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Stefania BERGAMINI
Ricercatore Universitario
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
- Bioactive Glasses in Periodontal Regeneration: Existing Strategies and Future Prospects—A Literature Review
[Articolo su rivista]
Cannillo, Valeria; Salvatori, Roberta; Bergamini, Stefania; Bellucci, Devis; Bertoldi, Carlo
abstract
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
- A Proteomic Analysis of Discolored Tooth Surfaces after the Use of 0.12% Chlorhexidine (CHX) Mouthwash and CHX Provided with an Anti-Discoloration System (ADS)
[Articolo su rivista]
Bergamini, Stefania; Bellei, Elisa; Generali, Luigi; Tomasi, Aldo; Bertoldi, Carlo
abstract
Chlorhexidine (CHX) is considered the gold standard for the chemical control of bacterial plaque and is often used after surgical treatment. However, CHX employment over an extended time is responsible for side effects such as the appearance of pigmentations on the teeth and tongue; the discoloration effects are less pronounced when using a CHX-based mouthwash with added an anti-discoloration system (ADS). The aim of this study was to evaluate, using one- and two-dimensional gel electrophoresis combined with mass spectrometry, the possible proteomic changes induced by CHX and CHX+ADS in the supragingival dental sites susceptible to a discoloration effect. The tooth surface collected material (TSCM) was obtained by curettage after resective bone surgery from three groups of patients following a supportive therapy protocol in which a mechanical control was combined with placebo rinses or CHX or a CHX+ADS mouthwash. The proteomic analysis was performed before surgery (basal conditions) and four weeks after surgery when CHX was used (or not) as chemical plaque control. Changes in the TSCM proteome were only revealed following CHX treatment: glycolytic enzymes, molecular chaperones and elongation factors were identified as more expressed. These changes were not detected after CHX+ADS treatment. An ADS could directly limit TSCM forming and also the CHX antiseptic effect reduces its ability to alter bacterial cell permeability. However, Maillard's reaction produces high molecular weight molecules that change the surface properties and could facilitate bacterial adhesion.
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 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
- 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
- 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
- Influence of tooth-brushing on early healing after access flap surgery: A randomized controlled preliminary study
[Articolo su rivista]
Bertoldi, C.; Generali, L.; Cortellini, P.; Lalla, M.; Luppi, S.; Tomasi, A.; Zaffe, D.; Salvatori, R.; Bergamini, S.
abstract
In the present study, the clinical outcomes obtained using three different protocols of post-operative plaque control for the 4 weeks after surgery were compared. Thirty healthy subjects, presenting at least one periodontal pocket requiring resective surgery, were selected and randomly distributed to three different groups corresponding to respective post-surgical protocols: (A) toothbrushes + chlorhexidine + anti-discoloration system (ADS + CHX); (B) toothbrushes + chlorhexidine (CHX); (C) only toothbrushes. The full-mouth plaque score (FMPS), full-mouth bleeding score (FMBS), probing pocket depth (PPD), recession depth (REC), clinical attachment level (CAL), and bleeding on probing (BoP) were measured in six aspects per tooth (mesio-buccal (MB), buccal (B), disto-buccal (DB), disto-lingual (DL), lingual (L), and mesio-lingual (ML)) at baseline, 3 months, and 6 months after surgery. FMPS and FMBS did not significantly change (p > 0.05), whereas PPD and CAL significantly decreased, and REC significantly increased in all groups during the study (p < 0.05). Clinical results were satisfactory in all cases, with no significant differences between groups 3 months after surgery. Six months after surgery, only PPD-MB was significantly different in the three groups (p < 0.05). Nevertheless, this value was not clinically relevant because the value of PPD-B (about 2 mm) in group C was physiologic. The mechanical plaque control was proven to be fundamental and sufficient in all the six aspects per tooth to guarantee an excellent clinical outcome without the need of chemical plaque control.
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
- 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
- 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.
