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Emanuela MONARI
Professore Associato Dipartimento Chirurgico, Medico, Odontoiatrico e di Scienze Morfologiche con interesse Trapiantologico, Oncologico e di Medicina Rigenerativa
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Pubblicazioni
2023
- Research of Prostate Cancer Urinary Diagnostic Biomarkers by Proteomics: The Noteworthy Influence of Inflammation
[Articolo su rivista]
Bellei, Elisa; Caramaschi, Stefania; Giannico, Giovanna A; Monari, Emanuela; Martorana, Eugenio; Reggiani Bonetti, Luca; Bergamini, Stefania
abstract
: Nowadays, in the case of suspected prostate cancer (PCa), tissue needle biopsy remains the benchmark for diagnosis despite its invasiveness and poor tolerability, as serum prostate-specific antigen (PSA) is limited by low specificity. The aim of this proteomic study was to identify new diagnostic biomarkers in urine, an easily and non-invasively available sample, able to selectively discriminate cancer from benign prostatic hyperplasia (BPH), evaluating whether the presence of inflammation may be a confounding parameter. The analysis was performed by two-dimensional gel electrophoresis (2-DE), mass spectrometry (LC-MS/MS) and Enzyme-Linked Immunosorbent Assay (ELISA) on urine samples from PCa and BPH patients, divided into subgroups based on the presence or absence of inflammation. Significant quantitative and qualitative differences were found in the urinary proteomic profile of PCa and BPH groups. Of the nine differentially expressed proteins, only five can properly be considered potential biomarkers of PCa able to discriminate the two diseases, as they were not affected by the inflammatory process. Therefore, the proteomic research of novel and reliable urinary biomarkers of PCa should be conducted considering the presence of inflammation as a realistic interfering element, as it could hinder the detection of important protein targets.
2022
- Diagnostic proteomic markers to detect kidney diseases
[Abstract in Rivista]
Ozben, T.; Bellei, E.; Monari, E.; Bergamini, S.; Ferrari, A.; Tomasi, A.
abstract
Objective
Early detection of kidney disorders based on selective biomarkers
could permit to diagnose patients at the initial stage of the disease,
where the therapy is still possible to stop or prevent occurrence of
advance disease. Urinary proteomics is primarily applied to the study
of renal and urogenital tract disorders. Here are reported two distinct
successful examples of this approach for the discovery of early urinary
biomarkers of kidney-related dysfunctions: diabetic nephropathy (DN),
a well-known complication of diabetes frequently leading to dialysis,
and drug-induced nephrotoxicity, a possible condition caused by medication-overuse headache (MOH).
Methods
Urine samples were first concentrated and desalted. Subsequently,
they were subjected to two-dimensional gel electrophoresis (2-DE) coupled to mass spectrometry (MS) for protein identification. Furthermore,
some proteins were verified by Western Blot and ELISA test.
Results
In diabetes-related study, 11 differentially expressed proteins were
detected (8 up-regulated and 3 down-regulated) in Type 2 Diabetic
(T2D) and Type 2 Diabetic Nephropathy (T2DN) patients compared to
the healthy control subjects. In MOH study, a total of 21 over-excreted
proteins was revealed in urine of non-steroidal anti-inflammatory drugs
(NSAIDs) and mixtures abusers versus controls. Particularly, 4 proteins
were positively validated by immunoblotting and ELISA.
Conclusion
Urinary proteomics allows non-invasive assessment of renal diseases
at an early stage by the identification of characteristic protein pattern.
2022
- Proteomics Disclose the Potential of Gingival Crevicular Fluid (GCF) as a Source of Biomarkers for Severe Periodontitis
[Articolo su rivista]
Bellei, Elisa; Bertoldi, Carlo; Monari, Emanuela; Bergamini, Stefania
abstract
: Periodontal disease is a widespread disorder comprising gingivitis, a mild early gum inflammation, and periodontitis, a more severe multifactorial inflammatory disease that, if left untreated, can lead to the gradual destruction of the tooth-supporting apparatus. To date, effective etiopathogenetic models fully explaining the clinical features of periodontal disease are not available. Obviously, a better understanding of periodontal disease could facilitate its diagnosis and improve its treatment. The purpose of this study was to employ a proteomic approach to analyze the gingival crevicular fluid (GCF) of patients with severe periodontitis, in search of potential biomarkers. GCF samples, collected from both periodontally healthy sites (H-GCF) and the periodontal pocket (D-GCF), were subjected to a comparison analysis using sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). A total of 26 significantly different proteins, 14 up-regulated and 12 down-regulated in D-GCF vs. H-GCF, were identified by liquid chromatography-tandem mass spectrometry (LC-MS/MS). The main expressed proteins were inflammatory molecules, immune responders, and host enzymes. Most of these proteins were functionally connected using the STRING analysis database. Once validated in a large scale-study, these proteins could represent a cluster of promising biomarkers capable of making a valuable contribution for a better assessment of periodontitis.
2021
- Comparison of pregnenolone sulfate, pregnanolone and estradiol levels between patients with menstrually-related migraine and controls: an exploratory study
[Articolo su rivista]
Rustichelli, Cecilia; Bellei, Elisa; Bergamini, Stefania; Monari, Emanuela; Lo Castro, Flavia; Baraldi, Carlo; Tomasi, Aldo; Ferrari, Anna
abstract
Background
Neurosteroids affect the balance between neuroexcitation and neuroinhibition but have been little studied in migraine. We compared the serum levels of pregnenolone sulfate, pregnanolone and estradiol in women with menstrually-related migraine and controls and analysed if a correlation existed between the levels of the three hormones and history of migraine and age.
Methods
Thirty women (mean age ± SD: 33.5 ± 7.1) with menstrually-related migraine (MM group) and 30 aged- matched controls (mean age ± SD: 30.9 ± 7.9) participated in the exploratory study. Pregnenolone sulfate and pregnanolone serum levels were analysed by liquid chromatography-tandem mass spectrometry, while estradiol levels by enzyme-linked immunosorbent assay.
Results
Serum levels of pregnenolone sulfate and pregnanolone were significantly lower in the MM group than in controls (pregnenolone sulfate: P = 0.0328; pregnanolone: P = 0.0271, Student’s t-test), while estradiol levels were similar. In MM group, pregnenolone sulfate serum levels were negatively correlated with history of migraine (R2 = 0.1369; P = 0.0482) and age (R2 = 0.2826, P = 0.0025) while pregnenolone sulfate levels were not age-related in the control group (R2 = 0.04436, P = 0.4337, linear regression analysis).
Conclusion
Low levels of both pregnanolone, a positive allosteric modulator of the GABAA receptor, and pregnenolone sulfate, a positive allosteric modulator of the NMDA receptor, involved in memory and learning, could contribute either to headache pain or the cognitive dysfunctions reported in migraine patients. Overall, our results agree with the hypothesis that migraine is a disorder associated with a loss of neurohormonal integrity, thus supporting the therapeutic potential of restoring low neurosteroid levels in migraine treatment.
2021
- Dehydroepiandrosterone Sulfate, Dehydroepiandrosterone,
5α-Dihydroprogesterone and Pregnenolone: Serum Analysis and Correlation Between Migraine and Non-headache Control Females
[Abstract in Atti di Convegno]
Rustichelli, Cecilia; Monari, Emanuela; Avallone, Rossella; Bellei, Elisa; Bergamini, Stefania; Tomasi, Aldo; Ferrari, Anna
abstract
Migraine is a very painful and disabling disorder of the nervous system (NS) affecting about 10% of the world's adult population, especially women and it is associated with a variety of comorbidities [1, 2]. Neuroactive steroids have pleiotropic actions on the NS. Alterations in their peripheral and central levels could be involved in the pathogenesis, still not fully understood, of migraine and its comorbidities [3]. The purpose of our exploratory study (approved by Modena Ethical Committee) was to determine serum levels of dehydroepiandrosterone sulfate (DHEAS), dehydroepiandrosterone (DHEA), 5α-dihydroprogesterone (DHP) and pregnenolone (PREGNE) in women suffering from migraine without aura (n=30) and age-matched non-headache control females (n=30). The patients were enrolled at the Headache Centre of Modena; controls were patients’ contacts. Calibrators and serum samples were spiked with the ISs solution and treated to deplete proteins and phospholipids. The obtained extracts were evaporated to dryness, derivatized and analysed by RP-LC-ESI-MS/MS in MRM mode. Analyte’s levels were determined by interpolation on the regression curves, generated from the analyte quantifier ion peak area to the corresponding IS. Migraine women presented significantly lower levels of DHEAS, DHEA and DHP compared to controls (P<0.05) and the found concentrations negatively correlated with migraine history, and migraine days in the last three months (P< 0.05). These results parallel to our previous studies showing reduced serum levels of allopregnanolone and pregnenolone sulfate in migraine women [4,5]. The low serum levels found for both inhibitory and excitatory neurosteroids indicate that migraine women may suffer from inadequate neuroprotection, anti-inflammation activity and pain modulation. These deficits could represent the link between migraine and its various comorbidities.
References
[1] Headache classification Committee of the International Headache Society (IHS). Cephalalgia 2018, 38,1–211.
[2] Yin JH., Lin YK., Yang CP., et al. Headache 2021. doi: 10.1111/head.14106. Online ahead of print.
[3] Yilmaz, C.; Karali, K.; Fodelianaki, G.; et al. Front Neuroendocrinol 2019, 55, 100788.
[4] Rustichelli, C.; Bellei, E.; Bergamini, S.; et al. Cephalalgia 2020, 40, 1355-1362.
[5] Rustichelli, C.; Bellei, E.; Bergamini, S.; et al. J Head Pain 2021, 22, 13.
2021
- Dehydroepiandrosterone sulfate, dehydroepiandrosterone, 5α-dihydroprogesterone and pregnenolone in women with migraine: analysis of serum levels and correlation with age, migraine years and frequency
[Articolo su rivista]
Rustichelli, Cecilia; Monari, Emanuela; Avallone, Rossella; Bellei, Elisa; Bergamini, Stefania; Tomasi, Aldo; Ferrari, Anna
abstract
Migraine is a very painful, disabling and extremely common disorder among the world's adult population, especially women, and it is associated with a variety of comorbidities. Neuroactive steroids exhibit pleiotropic actions on the nervous system. Alterations in their peripheral and central levels could be involved in the pathogenesis, still not fully understood, of migraine and its comorbidities. The purpose of our exploratory study was to determine and compare the serum levels of dehydroepiandrosterone sulfate (DHEAS), dehydroepiandrosterone (DHEA), 5α-dihydroprogesterone (DHP) and pregnenolone (PREGNE) between women suffering from migraine without aura (MO group, n=30) and age-matched non-headache women as controls (C group, n=30). Correlations with age, migraine years and frequency were also analyzed. The patients were enrolled at a headache centre; controls were patients’ contacts. Calibrators and serum samples were spiked with the internal standards (ISs) solution and treated to deplete proteins and phospholipids. The obtained extracts were evaporated to dryness, derivatized and analysed by LC-MS/MS in multiple reaction monitoring mode. Analytes’ levels were determined by interpolation on the regression curves, generated from the analyte quantifier ion peak area to the corresponding IS. MO group presented significantly lower levels of DHEAS, DHEA and DHP compared to C group (P <0.05, Student’t test) and the neurosteroid levels negatively correlated with years of migraine and migraine days/3 months (P <0.05, linear regression analysis). These results parallel to previous studies showing reduced serum levels of allopregnanolone and pregnenolone sulfate in women with migraine. The low serum levels found for both excitatory and inhibitory neurosteroids suggested that women with migraine might suffer from inadequate neuroprotection, anti-inflammation activity and pain modulation. These deficits might underlie the migraine chronification process and represent the link between migraine and its various comorbidities.
2021
- Diagnostic proteomic biomarkers to detect kidney diseases
[Abstract in Rivista]
Ozben, Tomris; Bellei, Elisa; Monari, Emanuela; Bergamini, Stefania; Pini, Luigi Alberto; Tomasi, Aldo
abstract
Urinary proteomics is primarily applied to the study of renal and urogenital tract disorders. Here are reported two distinct successful examples of this approach for the discovery of early urinary biomarkers of kidney related dysfunctions: diabetic nephropathy (DN), a well known complication of diabetes frequently leading to dialysis, and druginduced nephrotoxicity, a possible condition caused by medication overuse headache (MOH). Early detection of kidney disorders based on selective biomarkers could permit to diagnose patients at the initial stage of the disease, where the therapy is still possible to stop or prevent occurrence of advance disease. Urine samples were first concentrated and desalted. Subsequently, they were subjected to two-dimensional gel electrophoresis (2DE) coupled to mass spectrometry (MS) for protein identification. Furthermore, some proteins were verified by Western blot and ELISA test. In diabetes-related study, 11 differentially expressed proteins were detected (8 upregulated and 3 downregulated) in type 2 diabetic (T2D) and T2DN patients compared to the healthy control subjects. In MOH study, a total of 21 overexcreted proteins was revealed in urine of non-steroidal antiinflammatory drugs (NSAIDs) and mixtures abusers vs controls. Particularly, 4 proteins were positively validated by immunoblotting and ELISA. Urinary proteomics allows noninvasive assessment of renal diseases at an early stage by the identification of characteristic protein pattern.
2021
- Urinary proteomic profiles of prostate cancer with different risk of progression and correlation with histopathological features
[Articolo su rivista]
Bergamini, S.; Caramaschi, S.; Monari, E.; Martorana, E.; Salviato, T.; Mangogna, A.; Balduit, A.; Tomasi, A.; Canu, P.; Bellei, E.
abstract
Prostate cancer (PCa) is the most common tumor in men with extremely variable outcome, varying from latent or indolent form to very aggressive behavior. High grade tumors, expansions exceeding the prostatic capsule into the surrounding soft tissues and spreading through lymph vascular channels, represent the most consistent unfavorable prognostic factors. However, accuracy in the prediction of the disease progression is sometimes difficult. Along with new molecular diagnostic techniques and more accurate histopathological approaches, proteomic studies challenge to identify potential biomarkers predictive of PCa progression. In our study we analyzed the urinary proteomes of 42 patients affected by PCa through two-dimensional electrophoresis associated with mass spectrometry. Proteomic profiles were correlated to histopathological features including pTNM stage and tumor differentiation in order to provide new promising markers able to define more accurately the PCa aggressiveness and driving new therapeutic approaches.
2021
- Urinary proteomics reveals promising biomarkers in menstrually related and post-menopause migraine
[Articolo su rivista]
Bellei, Elisa; Bergamini, Stefania; Rustichelli, Cecilia; Monari, Emanuela; DAL PORTO, Michele; Fiorini, Alessandro; Tomasi, Aldo; Ferrari, Anna
abstract
Abstract: Migraine is an invalidating neuro-vascular disorder largely spread in the world population. Currently, its pathophysiology is not yet completely understood. The purpose of this study was to investigate the urinary proteome of women suffering from menstrually-related migraine (MM) and post-menopause migraine (PM) in comparison with non-headache women as controls, to search potential biomarkers of these migraine sub-types. Urine samples were analysed by mono-dimensional gel electrophoresis (SDS-PAGE) and two-dimensional gel electrophoresis (2DE) coupled to liquid chromatography-mass spectrometry (LC-MS/MS). Twenty-one urinary proteins were found significantly dysregulated in MM and PM (p<0.05). STRING Analysis database revealed interaction between 15 proteins, that resulted mainly involved in immune and inflammatory response. Seven of the most considerable proteins were further quantified by Western-blot: protein S100A8 (S10A8), up-regulated in MM, uromodulin (UROM), alpha-1-microglobulin (AMBP), gelsolin (GELS) and prostaglandin-H2 D-isomerase (PTGDS), over-expressed in PM, apolipoprotein A-I (APOA1) and transthyretin (TTHY), respectively down- and up-regulated in both migraineur groups vs controls. These candidate biomarkers might be involved in the neurophysiological network of MM and PM, thus helping to better understand the pathophysiology of these migraine forms. If validated in large-scale studies, this protein cluster could become a distinctive target for clinical applications in migraine diagnosis and treatment.
2020
- Clinical and histological reaction of periodontal tissues to subgingival resin composite restorations
[Articolo su rivista]
Bertoldi, C; Monari, E; Cortellini, P; Generali, L; Lucchi, A; Spinato, S; Zaffe, D
abstract
Objectives: To compare the clinical and histological response of supracrestal periodontal tissues to subgingival composite restorations versus natural root surfaces Material and methods: In 29 subjects with a single tooth requiring subgingival restorations, a deep margin elevation (DME) procedure with composite resin was applied. Full-mouth plaque score (FMPS), full-mouth bleeding score (FMBS), and focal probing depth (PD) were measured at baseline, before DME, and after 3 months. The distance between the coronal marked (CM) point to the apical margin of the composite reconstruction (AMR), at baseline, and to the tip of the periodontal probe inserted to reach the bottom of the sulcus (APP), 3 months later, was measured. An all-around secondary flap, harvested to ensure the subsequent single-crown prosthetic rehabilitation was histologically processed. The histological inflammation degree was evaluated in areas of gingival tissues adjacent to the composite (group B) and adjacent to the natural surface of each single tooth (group A). Results: Significant FMPS, FMBS, and PD decreases were observed (p < 0.05). CM-AMR and CM-APP were significantly different (p < 0.05), suggesting an attachment gain after 3-months. The inflammation level of gingival tissue was similar in groups A and B (p > 0.05). Conclusions: For the first time, this topic was clinically and histologically studied in humans. Subgingival restorations resulted compatible with gingival health, with levels similar to that of untreated root surfaces. Clinical relevance: Deep margin elevation procedure produces favorable clinical and histological outcomes allowing a routine utilization in reconstructive dentistry.
2020
- Heparin-induced lipoprotein precipitation apheresis in dyslipidemic patients: A multiparametric assessment
[Articolo su rivista]
Merolle, L; Marraccini, C; Latorrata, A; Quartieri, E; Farioli, D; Scarano, L; Fasano, T; Bergamini, S; Bellei, E; Monari, E; Tomasi, A; Di Bartolomeo, E; Baricchi, R; Pertinhez, Ta
abstract
Low-density lipoprotein (LDL) apheresis (LA) selectively eliminates lipoproteins containing apolipoprotein B 100 (ApoB100) on patients affected by severe dyslipidemia. In addition to lowering lipids, LA is thought to exert pleiotropic effects altering a number of other compounds associated with atherosclerosis, such as pro- and anti-inflammatory cytokines or pro-thrombotic factors.
2020
- Proteomic serum profile in menstrual-related and post menopause migraine
[Articolo su rivista]
Bellei, Elisa; Rustichelli, Cecilia; Bergamini, Stefania; Monari, Emanuela; Baraldi, Carlo; Lo Castro, Flavia; Tomasi, Aldo; Ferrari, Anna
abstract
The aim of this pilot study was to analyze the serum proteomic profile of women suffering from menstrual-related migraine (MM group, n = 15) and migraine in post-menopause (PM group, n = 15) in comparison with non-headache control females (C group, n = 15). Serum samples were subjected to two-dimensional gel electrophoresis (2-DE) followed by mass spectrometry (MS) analysis for protein identification. Based on 2D-gel maps and PDQuest 2-D software, 13 differentially expressed spots, corresponding to 12 unique proteins identified by Liquid Chromatography-Electrospray Ionization-Quadrupole-Time of Flight/tandem mass spectrometry (LC-ESI-QToF-MS/MS), were detected in the MM and PM groups vs C group. Five inflammatory and regulatory of vascular integrity proteins (prothrombin, serum amyloid P-component, Ig kappa chain C region, apolipoprotein A-I, serum amyloid A-4 protein) were found deregulated in both MM and PM groups compared to C group; MM group showed the upregulation of other inflammatory protein fragments (inter-alpha-trypsin inhibitor heavy chain H4 and complement C4-A) compared to C group; PM group, in comparison with C group, displayed a noteworthy upregulation of transthyretin and other deregulated proteins (tetranectin, alpha-1-antitrypsin, haptoglobin, apolipoprotein A-IV) playing a role in anti-inflammatory and reparative processes. In conclusion, proteomic analysis was able to reveal differences in protein expression between migraine sufferers and non-headache women; as in other neurological diseases characterized by neuroinflammation, the serum proteome of migraine women presents an abundance of proteins indicative of cellular damage, oxidative stress and inflammation. This relevant inflammatory status, if confirmed in larger series, could represent a target for menstrual-related migraine treatment.
2020
- Serum levels of allopregnanolone, progesterone and testosterone in menstrually-related and postmenopausal migraine: A cross-sectional study
[Articolo su rivista]
Rustichelli, Cecilia; Bellei, Elisa; Bergamini, Stefania; Monari, Emanuela; Baraldi, Carlo; Castro, Flavia Lo; Tomasi, Aldo; Ferrari, Anna
abstract
Background: Reduced blood or cerebrospinal fluid levels of allopregnanolone are involved in menstrual cycle-linked
CNS disorders, such as catamenial epilepsy. This condition, like menstrually-related migraine, is characterized by severe,
treatment-resistant attacks. We explored whether there were differences in allopregnanolone, progesterone and testosterone
serum levels between women with menstrually-related migraine (MM, n¼30) or postmenopausal migraine
without aura who had suffered from menstrually-related migraine during their fertile age (PM, n¼30) and non-headache
control women in fertile age (FAC, n¼30) or post-menopause (PC, n¼30).
