Diagnostic and prognostic significance of some inflammatory serum proteins in patients with precancerous diseases and cervical cancer

Background: The high social significance of cervical cancer, the shortcomings of the performed cervical screening is prerequisites for research in the field of mproving the diagnosis of this disease. We hypothesized that the systemic level of some inflammatory proteins could be used as a diagnostic criterion for cervical cancer.The aim of the study was to study the level of some vascular-inflammatory markers in the blood serum in patients with precancerous diseases and cervical cancer in order to improve their diagnosis and also to identify markers for predicting an unfavorable outcome in patients with cervical cancer.Materials and methods. A non-randomized prospective controlled study was carried out, the participants of which were patients with the diagnosis of cervical cancer (n = 49) and cervical intraepithelial neoplasia of the III degree (n = 13). The control group included 15 relatively healthy women. The following spectrum of inflammatory proteins was determined in blood serum by flow cytometry using the Human Vascular Inflammation Panel 1: myoglobin, calprotectin, lipocalin A, matrix metal peroxidase 2, osteopontin, myeloperoxidase, serum amyloid A, protein 4, which binds insulin-like growth factor cell-cell adhesion 1, vascular cell adhesion molecule, matrix metalloperoxidase 9, cystatin C. Statistical analysis was carried out by calculating the Mann-Whitney test with Bonferroni’s correction. The model was created using binary logistic regression to diagnose cervical cancer.Results. In the intergroup comparison of the protein`s spectrum in the blood serum, no significant differences were obtained. However, using the binary logistic regression method, an equation was drawn up to calculate the diagnostic coefficient of cervical cancer, which allows diagnosing cervical cancer with an accuracy of 82%, and in terms of information content is not inferior to cytological diagnostics. The developed coefficient can be used to predict an unfavorable outcome of cervical cancer after 1 year from the moment of diagnosis. Conclusion. The developed diagnostic coefficient makes it possible to  diagnose cervical cancer with high accuracy and can be used to predict cervical cancer

Potential Biomarkers for Noninfectious Scleritis Identified by Serum and Tear Fluid Proteomics

Purpose: Scleritis is an extremely painful and potentially blinding inflammation of the sclera with unknown pathogenesis and unpredictable course. To gain insight in its disease process and identify biomarker candidates, we performed extensive proteomics in serum and tear fluid. Design: Prospective multicenter cohort study. Participants: A total of 121 patients with noninfectious scleritis (of which 39 active cases), 30 healthy controls, and 23 disease controls (uveitis and rheumatoid arthritis) were enrolled in the Netherlands from 2020 to 2022. Methods: Serum, tear fluid of both eyes, and clinical data were gathered. The level of 368 inflammatory proteins was measured using proximity extension assays. Results were validated in an independent cohort of 15 patients with scleritis, and using addressable laser bead immunoassay, or enzyme-linked immunoassays. In addition, we studied an extended panel of matrix metalloproteinases in tear fluid of necrotizing scleritis with addressable laser bead immunoassay. Main Outcome Measures: Statistically significant differences in the level of inflammatory proteins between patients with scleritis and control groups.Results:Proteomics revealed 18 significantly upregulated or downregulated serum proteins in active scleritis cases compared with all control groups in both the discovery cohort and the validation cohort. The most upregulated protein was nuclear migration protein nudC (NudC; P = 0.0032), a protein involved in neurogenesis. The other significant hits included proteins involved in T-cell activation, apoptosis, epithelial barrier maintenance, and angiogenesis. Our tear fluid analysis showed matrix metalloproteinase 9 (MMP9) to be upregulated in the tear fluid of patients with scleral necrosis. Conclusions: The results of our proteomics analysis suggest a role for neurogenesis, T-cell activation, disruption of epithelial barrier, and angiogenesis in the pathogenesis of scleritis, and highlight MMP9 and NudC as biomarkers with potential clinical relevance. </p

Role of cachexia and fragility in the patient candidate for cardiac surgery

Frailty is the major expression of accelerated aging and describes a decreased resistance to stressors, and consequently an increased vulnerability to additional diseases in elderly people. The vascular aging related to frail phenotype reflects the high susceptibility for cardiovascular diseases and negative postoperative outcomes after cardiac surgery. Sarcopenia can be considered a biological substrate of physical frailty. Malnutrition and physical inactivity play a key role in the pathogenesis of sarcopenia. We searched on Medline (PubMed) and Scopus for relevant literature published over the last 10 years and analyzed the strong correlation between frailty, sarcopenia and cardiovascular diseases in elderly patient. In our opinion, a right food intake and moderate intensity resistance exercise are mandatory in order to better prepare patients undergoing cardiac operation

Erectile dysfunction: heritability and cognitive and physiological correlates of the subclinical vascular disorder

