (fold-changes 1.17-4.43) were linked to inflammation, coagulation, lipids metabolism, complement or immunoglobulins, and a third of them decreased their expression after treatment. We propose a set of candidate proteins for biomarkers of COVID-19 prognosis at the time of hospital admission. The study design employed is distinctive from previous works and aimed to optimize the chances of the candidates to be validated in confirmatory studies and, eventually, to play a useful role in the clinical practice

It is time to define an organizational model for the prevention and management of infections along the surgical pathway : a worldwide cross-sectional survey

Background The objectives of the study were to investigate the organizational characteristics of acute care facilities worldwide in preventing and managing infections in surgery; assess participants' perception regarding infection prevention and control (IPC) measures, antibiotic prescribing practices, and source control; describe awareness about the global burden of antimicrobial resistance (AMR) and IPC measures; and determine the role of the Coronavirus Disease 2019 pandemic on said awareness. Methods A cross-sectional web-based survey was conducted contacting 1432 health care workers (HCWs) belonging to a mailing list provided by the Global Alliance for Infections in Surgery. The self-administered questionnaire was developed by a multidisciplinary team. The survey was open from May 22, 2021, and June 22, 2021. Three reminders were sent, after 7, 14, and 21 days. Results Three hundred four respondents from 72 countries returned a questionnaire, with an overall response rate of 21.2%. Respectively, 90.4% and 68.8% of participants stated their hospital had a multidisciplinary IPC team or a multidisciplinary antimicrobial stewardship team. Local protocols for antimicrobial therapy of surgical infections and protocols for surgical antibiotic prophylaxis were present in 76.6% and 90.8% of hospitals, respectively. In 23.4% and 24.0% of hospitals no surveillance systems for surgical site infections and no monitoring systems of used antimicrobials were implemented. Patient and family involvement in IPC management was considered to be slightly or not important in their hospital by the majority of respondents (65.1%). Awareness of the global burden of AMR among HCWs was considered very important or important by 54.6% of participants. The COVID-19 pandemic was considered by 80.3% of respondents as a very important or important factor in raising HCWs awareness of the IPC programs in their hospital. Based on the survey results, the authors developed 15 statements for several questions regarding the prevention and management of infections in surgery. The statements may be the starting point for designing future evidence-based recommendations. Conclusion Adequacy of prevention and management of infections in acute care facilities depends on HCWs behaviours and on the organizational characteristics of acute health care facilities to support best practices and promote behavioural change. Patient involvement in the implementation of IPC is still little considered. A debate on how operationalising a fundamental change to IPC, from being solely the HCWs responsibility to one that involves a collaborative relationship between HCWs and patients, should be opened.Peer reviewe

Goya's artwork imaging with Terahertz waves

In this paper we use a Terahertz (THz) time-domain system to image and analyze the structure of an artwork attributed to the Spanish artist Goya painted in 1771. The THz images show features that cannot be seen with optical inspection and complement data obtained with X-ray imaging that provide evidence of its authenticity, which is validated by other independent studies. For instance, a feature with a strong resemblance with one of Goya"s known signatures is seen in the THz images. In particular, this paper demonstrates the potential of THz imaging as a complementary technique along with X-ray for the verification and authentication of artwork pieces through the detection of features that remain hidden to optical inspection

Protamine Characterization by Top-Down Proteomics: Boosting Proteoform Identification with DBSCAN

Protamines replace histones as the main nuclear protein in the sperm cells of many species and play a crucial role in compacting the paternal genome. Human spermatozoa contain protamine 1 (P1) and the family of protamine 2 (P2) proteins. Alterations in protamine PTMs or the P1/P2 ratio may be associated with male infertility. Top-down proteomics enables large-scale analysis of intact proteoforms derived from alternative splicing, missense or nonsense genetic variants or PTMs. In contrast to current gold standard techniques, top-down proteomics permits a more in-depth analysis of protamine PTMs and proteoforms, thereby opening up new perspectives to unravel their impact on male fertility. We report on the analysis of two normozoospermic semen samples by top-down proteomics. We discuss the difficulties encountered with the data analysis and propose solutions as this step is one of the current bottlenecks in top-down proteomics with the bioinformatics tools currently available. Our strategy for the data analysis combines two software packages, ProSight PD (PS) and TopPIC suite (TP), with a clustering algorithm to decipher protamine proteoforms. We identified up to 32 protamine proteoforms at different levels of characterization. This in-depth analysis of the protamine proteoform landscape of normozoospermic individuals represents the first step towards the future study of sperm pathological conditions opening up the potential personalized diagnosis of male infertility

Proteomic tools for the quantitative analysis of artificial peptide libraries: detection and characterization of target-amplified PD-1 inhibitors

We report a quantitative proteomics data analysis pipeline which, coupled to protein-directed dynamic combinatorial chemistry (DDC) experiments, enables the rapid discovery and direct characterization of protein-protein interaction (PPI) modulators. A low-affinity PD-1 binder was incubated with a library of >100 D-peptides under thiol-exchange favoring conditions, in the presence of the target protein PD-1, and we determined the S-linked dimeric species that resulted amplified in the protein samples versus the controls. We chemically synthesized the target dimer candidates and validated them by thermophoresis binding and protein-protein interaction assays. The results provide a proof-of-concept for using this strategy in the high-throughput search of improved drug-like peptide binders that block therapeutically relevant protein-protein interactions