2019
- The comparison of the proteomic profile of periodontal pocket and of corresponding gingival crevicular fluid to study periodontal disease biomarkers: feasibility study. biomarkers: feasibility study
[Abstract in Rivista]
Bertoldi, Carlo; Franceschetti, Filippo; Bergamini, Stefania; Giannetti, Luca; Generali, Luigi; Bellini, Pierantonio; Consolo, Ugo
abstract
Aim: Periodontitis is a set of inflammatory disorders characterized by periodontal attachment loss by periodontal pocket development, leading to tooth loss if remain untreated. The etiology and progress of periodontal disease is complex and remains mostly unknown. So, periodontal disease therapy has considerable limitations. The easy, reliable and correct early detection and control of the disease, markedly reduces biological and social costs. However, the diagnosis of periodontitis is established exclusively by clinical criteria based on probing to assess periodontal pockets, which are the pathognomonic expression of periodontal disease. The -omic sciences acquired substantial significance of late years and, in particular, proteomic seemed to be the more promising in this initial stage. Most proteomic analysis regarding periodontal diseases have been performed on saliva, crevicular fluid samples, peripheral blood or periodontal plaque samples which are more easily to harvest than the tissue of the periodontal pocket. However, they failed to provide reliable results for clinical applications. On the contrary, very few studies were directly performed on the periodontal pocket.
So, the aim of this study was to compare the proteomic profile of interproximal pocket tissues with that of GCF, and to analyze if they show a significant similarity in the proteomic profile.
Methods: in this preliminary study, we enrolled 3 healthy subjects affected by severe periodontitis needing of periodontal surgery. Immediately before the 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. All samples were immediately frozen at –80°C and maintained until further analysis. Tissue samples were mechanically disrupted and incubated in lysis buffer, while GCF was obtained incubating the collecting paper in phosphate buffered. In both cases, after centrifugation, the supernatant was precipitated in cold acetone overnight and protein content were pelleted by centrifugation and then dissolved in a rehydration buffer. Mono-dimensional gel electrophoresis was used to separate protein content. After staining gel images were acquired and compared. Liquid chromatography coupled to mass spectrometry (LC-MS/MS) analysis was performed to allow protein spot identification.
Results: 1-DE gels from periodontal pocket tissue and the correspondent GCF was analyzed by software Quantity One. Almost the same qualitative protein expression profile in pocket tissue and GCF was found from each patient. However, no statistical significant correlation between the quantitative proteomic profile of pocket tissue and GCF was found. Only one band (that of K immunoglobulin) resulted statistically significant between GCF and pocket tissue proteome in all patients.
Conclusions To date, this is the first study comparing the proteome of periodontal pocket tissue and corresponding GCF. The periodontal pocket and the GCF are similar as proteomic networks, but the protein network of the periodontal pocket does not influence significantly the GCF protein network and the other way around. So, with the limitations of this study, the preliminary results seem to indicate that the GCF does not seem suitable to study on the pathogenesis of periodontal disease explaining the reason for the failure of studies based only on GCF to control the periodontal disease in real-time.
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
- 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.
2018
- Validation of prostate cancer biomarkers and inflammation: a proteomic study
[Relazione in Atti di Convegno]
Ozben, T; Bergamini, S; Bellei, E; Cuoghi, A; Bianchi, G; Tomasi, A
abstract
Analysis of serum proteome to investigate possible confounding paramenters in the discrimination between prostate cancer nad benign prostatic hyperplasia
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
- Proteomic validation of biomarkers for discrimination of benign and malign prostatic hyperplasia
[Abstract in Rivista]
Ozben, T; Bergamini, S; Bellei, E; Cuoghi, A; Bianchi, G; Tomasi, A
abstract
Use of proteomic techniques for the identification and validation of discriminating protein biomarkers in prostate cancer and BPH.