Methods: Participants were women with migraine afferent to a headache centre; controls were female patients’
acquaintances. Serum samples obtained were analyzed by HPLC-ESI-MS/MS.
Results: In menstrually-related migraine and postmenopausal migraine groups, allopregnanolone levels were lower than
in the respective control groups (fertile age and post-menopause) (p<0.001, one-way analysis of variance followed by
Tukey-Kramer post-hoc comparison test) while progesterone and testosterone levels were similar. By grouping together
patients with migraine, allopregnanolone levels were inversely correlated with the number of years and days of migraine/
3 months (p 0.005, linear regression analysis).
Conclusion: Decreased GABAergic inhibition, due to low allopregnanolone serum levels, could contribute to
menstrually-related migraine and persistence of migraine after menopause. For the management of these disorders, a
rise in the GABAergic transmission by increasing inhibitory neurosteroids might represent a novel strategy.
2019
- Allopregnanolone serum levels in female migraineurs
[Abstract in Rivista]
Rustichelli, C.; Bellei, E.; Bergamini, S.; Monari, E.; Lo Castro, F.; Baraldi, C.; Cainazzo, M. M.; Tomasi, A.; Ferrari, A.
abstract
Background: Migraine and epilepsy are similar brain disorders in many aspects and for both, a neuronal hyperexcitability has been hypothesized. Cyclic changes in ovarian hormones are involved in exacerbating both migraine and epilepsy during perimenstrual period, leading to menstrually-related migraine and catamenial epilepsy, respectively. Ovarian hormones and derived neurosteroids can regulate important functions in neurons and glial cells in the brain; in particular, progesterone reduces seizure susceptibility partly through its conversion to allopregnanolone, a potent positive allosteric modulator of the GABA-A receptor [1]. In spite of their neuroprotective potential [2], the role of neurosteroids in migraine has not been thoroughly investigated. Therefore, we determined serum levels of testosterone, progesterone and allopregnanolone in three groups: women suffering from menstrually-related migraine (n=30), post-menopausal women suffering from migraine without aura (n=30) and non-headache control females (n=20).
Methods: The enrolled migraineurs were patients afferent to the Headache Centre of Modena University Hospital; the control females were friends or relatives of the above patients. All women gave their written consent and the Ethical Committee of the Province of Modena approved the study. The fasting blood specimens were processed and then analyzed by HPLC-ESI-MS/MS.
Results: Testosterone and progesterone levels were significantly higher in both non-headache control females and women suffering from menstrually-related migraine compared to post-menopausal women suffering from migraine without aura (P <0.005, t-test). Conversely, serum allopregnanolone levels were significantly lower in both women suffering from menstrually-related migraine (0.051 ng/mL; SD: 0.018) and post-menopausal women suffering from migraine without aura (0.025 ng/mL; SD: 0.013), compared to non-headache control females (0.078 ng/mL; SD 0.036, P <0.005, t-test).
Conclusion: Women suffering from migraine presented low serum levels of allopregnanolone, a neurosteroid that modulates GABAergic inhibition. Consequently, the reduced GABAergic inhibition could inadequately protect women suffering from migraine against inflammatory and algogenic stimuli. In particular, it could contribute to the severity and poor response to treatments of migraine attacks. According to our preliminary results, a raise in the GABAergic transmission achieved by drugs increasing the biosynthetic pathway of inhibitory neurosteroids or the use of synthetic analogs could represent a possible novel therapeutic strategy for migraine management.
[1] Meletti S., et al J. Neurochem. 2018; 147:275-284.
[2] Reddy D.S., et al. Trends Pharmacol Sci. 2016;37:543-561.
2019
- Comparative proteomic analysis between the gingival crevicular fluid and the corresponding periodontal pocket: a preliminary study
[Articolo su rivista]
Bertoldi, Carlo; Bergamini, Stefania; Ferrari, Monica; Lalla, Michele; Bellei, Elisa; Spinato, Sergio; Tomasi, Aldo; Monari, Emanuela
abstract
Introduction
The aim of this study was to compare the proteomic profile of periodontal pocket tissues with that of corresponding gingival crevicular fluid (GCF), and to search for similarities in their proteomic profile.
Material and Methods
Four patients suffering from moderate or severe chronic periodontitis, needing surgical periodontal treatment, were selected. Immediately before the periodontal surgery, GCF samples were taken by means of filter paper strips positioned in the gingival sulcus correspondent to periodontal pockets. Then, periodontal pocket tissue, harvested during surgery, was adequately stored for proteomic analyses.
Results
Using an image analysis software for proteomic data, we found almost the same protein expression profile in GCF and pocket tissue from each patient. Accordingly, in our patients we found no statistically significant correlation between the quantitative proteomic profile of GCF and pocket tissue. Only one band (that of K immunoglobulin) resulted statistically different between GCF and pocket tissue proteome in all patients (p=0.008).
Conclusions
The protein network of the periodontal pocket does not influence significantly the GCF protein network. The periodontal pocket and the GCF are similar as far as the proteomic networks are concerned, but the GCF does not seem suitable to study on the pathogenesis of periodontal disease.
2019
- Evaluation of potential cardiovascular risk protein biomarkers in high severity restless legs syndrome
[Articolo su rivista]
Bellei, E.; Bergamini, S.; Monari, E.; Tomasi, A.; Koseoglu, M.; Topaloglu Tuac, S.; Ozben, S.
abstract
Restless legs syndrome (RLS) is a common sensorimotor disorder that, in case of severe symptoms, can be very distressing and negatively interfere with quality of life. Moreover, increasing evidences associate RLS with higher risk of cerebrovascular and cardiovascular disease (CVD). The purpose of this study was to quantify two proteins, previously identified by proteomics and potentially linked with CVD risk, namely kininogen-1 (KNG1) and alpha-1-antitrypsin (A1AT), in primary RLS patients at high severity grade (HS-RLS) in comparison to healthy control subjects. Proteins were quantified through enzyme-linked immunosorbent assay (ELISA) in plasma samples from 14 HS-RLS patients and 15 control individuals. The two groups were closely matched for age and gender. The expression level of KNG1 resulted significantly higher (p < 0.001), while A1AT was significantly decreased (p < 0.05) in HS-RLS patients compared to controls, confirming the relationship between these proteins and the disease severity. Furthermore, in patients group the association between the protein concentrations and the following parameters was further evaluated: age, disease onset and diagnosis, scores obtained from the RLS rating scales (Epworth Sleepiness Scale, Pittsburgh Sleep Quality Index, Beck Depression Inventory) and smoking habit. All the considered variables resulted independent of protein levels, so the disease can be reasonably considered the main cause of protein changes. As emerged from the literature, high levels of KNG1 and low amounts of A1AT seem to be related with a highest probability to develop CVD. Consequently, these proteins may be reliable candidate biomarkers of CVD risk in patients with RLS at high severity grade.
2019
- Exploration of candidate serum biomarkers potentially related to the chronic pain condition in Medication-overuse headache
[Articolo su rivista]
Pellesi, Lanfranco; Bellei, Elisa; Guerzoni, Simona; Michela Cainazzo, Maria; Baraldi, Carlo; Monari, Emanuela; Pini, Luigi Alberto
abstract
Background
Medication Overuse Headache (MOH) is a prevalent and disabling disorder resulting from the overuse of analgesic drugs, triptans or other acute headache medications. In previous proteomic studies, several proteins have been found at high concentrations in the urine of MOH patients and in the serum of rats with neuropathic pain. The aim of this study was to compare the serum levels of lipocalin-type Prostaglandin D2 synthase (L-PGDS), Vitamin D-binding protein (VDBP), apolipoprotein E (APOE) and apolipoprotein A1 (APOA1) in MOH patients and healthy individuals, further exploring their relationship with cutaneous pain thresholds (CPTs) in the territories innervated by the trigeminal nerve.
Methods
69 MOH patients and 42 age- and sex-matched healthy volunteers were enrolled in the study. Von Frey-like filaments were applied to the skin territories innervated by the trigeminal nerve, to determine the CPTs. L-PGDS, VDBP, APOE and APOA1 were quantified in the serum by Enzyme-linked Immunosorbent Assay (ELISA). Clinical and laboratory data were collected. Comparisons between MOH patients and healthy individuals were performed using independent t test or χ2 test. To correlate serum proteins with CPTs, Pearson correlation coefficient or Spearman's rank correlation coefficient were used.
Results
CPTs were lower among MOH patients. L-PGDS, VDBP and APOE had significantly different serum concentrations between groups (p < 0.01), but no correlation was found with CPTs. APOA1 serum concentrations did not differ between patients and healthy individuals.
Conclusions
L-PGDS, VDBP and APOE had abnormal serum levels in MOH patients, confirming their alteration in some conditions of chronic headache and neuropathic pain. The in-depth study of target proteins represents a promising approach for a better understanding of MOH, as well as the detection of candidate biomarkers for chronic headache or the risks associated with overuse medications.
2019
- Proteomic serum profile of female migraineurs
[Abstract in Rivista]
Bellei, E; Bergamini, S; Monari, E; Rustichelli, C; Baraldi, C; Lo Castro, F; Tomasi, A; Ferrari, A
abstract
Background: Migraine is considered a complex disease, a variable disorder of nervous system function that has a genetic background, yet the final phenotypic outcome largely depends on the individual’s environment and lifestyle. In particular, there is a clear relationship between menstruation cycle and the onset of migraine. In fact, over 50% of migraine women suffer from perimenstrual attacks, that are more serious, lasting and resistant to the treatment than non-menstrual migraine attacks [1]. We hypothesized that serum proteome analysis could help to identify potential biomarkers of menstrually-related migraine (MM) and post-menopausal migraine (PMM).
Methods: We analyzed and compared the serum proteomic profile of three groups: women suffering from MM (n=15), post-menopausal women suffering from migraine without aura (n=15) and non-headache control females (n=14). The enrolled migraineurs were patients afferent to the Headache Centre of Modena University Hospital; the control females were friends or relatives of the above patients. All women gave their written consent and the Ethical Committee of Modena approved the study. Serum samples obtained from each study participant were subjected to bi-dimensional gel electrophoresis (2-DE) coupled to mass spectrometry (MS) analysis for protein identification. The 2D-gel maps were examined by the PDQuest software, to detect the differentially expressed protein spots between the different groups [2].
Results: A total of 13 significantly different protein spots were revealed in migraine women compared to controls. Of these proteins, most (n=10) resulted increased in migraineurs vs controls, while only 3 proteins were decreased. Specifically, the greater expression differences involved the up-regulation of transthyretin in PMM and the down-regulation of apolipoprotein A1 in MM. Other proteins, such as prothrombin, serum amyloid P-component and Ig-k-chain C region, were found significantly over-expressed in migraine sufferers in comparison to controls, while one spot, recognized as serum amyloid A-4 protein, resulted decreased.
Conclusion: The serum proteome of migraine women showed proteins characteristic of cell damage, oxidative stress and lipoperoxidation, as well as acute phase proteins and inflammation markers. This pilot study demonstrates the ability of proteomics to reveal differences in protein expression between women suffering from MM and post-menopausal women suffering from migraine without aura against non-headache women. Further analysis will be carried out to expand and confirm these preliminary results.
[1] Calhoun A.H. Headache 2018; 58:626-630.
[2] Bellei E., et al. Amino Acids 2011; 40:145-156.
2018
- Biomarkers of nutritional status in honeybee haemolymph: effects of different biotechnical approaches for Varroa destructor treatment and wintering phase
[Articolo su rivista]
Cabbri, R.; Ferlizza, E.; Nanetti, A.; Monari, E.; Andreani, G.; Galuppi, R.; Isani, G.
abstract
Oxalic acid achieves its maximum efficacy against Varroa destructor during the active season only when coupled with brood manipulation techniques like brood interruption and brood removal. This study aimed to assess the impact of these manipulations on the colony nutritional status and the subsequent wintering phase, focusing on selected haemolymph biomarkers: total proteins (TP), zinc (Zn), vitellogenin (VG) and apolipophorin (APO). Twenty-five days after the manipulations (T1), colonies that underwent brood interruption (BI) stored more TP and VG than colonies in the brood removal group (BR), with a lower APO percentage, suggesting a lower metabolic effort in summer. In winter, honeybee colonies of all groups reached similar concentrations of the above-mentioned parameters, but colonies in the BI group showed a higher population. TP, VG and APO are shown to be promising biomarkers of nutritional status of the colony. Basing on the results obtained, we suggest brood interruption coupled with oxalic acid as the preferred organic method for the control of V. destructor in summer.
2018
- Discovery of restless legs syndrome plasmatic biomarkers by proteomic analysis
[Articolo su rivista]
Bellei, Elisa; Monari, Emanuela; Ozben, Serkan; Koseoglu Bitnel, Mesrure; Topaloglu Tuac, Selma; Tomasi, Aldo; Bergamini, Stefania
abstract
Objectives: Restless legs syndrome (RLS) can lead to severe clinical consequences, thus negatively impacts on patients’ overall health and quality of life. Nevertheless, the pathophysiology of RLS is still unclear, resulting in underestimate, incorrect, or ignored diagnosis and in limited management and treatment. The aim of this study was to compare the plasma proteome of RLS patients and healthy controls, in the search of diagnostic biomarkers related to the disease severity. Materials and Methods: Two-dimensional gel electrophoresis coupled with liquid chromatography-mass spectrometry was employed to analyze plasma samples of 34 patients with primary RLS, divided into two subgroups according to the disease severity: MMS group (mild-moderate symptoms) and HS group (severe and very severe symptoms), and 17 age- and sex-matched control subjects. Sleep quality, daytime sleepiness, and the level of depression were also evaluated. Results: We identified eight upregulated spots, corresponding to five unique proteins, in both RLS group vs. controls (alpha-1B-glycoprotein, alpha-1-acid glycoprotein 1, haptoglobin, complement C4-A, and immunoglobulin kappa constant); five increased spots, consistent with three unique proteins, only in HS-RLS (kininogen-1, immunoglobulin heavy constant alpha 1, and immunoglobulin lambda constant 2); one downregulated spot in both patient's groups (complement C3) and another one only in HS-RLS (alpha-1-antitrypsin). Conclusions: The significantly different plasma proteins detected in RLS were mainly associated with inflammation, immune response, and cardiovascular disorders. Particularly, the gradual increasing in immunoglobulins could be indicative of the disease severity and evolution. Accordingly, these proteins may represent a valid set of useful biomarkers for RLS diagnosis, progression and treatment.
2018
- Gingival tissue reaction to direct adhesive restoration: A preliminary study
[Articolo su rivista]
Bertoldi, C.; Zaffe, D.; Generali, L.; Lucchi, A.; Cortellini, P.; Monari, E.
abstract
Introduction: It is debated whether composite resin marginal/submarginal direct restoration can be usefully performed without inflammatory consequences. This histological study is the first human analysis aimed to compare, in the same tooth, the gingival tissue close to composite resin restorations with gingival tissue close to hard tissue. Methods: Eight healthy patients with almost a residual strategic tooth needing endodontic therapy, and post-and-core restoration, then indirect prosthetic restoration, were selected. Direct margin relocation with composite resin was necessary to perform endodontic treatment. The crown lengthening with a secondary flap harvested was necessary to perform prosthetic rehabilitation. Three months after marginal relocation, the secondary flap was harvested, embedded in PMMA, 4-μm sectioned, and stained to analyze the inflammation degree. Results: All patients completed post-and-core reconstruction and the planned prosthetic therapy, maintaining the stringent hygienic protocol plan. The inflammation level comparison, slightly lower in gingiva close to the teeth (3.62 ± 0.38) than in gingiva close to the composite (3.75 ± 0.26), results in a p-value of 0.11 after Wilcoxon test. Conclusions: Results highlight a minimal, statistically not significant difference in the inflammation degree after margin relocation, conceivably due to patients, teeth and cases selection, together with adopted stringent methodological and supportive measures.
2018
- Serum changes of apolipoproteins in Medication Overuse Headache (MOH)
[Abstract in Rivista]
Pellesi, Lanfranco; Bellei, Elisa; Baraldi, Carlo; Guerzoni, Simona; Monari, Emanuela; Pini, Luigi Alberto
abstract
Background
Medication Overuse Headache (MOH) is a prevalent and disabling
disorder resulting from the overuse of analgesic drugs, triptans or
other acute headache medications. Previous proteomic studies have
identified some altered proteins, including different forms of apolipoproteins,
which are probably associated with the chronic painful
symptom and its consequences. The aim of the study was to explore
the relationship between cutaneous pain thresholds, Zung Self-
Rating Depression Scale (ZUNG-D) scores, Leeds Dependence Questionnaire
(LDQ) scores and serum levels of apolipoprotein A1
(APOA1) and apolipoprotein E (APOE) in patients with MOH.
Methods
69 patients with MOH and 42 healthy volunteers as control group
were enrolled in the study between September 2016 and January
2018. To investigate skin sensitivity, Von Frey-like filaments were
applied sequentially to the skin territories innervated by the divisions
of the trigeminal nerve, to determine cutaneous pain
thresholds. APOA1 and APOE, previously identified as potential
biomarkers candidates for the pathophysiology of chronic pain,
were quantified in the serum by Enzyme-linked Immunosorbent
Assay (ELISA).
Results
Cutaneous pain thresholds were lower among patients with MOH
than healthy controls. Serum APOE was significantly lower in patients
with MOH, compared to healthy volunteers (p < 0.01), but no differences
were found concerning serum APOA1 (Fig. 1). In patients with
MOH, serum APOE was positively related to Body Mass Index, albumin
and uric acid, whereas serum APOA1 was positively related to
creatinine. Serum APOA1 and APOE did not have any relationship
with cutaneous pain thresholds, ZUNG-D scores, LDQ scores and
other clinical or laboratory parameters.
Conclusions
Serum APOE is significantly altered in patients with MOH compared
to controls, but it is apparently not correlable with any aspect of the
disease. APOE may play a role in the pathophysiology of MOH and
the consequences associated with medication overuse; further studies
are needed to deepen this finding.
Ethics approval
This study was performed following the Helsinki Declaration principles
and approved by the local Ethical Committee (prot. 2073).
2018
- Studio degli effetti della LDL aferesi in pazienti affetti da ipercolesterolemia familiare
[Abstract in Rivista]
Merolle, L; Latorrata, A; Marraccini, C; Farioli, D; Scarano, L; Di Bartolomeo, E; Bergamini, S; Bellei, E; Monari, E; Fasano, T; Baricchi, R; Pertinhez, T. A.
abstract
La LDL-aferesi selettiva è un trattamento che riduce radicalmente l'LDL nei pazienti affetti da ipercolesterolemia familiare (FH) e al contempo va a modificare i livelli di altri composti associati al fenomeno dell'aterosclerosi.
2018
- Urinary proteomics in biomarker discovery of kidney-related disorders: Diabetic nephropathy and drug-induced nephrotoxicity in chronic headache
[Articolo su rivista]
Bellei, Elisa; Monari, Emanuela; Bergamini, Stefania; Pini, Luigi Alberto; Tomasi, Aldo; Ozben, Tomris
abstract
Objective Urinary proteomics is primarily applied to the study of renal and urogenital tract disorders. Here are reported two distinct successful examples of this approach for the discovery of early urinary biomarkers of kidney-related dysfunctions: diabetic nephropathy (DN), a well-known complication of diabetes frequently leading to dialysis, and drug-induced nephrotoxicity, a possible condition caused by medication-overuse headache (MOH). Early detection of kidney disorders based on selective biomarkers could permit to diagnose patients at the initial stage of the disease, where the therapy may be suspended or prevent disease advancement. Methods Urine samples were first concentrated and desalted. Subsequently, they were subjected to two-dimensional gel electrophoresis (2-DE) coupled to mass spectrometry (MS) for protein identification. Furthermore, some proteins were verified by Western blot and ELISA test. Results In diabetes-related study, 11 differentially expressed proteins were detected (8 up-regulated and 3 down-regulated) in type 2 diabetic (T2D) and T2DN patients compared to the healthy control subjects. In the MOH study, a total of 21 over-excreted proteins were revealed in urine of non-steroidal anti-inflammatory drugs (NSAIDs) and mixtures abusers vs controls. Particularly, 4 proteins were positively validated by immunoblotting and ELISA. Conclusion Urinary proteomics allows non-invasive assessment of renal diseases at an early stage by the identification of characteristic protein pattern.
