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

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

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

Furan warheads for covalent trapping of weak protein-protein interactions : cross-linking of thymosin β4 to actin

We describe furan as a triggerable 'warhead' for site-specific cross-linking using the actin and thymosin beta 4 (T beta 4)-complex as model of a weak and dynamic protein-protein interaction (PPI) with known 3D structure and with application potential in disease contexts. The identified cross-linked residues demonstrate that lysine is a target for the furan warhead. The presented in vitro validation of covalently acting 'furan-armed' T beta 4-variants provides initial proof to further exploit furan-technology for covalent drug design targeting lysines

Microwave spectrometry for the evaluation of the structural integrity of metallic stents

PURPOSE: To assess the feasibility of a method based on microwave spectrometry to detect structural distortions of metallic stents in open air conditions and envisage the prospects of this approach toward possible medical applicability for the evaluation of implanted stents. METHODS: Microwave absorbance spectra between 2.0 and 18.0 GHz were acquired in open air for the characterization of a set of commercial stents using a specifically design setup. Rotating each sample over 360°, 2D absorbance diagrams were generated as a function of frequency and rotation angle. To check our approach for detecting changes in stent length (fracture) and diameter (recoil), two specific tests were performed in open air. Finally, with a few adjustments, this same system provides 2D absorbance diagrams of stents immersed in a water-based phantom, this time over a bandwidth ranging from 0.2 to 1.8 GHz. RESULTS: The authors show that metallic stents exhibit characteristic resonant frequencies in their microwave absorbance spectra in open air which depend on their length and, as a result, may reflect the occurrence of structural distortions. These resonances can be understood considering that such devices behave like dipole antennas in terms of microwave scattering. From fracture tests, the authors infer that microwave spectrometry provides signs of presence of Type I to Type IV stent fractures and allows in particular a quantitative evaluation of Type III and Type IV fractures. Recoil tests show that microwave spectrometry seems able to provide some quantitative assessment of diametrical shrinkage, but only if it involves longitudinal shortening. Finally, the authors observe that the resonant frequencies of stents placed inside the phantom shift down with respect to the corresponding open air frequencies, as it should be expected considering the increase of dielectric permittivity from air to water. CONCLUSIONS: The evaluation of stent resonant frequencies provided by microwave spectrometry allows detection and some quantitative assessment of stent fracture and recoil in open air conditions. Resonances of stents immersed in water can be also detected and their characteristic frequencies are in good agreement with theoretical estimates. Although these are promising results, further verification in a more relevant phantom is required in order to foresee the real potential of this approach.Peer Reviewe