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 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
- 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
- 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 (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 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 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 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
- 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
- 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
- An unusual case of signet ring cell adenocarcinoma of the prostate
[Articolo su rivista]
REGGIANI BONETTI, Luca; Lupi, Massimo; Stauder, E.; Bergamini, Stefania; Scuri, M.; Maiorana, Antonino
abstract
We report the case of a 70-year-old man with symptoms of urinary obstruction and haematuria, with histological diagnosis of primary signet-ring cell carcinoma of the prostate. Almost 90% of the tumour cells contained characteristic intracytoplasmic vacuoles that positively stained with diastase-digested PAS, Alcian blue and mucicarmine. The positive immunostaining for PSA and PSAP confirmed the prostatic origin of the tumour. Although the patient received hormonal therapy, the disease progressed and the patient died 11 months after surgery.
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
- 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.
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
- Impairment of 8-iso-PGF(2ALPHA) isoprostane metabolism by dietary conjugated linoleic acid (CLA)
[Articolo su rivista]
Iannone, Anna; A., Petroni; E., Murru; L., Cordeddu; G., Carta; M. P., Melis; Bergamini, Stefania; DELLA CASA, Lara; L., Cappiello; R., Carissimi; M., O'Shea; D., Bell; E., De Santis; S., Banni
abstract
8-iso-PGF(2alpha) isoprostane (IP) is one of the most-used markers of lipid peroxidation in experimental models and humans. After its formation, it is promptly metabolized to 2,3 dinor (DIN) in peroxisomes. Conjugated linoleic acid (CLA) is preferentially beta-oxidized in peroxisomes which may compete with IP, and thereby may affect its metabolism. In order to verify whether CLA is able to influence IP formation and/or metabolism and to explain the mechanism, we challenged rats supplemented with CLA or with triolein (as a control fatty acid), with a single dose of carbon tetrachloride (CCl(4)) or of bacterial lipopolysaccharide (LPS). The results showed that IP and its precursor arachidonic acid hydroperoxide, as well as malondialdehyde (MDA), increase significantly in the liver of rats challenged with CCl(4), irrespective of the diet, while in LPS-treated rats only nitrites in liver and isoprostane in plasma increase. On the other hand, the peroxisomal beta-oxidation products of IP, the DIN, is significantly lower in the CLA group with respect to control and triolein groups. To further investigate whether this is due to competition between CLA and IP at the cellular level, we incubated human fibroblasts from healthy subjects or patients with adrenoleukodystrophy (ALD), with CLA and/or commercially available IP. The rationale of this approach is based on the deficient peroxisomal beta-oxidation of fibroblasts from ALD patients, leading to a reduced formation of DIN. In both normal and ALD cells, the presence of CLA significantly inhibits the formation of DIN from IP. We may conclude that both in vitro and in vivo studies strongly suggest that CLA may impair IP catabolism in peroxisomes. Consequently an increase of IP, as a sole result of CLA intake, cannot be considered as a marker of lipid peroxidation.