2017
- Serum protein changes in a rat model of chronic pain show a correlation between animal and humans
[Articolo su rivista]
Bellei, Elisa; Vilella, Antonietta; Monari, Emanuela; Bergamini, Stefania; Tomasi, Aldo; Cuoghi, Aurora; Guerzoni, Simona; Manca, Letizia; Zoli, Michele; Pini, Luigi Alberto
abstract
In previous works we showed the overexpression of some proteins in biological fluids from patients suffering chronic pain. In this proteomic study we analysed serum from a rat model of neuropathic pain obtained by the chronic constriction injury (CCI) of sciatic nerve, at two time intervals, 2 and 5 weeks after the insult, to find proteins involved in the expression or mediation of pain. Sham-operated and CCI rats were treated with saline or indomethacin. Two weeks after ligation, we identified three serum proteins overexpressed in CCI rats, two of which, alpha-1-macroglobulin and vitamin D-binding protein (VDBP), remained increased 5 weeks post-surgery; at this time interval, we found increased levels of further proteins, namely apolipoprotein A-I (APOA1), apolipoprotein E (APOE), prostaglandin-H2 D-isomerase (PTGDS) and transthyretin (TTR), that overlap the overexpressed proteins found in humans. Indomethacin treatment reversed the effects of ligation. The qPCR analysis showed that transcript levels of APOA1, APOE, PTGDS and VDBP were overexpressed in the lumbar spinal cord (origin of sciatic nerve), but not in the striatum (an unrelated brain region), of CCI rats treated with saline 5 weeks after surgery, demonstrating that the lumbar spinal cord is a possible source of these proteins.
2016
- Characterization of equine amniotic fluid by a proteomic approach
[Abstract in Atti di Convegno]
Isani, G; Ferlizza, E; Cuoghi, A; Bellei, E; Monari, E; Bianchin Butina, B; Castagnetti, C
abstract
This study succesfully applied SDS-PAGE coupled to MS for the identification of the most abundant proteins in equine AF.
2016
- Identification of the most abundant proteins in equine amniotic fluid by a proteomic approach
[Articolo su rivista]
Isani, Gloria; Ferlizza, Enea; Cuoghi, Aurora; Bellei, Elisa; Monari, Emanuela; Bianchin Butina, Barbara; Castagnetti, Carolina
abstract
Characterisation of the physiologic equine amniotic fluid (AF) proteome is a prerequisite to study its changes during diseases and discover new biomarkers. The aim of this study was to identify by a proteomic approach the most abundant proteins of equine AF. AF samples were collected at parturition from 24 healthy mares that delivered healthy foals. All samples were subjected to sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) on 4–12% gels. A pool of the 24 samples, after SDS-PAGE, was cut in 25 slices, trypsin-digested and analysed by mass spectrometry (MS) for protein identification. Mean AF protein concentration was 1.96 ± 1.12 g/L. Thirty-four proteins were successfully identified by MS and subsequently categorised according to Gene Ontology (GO). Twelve proteins (e.g. fibronectin, lumican, thrombospondin and fibulin) belonged to or interacted with the extracellular matrix (ECM) playing an important role in the development of foetal tissues. Most of the remaining proteins were classified as transport (e.g. albumin, major allergen Equ c1 and alpha-fetoprotein) delivering nutrients, ions and lipids essential for foetal growth and development. Among these proteins, major allergen Equ c1 is widely studied in human medicine because it induces Ig-E mediated type I allergic reaction. The absence of immunoglobulins in equine AF was also confirmed.
2016
- Influence of two different Varroa treatments on the most abundant proteins of honey bee hemolymph
[Poster]
Ferlizza, E; Cabbri, R; Andreani, G; Bellei, E; Cuoghi, A; Monari, E; Isani, G
abstract
Evaluation of the effect of different techniques on vitellogenin and other abundant hemolymph proteins to obtain successful wintering of the colonies
2016
- The protein expression network in periodontal pocket tissue: a preliminary study
[Abstract in Rivista]
Bertoldi, C; Monari, E; Generali, L; Franceschetti, F; Bellini, P; Salgarelli, Ac; Consolo, U
abstract
Background and Objective
The periodontal disease (PD) is caused by a set of inflammatory disorders characterized by periodontal pocket formation that lead to tooth loss if untreated.
Periodontitis diagnosis is only based on clinical assessment only, in the absence of a reliable pathogenic check based on appropriate interpretation of inflammation.
A modern pathogenic model based on a multilevel framework including disease-initiating and -resolving mechanisms is requested. Studies on PD utilizing proteomic analysis have been performed on saliva or crevicular fluid samples, peripheral blood or periodontal plaque samples, but not on the pathologic tissue of the periodontal pocket, which is the key lesion of the PD.
The aim of this work was to compare the proteomic profile of the pathologic interproximal gingival pocket tissue with the corresponding of interproximal gingival healthy tissue, obtained from sites where no periodontal-pathogenic bacteria were detectable.
Materials and methods
Twenty healthy subjects (T, test group), affected by chronic PD and twenty periodontally healthy subjects (C, control group), were enrolled in the study. T subjects underwent to the periodontal resective treatment, while C subjects underwent to the crown lengthening surgical treatment. To characterize the proteomic profile of periodontally-affected patients, their interproximal periodontal pocket tissue was compared with that of periodontally-healthy patients. Pocket-associated and healthy tissue samples, harvested during surgical therapy, were treated to extract the protein content. Tissues were always collected at sites where no periodontal-pathogenic bacteria were detectable. 2DE (Two-dimensional gel electrophoresis) and LC-MS/MS (Liquid Chromatography-tandem mass spectrometry) analysis were performed for T and C gingival tissue samples to separate and identify the proteins. Web-based bioinformatics tools (iPROClass and CateGOrize) were employed to investigate all potential localizations, molecular functions and biological processes of the identified proteins. After identification, proteins were selected for subsequent Western Blot quantitation both in pathological and healthy tissues.
Results
The identified proteins are mainly involved in metabolism (32 %), transport (13 %) and cell organization and biogenesis (13 %). A significant unbalance in protein expression between healthy and pathological sites was recorded. Thirty-two protein spots were overall identified, and four proteins, the protein S100-A9 (S100A9), heat shock protein beta-1 (HSPB1), Galectin-7 (LEG7) and 14-3-3 protein zeta/delta (14-3-3) were selected for Western blot analysis of both periodontally-affected and healthy patients. The four selected proteins resulted extremely over-expressed in periodontal pocket tissue when compared with the corresponding tissue of periodontally-healthy patients.
Discussion:
S100A9 is involved in the regulation of inflammatory processes, immune response, oxidant-scavenging and apoptosis-inducing activities. HSPB1 synthesis increases in response to a variety of stresses (e.g. elevated temperatures, heavy metals, toxins, oxidants, bacterial and viral infections) in order to minimize the attendant deleterious consequences. LEG7 expression contributes to the tissue remodeling processes following tissue damage and protects cell from death. The 14-3-3 proteins are involved in the control of several cell cycle checkpoint, connective tissue remodeling, apoptosis signaling, during inflammation response. The Western blot and proteomic analyses are congruent with the reaction of injured periodontal tissues in PD. The proteomic analysis was performed for the first time directly on periodontal pocket tissue. The proteomic network highlighted enhances the understanding of PD pathogenesis revealing a defensive protein strategy which, however, evidently fails in patients affected by PD. Further study are requested also for specific therapeutic strateg
2015
- A proteomics study in cats affected by chronic kidney disease: identification of putative biomarkers
[Abstract in Rivista]
Ferlizza, E; Campos, A; Cuoghi, A; Bellei, E; Monari, E; Dondi, F; Almeida, Am; Isani, G
abstract
Application of proteomic techniques for the identification of putative biomarkers of nephropathy in cats affected by chronic kidney disease.
2015
- Analysis of protein expression in periodontal pocket tissue: a preliminary study
[Articolo su rivista]
Monari, Emanuela; Cuoghi, Aurora; Bellei, Elisa; Bergamini, Stefania; Lucchi, Andrea; Tomasi, Aldo; Cortellini, Pierpaolo; Zaffe, Davide; Bertoldi, Carlo
abstract
The periodontal disease is caused by a set of inflammatory disorders characterized by periodontal pocket formation that lead to tooth loss if untreated. The proteomic profile and related molecular conditions of pocket tissue in periodontally-affected patients are not reported in literature. To characterize the proteomic profile of periodontally-affected patients, their interproximal periodontal pocket tissue was compared with that of periodontally-healthy patients. Pocket-associated and healthy tissue samples, harvested during surgical therapy, were treated to extract the protein content. Tissues were always collected at sites where no periodontal-pathogenic bacteria were detectable. Proteins were separated using two-dimensional gel electrophoresis and identified by liquid chromatography/mass spectrometry. After identification, four proteins were selected for subsequent Western Blot quantitation both in pathological and healty tissues.
2015
- Erratum to: Proteomic analysis of protein extraction during hemofiltration with on-line endogenous reinfusion (HFR) using different polysulphone membranes [JMater Sci:MaterMed, 25, (2014) 2691-2698, DOI 10.1007/s10856-014-5290-5]
[Articolo su rivista]
Monari, E.; Cuoghi, A.; Bellei, E.; Bergamini, S.; Caiazzo, M.; Aucella, F.; Loschiavo, C.; Corazza, L.; Palladino, G.; Sereni, L.; Atti, M.; Tomasi, A.
abstract
2015
- New horizon in dialysis depuration: Characterization of a polysulfone membrane able to break the 'albumin wall'
[Articolo su rivista]
Cuoghi, Aurora; Caiazzo, Marialuisa; Monari, Emanuela; Bellei, Elisa; Bergamini, Stefania; Sereni, L; Aucella, F; Loschiavo, C; Atti, M; Tomasi, Aldo
abstract
The uremic syndrome is attributed to the progressive retention of a large number of toxins, which under normal conditions are excreted by the healthy kidneys. Standard dialytic membranes do not purify middle-high molecular weight toxins. Haemodiafiltration with endogenous reinfusion coupled with a highly permeable membrane could break the limit of the 'albumin wall' improving the dialytic depuration without loss of important nutrients. The aim of this study was to evaluate the performance of a new polysulfone membrane, Synclear 0.2, to remove uremic molecules. Surface Enhanced Laser Desorption Ionization-Time of Flight was employed to evaluate the proteomic profile of ultrafiltrate and Electrospray Ionization-Quadruple-ToF coupled with on-chip elution was used for proteins identification. A high and specific permeability for middle-high molecular weight molecules was revealed by mass spectrometry for the investigated membrane. The identified proteins are mostly uremic toxins: their relative abundance, estimated in the ultrafiltrate by exponentially modified protein abundance index, showed a high purification efficiency of the new membrane when compared with conventional ones. In conclusion, Synclear 0.2, used as convective membrane in hemodiafiltration with endogenous reinfusion treatment, permits to break the 'albumin wall', clearing middle-high molecular weight uremic toxins, improving the dialytic treatment purification efficiency.
2015
- Proteomic analysis of proteins adsorbed by resin cartridge filter during hemodiafitration with online
endogenous reinfusion.
[Abstract in Rivista]
Monari, Emanuela; Bergamini, Stefania; Cuoghi, Aurora; Bellei, Elisa; Solano, Francesco; Bruni, Francesco; Ozben, Tomris; Tomasi, Aldo
abstract
BACKGROUND-AIM
Background: Lupus nephritis (LN) is kidney inflammation caused by systemic lupus erythematous (SLE), that lead to
end stage renal disease and consequently to dialytic therapy. Inflammation mediators over-expression play a key
role in disease initiation and progression. Immuno-complexes and/or autoantibodies deposition in the kidney induce
cytokines production in renal resident cells, which may further amplify inflammatory processes. Hemodiafiltration
with Endogenous Reinfusion (HFR) dialysis treatment with super high flux membrane Synclear 02 (SUPRA) is a dialytic
method, which combines the diffusion and convection processes with adsorption by a resin cartridge filter. Proteomic
approach was applied for protein separation and identification in order to evaluate the quality of proteins retained by
resin bed during dialytic treatment.
METHODS
Methods: Plasma and ultrafiltrate (UF) samples of three patients with LN, treated with SUPRA HFR (Bellco, Italy), were
collected at 15 and 235 min of two different dialytic sessions. The utilized cartridges, containing styrenic resin, were
opened and the proteins kept by the resin were eluted. Gel electrophoresis was used to separate protein content
before protein identification by ESI-QTOF-MS (Electrospray Ionization-Quadrupole Time-of-Flight-Mass Spectrometry)
analysis
RESULTS
Results: The comparison of proteomic profiles of plasma, UF and eluted samples demonstrate the removal of several
protein species by the resin bed. ESI-QTOF analysis allowed to identify several biomarker of kidney injury, such as:
Retinol binding protein 4, Neutrophil gelatinase-associated lipocalin (NGAL), Prostaglandin-H2 D-isomerase, Cystatin-
C, Serotransferrin, Alpha-1-acid glycoprotein (A1AG1), Transthyretin and several fragments of Immunoglobulins.
Moreover, Beta-2-glycoprotein 1 (APO-H), involved in antiphospholipid syndrome, a disorder that manifests clinically
as recurrent venous or arterial thrombosis, was identified.
CONCLUSION
Conclusions: The proteomic approach was used in this study to evaluate the performance of styrenic resin to retain
proteins implicated in the LN pathogenesis and pathophisiology. The treatment with SUPRA-HFR demonstrate to be
suitable to reduce inflammatory status, uremic toxin level and antiphospholipid syndrome in LN patients.
2015
- Proteomic research of proteins involved in pain expression in an animal model of chronic pain
[Articolo su rivista]
Bellei, Elisa; Bergamini, Stefania; Monari, Emanuela; Cuoghi, Aurora; Zoli, Michele; Tomasi, Aldo; Cainazzo, Maria Michela; Guerzoni, Simona; Pini, Luigi Alberto
abstract
nd
2015
- Proteomics in the clinic: the search for biomarkers
[Abstract in Atti di Convegno]
Tomasi, A; Bellei, E; Bergamini, S; Cuoghi, A; Monari, E; Ozben, T
abstract
Biomarkers are molecules that exist naturally in the body; they can help to predict or reflect the presence of a disease, the relapse risk of the disease, and/or response to treatment.
2015
- The effect of chronic kidney disease on the urine proteome in the domestic cat (Felis catus)
[Articolo su rivista]
Ferlizza, E.; Campos, A.; Neagu, A.; Cuoghi, Aurora; Bellei, Elisa; Monari, Emanuela; Dondi, F.; Almeida, A. M.; Isani, G.
abstract
Chronic kidney disease (CKD) is a major cause of mortality in cats, but sensitive and specific biomarkers for early prediction and monitoring of CKD are currently lacking. The present study aimed to apply proteomic techniques to map the urine proteome of the healthy cat and compare it with the proteome of cats with CKD. Urine samples were collected by cystocentesis from 23 healthy young cats and 17 cats with CKD. One-dimensional sodium-dodecyl-sulfate polyacrylamide gel electrophoresis (1D-SDS-PAGE) was con- ducted on 4–12% gels. Two-dimensional electrophoresis (2DE) was applied to pooled urine samples from healthy cats (n = 4) and cats with CKD (n = 4), respectively. Sixteen protein bands and 36 spots were cut, trypsin-digested and identified by mass spectrometry.
1D-SDS-PAGE yielded an overall view of the protein profile and the separation of 32 ± 6 protein bands in the urine of healthy cats, while CKD cats showed significantly fewer bands (P < 0.01). 2-DE was es- sential in fractionation of the complex urine proteome, producing a reference map that included 20 proteins. Cauxin was the most abundant protein in urine of healthy cats. Several protease inhibitors and trans- port proteins that derive from plasma were also identified, including alpha-2-macroglobulin, albumin, transferrin, haemopexin and haptoglobin. There was differential expression of 27 spots between healthy and CKD samples (P < 0.05) and 13 proteins were unambiguously identified. In particular, increased ex- pression of retinol-binding protein, cystatin M and apolipoprotein-H associated with decreased expression of uromodulin and cauxin confirmed tubular damage in CKD cats suggesting that these proteins are can- didate biomarkers.
2015
- The importance of inflammation in the search of prostate cancer biomarkers
[Poster]
Bergamini, S; Bellei, E; Monari, E; Cuoghi, A; Reggiani Bonetti, L; Borelli, F; Sighinolfi, C; Bianchi, G; Ozben, T; Tomasi, A
abstract
The importance of inflammation in the search of prostate cancer biomarkers by proteomics.
2015
- Validation of potential candidate biomarkers of drug-induced nephrotoxicity and allodynia in medication-overuse headache
[Articolo su rivista]
Bellei, Elisa; Monari, Emanuela; Bergamini, Stefania; Cuoghi, Aurora; Tomasi, Aldo; Guerzoni, Simona; Ciccarese, Michela; Pini, Luigi Alberto
abstract
Medication-overuse headache (MOH) is a chronic disorder that results from the overuse of analgesics drugs, triptans or other acute headache compounds. Although the exact mechanisms underlying MOH remain still unknown, several studies suggest that it may be associated with development of "central sensitization", which may cause cutaneous allodynia (CA). Furthermore, the epidemiology of drug-induced disorders suggests that medication overuse could lead to nephrotoxicity. The aim of this work was to confirm and validate the results obtained from previous proteomics studies, in which we analyzed the urinary proteome of MOH patients in comparison with healthy non-abusers individuals.
2014
- A 2DE map of the urine proteome in the cat: effect of Chronic Kidney Disease
[Relazione in Atti di Convegno]
Ferlizza, E; Campos, A; Cuoghi, A; Bellei, E; Monari, E; Dondi, F; Almeida, Am; Isani, G
abstract
The aim of this study was to produce a preliminary map of the urine proteome of healthy cats (Felis catus) and to compare it with the proteome of cats affected by chronic kidney disease.
2014
- Analisi tissutale proteomica della tasca parodontale. Uno studio pilota
Periodontal pocket tissue analysis using proteome. A pilot study
[Articolo su rivista]
Chiara, Pellacani; Monari, Emanuela; Zaffe, Davide; Cuoghi, Aurora; Bellei, Elisa; Andrea, Lucchi; Bergamini, Stefania; Tomasi, Aldo; Bertoldi, Carlo
abstract
Obbiettivo: Scopo dello studio è analizzare nello stesso soggetto, in siti in cui non erano rilevabili batteri parodontopatogeni, il tessuto inteprossimale, sia associato alla tasca parodontale sia sano, al fine di determinare un quadro proteico associabile al danno parodontale.
Materiali & Metodi: Nello studio sono stati inclusi quindici soggetti sistemicamente sani, affetti da moderata-avanzata parodontite cronica, che presentavano almeno un difetto intraosseo prossimo ad un analogo sito senza danno parodontale clinicamente evidente. I pazienti sono stati trattati mediante terapia resettiva. Durante la fase chirurgica i tessuti associati alla lesione parodontale e quelli clinicamente sani sono stati prelevati per l’analisi proteomica.
Risultati: Confrontando i profili proteici relativi al danno parodontale con quelli clinicamente sani, sono state identificate 19 proteine differentemente espresse. In particolare, in tutti i pazienti 8 proteine sono risultate sovra-espresse nel tessuto patologico (Anexina A2- ANX A2; Actina citoplasmatica 1 (spot 14 e 15)- ACTB; Anidrasi carbonica 1 - CAH1; Anidrasi carbonica 2- CAH2; Ig catena Kappa regione C (spot 17 e 18)- IGKC e flavina reduttasi- BLVRB) mentre 11 proteine sono risultate sotto-espresse (Tropomiosina catena -4- TPM3; proteina 14-3-3 - 1433S; proteina / 14-3-3 - 1433Z; -enolasi - ENOA; Heat shock proteina -1 (spot 5 e spot 6) - HSPB1; Triosofosfatoisomerasi - TPIS; Perossiredoxina-1 - PRDX1; Proteina epidermica legante acidi grassi - FABP5; Proteina S100-A9 - S10A9 e Galectina -7 - LEG7).
Conclusioni: Dai dati preliminari ottenuti risulta evidenziata l’espressione differenziale, tra tessuto clinicamente sano e relativo al danno parodontale, di proteine che possono giocare un ruolo importante nella prevenzione del danno cellulare da stress, nella mediazione delle risposte immunitarie, nonché nei meccanismi di rigenerazione tissutale. Lo studio del profilo profilo proteomico del tessuto della tasca parodontale potrebbe essere cruciale sia per la conoscenza della patogenesi che per la terapia della malattia parodontale.
Objective: To analyze in the same subject, in sites where no periodontopathogenic bacteria were detectable, pocket-associated and neighboring healthy interproximal tissues to qualify proteins associated with the periodontal damage.
Matherials & Methods: Fifteen healthy patients, affected by moderate to advanced chronic periodontitis and presenting at least one intrabony defect and a neighboring not-damaged interproximal site were enrolled. Patients underwent osseous resective surgery. During surgery pocket-associated and clinically healthy tissues were harvested for proteomic analyses.
Results: In both pocket-associated and clinically healthy tissues, nineteen differently expressed proteins were successfully identified. In particular, 8 proteins (Annexin A2; Actin cytoplasmic 1 (2 spots); Carbonic anhydrase 1; Carbonic anhydrase 2; Ig kappa chain C region (2 spots) and Flavinreductase) were over-expressed, while 11 proteins (Tropomyosin alpha-4 chain; 14-3-3 protein sigma; 14-3-3 protein zeta/delta; Alpha-enolase; Heat shock protein ß-1 (2 spots); Triosophosphateisomerase; Peroxiredoxin-1; Fatty acid-binding protein-epidermal; Protein S100-A9 and Galectin-7) were under-expressed in the pathological tissue of all patients.
Conclusions: The preliminary data indicate differentially expression of proteins that may play important roles in the prevention of cellular damage by stress, in mediating the immune response as well as in tissue regeneration. The proteomic profile study of pocket tissue would be crucial both to appreciate the pathogenesis and the therapy of periodontitis.