The broad goals of this dissertation were to explore possible causes of erectile dysfunction (ED), and examine outcomes associated with the condition. To address these objectives, data from the Vietnam Era Twin Study of Aging (VETSA), a longitudinal study of cognitive aging, were utilized. Previous research demonstrated moderate heritability of ED in middle-aged men, but questions remain about the replicability of these findings given that only one study exists on the topic and it utilized an unvalidated ED measure. Additionally, it is unknown to what extent genetic factors account for stability and change of ED symptomatology over time. In the first study, we conducted a longitudinal assessment of ED using the Sexual Health Inventory for Men (SHIM) administered to 668 male twins. Results revealed that the heritability (proportion of variance explained by genetic factors) of ED at both time points is moderate (0.3 –0.6). Over the past two decades ED has been recognized as primarily a disorder of vascular dysfunction. Inflammation is an important correlate of vascular dysfunction. Chapter 2 examined the association of ED with four inflammatory serum protein indices [albumin, globulin, albumin/globulin ratio (AGR), and the albumin*globulin interaction (AGI)] in 388 men free from cardiovascular disease (CVD). Results indicated that the AGR, albumin, and globulin protein levels are significantly associated with ED, even after accounting for other risk factors. Intriguingly, AGR and globulin levels were found to be more strongly associated with ED than conventional vascular risk factors, such as hypertension, high cholesterol, obesity, and smoking. As such, globulin and AGR may be useful biomarkers when diagnosing and managing ED. The simplicity and affordability of these assays make these markers particularly valuable. Given the negative impact that vascular disease has on cognition, Chapter 3 explored whether men with ED are at greater risk for experiencing cognitive decline compared to healthy peers. In total, 485 men free from CVD were administered cognitive tests across 13 domains. Tests were readministered 5 years later. Results revealed that men with ED were more likely to show significant decline on tasks of set-shifting, tool/mechanical knowledge, vocabulary, and verbal fluency

Inhaled multiwalled carbon nanotubes induce systemic inflammation and endothelial dysfunction via MMP-9 induction

Assessing the mechanisms underlying adverse cardiovascular effects induced by inhaled toxins presents a substantial research challenge. We propose that blood carries an as yet unknown inflammatory potential consisting of modified proteins or other biomolecules and reaction byproducts that affects a pathological bioactivity which can be assessed using naïve endothelial cells and blood vessels. The approach involves applying serum from exposed animals to cultured primary endothelial cells or ex vivo isolated arteries. Mice were exposed to multi-walled carbon nanotubes (MWCNT; 0, 10 or 40 μg) or other pollutants via pharyngeal aspiration and serum was collected at 4 and 24 h post-exposure. Serum from exposed mice increased endothelial cell surface vascular cell adhesion molecule-1 (VCAM-1) and intracellular adhesion molecule-1 (ICAM-1) expression and proinflammatory transcripts, and decreased ATP-stimulated nitric oxide (NO) production. The functional impact of this loss of NO bioavailability was confirmed via myography, in which serum from pollutant-exposed mice significantly impaired vasodilation to acetylcholine. In addition, serum from pollutant-exposed mice reduced cell migration in a traditional scratch assay experiment. In vivo MWCNT exposure was able to both increase the permeability of the Blood Brain Barrier (BBB), as well as induce transcription of pro- inflammatory cytokines in the hippocampus and frontal cortex regions of the brain. This affect was abolished with the treatment of fasudil, or the absence of CD36. There was also evidence of astrocyte activation in the short and microglia activation at 24 hours. CD 36 has also been identified as playing a key role in mediating loss of vasodilatory properties ex vivo. In conclusion, pulmonary exposure to MWCNT dynamically alters circulating factors, which promotes endothelial cell activation, decreased NO bioavailability, and altered functionality all directionally predicting adverse cardiovascular outcomes

The role of inflammation in retinal ischaemia

Purpose: The immune cascade is known to contribute to the pathology seen in oxygen induced retinopathy (OIR), however its exact role is poorly defined. We hypothesised that immune cell activation worsens hypoxia and exacerbates the neovascular sequelae seen in ischaemic retinopathy. Methods: We assess the effect of intraperitoneal lipopolysaccharide (IP LPS) injection in OIR mice. Neovascularisation was assessed by measuring the avascular area and neovascular tuft area at P17. Hypoxia was assessed using vessel tortuosity and EF5 hypoxia staining at P14. The activated retinal inflammatory cell population was characterised using immunohistochemistry and flow cytometry. Transgenic mice were bred to delete different subpopulations of inflammatory cells to define their role in hypoxia modulation. RNA sequencing performed on retinal tissue analysed the effect of systemic LPS on retinal cytokines and angiogenesis markers. Results: IP LPS injection at P12 in OIR mice significantly reduced neovascularisation at P17 and hypoxia at P14. Immunohistochemistry revealed an influx of round, CD11b+, lectin stained cells into the retina of LPS treated mice, which on flow cytometry were identified as myeloid cells, being predominantly Cd11b+Ly6Ghi neutrophils. Experimental depletion of the myeloid lineage was achieved using ROSA26eGFP-DTALysMcre, CCL2 and CCR2 knockout mouse lines and anti-CCR2 antibody MC21. However, when these mice were LPS treated, the effects on hypoxia readouts caused by LPS were still seen. Transcriptional profiling of the retina (using RNAseq) revealed a large upregulation in IL1β in the central, hypoxic retina of LPS treated mice. Injection of IL1β at P12 also mimicked the effects of LPS, suggesting that IL1β is a key mediator of the hypoxia reducing effect LPS has in the OIR model. Conclusions: These findings are counterintuitive to the current literature and provide new insight into the role the immune system has on regulating oxygen demand in the retina. This novel approach to reducing hypoxia has the potential to lead to novel therapies targeting hypoxia and preventing neovascularisation in ischaemic eye disease