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 AS DIAGNOSTIC MARKERS
[Abstract in Rivista]
Tomasi, Aldo; Bellei, Elisa; Bergamini, Stefania; Cuoghi, Aurora; Monari, Emanuela
abstract
non disponibile
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
2008
- Long-term n-acetylcysteine and l-arginine administration reduces endothelial activation and systolic blood pressure in hypertensive patients with type 2 diabetes
[Articolo su rivista]
V., Martina; A., Masha; V. R., Gigliardi; L., Brocato; E., Manzato; A., Berchio; P., Massarenti; F., Settanni; DELLA CASA, Lara; Bergamini, Stefania; Iannone, Anna
abstract
OBJECTIVE: Reactive oxygen and nitric oxide (NO) have recently been considered to be involved in the cardiovascular complications of patients with type 2 diabetes, as NO is thought to lose its beneficial physiological effects in the presence of oxygen radicals. For this reason, we tested the effects of l-arginine (ARG) and N-acetylcysteine (NAC) administration in increasing NO bioavailability by reducing free radical formation. RESEARCH DESIGN AND METHODS: A double-blind study was performed on 24 male patients with type 2 diabetes and hypertension divided into two groups of 12 patients that randomly received either an oral supplementation of placebo or NAC + ARG for 6 months. RESULTS: The NAC + ARG treatment caused a reduction of both systolic (P < 0.05) and diastolic (P < 0.05) mean arterial blood pressure, total cholesterol (P < 0.01), LDL cholesterol (P < 0.005), oxidized LDL (P < 0.05), high-sensitive C-reactive protein (P < 0.05), intracellular adhesion molecule (P < 0.05), vascular cell adhesion molecule (P < 0.01), nitrotyrosine (P < 0.01), fibrinogen (P < 0.01), and plasminogen activator inhibitor-1 (P < 0.05), and an improvement of the intima-media thickness during endothelial postischemic vasodilation (P < 0.02). HDL cholesterol increased (P < 0.05). No changes in other parameters studied were observed. CONCLUSIONS: NAC + ARG administration seems to be a potential well-tolerated antiatherogenic therapy because it improves endothelial function in hypertensive patients with type 2 diabetes by improving NO bioavailability via reduction of oxidative stress and increase of NO production. Our study's results give prominence to its potential use in primary and secondary cardiovascular prevention in these patients.
2007
- Metabolism of 8-iso-PGF2 alpha and conjugated linoleic acid (CLA) in vivo and in X-adrenoleukodystrophy fibroblast.
[Abstract in Rivista]
Petroni, A; Iannone, Anna; Cordeddu, L; Murru, E; Carta, G; Melis, Mp; Bergamini, Stefania; Blasevich, M; Carissimi, R; Oshea, M; DE SANTIS, E; Banni, S.
abstract
not available
2007
- Metabolism of 8-iso-PGF2 alpha isoprostane is impaired by conjugated linoleic acid (CLA)
[Abstract in Rivista]
Oshea, M; Iannone, Anna; Petroni, A; Murru, E; Cordeddu, L; Carta, G; Melis, Mp; Bergamini, Stefania; DELLA CASA, Lara; Blasevich, M; Carissimi, R; DE SANTIS, E; Banni, S.
abstract
not available
2006
- Oxidative stress in fibroblasts from patients with pseudoxanthoma elasticum: possible role in the pathogenesis of clinical manifestations
[Articolo su rivista]
Ronchetti, Ivonne; MI Garcia, Fernandez; Boraldi, Federica; Quaglino, Daniela; Gheduzzi, Dealba; De Vincenzi Paolinelli, C.; Tiozzo, Roberta; Bergamini, Stefania; D., Ceccarelli; U., Muscatello
abstract
Pseudoxanthoma elasticum (PXE) is a genetic disease characterized by calcification and fragmentation of elastic fibres of the skin, cardiovascular system and eye, caused by mutations of the ABCC6 gene, which encodes the membrane transporter MRP6. The pathogenesis of the lesions is unknown. Based on studies of similar clinical and histopathological damage present in haemolytic disorders, our working hypothesis is that PXE lesions may result from chronic oxidative stress occurring in PXE cells as a consequence of MRP6 deficiency. Our results show that PXE fibroblasts suffer from mild chronic oxidative stress due to the imbalance between production and degradation of oxidant species. The findings also show that this imbalance results, at least in part, from the loss of mitochondrial membrane potential (Delta Psi(m)) with overproduction of H2O2. Whether mitochondrial dysfunction is the main factor responsible for the oxidative stress in PXE cells remains to be elucidated. However, mild chronic generalized oxidative stress could explain the great majority of structural and biochemical alterations already reported in PXE.