2014
- Evaluation of proteomic profile in menstrual-related migraine
[Abstract in Rivista]
Cuoghi, Aurora; Monari, Emanuela; Bellei, Elisa; Bergamini, Stefania; Tomasi, Aldo; Ferrari, Anna
abstract
nd
2014
- Evalutation of proteomic profile in menstrually-related migraine
[Abstract in Rivista]
Cuoghi, A; Monari, E; Bellei, E; Bergamini, S; Tomasi, A; Ferrari, A.
abstract
Valutazione del profilo proteomico (sierico e urinario) nelle donne con emicrania correlata alle mestruazioni
2014
- Inflammation: an important parameter in the search of prostate cancer biomarkers
[Articolo su rivista]
Bergamini, Stefania; Bellei, Elisa; REGGIANI BONETTI, Luca; Monari, Emanuela; Cuoghi, Aurora; Francesco, Borelli; Sighinolfi, Maria Chiara; Bianchi, Giampaolo; T., Ozben; Tomasi, Aldo
abstract
Background
A more specific and early diagnostics for prostate cancer (PCa) is highly desirable. In this study, being inflammation the focus of our effort, serum protein profiles were analyzed in order to investigate if this parameter could interfere with the search of discriminating proteins between PCa and benign prostatic hyperplasia (BPH).
Methods
Patients with clinical suspect of PCa and candidates for trans-rectal ultrasound guided prostate biopsy (TRUS) were enrolled. Histological specimens were examined in order to grade and classify the tumor, identify BPH and detect inflammation. Surface Enhanced Laser Desorption/Ionization-Time of Flight-Mass Spectrometry (SELDI-ToF-MS) and two-dimensional gel electrophoresis (2-DE) coupled with Liquid Chromatography-MS/MS (LC-MS/MS) were used to analyze immuno-depleted serum samples from patients with PCa and BPH.
Results
The comparison between PCa (with and without inflammation) and BPH (with and without inflammation) serum samples by SELDI-ToF-MS analysis did not show differences in protein expression, while changes were only observed when the concomitant presence of inflammation was taken into consideration. In fact, when samples with histological sign of inflammation were excluded, 20 significantly different protein peaks were detected. Subsequent comparisons (PCa with inflammation vs PCa without inflammation, and BPH with inflammation vs BPH without inflammation) showed that 16 proteins appeared to be modified in the presence of inflammation, while 4 protein peaks were not modified. With 2-DE analysis, comparing PCa without inflammation vs PCa with inflammation, and BPH without inflammation vs the same condition in the presence of inflammation, were identified 29 and 25 differentially expressed protein spots, respectively. Excluding samples with inflammation the comparison between PCa vs BPH showed 9 unique PCa proteins, 4 of which overlapped with those previously identified in the presence of inflammation, while other 2 were new proteins, not identified in our previous comparisons.
Conclusions
The present study indicates that inflammation might be a confounding parameter during the proteomic research of candidate biomarkers of PCa. These results indicate that some possible biomarker-candidate proteins are strongly influenced by the presence of inflammation, hence only a well-selected protein pattern should be considered for potential marker of PCa.
2014
- Protein survey in periodontal pockets using proteome analysis.
[Relazione in Atti di Convegno]
Bertoldi, Carlo; M., Martani; Monari, Emanuela; D., Ferrari; C., Pellacani; P., Cortellini; Consolo, Ugo
abstract
Aim: To reveal proteins associated with periodontal disease in sites where periodontopathogenic bacteria were not detectable comparing the proteomic profile of interproximal pocket tissues with interproximal healthy tissues in the same subject
Methods: 39 patients, with moderate to advanced chronic periodontitis and presenting with at least one intrabony defect next to a healthy interproximal site were enrolled. The periodontal defects were treated with osseous resective surgery, and the flap design included both the periodontal pockets and the neighboring interproximal healthy sites. Pocket-associated and healthy tissues were harvested for proteomic analyses. The results were processed using standard proteomic and statistical analyses to size and density of the spots.
Results: 15 proteins resulted differently expressed between pathological and healthy tissues. Particularly, annexin A2, actin cytoplasmic 1, carbonic anhydrase 1 & 2; Ig kappa chain C region (2 spots) and flavinreductase were overexpressed, whereas 14-3-3 protein sigma and zeta/delta, heat shock protein beta -1 (2 spots), triosophosphateisomerase, peroxiredoxin-1, fatty acid-binding protein-epidermal, and galectin-7 were underexpressed in pathological tissue.
Conclusions: The unbalanced functional network of proteins involved could hinder adequate tissue response to pathogenic noxa through different mechanisms, ranging from a lack of control of cellular stresses to an alteration of immune and apoptotic capacity. In particular it seems that it is more significant deficiency more than the over-expression of proteins to justify the pathodynamic of the periodontal disease. The study of periodontal pocket tissue proteomic profile would be crucial to better understand the pathogenesis of and the therapeutic strategies for periodontitis.
2014
- Proteomic analisys of protein extraction during hemofiltration with on-line endogenous reinfusion (HFR) using different polysulphone membranes
[Articolo su rivista]
Monari, Emanuela; Cuoghi, Aurora; Bellei, Elisa; Bergamini, Stefania; Marialuisa, Caiazzo; Filippo, Aucella; Carmelo, Loschiavo; Luca, Corazza; Giuseppe, Palladino; Luisa, Sereni; Mauro, Atti; Tomasi, Aldo
abstract
In end-stage renal disease patients, extracor- poreal dialytic therapy is not able to prevent the accumu- lation of toxins related to the uremic syndrome, a severe complication that increases morbidity and mortality rate. In this paper, hemoFiltration with on-line Reinfusion (HFR) architecture is used to evaluate the effect of a more per- meable membrane on the extraction of medium–high molecular weight molecules. The aim of this study was to compare two polysulphone membranes for convective chamber: polyphenylene High Flux (pHF) and polyphen- ylene Super High-Flux (pSHF). Fourteen patients were subjected to HFR with pHF and pSHF membranes and ultra filtrate (UF) samples were collected to evaluate molecular weight cut-off (MWCO) and to identify extracted proteins. Furthermore, image analysis software was used in order to evaluate change in protein extraction during the dialysis. The quantification of four proteins by immunoassay dem- onstrates a higher permeability of pSHF membrane. Two- dimensional electrophoresis (2-DE) gels showed, for both
Electronic supplementary material The online version of this article (doi:10.1007/s10856-014-5290-5) contains supplementary material, which is available to authorized users.
M. Emanuela (&) ? C. Aurora ? B. Elisa ? B. Stefania ? T. Aldo Department of Diagnostic, Clinical and Public Health Medicine,
University of Modena and Reggio Emilia, Via del Pozzo, 71-41124 Modena, Italy e-mail: emanuela.monari@unimore.it
C. Marialuisa ? C. Luca ? P. Giuseppe ? S. Luisa ? A. Mauro Scientific Affairs, Bellco s.r.l, Mirandola, Modena, Italy
A. Filippo IRCCS Hospital CSS, San Giovanni Rotondo, Foggia, Italy
L. Carmelo Division of Nephrology and Dialysis, Legnago, Verona, Italy
membranes, the greater number of protein spots at 235 min. Some of the identified proteins, involved in nephropathic disease complications, were compared to assess differences in extraction during dialytic treatment by PDQuest analysis. UF proteomic analysis demonstrated a different behavior for the two membranes; pHF membrane was more permeable at the beginning of HFR treatment (15 min), while pSHF membrane at the end of treatment (235 min). Proteomic analysis is a suitable approach to investigate the behavior of different membranes during dialysis. Results indicated that pSHF membrane offers the higher permeability, and showed higher efficiency in removal of middle molecules related to uremic syndrome.
2014
- Proteomic profile of retained proteins from hemodiafiltration with on-line endogenous reinfusion (SUPRA) cartridge.
[Abstract in Rivista]
Cuoghi, Aurora; Bellei, Elisa; Monari, Emanuela; Bergamini, Stefania; Tomasi, Aldo; Atti, Mauro; Caiazzo, Marialuisa; Palladino, Giuseppe; Bruni, Francesco
abstract
nd
2014
- Validation of prostate cancer biomarkers and inflammation: a proteomic study
[Abstract in Atti di Convegno]
Bergamini, S; Bellei, E; Monari, E; Cuoghi, A; Reggiani Bonetti, L; Borelli, F; Sighinolfi M., C; Bianchi, G; Ozben, T; Tomasi, A
abstract
Validation of prostate cancer biomarkers and inflammation by a proteomic approach
2013
- Detection of predictive urinary biomarkers of nephropathy in type 1 and type 2 diabetes by proteomic analysis
[Abstract in Rivista]
Bellei, Elisa; Cuoghi, Aurora; Monari, Emanuela; Bergamini, Stefania; Ligabue, Giulia; Cappelli, Gianni; Ozben, Tomis
abstract
Background: Nephropathy associated with diabetes is a severe
complication that cause slow kidneys deterioration, leading to
end-stage renal disease. Renal involvement during diabetes
mellitus may affect all the structural components of the kidneys,
causing functional and organic alterations frequently
associated with inflammatory processes, that give rise to
multiple clinical manifestations. Currently, despite rapid
research progress, predictors able to assess prospectively and
with high precision the risk to develop diabetic nephropathy
(DN) are still lacking.
Methods: The aim of this project was to identify differences in
urinary protein excretion, both in type 1 (T1D) and type 2
diabetic (T2D) patients, in comparison with healthy control
subjects. Ninety diabetic patients were recruited and divided in
3 groups (for each diabetes type), according to the level of
albuminuria: normoalbuminuric, with microalbuminuria (MA)
and with overt proteinuria. Second void morning urine samples
were collected and centrifuged to remove cell debris and
contaminations. Urinary proteins were separated by twodimensional
electrophoresis (2-DE) and identified by mass
spectrometry analysis (MS).
Results: Comparing the patients proteomic profiles with those
of normal subjects, firstly we noted a significant increase of
alpha-1-antitrypsin and albumin, also in the form of numerous
fragments, in urine of diabetic subjects. Particularly, statistical
analysis and spot quantification by PDQuest image software
revealed several proteins differentially expressed in diabetes
condition. Some proteins resulted increased in urine of both
T1D and T2D patients with MA, such as transthyretin,
apolipoprotein-A1 and transferrin, while the majority of the overexcreted
proteins were found in T2D patients with proteinuria,
e.g. vitamin-D-binding protein, protein AMBP, zinc-alpha-2-
glycoprotein, fetuin-A and ganglioside GM2 activator.
Conclusions: This protein pattern might represent a potential
tool for a better understanding of DN and could help to identify
patients at increased risk of renal disease progression.
Therefore, in diagnostic field, 2-DE and MS proteomic analysis
could be a suitable approach to discover early and predictive
biomarkers of DN in urine of diabetic patients.
2013
- Discovery by a proteomic approach of possible early biomarkers of drug-induced nephrotoxicity in medication-overuse headache
[Articolo su rivista]
Bellei, Elisa; Monari, Emanuela; Cuoghi, Aurora; Bergamini, Stefania; Guerzoni, Simona; Ciccarese, Michela; Ozben, T.; Tomasi, Aldo; Pini, Luigi Alberto
abstract
BACKGROUND:
Medication-overuse headache (MOH) is a chronic headache condition that results from the overuse of analgesics drugs, triptans, or other antimigraine compounds. The epidemiology of drug-induced disorders suggests that medication overuse could lead to nephrotoxicity, particularly in chronic patients. The aim of this work was to confirm and extend the results obtained from a previous study, in which we analyzed the urinary proteome of 3 MOH patients groups: non-steroidal anti-inflammatory drugs (NSAIDs), triptans and mixtures abusers, in comparison with non-abusers individuals (controls).
METHODS:
In the present work we employed specialized proteomic techniques, namely two-dimensional gel electrophoresis (2-DE) coupled with mass spectrometry (MS), and the innovative Surface-Enhanced Laser Desorption/Ionization Time-of-Flight mass spectrometry (SELDI-TOF-MS), to discover characteristic proteomic profiles associated with MOH condition.
RESULTS:
By 2-DE and MS analysis we identified 21 over-excreted proteins in MOH patients, particularly in NSAIDs abusers, and the majority of these proteins were involved in a variety of renal impairments, as resulted from a literature search. Urine protein profiles generated by SELDI-TOF-MS analysis showed different spectra among groups. Moreover, significantly higher number of total protein spots and protein peaks were detected in NSAIDs and mixtures abusers.
CONCLUSIONS:
These findings confirm the presence of alterations in proteins excretion in MOH patients. Analysis of urinary proteins by powerful proteomic technologies could lead to the discovery of early candidate biomarkers, that might allow to identify MOH patients prone to develop potential drug overuse-induced nephrotoxicity.
2013
- Non-bacterial protein expression in periodontal pockets by proteome analysis
[Articolo su rivista]
Bertoldi, Carlo; Bellei, Elisa; Chiara, Pellacani; Davide, Ferrari; Andrea, Lucchi; Aurora, Cuoghi; Bergamini, Stefania; Pierpaolo, Cortellini; Tomasi, Aldo; Zaffe, Davide; Monari, Emanuela
abstract
Objectives: To compare the proteomic profile of inter-proximal pocket tissues
with inter-proximal healthy tissues in the same subject to reveal proteins associated
with periodontal disease in sites where periodontopathogenic bacteria were
not detectable.
Methods: Twenty-five healthy patients, with moderate-to-advanced chronic periodontitis
and presenting with at least one intra-bony defect next to a healthy
inter-proximal site were enrolled. The periodontal defects were treated with osseous
resective surgery, and the flap design included both the periodontal pockets
and the neighbouring inter-proximal healthy sites. Pocket-associated and healthy
tissues were harvested for proteomic analyses.
Results: Fifteen proteins were differently expressed between pathological and
healthy tissues. In particular, annexin A2, actin cytoplasmic 1, carbonic anhydrase
1 & 2; Ig kappa chain C region (two spots) and flavinreductase were overexpressed,
whereas 14-3-3 protein sigma and zeta/delta, heat-shock protein beta -1
(two spots), triosephosphateisomerase, peroxiredoxin-1, fatty acid-binding
protein-epidermal, and galectin-7 were underexpressed in pathological tissue.
Conclusions: The unbalanced functional network of proteins involved could hinder
adequate tissue response to pathogenic noxa. The study of periodontal pocket
tissue proteomic profile would be crucial to better understand the pathogenesis of
and the therapeutic strategies for periodontitis.
2013
- Proteomic Analysis of PTCH1+/- Fibroblast Lysate and Conditioned Culture Media Isolated from the Skin of Healthy Subjects and Nevoid Basal Cell Carcinoma Syndrome Patients.
[Articolo su rivista]
Ponti, Giovanni; Bertazzoni, G; Pastorino, L; Monari, Emanuela; Cuoghi, Aurora; Bergamini, Stefania; Bellei, Elisa; Benassi, Luisa; Azzoni, Paola; Petrachi, Tiziana; Magnoni, Cristina; Pellacani, Giovanni; Loschi, P; Pollio, A; Witkowski, Am; Tomasi, Aldo
abstract
Background. The pathogenesis underlying the increased predisposition to the development of basal cell carcinomas (BCCs) in the context of Gorlin-Goltz syndrome is linked to molecular mechanisms that differ from sporadic BCCs. Patients with Gorlin syndrome tend to develop multiple BCCs at an early age and present with tumors of non-sun-exposed skin. The aim of this study was to compare the proteomic profile of cultured fibroblast and fibroblast conditioned culture media of PTCH1+ and nonmutated fibroblasts. Results. Proteomic analysis was performed using Surface-Enhanced Laser Desorption/Ionization Time-of-Flight mass spectrometry in PTCH1+ fibroblast conditioned media isolated from not affected sun-protected skin areas of Gorlin patients and from healthy subjects. 12 protein cluster peaks, >5 kDa, had significant differences in their peak intensities between PTCH1+ and PTCH1- subject groups. We detected a strongly MMP1 overexpression in PTCH1+ fibroblasts obtained from NBCCS patients with respect to healthy donors. Conclusion. Protein profiles in the fibroblast conditioned media revealed statistically significant differences between two different types (missense versus nonsense) of PTCH1 mutations. These differences could be useful as signatures to identify PTCH1 gene carriers at high risk for the development of NBCCS-associated malignancies and to develop novel experimental molecular tailored therapies based on these druggable targets.
2013
- Proteomic analysis of PTCH1+/- fibroblast lysate and conditioned culture media isolated from the skin of healthy subjects and Nevoid Basal Cell Carcinoma Syndrome (NBCCS) patients.
[Poster]
Pastorino, L.; Bertazzoni, G.; Monari, Emanuela; Cuoghi, A.; Bergamini, Stefania; Bellei, E.; Ruini, C.; Ghiorzo, P.; Bianchi Scarrà, G.; Pellacani, Giovanni; Loschi, P.; Pollio, A.; Tomasi, Aldo; Farnetani, F.; Ponti, Giovanni
abstract
PTCH1 mutations lead to complex syndromes such as the Gorlin Syndrome (GS) also named Nevoid basal Cell Carcinoma Syndrome (NBCCS, OMIM #109400) is a rare autosomal dominant disorder characterized by striking predisposition to the development of multiple basal cell carcinomas (BCCs), keratocystic odontogenic tumors (KOCTs) of the jaws, palmar and/or plantar pits and developmental defects. A variety of other benign or malignant tumors i.e., ovarian fibroma, medulloblastoma, rhabdomyosarcoma, cardiac fibroma and ameloblastoma can be found.
The mechanisms underlying the increased predisposition to the development of BCCs in the context of Gorlin-Goltz syndrome is linked to molecular pathways that differ from sporadic cases. Patients with Gorlin syndrome tend to develop multiple BCCs at an early age: moreover, the tumors typically arise on non-sunexposed skin. Fibroblasts of patients with Gorlin Syndrome may display properties determining BCC development. The aim of this study was to compare the proteomic profile of cultured fibroblast and fibroblast conditioned culture media of PTCH1+ and non-mutated fibroblasts. Proteomic analyses was performed using Surface-Enhanced Laser Desorption/Ionization Time-of-Flight mass spectrometry in PTCH1+ fibroblast conditioned media isolated from not affected sun-protected skin areas of Gorlin patients and from healthy subjects. The protein profiles were obtained using Copper pre-activated IMAC30 ProteinChip array.
12 protein cluster peaks, >5 kDa, had statistically significant differences in their peak intensities between PTCH1+ and PTCH1- subject groups (p<0.05). Protein profiles in the fibroblast conditioned media revealed statistically significant differences between two different types (missense vs nonsense) of PTCH1 mutations. These differences could be useful as signatures to identify PTCH1 gene carriers at high risk for the development of NBCCS-associated malignancies, and to develop novel experimental molecular tailored therapies based on these druggable targets.
These protein peaks profiles provided better understanding of the complex skin cancer microenvironment and could be useful to select patients at risk to develop multiple and aggressive BCCs and/or other NBCCS-associated malignancies.
2013
- Proteomic profile of retained proteins from hfr cartridge
[Abstract in Rivista]
Caiazzo, Marialuisa; Cuoghi, Aurora; Monari, Emanuela; Bellei, Elisa; Bergamini, Stefania; Palladino, Giuseppe; Atti, Mauro; Bruni, Francesco; Tomasi, Aldo
abstract
Hemodiafiltration with on-line endogenous
reinfusion (HFR) is a dialytic method, which combines the
processes of diffusion, convection and adsorption. The
performance of this system is linked to the optimal combination
of the membrane permeability and cartridge resin bed. Lupus
nephritis (LN) remains one of the most severe manifestations
of systemic lupus erythematous (LES), associated with
considerable morbidity and mortality. In this preliminary study,
ESI-QTOF Mass Spectrometer was used for identification of
protein ultrafiltrate (UF) and for the protein captured by resin
bed, obtained from one dialysed patient with LN.
Methods: Plasma and UF (pre and post cartdrige) samples of
one patient with LN treated with HFR, were collected at 15 min
and at 235 min of the dialytic session. The cartridge utilized
during treatment, containing styrenic resin, was then opened
and the proteins kept by the resin were eluted by incubation
O/N with 60% ACN and 1%TFA. Samples were desalted and
separated by SDS-page, interesting bands were picked and
“in-gel” tryptic digested before ESI-QTOF mass spectrometer
analysis.
Results: ESI-QTOF results of the retained proteins allowed to
identifies several biomarker of kidney injury in LN, such as
Retinol binding protein 4, Neutrophil gelatinase-associated
lipocain, and Cystatin-C (and also TRFE, A1AG1, PTGDS,
TTHY). Moreover we identify several fragments of
Immunoglobulin, which are implicated in the etiopathogenesis
of LES.
Conclusion: The results of this preliminary study demonstrate
that styrenic resin retain several proteins implicated in the LN
pathogenesis, in fact the corresponding bands in the UF precartdrige
at 240 min disappear confirming the removal of this
proteins from the cartridge.
2013
- Superhighflux therapies for hemodialysis: ultrafiltrate proteomic profile and protein identification by on-chip elution.