Syndrome métabolique affectant les survivants de la leucémie lymphoblastique aiguë pédiatrique : rôle et dysfonctions des lipoprotéines « HDL »

La leucémie lymphoblastique aiguë est le cancer le plus fréquent dans la population pédiatrique. L’amélioration des traitements a permis d’atteindre une survie à 5 ans post- diagnostic surpassant les 90%. Cette réussite de la médecine moderne s’accompagne toutefois d’importants effets à long terme. Ainsi, plus de 65% des survivants présentent au moins une complication sévère comme le syndrome métabolique et les maladies cardiovasculaires. Dans ce contexte, une importante proportion de survivants de cancers pédiatriques présente un taux abaissé du contenu en cholestérol des lipoprotéines de haute densité (HDL-Cholestérol). Les causes d’un tel désordre ne sont pas connues. Les HDLs sont des particules multifonctionnelles primordiales pour le maintien de la santé cardiométabolique, entre autres, de par leurs nombreuses actions antioxydantes, anti-athérosclérotiques, anti-inflammatoires et anti- thrombotiques. L’étude de la fonctionnalité des HDLs dans plusieurs pathologies a révélé des dysfonctionnements importants liés à des altérations de leur composition. L’objectif de cette étude était de caractériser les HDLs isolées de survivants de la leucémie lymphoblastique aiguë pédiatrique au niveau de leur composition et fonctionnalité. Les HDLs des survivants contiennent moins de cholestérol que les HDLs de sujets contrôles. L’analyse du protéome a révélé un enrichissement des HDLs des survivants métaboliquement non sains en protéines pro-thrombotiques (ex : fibrinogène) et pro- inflammatoires (ex : amyloïde sérique A). La capacité à promouvoir l’efflux de cholestérol ainsi que la capacité antioxydante de ces HDLs n’en était pas affectée. Les résultats de ces travaux indiquent une altération de la composition et du protéome des HDLs de survivants de la leucémie lymphoblastique aiguë pédiatrique. Bien que plus d’études soient nécessaires pour la validation de la fonctionnalité de ces HDLs, ces travaux semblent pertinents pour la santé des survivants étant donné la révélation de biomarqueurs potentiels du métabolisme et de la fonctionnalité des HDLs, et donc de l’athérosclérose.Acute lymphoblastic leukemia is the most frequent malignancy in children. With the use of more modern, efficient treatments, 5-year survival has reached more than 90% in this population. However, this achievement comes with many long-term effects: more than 65% of the survivors experience at least one severe complication, including the metabolic syndrome and cardiovascular diseases. In this context, a significant proportion of pediatric cancer survivors have low HDL-Cholesterol levels. The causes of such an abnormality remain so far unknown. HDLs are multifunctional particles, which are primordial in maintaining cardiovascular health given their antioxidative, anti-atherosclerosis, anti-inflammatory and anti- thrombotic capacities, to name but a few functions. Accordingly, studies on HDL functionality in diverse pathologies have revealed some important losses of functions linked to an altered composition. The main objective of the present work was to characterize the composition and functionality of HDLs isolated from pediatric acute lymphoblastic leukemia survivors. The HDL fractions from the survivors contained less cholesterol than the controls. In addition, proteomic analyses using mass spectrometry revealed an enrichment of pro-thrombotic (e.g. fibrinogen) and pro-inflammatory (e.g. serum amyloid A) proteins in the HDLs deriving from metabolically unhealthy survivors. The cholesterol efflux and antioxidant capacities of these HDLs were similar to those of the controls. These results indicate an alteration in the composition of lipid and protein content of HDLs from childhood acute lymphoblastic leukemia survivors with metabolic disorders. Although more work is needed to validate the functionality of these HDLs, the data seem relevant for survivor health given the detection of potential biomarkers related to HDL metabolism and functionality in cancer