2006
- Role of oxidative stress in progressive kidney failure
[Abstract in Rivista]
Tomasi, Aldo; S., Uggeri; Bergamini, Stefania; DELLA CASA, Lara; Albertazzi, Alberto; L., Lucchi; Iannone, Anna
abstract
not available
2005
- Comparison between hydroperoxides and malondialdehyde as markers of acute oxidative injury during hemodialysis
[Articolo su rivista]
L., Lucchi; Iannone, Anna; Bergamini, Stefania; L., Stipo; S., Perrone; S., Uggeri; V., Gatti; F., Ferrari; Tomasi, Aldo; Albertazzi, Alberto
abstract
An increased free-radical production has been documented during hemodialysis (HD) particularly when bio-incompatible membranes are utilized. These highly reactive free radicals can cause damage through several pathways, one of the best known being lipid peroxidation. Malondialdehyde (MDA) is a product of lipid peroxidation, which can partly be removed by HD due to its low molecular weight and water solubility. Hydroperoxides are predominantly found in lipid substances, and therefore their removal by HD could be difficult. We evaluated the behavior of these two by-products of lipid peroxidation during HD, comparing their behavior in three different membranes, in order to study their reliability as markers of acute oxidative injury. Fifteen stable HD patients were dialyzed with each of the following membranes: cuprophan, polyamide, and polysulfone, three sessions for every membrane. MDA and hydroperoxides were measured pre-HD and then both from the arterial and venous line at 8, 15, 30, and 240 min. During HD with cuprophan membrane MDA decreased significantly in the venous line compared with the arterial line at 8, 15, and 30 min (P < 0.05). At the end of HD, MDA was significantly reduced compared with MDA pre-HD (P < 0.05). Plasma hydroperoxides increased significantly in the venous line compared with the arterial line at 8, 15, 30, and 240 min (P < 0.05). At the end of HD, hydroperoxides had increased significantly as compared with pre-HD (P < 0.05). When the polyamide and polysulfone membranes were used, the behavior of MDA was similar to that found with cuprophan. Hydroperoxides were unchanged during HD using both membranes. MDA is not a reliable marker of acute oxidative injury during HD as it is removed during HD. Hydroperoxide measurement is a better marker of acute oxidative injury during HD.
2005
- Effetto della N-acetilcisteina sulla progressione dell'insufficienza renale cronica nel ratto
[Poster]
Bellei, E; Bergamini, S; Ligabue, G; Ballestri, M; Albertazzi, A; Tomasi, A; Iannone, A.
abstract
L’insufficienza renale cronica porta nel tempo ad un progressivo deterioramento della funzionalità renale, sino ad arrivare a quella che viene definita come “end-stage renal disease” (ESRD).
I meccanismi coinvolti in questo processo multifattoriale sono mediati dall’azione di diverse sostanze, fra cui: angiotensina II, TGF, endotelina, monossido d’azoto (NO), radicali liberi dell’ossigeno e citochine.
Si è voluto indagare l’effetto di una dieta arricchita con una sostanza antiossidante e anti-infiammatoria, la NAC, sulla progressione dell’insufficienza renale cronica indotta in ratti mediante asportazione chirurgica dei 5/6 del parenchima renale.