[Abstract in Rivista]
Monari, Emanuela; Cuoghi, Aurora; Caiazzo, Marialuisa; Bellei, Elisa; Bergamini, Stefania; Palladino, Giuseppe; Ozben, Tomris; Tomasi, Aldo
abstract
between serum levels of symmetric dimethylarginine (SDMA)
and N-terminal natriuretic propeptide B-type (NT-proBNP ) and
the degree of estimated glomerular filtration rate (eGFR).
Methods: This study involved 98 chronic kidney disease (CKD)
patients and 44 renal transplant (RT) recipients. Measurement
of SDMA was determined using ELISA (DLD Diagnostica
GMBH) and NT-proBNP was determined with ELFA
(bioMerieux). GFR was calculated according to the Modified
Diet in Renal Disease (MDRD) equation.
Results: To evaluate the changes of examined parameters
according to renal function we stratified CKD and RT patients
to GFR categories. All patients have moderate (GFR 30–59
ml/min/1.73 m2) to severe (GFR <30 mL/min/1.73 m2) impaired
renal function. Both NT-proBNP and SDMA were significantly
higher in CKD and RT patients with severe than moderate
impaired renal function. In CKD group, NT-proBNP values rose
from 88,09 ng/L in patients with moderate eGFR to 988,78 pg/L
in patients with severe eGFR (P <0.001) and from 197,05 ng/L
to 996,87 ng/L in RT group (P=0.003). Obtained values for
SDMA between moderate and severe eGFR were 1,09 μmol/L
vs 1,62 μmol/L in CKD group and 0,58 μmol/L vs 1,18 μmol/L
in RT group (P <0.001).
Conclusion: NT-proBNP and SDMA are usefull for detection of
severity of renal disfunction in CKD patients and renal
transplant recipients.
2013
- The influence of inflammation in the search of discriminatory biomarkers for prostate cancer: a proteomic study
[Abstract in Rivista]
Bergamini, Stefania; REGGIANI BONETTI, Luca; Monari, Emanuela; Bellei, Elisa; Cuoghi, Aurora; Majorana, Antonino; Ozben, Tomis; Micali, Salvatore; Sighinolfi, Maria Chiara; Tomasi, Aldo; Bianchi, Giampaolo
abstract
background: Despite the improvements in clinical and surgical practice, prostate cancer (PCa) remains one of the most
widespread cancer in male. The serum marker currently used
for the diagnosis of PCa is the prostate-specific antigen (PSA),
but its increase does not discriminate benign prostatic
hyperplasia (BPH) from PCa. In our study, we investigated the
serum protein expression of BPH compared to PCa, in order to
identify by Surface Enhanced Laser Desorption/Ionization -
Time of Flight - Mass Spectrometry (SELDI-ToF-MS) analysis
distinctive protein profiles able to unquestionably discriminate
patients with a benign prostate condition from those with a
malignant situation. Moreover, we considered these conditions
focusing on the co-existence of inflammation.
Methods: Patients with clinical suspect of PCa (PSA elevation
and/or palpable mass at digital rectal exploration) and
candidates for trans-rectal ultrasound guided prostate biopsy
were enrolled. The analysis of protein profile of 30 patients with
PCa and 30 subjects with BPH was carried out. All histological
specimens were examined in order to graduate and classify
the tumor and to recognize the BPH condition and presence
of inflammation, that was classed in chronic and acute and then
graduated in mild, moderate and severe. Serum was depleted
of the 6 high-abundance proteins by immunoaffinity
chromatography prior to SELDI-ToF-MS analysis.
Results: The comparison between protein spectra from PCa
and BPH considering the inflammation parameter and
excluding samples with moderate and/or severe inflammation,
identified 17 differentially expressed protein peaks using H50
ProteinChip Array.The analysis of protein profile in presence of
inflammation showed different protein peaks in the two groups,
some of which overlapped with those found also in the
comparison between PCa and BPH in absence of
inflammation.
Conclusions: The inflammation seems to lead a crucial
contribution in the protein profile assessments of these
conditions. On the basis of our results, we believe that certain
different protein peaks could be reasonably associated to
inflammation rather than to cancer. Therefore, inflammation
might be a confounding parameter in the search of specific
biomarkers to discriminate PCa from BPH.
2012
- Evaluation of a possible relationship between medication-overuse headache and potential renal dysfunctions by a proteomic study on urine samples
[Abstract in Rivista]
Bellei, Elisa; Monari, Emanuela; Bergamini, Stefania; Cuoghi, Aurora; Tomasi, Aldo; Guerzoni, Simona; A., Bazzocchi; Pini, Luigi Alberto
abstract
This proteomic study confirms the previous finding of alterations
in urinary proteins excreted in MOH patients. Some of these proteins, identified as over-expressed particularly in NSAIDs abusers, were related to different renal dysfunctions and, probably in the development of CA. Proteomic analysis of urine proteins by the combination of 2-DE and MS could improve the knowledge of the pathophysiology of the MOH condition and identify early biomarkers to prevent the potential drug overuse-induced nephrotoxicity
2012
- Proteomic analysis of urine in medication-overuse headache patients: possible relation with renal damages
[Articolo su rivista]
Bellei, Elisa; Cuoghi, Aurora; Monari, Emanuela; Bergamini, Stefania; Fantoni, Luca Isaia; Zappaterra, Maurizio; Guerzoni, Simona; Bazzocchi, Annalisa; Tomasi, Aldo; Pini, Luigi Alberto
abstract
Medication-overuse headache (MOH) is a chronic disorder associated with overuse of analgesic drugs, triptans, non-steroidal anti-inflammatory drugs (NSAIDs) or other acute headache compounds. Various epidemiologic investigations proved that different drug types could cause nephrotoxicity, particularly in chronic patients. The aim of the present work was to analyze, by aproteomic approach, the urinary protein profiles of MOH patients focusing on daily use of NSAIDs, mixtures and triptans that could reasonably be related to potential renal damage. We selected 43 MOH patients overusing triptans (n = 18), NSAIDs (n = 11), and mixtures (n = 14), for 2–30 years with a mean daily analgesic intake of 1.5 ± 0.9 doses, and a control group composed of 16 healthy volunteers. Urine proteins were analyzed by monodimensional gel electrophoresis and identified by mass spectrometry analysis. Comparing the proteomic profiles of patients and controls, we found a significantly different protein expression, especially in the NSAIDs group, in which seven proteins resulted over-secreted from kidney (OR = 49, 95% CI 2.53–948.67 vs. controls; OR = 11.6, 95% CI 0.92–147.57 vs. triptans and mixtures groups). Six of these proteins (uromodulin, a-1-microglobulin, zinc-a-2- glycoprotein, cystatin C, Ig-kappa-chain, and inter-a-trypsin heavy chain H4) were strongly correlated with various forms of kidney disorders. Otherwise, in mixtures and in triptans abusers, only three proteins were potentially associated to pathological conditions (OR = 4.2, 95% CI 0.33–53.12, vs. controls). In conclusion, this preliminary proteomic study allowed us to define the urinary protein pattern of MOH patients that is related to the abused drug. According with the obtained results, we believe that the risk of nephrotoxicity should be considered particularly in MOH patients who abuse of NSAIDs.
2012
- Proteomic profile of non-bacterial proteins in periodontal pockets: a pilot study.
[Abstract in Rivista]
Simonazzi, L.; Monari, Emanuela; Ferrari, Davide; Bellei, Elisa; Melpignano, F.; Cuoghi, Aurora; Bergamini, Stefania; Pellacani, C.; Campedelli, F.; Tomasi, Aldo; Bertoldi, Carlo
abstract
Aim: In this study, we attempted to identify the proteins
involved in periodontitis comparing the proteomic profile of
interproximal pocket tissue affected by the periodontal lesion
with interproximal healthy tissue in the same subject, in sites
where no periodontopathogenic bacteria were detectable.
Material and Methods: Using specific inclusion and exclusion
criteria, we enrolled 15 subjects affected by severe chronic
periodontitis. The subjects presenting at least one intrabony
defect (next to a healthy interproximal site to be included in the
flap design) suitable for treatment by osseous respective surgery,
were considered eligible for this study. Biopsies of connective
tissue were harvested from the intrabony component of the
defect and from healthy tissue (from the secondary flap) and
immediately frozen at –80°C. A two-dimensional gel electrophoresis
system was used to detect differences in protein expression
between pathologic and healthy tissue and protein identification
by LC-MS/MS analysis was performed.
Results: Six proteins (tropomyosin α-4 chain, 14-3-3 protein
z/δ, putative heat shock protein HSP 90-β, peroxiredoxin-1, fatty
acid binding protein, S100-A9) displayed a higher while five a
lower (annexin A1, actin cytoplasmatic-1, carbonic anhydrase-2,
Ig K chain C region and flavin reductase) protein expression level
in pathologic tissue compared with healthy tissue.
Conclusion: The highlight of proteins involved in the
immunological and cytokine signaling cascade mediation or
in connective and epithelial regeneration and differentiation
shows that the pathogenic process is active in periodontal sites
also in absence of periodontopathogen microbiota. Besides,
proteomic analysis could be considered very useful in studying
the pathogenesis of periodontitis.
Abstract N: P0224
ISSN:1600-051X (Electronic).
2012
- Quantification of P-cresol sulphate in human plasma of uremic patients by MRM
[Abstract in Rivista]
Cuoghi, Aurora; Bellei, Elisa; Caiazzo, Marialuisa; Bergamini, Stefania; G., Palladino; Monari, Emanuela; Tomasi, Aldo
abstract
n/a
2012
- Quantification of p-cresol sulphate in human plasma by selected reaction monitoring.
[Articolo su rivista]
Cuoghi, Aurora; M., Caiazzo; Bellei, Elisa; Monari, Emanuela; Bergamini, Stefania; G., Palladino; T., Ozben; Tomasi, Aldo
abstract
Chronic renal failure patients accumulate in the blood molecules that are normally excreted into the urine. p-Cresol Sulphate (pCS), the most representative retained toxin, shows a high level of toxicity. Therefore, its quantification could represent a prediction factor to determine the risk of endothelial dysfunction and cardiovascular complication and response to the haemodialysis treatment. The aim of this study was to evaluate the suitability of the multiple reaction monitoring (MRM) technique in order to improve the sensibility, the selectivity and the timing of pCS detection in a small amount of plasma. Deproteinized plasma of uremic patients was concentrated and dissolved in liquid chromatography (LC) mobile phase solution. pCS was quantified by LC coupled to tandem mass spectrometry (LC-MS/MS) on a triple-quadrupole mass spectrometer. Selective and sensitive detection of pCS was achieved by selecting the specific parent ion and monitoring two specific fragment ions. The MRM assay was carried out using the following transitions: m/z 187 → 80.00 and m/z 187 → 107.00. A good linearity was observed for each calibration curve. The intra-day and inter-day results showed a good precision and repeatability. The percentage recoveries indicate an optimal selectivity of the analytical method. The MRM assay to quantify pCS in a small amount of human plasma is rapid, highly sensitive, selective and with a good repeatability.
2012
- Quantificazione del p-cresol solfato nel plasma di pazienti uremici mediante MNR.
[Abstract in Atti di Convegno]
Cuoghi, Aurora; Bellei, Elisa; Caiazzo, Marialuisa; Bergamini, Stefania; G., Palladino; Tomasi, Aldo; Monari, Emanuela
abstract
n/a
2012
- Steps Study: Superhighflux therapies for hemodialysis. A proteomic approach.
[Abstract in Rivista]
Caiazzo, Marialuisa; Monari, Emanuela; Cuoghi, Aurora; Bellei, Elisa; Bergamini, Stefania; G., Palladino; Tomasi, Aldo
abstract
n/a
2012
- Studio STEPS: Superior Therapies for hemodialysis. Approccio proteomico.
[Abstract in Atti di Convegno]
Caiazzo, Marialuisa; Cuoghi, Aurora; Monari, Emanuela; Bellei, Elisa; Bergamini, Stefania; G., Palladino; Tomasi, Aldo
abstract
n/a
2011
- Can prostatitis to be a confounding parameter in prostatic proteomic profile designation?
[Abstract in Rivista]
Bergamini, Stefania; REGGIANI BONETTI, Luca; Monari, Emanuela; Bellei, Elisa; Maiorana, Antonino; Ozben, T.; Tomasi, Aldo; Micali, Salvatore; Bianchi, Giampaolo
abstract
N/A
2011
- Detection of serum protein profile changes in atherosclerosis
[Abstract in Rivista]
Dursun, Evrim; Ozben, Beste; Monari, Emanuela; Cuoghi, Aurora; Bergamini, Stefania; Tomasi, Aldo; Ozben, Tomris
abstract
nd
2011
- Effect of nebivolol treatment on proteomic profiling during atherosclerosis progression.
[Abstract in Atti di Convegno]
Ozben, T.; Ozben, B.; Dursun, E.; Monari, Emanuela; Cuoghi, Aurora; Bellei, Elisa; Bergamini, Stefania; Tomasi, Aldo
abstract
na
2011
- Enriched sera protein profiling for detection of non-small cell lung cancer biomarkers.
[Articolo su rivista]
Monari, Emanuela; Casali, Christian; Cuoghi, Aurora; Nesci, Jessica; Bellei, Elisa; Bergamini, Stefania; Fantoni, Luca Isaia; Natali, Pamela; Morandi, Uliano; Tomasi, Aldo
abstract
BackgroundNon Small Cell Lung Cancer (NSCLC) is the major cause of cancer related-death. Many patients receive diagnosis at advanced stage leading to a poor prognosis. At present, no satisfactory screening tests are available in clinical practice and the discovery and validation of new biomarkers is mandatory. Surface Enhanced Laser Desorption/Ionization Time-of-Flight Mass Spectrometry (SELDI-ToF-MS) is a recent high-throughput technique used to detect new tumour markers. In this study we performed SELDI-ToF-MS analysis on serum samples treated with the ProteoMinerTM kit, a combinatorial library of hexapeptide ligands coupled to beads, to reduce the wide dynamic range of protein concentration in the sample. Serum from 44 NSCLC patients and 19 healthy controls were analyzed with IMAC30-Cu and H50 ProteinChip Arrays.ResultsComparing SELDI-ToF-MS protein profiles of NSCLC patients and healthy controls, 28 protein peaks were found significantly different (p<0.05), and were used as predictors to build decision classification trees. This statistical analysis selected 10 protein peaks in the low-mass range (2-24 kDa) and 6 in the high-mass range (40-80 kDa). The classification models for the low-mass range had a sensitivity and specificity of 70.45% (31/44) and 68.42% (13/19) for IMAC30-Cu, and 72.73% (32/44) and 73.68% (14/19) for H50 ProteinChip Arrays.ConclusionsThese preliminary results suggest that SELDI-ToF-MS protein profiling of serum samples pretreated with ProteoMinerTM can improve the discovery of protein peaks differentially expressed from NSCLC patients and healthy subjects, useful to build classification algorithms with high sensitivity and specificity. However, identification of the significantly different protein peaks needs further study in order to provide a better understanding of the biological nature of these potential biomarkers and their role in the underlying disease process.
2011
- High-abundance proteins depletion for serum proteomic analysis: concomitant removal of non-targeted proteins.
[Articolo su rivista]
Bellei, Elisa; Bergamini, Stefania; Monari, Emanuela; Fantoni, Luca Isaia; Cuoghi, Aurora; Ozben, T; Tomasi, Aldo
abstract
In clinical and pharmaceutical proteomics, serum and plasma are frequently used for detection of early diagnostic biomarkers for therapeutic targets. Although obtaining these body fluid samples is non-invasive and easy, they contain some abundant proteins that mask other protein components present at low concentrations. The challenge in identifying serum biomarkers is to remove the abundant proteins, uncovering and enriching at the same time the low-abundance ones. The depletion strategies, however, could lead to the concomitant removal of some non-targeted proteins that may be of potential interest. In this study, we compared three different methods aimed to deplete high-abundance proteins from human serum, focusing on the identification of non-specifically bound proteins which might be eventually removed. A Cibacron blue-dye-based method for albumin removal, an albumin and IgG immunodepletion method and an immunoaffinity column (Multiple Affinity Removal System) that simultaneously removes a total of six high-abundance proteins, were investigated. The bound proteins were eluted, separated by two-dimensional gel electrophoresis and identified by Nano LC-CHIP-MS system. Flow-through fractions and bound fractions were also analysed with the ProteinChip technology SELDI-TOF-MS. Our results showed that the methods tested removed not only the targeted proteins with high efficiency, but also some non-targeted proteins. We found that the Multiple Affinity Removal Column improved the intensity of low-abundance proteins, displayed new protein spots and increased resolution. Notably, the column showed the lowest removal of untargeted proteins, proved to be the most promising depletion approach and a reliable method for serum preparation prior to proteomic studies.
2011
- Optimizing protein recovery yield from serum samples treated with beads technology.
[Articolo su rivista]
Bellei, Elisa; Monari, Emanuela; Bergamini, Stefania; Ozben, T.; Tomasi, Aldo
abstract
Proteomics studies are often complicated by the wide dynamic range of the biologicalfluids, in which few highly abundant proteins obscure the signal of low abundant ones.To overcome this problem, several techniques have been developed on the basis of‘‘depletion principles,’’ namely immuno-subtraction with specific antibodies against themost-abundant proteins. Unfortunately, the probability of codepletion is a noteworthydrawback associated with these strategies. The ProteoMinerTM (PM) technology is anovel approach, consisting of a combinatorial library of hexapeptide ligands coupled tobeads, that allows the capture of all species present in a proteome, but at much reducedprotein concentration differences, simultaneously enhancing the concentration of themost dilute species. In this study, we evaluated the compatibility of the PM kit’s elutionreagent with 2-DE analysis, comparing five different purification methods on serumsamples eluted from the beads: the ‘‘ReadyPrep 2-D Clean-up kit’’ and precipitation withorganic solvents, as acetone/methanol, TCA/acetone, ACN, and chloroform/methanol.Considering protein recovery yield (quantity) and 2-DE spot pattern (quality), precipitationwith ACN offered the most promising approach, showing the best spot resolution inall regions of the pH gradient and the greatest number of protein spots visualized on 2-Dgels.
2011
- Proteomic identification of periodontal pocket involved proteins: a pilot study.
[Abstract in Rivista]
Bertoldi, Carlo; Bellei, Elisa; Cuoghi, Aurora; Pellacani, C.; Tomasi, Aldo; Monari, Emanuela
abstract
Vedi allegato
2011
- Urine proteomic analysis in patients with cronic headache and overuse of analgesic drugs: possible relation with renal damage
[Abstract in Rivista]
Bellei, Elisa; Bergamini, Stefania; Cuoghi, Aurora; Monari, Emanuela; Tomasi, Aldo; Zappaterra, Maurizio; Bazzocchi, Annalisa; Guerzoni, Simona; Pini, Luigi Alberto
abstract
NA
2010
- POTENTIAL ROLE OF PROTEOMIC ANALYSIS IN PROSTATE CANCER DIAGNOSIS
[Abstract in Rivista]
Bianchi, Giampaolo; Sighinolfi, Maria Chiara; Bergamini, Stefania; Bellei, Elisa; Monari, Emanuela; Cuoghi, Aurora; Micali, Salvatore; DE STEFANI, Stefano; Tomasi, Aldo
abstract
not available
2010
- Proteomic profiling during atherosclerosis progression using SELDI-TOF-MS: Effect of darbepoetin treatment
[Articolo su rivista]
Dursun, E; Monari, Emanuela; Cuoghi, Aurora; Bergamini, Stefania; Ozben, B; Suleymanlar, G; Tomasi, Aldo; Ozben, T.
abstract
Narrowing of the arteries due to atherosclerosis may lead to congestive heart failure (CHF). It is advantageous to perform atherosclerosis studies in apolipoprotein E-deficient (Apo E(-/-)) mice models, which develop atherosclerosis very rapidly in comparison to humans. Darbepoetin is a synthetic erythropoietin analogue and stimulates erythropoiesis. The aim of this study was to explore the effect of 16 weeks of darbepoetin treatment on serum protein profiles in Apo E(-/-) mice during atherosclerosis progression. Serum proteomic analyses were performed using surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF-MS) in the darbepoetin-treated and non-treated (control) Apo E(-/-) mice groups. The protein profiles obtained using three different chips, CM-10 (weak cation exchange), H50 (reversed-phase) and IMAC-30 (immobilized metal affinity capture), were statistically analyzed using the ProteinChip data manager 3.0 program. At the end of 16 weeks of darbepoetin treatment, there was no significant difference in the size and degree of atherosclerotic lesions between the darbepoetin and control mice groups. In contrast, 145 protein/peptide-clustering peaks, >5kDa, had statistically significant differences in their peak intensities between the darbepoetin and control mice groups (p<0.05). That the proteomic profiles of darbepoetin-treated Apo E(-/-) mice were found to differ from those of the control group indicates a potential beneficial role of darbepoetin in atherosclerosis. Our study contributes to understanding the effects of darbepoetin on protein/peptide expressions during atherosclerosis development.
2010
- Proteomic profiling in apolipoprotein E-deficient mice during atherosclerosis progression
[Articolo su rivista]
E., Dursun; B., Ozben; Monari, Emanuela; Cuoghi, Aurora; Tomasi, Aldo; T., Ozben
abstract
Atherosclerosis and related complications are a major worldwide cause of human morbidity and mortality. It is advantageous to perform atherosclerosis studies in the apolipoprotein E-deficient (Apo E−/−) mouse model, which develops atherosclerosis very fast in comparison to humans. The aim of this study was to compare serum protein profiles in Apo E−/− mice during atherosclerosis progression with the data of control C57BL/6 mice. Serum proteomic analyses were performed using surface-enhanced laser desorption/ionization-time of flight mass spectrometry (SELDI-TOF-MS). The protein profiles obtained using three different chips, CM-10 (weak cation exchange), H50 (reversed-phase) and IMAC-30 (immobilized metal affinity capture) were statistically analyzed using the ProteinChip data manager 3.0 program. At 20 weeks of age, all Apo E−/− mice developed early atherosclerotic lesions. The peak intensities of 742 serum protein/peptide clusters were found to be different between the atherosclerotic and control mice groups, and the differences reached statistical significance for 107 serum protein/peptide clusters (p<0.05). This study contributes to understanding the changes in serum protein/peptide profiles during atherosclerosis development and may inform discovery of new protein biomarkers for early diagnosis of atherosclerosis.
2009
- Analisi proteo mica nella neoplasia prostatica: Risultati preliminari e nuove prospettive.
[Poster]
Bianchi, Giampaolo; Sighinolfi, M. C.; Micali, Salvatore; DE STEFANI, Stefano; Bergamini, Stefania; Bellei, E.; Monari, Emanuela; Cuoghi, A.; Tomasi, Aldo
abstract
Analisi proteo mica nella neoplasia prostatica: Risultati preliminari e nuove prospettive.
2009
- Biomarkers discovery of renal alterations in urine of diabetic patients by proteomic analysis
[Abstract in Rivista]
Tomasi, A; Bellei, E; Bergamini, S; Monari, E; Cuoghi, A; Albertazzi, A; Lucchi, L
abstract
Discovery of potential biomarkers correlated to renal alterations in urine of diabetic patients by proteomic analysis.
2009
- Free radical induced oxidative protein modification in kidney diseases: proteomic detection
[Abstract in Atti di Convegno]
Bergamini, S; Bellei, E; Monari, E; Cuoghi, A; Fantoni, Li; Lucchi, L; Tomasi, A
abstract
Proteomic detection of free radical induced oxidative protein modification in kidney diseases
2009
- PROTEOMICS AS DIAGNOSTIC MARKERS
[Abstract in Rivista]
Tomasi, Aldo; Bellei, Elisa; Bergamini, Stefania; Cuoghi, Aurora; Monari, Emanuela
abstract
non disponibile
2009
- Proteomic analysis to discover early urinary biomarkers of type 2 diabetic nephropathy
[Poster]
Bellei, E; Cuoghi, A; Monari, E; Lucchi, L; Bergamini, S; Tomasi, A
abstract
Objectives. Nephropathy associated with type 2 diabetes mellitus (T2D) is the most frequent cause of end stage renal disease (ESRD) in all Western countries, and a major healthcare problem. In the United States, the incidence of diabetic nephropathy (DN) has increased by 150% in the past 10 years: a similar trend has been reported also in Europe.
Despite rapid research progress, predictors able to assess prospectively with high precision the risk for DN in individuals with diabetes are still lacking. Thus, at present the available therapies are usually initiated at more advanced stages of DN, characterized by evident clinical manifestations and progressive decline in glomerular filtration rate (GFR), leading toward both ESRD and cardiovascular events.
In this study we made a comparison between the proteomic pattern of a group of normoalbuminuric T2D patients with those of a group of T2D patients with DN (T2DN). Both were compared with a group of healthy subjects, in the search of a diagnostic proteomic pattern that could help to identify patients at increased risk of kidney disease progression.
Methods. We used two different proteomic approaches: the Surface-Enhanced Laser Desorption Ionization-Time of Flight technique (SELDI-TOF) and the two-dimensional gel electrophoresis (2-DE) coupled to mass spectrometry (MS).
We performed urinary proteomic analysis of 10 T2D patients with microalbuminuria in the normal range, 12 T2DN patients, and 12 healthy subjects as normal controls.
For protein profiling with SELDI-TOF, urine samples were analyzed using immobilized metal affinity capture (IMAC30), weak cation exchanger (CM10) and reverse phase (H50) ProteinChip Arrays. Statistical analysis was performed in the obtained spectra (Ciphergen Express 3.0) in order to identify peaks that showed significant differences among groups.
Furthermore, urinary proteins were separated and visualized by 2-DE; the differentials spot were selected and identified by MS, using an ESI-Q-TOF mass spectrometer.
Results. Comparing the patients proteomic profiles with those of normal controls, we identified 11 progressively and differently expressed proteins. The majority of these proteins resulted significantly increased in both patients groups, and corresponded to: transthyretin precursor, plasma retinol-binding protein, -2-microglobulin, carbonic anhydrase, -2-glycoprotein 1 (or apolipoprotein H), Ig kappa chain C region, Ig kappa chain V-III region SIE and Ig kappa chain V-II region Cum (or Bence-Jones protein). Only 3 proteins were gradually decreased in patients groups, including the prostatic acid phosphatase precursor, the ribonuclease and the kallikrein-3.
Conclusions. The proteomic analysis allowed us to identify several increased urinary proteins, not only in T2DN, but also in T2D patients in which the microalbuminuria was in the normal range. These patterns of urinary proteins might represent a potential tool for a better understanding of diabetic renal damage and could help to identify the diabetic patients prone to progress toward DN.
2009
- Proteomic profiling during atherosclerosis progression: Effect of nebivolol treatment
[Articolo su rivista]
Ozben, B; Dursun, E; Monari, Emanuela; Cuoghi, Aurora; Bergamini, Stefania; Tomasi, Aldo; Ozben, T.
abstract
There is a great need for the identification of biomarkers for the early diagnosis of atherosclerosis and the agents to prevent its progression. The aim of this study was to explore the effect of 24 week of nebivolol (a third-generation vasodilatory beta-blocker) treatment on serum protein profiles in Apo E(-/-) mice during atherosclerosis progression. Nebivolol treated and non-treated (the control group) groups consisted of 10 genetically modifiedhomozygous Apo E(-/-) mice. Proteomic analyses were performed using surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF MS) in the serum samples from the nebivolol treated and non-treated Apo E(-/-) mice. The protein profiles obtained using three different chips, CM10 (weak cation-exchange), H50 (reverse phase), and IMAC30-Cu(2+) (immobilized metal affinity capture) were statistically analyzed using the ProteinChip data manager 3.0 program. At the end of 24 week of nebivolol-treatment period, a total of 662 protein/peptide clustering peaks were detected using 12 different conditions and reading with high and low intensity laser energy. The highest total number of protein/peptide clusters was found on H50 chip array. The peak intensities of 95 of the 662 protein/peptide clusters were significantly different in the nebivolol-treated atherosclerotic group in comparison to the non-treated control mice groups (P < 0.05). Forty-three protein/peptides were up-regulated (high signal intensity) while 52 protein/peptides had lower signal intensity (down-regulated) in the nebivolol-treated atherosclerotic group. The proteomic profiles of nebivolol-treated Apo E(-/-) mice were different than the control group indicating a potential role of nebivolol in atherosclerosis. Our study contributes to understand the efficacy of nebivolol on serum protein/peptide profiles during atherosclerosis development.
2009
- Proteomics to identify free radical induced oxidative protein modifications as biomarkers associated to type 2 diabetic nephropathy
[Abstract in Atti di Convegno]
Bellei, E; Bergamini, S; Lucchi, L; Monari, E; Cuoghi, A; Fantoni, Li; Tomasi, A
abstract
Proteomics as a tool to identify free radical induced oxidative protein modifications as candidate biomarkers associated to type 2 diabetic nephropathy
2007
- Detection of type 2 diabetic nephropathy-associated biomarkers using proteomic tachniques
[Abstract in Rivista]
Tomasi, Aldo; Bellei, Elisa; Albertazzi, Alberto; Monari, Emanuela; Lucchi, L.
abstract
not available
2007
- Urinary protein profiling for biomarker discovery in diabetic nephropathy
[Abstract in Rivista]
Tomasi, Aldo; Bellei, Elisa; Monari, Emanuela; Albertazzi, Alberto; Lucchi, L.
abstract
Renal disease in patients with Type II diabetes is the leading cause of terminal renal failure and a major healthcare problem. Diabetic renal damage might be reflected by a change in urinary proteins and polypeptides excretion at a very early stage. The characterization of urinary polypeptides is of great clinical interest and significance, yielding to a better understanding of the changes within the kidney during the development of diabetic nephropathy (DN).Proteomics has now become an emerging field in the post-genomic area and offers the opportunity of large-scale protein analysis. Two-dimensional polyacrylamide gel electrophoresis (2DE-PAGE) combined with mass spectrometry (MS) is commonly used for protein separation and to determine the molecular weight of polypeptides and proteins. Surface-Enhanced Laser Desorption Ionization-Time of Flight-Mass Spectrometry (SELDI-TOF-MS), which represents the succesful combination of retentate chromatography and MS, is an alternative technology employed for high-throughput analysis of biological samples.The aim of this study was to identify urinary protein patterns in Type II diabetic subjects, that may serve as diagnostic tool for the early recognition and development of DN.In our study, we included 25 Type II diabetes mellitus patients with no nephropathy, 25 Type II diabetes mellitus patients with nephropathy (identified by serum creatinine level < 2mg/dl and microalbuminuria) and 20 healthy volunteers. Morning midstream urine samples were collected, centrifuged to remove cell debris and precipitated with acetone/methanol at -20°C. After centrifugation, the protein pellet was resuspended in a specific solubilization buffer and finally 100 micrograms of protein were subjected to 2DE-PAGE. Silver-stained gels were analyzed by the PDQuest® Image Analysis software, differential proteins were cut and identified by LC-ESI-MS/MS.For protein profiling with SELDI-TOF-MS, urine samples were analyzed using immobilized metal affinity capture (IMAC30), strong anionic exchanger (Q10), weak cation exchanger (CM10), reverse phase (H50) ProteinChip® Arrays and sinapinic acid (SPA) as matrix, according to the manufacturer’s instructions (Ciphergen, USA). Urinary protein profiles were collected in the range 0-200.000 m/z, using different acquisition protocols and different laser energies. Statistical analysis was performed in the obtained spectra (Ciphergen Express 3.0) in order to identify peaks that showed significant differences among groups.Two characteristics proteins (prostaglandin-H2 D-isomerase and antithrombin-III) were identified in the urine of Type II diabetes patients compared to healthy individuals using 2DE-PAGE coupled to MS. In addition to these two proteins, the following proteins were identified in urine of patients with Type II diabetes mellitus with nephropathy: vitamin-D binding protein, gelsolin, apolipoprotein A-IV, plasma retinol binding protein and lipocalin-1, with an increased amount of albumin.In conclusion, comparative data analysis discovered distinct proteins characteristic for diabetic patients and leads to the establishment of possible individual risk factors for the development of DN, which may contribute to clinical staging in the future. These techniques may also enable the identification of novel proteins related to the pathophysiology of DN, which may serve as therapeutical targets
2006
- Bone Augumentation With Bioactive Glass In Three Cases of Dental implant Placement
[Articolo su rivista]
Gatti, Antonietta; Simonetti, L. A.; Monari, Emanuela; Guidi, S.; Greenspan, D.
abstract
This study reports the clinical use of a bioactive bone graft material, PerioGlas, in the treatment of dental extraction sites before dental implant placement, to effect bone regeneration and to give early fixation to the implant.PerioGlas, granules, ranging from 90 to 710 mm, are implanted after tooth extraction in three patients; after 6 months bone biopsies were performed in the site of the glass implantation and observed under Electron Scanning Microscopy.All the granules showed a biodegradation involving precipitation of calcium phosphate that worked as a scaffold for osteoblasts colonization. All cases examined showed the bioactivity of PerioGlas granules resulting in new bone formation and biodegradation of the glass. After a two-year clinical follow-up all the implants were successfully loaded and appeared stable.
2006
- Ricerca di biomarkers nel tumore ovarico mediante analisi proteomica
[Abstract in Atti di Convegno]
Rossi, E; Cortesi, L; Barchetti, A; Bellei, E; Monari, E; Tomasi, A; Iannone, A; Abrate, M; Federico, M.
abstract
Il carcinoma ovarico rappresenta la quarta causa di morte nel sesso femminile in Europa e negli Stati Uniti. Nell’80 % dei casi infatti, la diagnosi avviene in uno stadio avanzato poiché la maggior parte delle neoplasie ovariche rimane clinicamente asintomatica negli stadi I e II. La sola terapia chirurgica assicura una sopravvivenza a cinque anni del 90 % delle pazienti diagnosticate allo stadio I, percentuale che si riduce al 35% negli stadi più avanzati. Il nostro obbiettivo è quello di utilizzare l’elettroforesi bidimensionale (2DE) e la spettrometria di massa per identificare e caratterizzare nuovi specifici biomarkers tumorali ovarici che permettano di effettuare una diagnosi precoce, di monitorare la progressione tumorale e di impostare una terapia mirata.
Le indagini preliminari sono state effettuate su campioni bioptici di tessuto ovarico sano e tumorale, e su liquido proveniente da lavaggio peritoneale.
Le proteine sono state estratte da circa 100 mg di tessuto congelato in azoto liquido, polverizzato in un mortaio e risospeso in tampone di lisi contenente urea, tiourea, CHAPS, Tris-HCl, ampholine pH 3-10 e inibitori delle proteasi. Dopo incubazione a temperatura ambiente per un’ora, i campioni sono stati sonicati e centrifugati. Il pellet, ottenuto dopo precipitazione in acetone freddo, è stato risospeso in un buffer di reidratazione (urea, tiourea, CHAPS, DTT e ampholine) e il contenuto proteico totale misurato con il metodo Bradford. L’isoelettrofocalizzazione (IEF) è stata effettuata caricando 80 g di proteine totali su IPG strips a range di pH 3-10 non lineare, fino al raggiungimento di 70000 Vh finali.
Le proteine focalizzate sono state ridotte con DTT e alchilate con Iodoacetamide in tampone di equilibrazione (Urea, Tris-HCl, Glicerolo, SDS, Blu di Bromofenolo) prima della separazione elettroforetica su gels di poliacrilamide (SDS–PAGE), a concentrazione omogenea 10% e a gradiente 8-16%. I gel sono stati infine colorati con metodica Silver-staining massa-compatibile. L’analisi con il programma PDQuest ha evidenziato la presenza di numerosi spot proteici differenzialmente espressi nei campioni tumorali rispetto ai controlli sani; la spettrometria di massa ESI-Q-TOF-MS/MS porterà all’identificazione delle proteine di interesse.
Per quanto riguarda il liquido derivato da lavaggio peritoneale, prima dell’analisi 2DE le aliquote prelevate sono state concentrate con filtri Amicon aventi cut-off di PM 5 KDa.
Parallelamente alla 2DE, sui campioni tissutali è stata effettuata anche l’analisi con tecnologia SELDI-TOF (Surface Enhanced Laser Desorption Ionisation-Time of Flight).
I risultati ottenuti hanno evidenziato variazioni significative dell’espressione di numerose proteine, soprattutto a basso peso molecolare (<20 KDa). Lo scopo dello studio ha la finalità di giungere all’identificazione di biomarcatori specifici del tumore ovarico che possono essere evidenziati anche nel liquido peritoneale ed eventualmente nel siero, fungendo da fattori prognostici della malattia ed anche predittivi di risposta al trattamento con derivati del platino.
2005
- Nitinol Carbofilm coated stents for peripheral applications: Study in the porcine model
[Articolo su rivista]
M., Prunotto; C., Isaia; Gatti, Antonietta; Monari, Emanuela; E., Pasquino; M., Galloni
abstract
Testing the safety and foreign body reaction (FBR) of a nitinol self-expandable carbon-coated stent system in the porcine animal model at different follow-up (FU) periods. Methods: Fifteen minipigs received 30 carbon-coated self-expandable nitinol stents in iliac arteries. Explants were carried out at 7 (3 animals), 30 (4 animals), 90 (4 animals) and 180 (4 animals) day FU, for evaluation of acute, sub-acute and chronic biological response to the implanted devices. Histological, immunohistochemical, histomorphometric and scanning electron microscopy (SEM) analyses were performed to assess inflammatory reaction, endothelialization process, neointimal growth and cellular composition. Results: Thirty stents were successfully implanted. No mural thrombi were observed at gross examination or by angiography. Histologically no significant inflammatory reaction was detected: the stents appeared covered by a thin monolayer of endothelial cells even at 7 day FU. The neointima presented homogeneous growth and moderate thickness after 30, 90 and 180 days explants (0.38 +/- 0.36 mm, 0.33 +/- 0.30 mm, 0.27 +/- 0.25 mm respectively). Internal and external elastic laminae were intact in 95% of stented arteries. Histological data validations of vessel endothelialization was obtained with SEM for the seven day follow-up group. Conclusions: This study showed good remarkable technical performances, minimal FBR and biocompatibility comparable with other available pre-clinical experimentations.
2005
- Nitinol stents for peripheral applications: comparative study in the porcine model.
[Abstract in Atti di Convegno]
M., Galloni; M., Prunotto; C., Isaia; Gatti, Antonietta; Monari, Emanuela; E., Pasquino
abstract
N/A
2004
- Alteration of innate defence cell functions by mineral dust nano-particles
[Relazione in Atti di Convegno]
D., Boraschi; M., Lucarelli; Monari, Emanuela; Gatti, Antonietta
abstract
n/a
2004
- Clinical investigation on bioactive glass particles for dental bone defects
[Relazione in Atti di Convegno]
Gatti, Antonietta; Monari, Emanuela; L., Simonetti
abstract
The research investigates the bioactive behaviour of a glass used as granules in dental bone defects. The materials was inserted in post-extractive defects to induce a bone growth for following dental implants. During the surgical manouvre of implantation a bone biopsy was extracted and analyzed hystologically and under Electron Scanning Microscope. The observations showed the integration of the granules in a new bone grown environment. Some of them appeared already degraded after six months from implantation.
2004
- Detection of CrCo particles in the spleen of guinea pigs six weeks after their intra-osseus implantation
[Abstract in Atti di Convegno]
P. A., Revell; Gatti, Antonietta; Gambarelli, Andrea; Monari, Emanuela; B., Hercus; S., Saeed; T., Macinnes
abstract
n/a
2004
- Detection of micro- and nano-sized biocompatible particles in the blood.
[Articolo su rivista]
Gatti, Antonietta; S., Montanari; Monari, Emanuela; Gambarelli, Andrea; F., Capitani; B., Parisini
abstract
The research deals with new scanning electron microscopic evaluations of the interface between blood and explanted temporary vena cava filters from patients affected by blood disorders. The biological tissues adherent to the filter and the small thrombi formed in vivo were detached from the metallic structure of the device, fixed, dehydrated and prepared for the histological and the electron microscopy. The analyses showed that both samples (thrombus and newly formed tissue) contained foreign, in some cases nano-sized, bodies. The chemistry of these particles was different and varied, and unusual compounds containing non-biocompatible elements like bismuth, lead, wolfram, tungsten were also detected. The interaction between these debris travelling in the blood stream and the blood itself leads to suspect that the formation of the thrombus can originate from these inorganic and inert foreign bodies that act as triggering agent of the blood coagulation.
2004
- Effects of nano-scaled on endotelial cell functions in vitro
[Relazione in Atti di Convegno]
K., Peters; R. E., Unger; Gatti, Antonietta; Monari, Emanuela; C. J., Kirkpatrik
abstract
n/a
2004
- Effects of nano-scaled particles on endothelial cell function in vitro: studies on viability, proliferation and inflammation.
[Articolo su rivista]
Kirsten, Peters; Ronald E., Unger; C., James Kirkpatrick; Gatti, Antonietta; Monari, Emanuela
abstract
Recent studies give support for a connection between the presence of inorganic particles (of microm and nm size) in different organs and tissues and the development of inflammatory foci, called granulomas. As the potential source of particles (e.g. porcelain dental bridges) and the location of particle detection were topographically far apart, a distribution via the blood stream appears highly probable. Thus, endothelial cells, which line the inner surface of blood vessels, would come into direct contact with these particles, making particle-endothelial interactions potentially pathogenically relevant. The objective of this study was to evaluate the effects that five different nano-scaled particles (PVC, TiO2, SiO2, Co, Ni) have on endothelial cell function and viability. Therefore, human endothelial cells were exposed to different amounts of the above-mentioned particles. Although most particle types are shown to be internalised (except Ni-particles), only Co-particles possessed cytotoxic effects. Furthermore, an impairment of the proliferative activity and a pro-inflammatory stimulation of endothelial cells were induced by exposure to Co- and, to a lesser extent, by SiO2-particles. If a pro-inflammatory stimulation of endothelial cells occurs in vivo, a chronic inflammation could be a possible consequence.
2004
- Innate defence functions of macrophages can be biased by nano-sized ceramic and metallic particles.
[Articolo su rivista]
Marilena, Lucarelli; Gatti, Antonietta; Graziana, Savarino; Paola, Quattroni; Lucia, Martinelli; Monari, Emanuela; Diana, Boraschi
abstract
Nano-sized particles of ceramic and metallic materials are generated by high-tech industrial activities, and can be generated from worn-out replacement and prosthetic implants. The interaction with the human body of such nanoparticles has been investigated, with a particular emphasis on innate defence mechanisms. Human macrophages (PMA-differentiated myelomonocytic U-937 cells) were exposed in vitro to non-toxic concentrations of TiO(2), SiO(2), ZrO(2), or Co nanoparticles, and their inflammatory response (expression of TLR receptors and co-receptors, and cytokine production) was examined. Expression of TLR receptors was generally unaffected by exposure to the different nanoparticles, except for some notable cases. Exposure to nanoparticles of ZrO(2) (and to a lesser extent TiO(2)), upregulated expression of viral TLR receptors TLR3 and TLR7. Expression of TLR10 was also increased by TiO(2) and ZrO(2) nanoparticles. On the other hand, TLR9 expression was decreased by SiO(2) nano-particles, and expression of the co-receptor CD14 was inhibited by Co nanoparticles. Basal and LPS-induced production of cytokines IL-1beta, TNF-alpha, and IL-1Ra was examined in macrophages exposed to nanoparticles. SiO(2) nanoparticles strongly biased naive macrophages towards inflammation (M1 polarisation), by selectively inducing production of inflammatory cytokines IL-1beta and TNF-alpha. SiO(2) nanoparticles also significantly amplified the inflammatory phenotype of LPS-polarised M1 macrophages. Other ceramic nanoparticles had little influence on cytokine production, either in resting macrophages, or in LPS-activated cells. Generally, Co nanoparticles had an overall pro-inflammatory effect on naive macrophages, by reducing anti-inflammatory IL-1Ra and inducing inflammatory TNF-alpha. However, Co nanoparticles reduced production of IL-1beta and IL-1Ra, but not TNF-alpha, in LPS-polarised M1 macrophages. Thus, exposure to different nanoparticles can modulate, in different ways, the defence/inflammatory capacities of macrophages. A thorough analysis of these biasing effects may shed light on the mechanisms of pathogenesis of several diseases based on dysregulation of the immune response (allergies, autoimmunity, tumours).
2004
- Modulation of defense cell functions by nano-particles in vitro
[Relazione in Atti di Convegno]
Lucarelli, M; Monari, Emanuela; Gatti, Antonietta; Boraschi, D.
abstract
The interaction of nano-particles derived from ceramic objects and handicrafts with the human body has been investigated with a particular emphasis to defense mechanisms. Macrophage (professional) and epithelial (non-professional) cells were exposed in vitro to TiO2, SiO2, or ZrO2 nano-particles, and their vitality and inflammatory/immune activation checked. All nano-particles showed effects on cell activation, although different in nature and extent. In particular, SiO2 nano-particles directly induced significant inflammatory activation of macrophages, as judged by production of the inflammatory cytokines IL-1beta and TNF-alpha and TLR modulation. Thus, it is suggested that exposure to ceramic nano-particles can contribute to the induction of inflammation-related pathologies.
2003
- Carbon-coated stents implanted in porcine iliac and renal arteries: Histologic and histomorphometric study
[Articolo su rivista]
M., Galloni; M., Prunotto; A., Santarelli; F., Laborde; A., Dibie; I., Isaia; Monari, Emanuela; Gatti, Antonietta; E., Pasquino
abstract
PURPOSE: To test in a pig model the biocompatibility and effectiveness of carbon-coated renal and iliac artery stent systems during implantation procedures and at different follow-up periods. MATERIALS AND METHODS: Twenty-two miniature pigs received carbon-coated balloon-expandable stainless-steel stents in their renal and iliac arteries. Animals were killed at 7, 30, or 180 days for evaluation of acute, subacute, and chronic biologic response to the implanted devices. Histologic, histomorphometric, and scanning electron microscopy (SEM) analyses were performed to assess inflammatory reaction, endothelialization process, and neointimal growth. RESULTS: Forty-four iliac stents and 42 renal stents were successfully implanted. Mural thrombi were not observed by angiography or histologic examinations. Histologically, no significant inflammatory reaction was detected: the stents appeared covered by a thin monolayer of endothelial cells even at 7-day follow-up. The neointima showed homogeneous growth and moderate thickness at 30-day and 180-day explantations (0.09 mm +/- 0.06 and 0.15 mm +/- 0.13, respectively, for renal arteries; 0.12 mm +/- 0.04 and 0.21 mm +/- 0.12, respectively, for iliac arteries). Internal and external elastic laminae were intact in 82 of 86 cases (95%) of stent-implanted arteries. Histologic validations were obtained with SEM observations for each follow-up group. CONCLUSIONS: This study showed good technical results of deployment of carbon-coated renal and iliac stents and very satisfactory biologic behavior in terms of tissue and hematologic compatibility. The devices do not induce thrombus formation.
2002
- Assesment of cytotoxicity and skin sensitization of stainless steel and alloys accordin to European Standard Method.
[Relazione in Atti di Convegno]
Gatti, Antonietta; A., Tamba; Monari, Emanuela
abstract
n/a
2002
- In vitro cytocompatibility of experimental calcium-releasing cements
[Abstract in Atti di Convegno]
Monari, Emanuela; Gatti, Antonietta; R. Z., Legeros; S., Lin
abstract
n/a
2001
- Assesment of the cytotoxicity of experimantal calcium releasing cements.
[Abstract in Rivista]
Gatti, Antonietta; R. Z., Legeros; Monari, Emanuela; S., Lin; Magnoni, Cristina; Cazzella, Francesco
abstract
The purpose of this study is to evaluate the biocompatibility in rabbit of calcium phosphates. The materials differd in the degree of biodegradation and can therefore be used for application requiring rapid or slow biodegradation.
2001
- Clinical long-term evaluation of hydroxyapatite granules implantation in periodontal defects
[Relazione in Atti di Convegno]
Gatti, Antonietta; Monari, Emanuela; Poli, Giorgio; Galli, Ermanno
abstract
Materials foreign to the patient's body are routinely used in medicine. In dentistry, in order to maintain the stability of dental elements, dentists fill the periodontal pockets with allograft or xenograft materials, that can work to keep the room open until the bony tissue fills it again, and/or to stimulate the bone growth itself. According to the function of their activity in the biological tissues they can be divided into two classes: osteo-conductive or osteo-inductive materials. Calcium-phosphates are considered in general to be osteo-inductive, since, when degrading, they release calcium and phosphate ions capable of inducing new-bone growth. This paper deals with the clinical long-term results of hydroxyapatite granules implanted in an anterior maxillar bone defect. Through physical and biological analyses carried out on the explanted biopsy, the Authors conclude that that type of material was only osteo-conductive.
2001
- Correlation among dental ceramics and systemic pathologies
[Abstract in Rivista]
Gatti, Antonietta; Monari, Emanuela; Ballestri, M; Rapana, R.
abstract
non disponibile
2000
- Conditions of non-compatibility of dental porcelains
[Abstract in Atti di Convegno]
Gatti, Antonietta; M., Ballestri; Monari, Emanuela; A., Bagni
abstract
n/a
2000
- Different calcium phosphates implanted in rabbits
[Abstract in Rivista]
Gatti, Antonietta; Legeros, Rz; Monari, Emanuela; Tanza, D.
abstract
non disponibile
2000
- In-vivo biodegradation of glasses: images of bioactivity
[Relazione in Atti di Convegno]
Gatti, Antonietta; Monari, Emanuela; L. L., Hench; D. C., Greenspan
abstract
The work takes into consideration the in-vivo degradative aspects of three different glasses, showing some aspects of their bioactivity. Sterilized granules of Bioglass, S53P4 and A-WG glass granules were implanted in rabbit dorsal muscles and in sheep’s mandible. Aher 60 days of implantation, the materials with the living tissues were explanted, embedded in PPMA and sectioned for the histologic and scanning electron microscopic observations.
Elemental microanalyses with an Energy Dispersive System were carried out to check the degradation of the materials through the topographic distribution of the elements. The three different glasses exhibited different states of degradation with different interactions with the soli and hard tissues for the same implatation tissues.
2000
- Long-term biocompatibility of silicon breast implant
[Abstract in Atti di Convegno]
Monari, Emanuela; Gatti, Antonietta; F., Ferrara; G., Barbolini
abstract
n/a
2000
- Transformation of different calcium-phopshates after different implantation
[Relazione in Atti di Convegno]
Gatti, Antonietta; Legeros, R.; Monari, Emanuela; Tanza, D.
abstract
The biocompatibility, biodegradation and in vivo transformation of four calcium phosphates (Ca-P) after implantation were investigated. The Ca-P materials (carbonatehydroxyapatite, CHA; F-substituted apatite, FI-IA; calcium-deficient apatite, AP; and magnesium-substituted tricalcium phosphate, beta -TCMP) were prepared and characterized using X-ray diffraction, FTIR and scanning electron microscopy (SEM). Dissolution properties were investigated ill acidic buffer (0.1 M KAc, pH 6, 37 degreesC). Sterilized materials were implanted in surgically created defects ill femoral diaphyses of rabbits for 30 days. Retrieved implants were examined histologically and by back-scattered SEM with energy dispersive system (EDAX).All materials showed biocompability and intimate interface with the hard tissues. FA showed the least, and CHA, the most extent of biodegradation. The Ca/P of the surfaces of implanted material were similar to that of bone apatite regardless of their original Ca/P. Conclusion: Ca-P materials differing in composition also differ in biodegradation. These materials partially transform to bone-apatite like regardless of their original composition.
1999
- Biocompatibilità di Leghe auree per odontoiatria
[Relazione in Atti di Convegno]
Gatti, Antonietta; Monari, Emanuela; F., Mintrone; Salvatori, Roberta
abstract
n/a
1999
- Biocompatibilità di leghe odontoiatriche
[Abstract in Atti di Convegno]
Gatti, Antonietta; Monari, Emanuela; Salvatori, Roberta
abstract
n/a
1999
- In-vivo biodegradation of alumino-silicates in Dentistry
[Abstract in Atti di Convegno]
Gatti, Antonietta; Monari, Emanuela; M., Ballestri; R., Rapanà
abstract
n/a
1999
- Trattamenti di difetti paradontali e perimplantari col Perioglass
[Abstract in Atti di Convegno]
Gatti, Antonietta; Monari, Emanuela; L., Simonetti
abstract
n/a
1999
- Valutazione della biocompatibilità del fisiograft per difetti ossei
[Articolo su rivista]
Gatti, Antonietta; Monari, Emanuela; Tanza, D.; Betti, V.
abstract
BACKGROUND: This paper examines a new material composed of polylactic and polyglycolic acids mixed with destrane easy to handle in bone defect reconstruction in periodontology and implantology: its name is fisiograft. METHODS: According to the European directives, fisiograft was checked for some biological tests selected for its specific use. In particular, tests of cytotoxicity, genotoxicity, sensitization and implantation in rabbit's femurs were carried out. RESULTS: The results show that the composite material is not cytotoxic when it is in direct and indirect contact with murine fibroblasts. It is not genotoxic and not allergenic. The implantation in rabbit, already after 30 days, shows that around the material there is a rapid bone growth and the material is reabsorbing. The degradation is suitable for its clinical use. CONCLUSIONS: Fisiograft showed to be biocompatible, easy to handle and its degradation kinetics in bone is appropriate.
1999
- Valutazione d’espianti di protesi mammarie in silicone a lungo termine
[Relazione in Atti di Convegno]
Gatti, Antonietta; Ferrara, F.; Guicciardi, L.; Barbolini, C. M; Galli, Ermanno; Monari, Emanuela
abstract
n/a
1998
- Bioactivity of the active glasses
[Relazione in Atti di Convegno]
Gatti, Antonietta; Monari, Emanuela
abstract
Medicine requires newer and newer materials to repair or substitute tissues or replace functions. Active galsses have been developed for 20 years in order to help bone in reconstruction.
many researcher have syntetized glasses that during implantation degrade releasing ions used by cells for their metabolism. Bone reconstruction is helped by these ions both in tems of quantity and in time. At the bone-glass interface the outer glass layer, rich in Calcium and Phosphorous, chemically bonds bone to the material. The glass degradation and the interface developed after implantation depend principally on the chemical composition of the material. The compositions of some glasses are presented with the interfaces created with bone or soft tissues during the in-vivo implantation. The morphology of the embedded cross-sections under scanning Electron Microscope is shown with the relevant X-ray dot maps for principal elements. An Energy Dispersive System applied to a SEM detects the inorganic elements present in the glass and their topographic distribution in the material before and after implantation giving the image of degradation of the glass and showing its bioactivity.
1998
- Biocompatibilità di un nuovo materiale per difetti parodontali
[Abstract in Atti di Convegno]
Gatti, Antonietta; Monari, Emanuela; Salvatori, Roberta; V., Betti
abstract
n/a
1998
- NEW PUTTY COMPOSITE MATERIALS FOR BONE DEFECTS IN DENTISTRY
[Relazione in Atti di Convegno]
Gatti, Antonietta; L. L., Hench; Monari, Emanuela; J., Thompson; D., Tanza
abstract
Two types of composite materials for bone repair were produced at Imperial College of London, one of Bioglass® dextran resorbable matrix and the second of Biog|ass® and a non resorbable polysulphone polymer matrix. Three samples ofthe first and a fourth sample ofthe second were sterilized and implanted in drilled holes in a sheep's mandible and also in tibial defects. After 3 months the materials with the surrounding tissues were explanted, fixed and radiographed in order to have the macroscopic relationship between the material and bone. Sections were prepared for scanning electron microscopic observations and X-ray microanalyses to detect any resorption of the materials and the interface developed with bone, Results show good biocompatibility for all four materials. The polysulphone/glass showed a small surface degradation. The bioactive glass-dextran composite showed maximum resorption and at times it was difficult to recognize the site of implantation. The dextran-glass composite was very promising in mandibular bone augmentation or repair of bone defects. The polysulphone- bioactive glass composite may be more useful for long term implants.
1998
- Strategie chirurgiche nei difetti ossei.
[Abstract in Atti di Convegno]
Gatti, Antonietta; Monari, Emanuela; Bertoldi, Carlo; Simonetti, L.
abstract
Per la correzione di difetti ossei possono essere impiegati materiali sintetici. Questi, a seconda, possono comportarsi biocompatibilmente in modo inerte, osteoconduttivo od osteoinduttivo.
Nel presente studio abbiamo voluto valutare il comportamento del biovetro Perioglass* sviluppato dal professor Hench presso l'Università della Florida (USA) ed indicato addirittura come provvisto di effetti osteoinduttivi.
Tale materiale è stato dapprima testato su modelli animali (pecora e coniglio) al fine di verificarne la biocompatibilità e l'effetto sull'accrescimento osseo in tali sedi. Tali studi sono stati eseguiti mediante valutazioni cliniche, analisi al microscopio elettronico a scansione (SEM) dotato di microsonda a raggi X e su preparati istologici.
Verificati i prescritti requisiti di biocompatibilità, granuli di Perioglass* sono stati impiantati in due gruppi di pazienti con tipologia di difetto osseo, rispettivamente peri-implantare a "3 pareti" e di tipo granulomatoso-cistico.
Il biovetro è stato innestato a diretto contatto con l'osso perilesionale.
Il monitoraggio longitudinale dei pazienti, sino ad un anno dall'intervento, è avvenuto tramite valutazioni radiografiche, avvalendoci di un craniostato e del centratore. Le radiografie, quindi, venivano passate all'analisi d'immagine.
I risultati ottenuti indicano che il biovetro impiegato risulta provvisto di un buon effetto osteoconduttivo, mentre sull'osteoinduttività permangono dubbi dati i tempi (piuttosto lunghi) di ossificazione ed il livello di ossificazione raggiunto.
1997
- Indagine morfologico-strumentale su espianti di placche e viti per fissazione interna rigida
[Relazione in Atti di Convegno]
Bertoldi, Carlo; Gatti, Antonietta; Monari, Emanuela
abstract
In questi ultimi anni si è assistito ad un sempre più diffuso impiego, in chirurgia maxillo-facciale, dell'osteosintesi tramite Fissazione Interna Rigida (FIR) per diversi motivi tra cui indubbiamente il principale si identifica nella capacità e nell'efficienza di questa metodica nel produrre una fissazione spaziale precisa e stabile dei monconi di frattura. Essa, quindi, se ben condotta, consente la guarigione per prima intenzione, un ridotto rischio infettivo locale, tempi di recupero minori, una maggiore stabilità del risultato ottenuto ed una superiore possibilità di igiene orale.
Tra i mezzi, tuttavia, più comunemente impiegati in tali procedure vi sono i metalli e le loro leghe la cui interazione specifica tra di essi e i tessuti biologici può dare origine a fenomeni di depauperazione di massa per usura o corrosione. In special modo il secondo meccanismo è stato molte volte accusato come potenziale induttore di fenomeni infiammatori locali, di patologie locali e sistemiche da accumulo e di attivazione specifica od aspecifica del comparto immunitario.
Inutile precisare la ricaduta patologica pesante che, se tali meccanismi venissero confermati, sarebbero conseguenti ad un inappropriato impiego di materiali.
Lo scopo di questo lavoro è quello di verificare, nell'ambito dei modelli metallurgici più diffusi nelle placche, la reale potenzialità corrosiva di questi mezzi, quando posti a contatto dell'ambiente biologico in corrispondenza del sito fratturativo dove essi sono chiamati ad agire.
A 5 pazienti, dai 20 ai 60 anni, sono state espiantate 9 placche, dopo periodi di permanenza in ambito biologico sino ai 13 mesi. Tali soggetti presentavano indici clinico-strumentali normali ed erano stati accuratamente informati (ed avevano liberamente acconsentito) sugli scopi della ricerca.
I 5 soggetti studiati non presentavano neppure localmente segni di flogosi.
Sono state asportate 9 placche di cui 5 erano in lega di titanio, 2 in acciaio AISI 316L ed 1 in Vitallium*.
Le placche in acciaio e Vitallium* hanno rivelato possedere diverse aree interpretabili come siti di corrosione , mentre le placche in lega di titanio, sebbene più "rovinate" dalle manovre chirurgiche (in quanto il titanio è più morbido delle altre due leghe), hanno rivelato un numero assai più modesto di siti potenzialmente corrosivi.
Poco sappiamo degli effetti sulla salute di alcuni costituenti (come, ad esempio, l'alluminio) contenuti nelle leghe suddette (anche in alcune leghe al titanio).
La comprovata possibilità di diffusione ionica dei costituenti metallici dovrebbe far riflettere sul possibile effetto patologico esercitato da questi mezzi osteosintetici e spingere la ricerca a produrre sistemi per FIR che uniscano i benefici insiti in tale metodica con sempre minori possibili effetti collaterali.
Codice: IT\ICCU\BVE\0121041
1997
- Le nuove frontiere dei trattamenti parodontologici: i materiali bioattivi.
[Relazione in Atti di Convegno]
Gatti, Antonietta; Monari, Emanuela; S., Bulgarelli
abstract
In campo parodontologico sono da anni già in uso materiali che inseriti nelle tasca parodontali, evitano ai tessuti molli di occupare tale spazio e talvolta agiscono come elemento›ponte per l'osso, così da ripristinare la situazione precedente alla patologia. Il presente studio ha verificato il comportamento in vivo su osso mandibolare di pecora di tre vetri bioattivi con diverso contenuto in silice, valutando e l’accrescimento osseo e la loro degradazione mediante microscopia elettronica a scansione e con una microsonda a raggi X. I tre vetri hanno dimostrato delle caratteristiche diverse: il vetro meno degradabile si è mostrato essere il 55S seguito dal 525 e dal Bioglass®. Nel caso del vetro Bioglass® la biodegrazione è molto veloce, sicuramente più veloce della osteogenesi, ma ugualmente ne influenza la velocità; pertanto si presta a riempire difetti parodontali e cisti ossee. Il vetro 52S,invece potrebbe aiutare nei rialzi di creste ossee ove la quantità d”osso in gioco è maggiore e quindi occorre avere un materiale bioattivo, ma che rimane in sede per un tempo più lungo. Questi primi risultati non sembrano particolarmente favorevoli al vetro 55S, anche se si è vista una certa precipitazione di fosfati di calcio, probabilmente non sufficienti per l’osteostimolazione.
1997
- TEST OF BIOACTIVITY IN FOUR DIFFERENT GLASSES
[Relazione in Atti di Convegno]
Gatti, Antonietta; L. L, Hench; Monari, Emanuela; F., Gonella; F., Caccavale
abstract
A special preparation was carried out on granules of the glasses under investigation before their implantation in rabbit's muscles, so as to understand how their bioactivity evolves, to know what the ions involved in the diftirsion are and how the diflirsion is directed. At the explantation the materials and the surrounding tissues were fixed and prepared for the scanning electron microscopic examinations and the energy dispersive analyses. The results show the ditference in degradation of the glasses that induce some considerations on the mechanism of bioactivity.
1997
- Valutazione della biocompatibilità di una lega non nobile per uso dentale.
[Relazione in Atti di Convegno]
Monari, Emanuela; Bertoldi, Carlo; Gatti, Antonietta
abstract
In una fase preliminare rispetto all'utilizzo di dispositivi biomedici, deve essere previsto che la loro libera distribuzione avvenga solo dopo averne accertato la sicurezza sul paziente.
Il presente lavoro vuole cercare di individuare quello che potrebbe essere un modello di validazione "in vitro" ed "in vivo", preliminare all'impiego sull'uomo, al fine di poter scongiurare la possibile presenza di effetti citotossici, irritativi e di intolleranza integrativa tissutale.
Nella fattispecie viene valutata una lega metallica per protesi scheletrate in Cr-Mo-Ni.
Vengono compiuti in triplo i test di citotossicità, per contatto diretto ed indiretto, utilizzando la linea immortalizzata murina L-929.
Vengono impiegati come confronti "positivi" e "negativi", per i test per contatto diretto, rispettivamente, dei dischi di rame e il disco di polietilene, per i test per contatto indiretto, rispettivamente, una soluzione di fenolo ed il terreno di coltura.
Ai normali conteggi si aggiunge lo svolgimento dei test con rosso neutro e dei test di riduzione dei sali di tetrazolio (MTT test) per valutare sotto diversi aspetti la vitalità cellulare.
Si eseguono, poi, su conigli New Zeland i test di irritazione cutanea, di sensibilizzazione cutanea e d'impianto.
Nella fattispecie la lega studiata ha mostrato buona biocompatibilità nei test di irritazione e di impianto. Risultati un po' meno lusinghieri, ma sempre accettabili, si sono ottenuti nelle valutazioni della citotossicità e di sensibilizzazione cutanea.
Ciò potrebbe indicare un effetto di passivizzazione che la lega subisce a contatto con l'ambiente esterno, anche se questo effetto desiderato pare essere tutt'altro che completo.
In ogni caso un siffatto studio multicentrico in fase preclinica ci parrebbe pressochè completo e doveroso, poichè in grado di porre in evidenza un buon numero di pregi, ma anche possibili difetti dei materiali impiegabili che spesso sono stati incolpati quale origine di svariate patologie orali e sistemiche.
Codice: IT\ICCU\BVE\0121041
1997
- Valutazione dell’interazione biologica di un composito degradabile per difetti ossei.
[Relazione in Atti di Convegno]
S., Bulgarelli; Gatti, Antonietta; Monari, Emanuela
abstract
Negli ultimi anni è nata l'esigenza di ricercare materiali biocompatibili per il riempimento di difetti ossei. La presente ricerca è volta, appunto, allo studio in-vivo di un nuovo materiale ”Fin-Biograft” (Finceramica Faenza) composito per difetti ossei. Allo scopo di valutarne la biocompatibilità, due campioni di tale materiale sono stati impiantati per un mese in femore di coniglio, seguendo le norme europee CEN, ed all’espianto si è studiata l`interfacie materiale-osso tramite osservazioni morfologiche al microscopio elettronico a scansione. Dalle analisi eseguite si è potuto evidenziare che il materiale composito non ha dato reazioni infiammatorie macroscopiche per il tempo d`impianto durante il quale è stato fatto l’esperimento. Il buon comportamento riscontrato fa supporre che tale composito svolga un azione di osteo-stimolazione nei confronti del tessuto osseo.
1996
- Active materials inducing calcium-phosphate precipitation for the treatment of bone defects.
[Capitolo/Saggio]
Gatti, Antonietta; Monari, Emanuela; Chiarini, Luigi; D., Tanza
abstract
Active glasses are materials that, when implanted in bone, degrade releasing ions that can positively interact with the bony cells stimulating the osteogenesis. This phenomenon regards two different aspects: the biological and the chemical-physical one. Glassy granules of different compositions were implanted in rabbit’s muscles and sheep’s mandibular bone at different times. After the animals’ sacrifice particles with the surrounding tissues were explanted, fixed and embedded in PMMA.
Sections were prepared for the scanning electron microscopy. The
observations showed that the glass with the less amount of silicon
degrades in a shorter time than that with the higher content. In sheep`s mandibular bone, the glassy particles with 52 and 55% molar weight content in silicon, degrade and on the surface of glass little needles of precipitates appear. The EDS analyses show that they are composed of Calcium and Phosphorus with a P content greater than that of HA of bone. New bone trabeculae surrounds the granules.
1996
- In-vivo leaching of A-WG particles for dental use.
[Relazione in Atti di Convegno]
Gatti, Antonietta; T., Kokubo; Monari, Emanuela; Chiarini, Luigi; D., Tanza
abstract
Bioactive glass granules are used as filler for bone defects in Dentistry.
Present work studies the in-vivo behaviour of a glass-ceramic material (A-WG) used as granules of three different sizes. After implantation in soft and hard tissues, the morphology of granules is studied with Scanning Electron Microscopy, and the chemical
degradation is detected with an Energy Dispersive System. The results show the biocompatibility of the glass and the best size in order to optimize the interaction with bone.
1996
- Pre-clinical evaluation of a new pacing lead with the electrodes and tines coated with a thin film of turbostratic carbon (Carbofilm TM)
[Abstract in Rivista]
M., Galloni; Gatti, Antonietta; G., Noera; V., Lolli; Monari, Emanuela; S., Bulgarelli; L., De Benedetti; G., De Giorgi; E., Pasquino; F., Renesto; G., Rolando
abstract
n/a
1995
- In vivo study of the degradation of 4 different composition active glass
[Relazione in Atti di Convegno]
Gatti, A. M.; Hench, L.; Monari, Emanuela; Chiarini, Luigi; Bulgarelli, S.; Tanza, D.
abstract
The present study investigated the behavior of 4 different glasses. used as granules for bone repair, to understand the influence of the composition in bone stimulation and their degradation in a biological environment.
1995
- Influence of granule size on the reactions of a bioactive glass for repair of bone defects.
[Relazione in Atti di Convegno]
Gatti, Antonietta; O. H., Andersson; G., Valdrè; Chiarini, Luigi; D., Tanza; S., Bulgarelli; Monari, Emanuela
abstract
the present work analyses the in-vivo behaviour of a bioactive glass that can be used for osteostimulation in bone defects. The glass called SP53P4 was implanted as granules of 3 different sizes in pockets created in rabbit' dorsal muscles to investigate the reactions in the biological fluids. Before the implantation the granules were coated with a layer of gold on one side in order to obtain two sides with different permeability in the same granule and thus two different velocities of diffusion.
After 2 months bone and granules were explanted and processed for observation under Scanning Electron Microscope with an X_ray microprobein ordet to study the changed morphology of the granules and the distribution of the elements.
The granules were partly leached but there were no sign of total glass breakdown and resorption.
1995
- Studio in vivo di un nuovo impianto protesico
[Abstract in Rivista]
Gatti, Antonietta; Monari, Emanuela; Chiarini, Luigi; S., Bulgarelli; D., Tanza
abstract
n/a
1995
- Valutazioni cliniche e sperimentali di una nuova linea implantoprotesica: ricerca sperimentale.
[Articolo su rivista]
Gatti, Antonietta; Monari, Emanuela; S., Bulgarelli; Chiarini, Luigi; D., Tanza
abstract
La ricerca in oggetto è consistita in un test d'impianto su animale di perni in titanio e della successiva analisi sia del materiale che dell'interfaccia creata coi tessuti biologici.
1994
- Healing of periodontal ligament after implantation of bioactive glass in surgically created periodontal defects: a pilot study.
[Relazione in Atti di Convegno]
Gatti, Antonietta; O. H., Andersson; A., Piattelli; P., Trisi; Monari, Emanuela; Chiarini, Luigi
abstract
Granules of a bioactive glass were implanted in a surgically created hole in a sheep's mandibular bone. After one year the bone with the glass was explanted, processed and examined under optical and scanning electron microscope. All particle were surrounded by a layer of minerilized tissue with histological features very similar to root cement; serial sections revealed that a small cementum bridge connected this mineralized tissue directly to root cementum. Under polarized light it was possible to observe oriented collagen fibers similar to the periodontal ligament inserting on the mineralized tissue.
1994
- Importance of the physical parameters in the bone stimulation induced by active- surface glasses.
[Capitolo/Saggio]
Gatti, Antonietta; O. H., Andersson; G., Valdrè; Chiarini, Luigi; D., Tanza; S., Bulgarelli; Monari, Emanuela
abstract
The present work analyses the in vivo behaviour of a bioactive glass that can be used for osteostimulation in bone defects. the glass called S53P4 was implanted as granules of 3 different sizes in pokets created in rabbit's dorsal muscles to investigate the reactions in the biological fluids. Before the implantation the granules were coated on one side with a layer of gold in order to obtain two sides with different permeability in the same granule and thus two differen velocities of diffusion. After 2 months granules were explanted and processed for observation under Scanning Electron Microscope with an X-ray microprobe in order to study the changed morphology of the granules and the distribution of the elements. The granules were partly leached but there were no signs of total glass breackdown and resorption.
1994
- In-vivo study of the interface developed by three different grain size of a bioactive glass-ceramics AW-G
[Abstract in Atti di Convegno]
Gatti, Antonietta; T., Kokubo; Monari, Emanuela; Chiarini, Luigi; S., Bulgarelli; D., Tanza
abstract
n/a
1994
- Influence on the granule size on the bioactivity of a bioactive glass for the repair of bone defects
[Abstract in Atti di Convegno]
Gatti, Antonietta; O. H., Andersson; G., Valdrè; Chiarini, Luigi; Monari, Emanuela; D., Tanza
abstract
n/a
1994
- PHYSICAL CHARACTERIZATION OF A NEW BIOMATERIAL FOR WOUND MANAGEMENT
[Articolo su rivista]
Gatti, Antonietta; P., Pinchiorri; Monari, Emanuela
abstract
The aim of this work was to evaluate some physical characteristics of a new biosynthetic membrane (Bioprocess(R)) used as temporary skin substitute for wound management. The tested characteristics, important for the clinical use of the membrane, were: permeability to some gases (O2, CO2 and N2) and water vapour, and absorption of fluids simulating a physiological-like environment. The tests were completed by scanning electron microscope (SEM) observations in order to verify the surface morphology before and after the tests. The results confirmed the good in vivo performance of this material.
1993
- Development and in-vivo evaluation of a system for power transfer through intact skin for artificial hearth.
[Articolo su rivista]
Gatti, Antonietta; Noera, G.; Monari, Emanuela; Fattori, G.; Incerti Medici, A.; Lolli, V.; Fumero, R.; Pizzutilo, F.; Strini, G.
abstract
Purpose of the present study is to evaluate the possibility to transfer power from outside to inside of human body, in order to feed a long term implanted prosthesis as the artificial heart. The system adopted was the medium-low frequency electromagnetic coupling between a special primary coil to be placed on the patient's body for a relatively short period 8e.g 8 hours per night) and a permanent implanted coil for supplying an internal accumulator. The construction details of the implanted circuit are presented and the results about the in-vivo testing measurements, in particular the histological analyses of the skin after many energy transfer processes, are shown.
1993
- Effect of the type of sterilization on the structure of polymers used in vad
[Abstract in Atti di Convegno]
Gatti, Antonietta; M. C., Tanzi; Monari, Emanuela; P., Pinchiorri; G., Ambrosio
abstract
n/a
1993
- Galss corrosion layers on biactive glass granules of uniform size affect cellular function
[Relazione in Atti di Convegno]
Gatti, a. M.; Ducheyne, P.; Piattelli, A.; Schepers, E.; Trisi, P.; Chiarini, Luigi; Monari, Emanuela
abstract
A bioactive glass called Biogran was implanted as granules of very narrow size range (300-355mm) in rabbit's dorsal muscles and in sheep's mandibular bone in order to determine the glass degratation in biological fluids and its influence on cellular function.Afert 1 month implantation in rabbit and 3 months in sheep , the granules with the surrounding tissue were resected and prepared for the optical and scanning electron microscopy.the results in dorsal muscle tissue showed tha the glass granules were uniformily surrounded by a highly cellular reactive tissue.In bone, granules were found extensively surrounded by bone tissue, in a almost-obliterated defect.
1993
- IN-VIVO EVALUATION OF AN EXPERIMENTAL DENTAL HA-TI IMPLANT
[Articolo su rivista]
Gatti, Antonietta; Chiarini, Luigi; Monari, Emanuela; D., Tanza
abstract
A new design of Titanium dental implant, coated with Titanium oxide and with a Hydroxyapatite apex, was experimented in a Beagle dog's jaw for one year to evaluate the biocompatibility of the materials used and the biomechanical efficiency of the design. The explanted prosthesis and the surrounding bone were embedded in methylmethacrylate resin and the sections observed under Scanning Electron Microscopy (SEM), with an X-ray microprobe to check the interface with bone and the occurrence of ionic release. The results showed that the pin was well accepted by the bone. New bone grew on the metal surface developing a mechanical bond with the prosthesis. No ionic release of the metal was seen.
1992
- Attività osteoconduttrice del Bop: risultati sperimentali.
[Relazione in Atti di Convegno]
Gatti, Antonietta; Chiarini, Luigi; Tanza, D.; Monari, Emanuela; Giacobbi, F.
abstract
n/a
1992
- Crystallizability and degradation of different poly(lactic acid) samples
[Abstract in Atti di Convegno]
Migliaresi, ; A., De Lollis; L., Fambri; Monari, Emanuela; D., Cohn
abstract
n/a
1992
- In vivo study of a new polymeric material.
[Abstract in Atti di Convegno]
Monari, Emanuela; Gatti, Antonietta; Chiarini, Luigi
abstract
n/a
1992
- Risposte dell'osso in presenza di alcuni bioceramici attivi in forma granulare.
[Relazione in Atti di Convegno]
Gatti, Antonietta; Chiarini, Luigi; Monari, Emanuela; D., Tanza
abstract
Tre tipi di materiali a diversa bioattività sono stati inseriti in nicchie create chirurgicamente in mandiboila di pecora. Dopo 5 mesi i materiali sono stati espiantati e trattati per l'osservazione alla microscopia elettronica a scansione e alla microsonda a raggi X. Si sono riscontrate tre diverse interfacie con l'osso e tre diversi comportamenti bioconduttivi.
1992
- Thromboresistance of Ti6Al4vm coated with a thin film of turbostratic carbon for cardiovascular applications.
[Relazione in Atti di Convegno]
Gatti, Antonietta; Monari, Emanuela; M., Dondi; G., Noera; G., Fattori; F., Vallana; S., Rinaldi; E., Pasquino
abstract
n/a
1992
- Tissue reaction to subcutaneously implanted polymers coated with a pyrolytic carbon film (CARBOFILM).
[Abstract in Rivista]
E., Pasquino; Gatti, Antonietta; Monari, Emanuela; M., Galloni; V., Cassolaro; S., Pascale; S., Rinaldi
abstract
n/a
1992
- Unsaturated tertiary arylamines as low toxicity activators in the formulation of biomedical acrylic resins.
[Relazione in Atti di Convegno]
Tanzi, M. C.; Levi, M.; Gatti, Antonietta; Monari, Emanuela
abstract
Two unsaturated tertiary aromatic amines, namely acryloyl- (ArcNP) and methacryloyl-N-phenyl piperazine are proposed as potential substitutes of dimetyl-p toluidine (DMPT), in the ''coldt curing of biomedical acrylic resins. The paper give an overview of the results so far obtained with the new compounds, with particular reference to some unpublished data on their efficacy as activators in the MMA/PMMA cold curing, and on their leachability from MMA cured samples.
AcrNP and MNP showed good efficacy as, accelerators, with similar gel times but lightly prolonged setting times in comparison with DMPT; both compounds (AcrNP in particular), produced PMMA-based cements with similar mechanical properties, but less accelerated ageing than DMPT; differently from MNP, AcrNP showed the ability to copolymerize with MMA, and to be hardly leachable from cured PMMA samples. Above all, the last property makes AcrNP preferable to MNP as red-ox activator, and good potential substitute of DMPT.
1991
- CALCIFICATION OF CHEMICALLY TREATED BOVINE PERICARDIUM
[Articolo su rivista]
Gatti, Antonietta; M., Galloni; Monari, Emanuela; G., Noera; E., Pasquino; F., Vallana
abstract
One of the most important problems arising in cardiac bioprostheses made with bovine pericardium and, more generally, with biologically-derived tissues is tissue calcification. The present study assessed four chemical treatments on patches of bovine pericardium, intended to avoid or minimize calcification. Pericardium specimens were treated with: A) 0.5% glutaraldehyde; B) 0.5% glutaraldehyde + 4% formaldehyde; C) same as A, but with a further neutralization treatment; D) acylation of fresh bovine pericardium. Circular samples of 1 cm diameter were subcutaneously implanted in the abdominal region of three groups of six rats. The explants were retrieved after 2, 4 and 8 weeks. The calcium content and the histological results showed better behaviour for C and D samples than with the commonly used fixation methods (A and B). The lowest calcification was observed with treatment D, even though its morphological structures were somewhat modified with homogenation of collagen bundles. Among the glutaraldehyde-based treatments, treatment C appears to be the most promising because the pericardium shows slower calcium accumulation with a diffusive pattern.
1991
- Physical characterization of acrylic bone cement cured with new accelerator systems
[Articolo su rivista]
Tanzi, Mc; Sket, I; Gatti, Antonietta; Monari, Emanuela
abstract
In the attempt to substitute dimethyl-p-toluidine (DMPT), a toxic tertiary aryl-amine accelerator, into the formulation of acrylic cements, less toxic accelerator systems are developed. These systems consist of benzoyl peroxide (BPO) and unsaturated tertiary-aryl-amines, such as acryloyl- (ANP) and methacryloyl-(MNP) N-phenylpiperazine, which can be chemically incorporated in the polymerizing resin or, at least, result in less leaching from cured materials. In this work compressive mechanical properties and ageing tests for colour stability of acrylic cement cured with BPO and ANP or MNP have been considered. For compressive tests, cylindrical specimens were cured with BPO and equivalent molecular amounts of DMPT, ANP and MNP. Compressive yield stress (sigma y), strain at yield (epsilon y, %) and elastic modulus (E) gave very similar results for samples cured with DMPT and ANP, and slightly lower results for samples cured with MNP. In colour stability tests, the samples (disks of 1.5 cm diameter) were exposed to UV light at different irradiation times (up to 42 h). The evaluation of the colour change was performed with a digital analyser for images, and observed under scanning electron microscopy. From the obtained results, ANP appeared to be the best candidate as accelerator in the preparation of biomedical acrylic resins and composites.
1991
- Programma di valutazione in vivo della valvola bicarbon: risultati a lungo termine.
[Articolo su rivista]
E., Pasquino; A., Incerti Medici; V., Lolli; Monari, Emanuela; Gatti, Antonietta
abstract
n/a
1991
- Studio in vitro di un nuovo composito per restauri di denti posteriori
[Articolo su rivista]
Gatti, Antonietta; Monari, Emanuela
abstract
E' stato preso in considerazione un nuovo composito per restauri di denti posteriori, l'Adaptic II della J &J, e si sono valutate alcune caratteristiche fisiche del materiale importanti per il buon andamento una volta applicato sul paziente, quali ad esempio: l'assorbimento di liquidi e l'invecchiamento simulato. Tali indagini hanno dimostrato che il materiale presenta caratteristiche accettabili per l'utilizzo.
1991
- Unsaturated tertiary arylamines as low toxicity activators in the formulation of biomedical acrylic resins
[Abstract in Atti di Convegno]
M. C., Tanzi; M., Levi; Gatti, Antonietta; Monari, Emanuela
abstract
n/a
1990
- Biocompatibility of chemically treated bovine pericardium in rats
[Abstract in Rivista]
Gatti, Antonietta; M., Galloni; Monari, Emanuela; G., Noera; E., Pasquino; F., Vallana
abstract
n/a
1990
- Physical characterization of acrylic bone cement cured with a new accelerator system
[Abstract in Atti di Convegno]
M. C., Tanzi; I., Sket; Gatti, Antonietta; Monari, Emanuela
abstract
n/a
1989
- Development od the crustacean copepod lernaea cyprinacea L.
[Abstract in Rivista]
I., Benedetti; Mola, Lucrezia; Monari, Emanuela; Sabatini, Maria Agnese; Marini, Milena; B., Fratello
abstract
The cleavage, gastrulation and naupliar stage of the copepod L. cyprinacea are described. the naupliar stages is considered an embryonal stage for the lack of a mouth or developing gut.
1989
- Preliminary findings on the development of the parasitic copepod Lernaea cyprinacea L. from cleavage to hatching
[Articolo su rivista]
Benedetti, I; Mola, Lucrezia; Monari, Emanuela; Sabatini, Maria Agnese; Marini, Milena; Fratello, Bernardo
abstract
We report preliminary findings on the development of Lernaea cyprinacea from cleavage to hatching. The eggs are centrolecithal with superficial cleavage. Nucleoli were found in the nuclei of the perilecithal syncytium. Gastrulation occurs by in situ aggregation of cells, without evident morphogenetic movements.
1989
- Small diameter vascular graft evaluation by replacement of shee carotid artery: short term results with carbon coated prostheses.
[Articolo su rivista]
G., Noera; G., Fattori; A., Incerti Medici; Monari, Emanuela; Lodi, Renzo; Gatti, Antonietta; M., Galloni; E., Pasquino; F., Vallana
abstract
not available