2005
- Erythrocyte susceptibility to oxidative stress in chronic renal failure patients under different substitutive treatments
[Articolo su rivista]
Lucchi, L; Bergamini, Stefania; Iannone, Anna; Perrone, S; Stipo, L; Olmeda, F; Caruso, F; Tomasi, Aldo; Albertazzi, Alberto
abstract
An increased oxidative stress is now considered one of the major risk factors in chronic renal failure (CRF) patients that may be exacerbated by dialysis. It has been postulated that this increased oxidative stress might cause an augmented red blood cell (RBC) membrane lipid peroxidation with the consequent alteration in membrane deformability. The aim of this study was to evaluate RBC susceptibility to an in vitro induced oxidative stress and RBC antioxidant potential in different groups of CRF patients undergoing different substitutive treatment modalities. Fifteen end-stage CRF patients were evaluated in conservative treatment, 23 hemodialysis (HD) patients, 15 continuous ambulatory peritoneal dialysis (CAPD) patients, 15 kidney transplanted patients, and 16 controls. Their RBCs were incubated with the oxidative stress-inducing agent tert-butylhydroperoxide both in the presence and in the absence of the catalase inhibitor sodium azide, and the level of malondialdehyde (MDA) (a product of lipid peroxidation), was measured at 0, 5, 10, 15, and 30 min of incubation. In addition, the RBC content of reduced glutathione (GSH) was measured by HPLC. As opposed to the controls, RBCs from end-stage CRF patients exhibited an increased sensitivity to oxidative stress induced in vitro, both in the absence and presence of a catalase inhibitor, as demonstrated by a significantly higher level of MDA production at all the incubation times (P < 0.05). Different substitutive treatments had different impacts on this phenomenon; CAPD and kidney transplantation were able to normalize this alteration while HD was not. GSH appeared to be related to the increase in RBC susceptibility to oxidative stress; its content being significantly elevated in end-stage CRF and HD patients as compared with CAPD and transplanted patients and controls (P < 0.05). No significant changes were observed in the RBC glutathione content during the HD session. The increase of GSH in RBCs of end-stage CRF and HD patients seems to indicate the existence of an adaptive mechanism under increased oxidative stress occurring in vivo. Unlike HD, the beneficial effect of CAPD on the anemia of dialysis patients might partly be due to a condition of lower oxidative stress that might in addition counterbalance the cardiovascular negative effects of dislipidemia of CAPD patients.
2005
- New insights on the pathogenesis of clinical manifestations in Pseudoxathoma elasticum.
[Abstract in Rivista]
Ronchetti, Ivonne; M. I., GARCIA FERNANDEZ; Boraldi, Federica; Quaglino, Daniela; Gheduzzi, Dealba; PAOLINELLI DEVINCENZI, Chiara; Tiozzo, Roberta; Bergamini, Stefania; Ceccarelli, Daniela; Muscatello, Umberto
abstract
Data are presented on the presence of oxidative stress in fibroblasts from PXE patients and the potential role in PXE pathogenesis is discussed.
2005
- No evidences of oxidative injury associated with slow intravenous administration of ferric gluconate during haemodialysis
[Relazione in Atti di Convegno]
Bergamini, Stefania; DELLA CASA, Lara; Iannone, Anna; Albertazzi, Alberto
abstract
No evidences of oxidative injury associated with slow intravenous administration of ferric gluconate during haemodialysis
2005
- Oxidative stress and experimental shock
[Abstract in Rivista]
Tomasi, Aldo; Bergamini, Stefania; Bellei, Elisa; Bazzani, Carla; Bertolini, Alfio; Guarini, Salvatore; Iannone, Anna
abstract
In this introductory presentation, some of the recent results of our and other groups on the oxidative stress theory in the development of experimental shock, will be presented and discussed.
2005
- Oxidative stress in renal patients
[Abstract in Rivista]
Iannone, Anna; Bergamini, Stefania; Bellei, Elisa; Rota, Cristina; DELLA CASA, Lara; Tomasi, Aldo
abstract
non disponibile
2004
- Chronic inflammation in end stage renal disease: markers of oxidative stress and redox modification
[Abstract in Atti di Convegno]
Iannone, A; Bergamini, S; Bellei, E; Rota, C; Tomasi, A
abstract
Markers of oxidative stress and redox modification in patients with end stage renal disease and chronic inflammation
2004
- Effect of alpha-tocopherol and N-acetylcysteine on benzoyl peroxide toxicity in human keratinocytes
[Articolo su rivista]
Bellei, Elisa; Rota, Cristina; Bergamini, Stefania; P., Manfredini; Albertazzi, Alberto; Tomasi, Aldo; Iannone, Anna
abstract
Benzoyl peroxide is a free-radical generating compound widely used in the polymer industry and also in pharmaceuticals as antimicrobial agent to treat acne. However, benzoyl peroxide causes irritation and contact dermatitis in about 1% of patients. Concern over the use of this compound is motivated by the demonstration that it can also act as skin tumor promoter in mice. In addition, benzoyl peroxide induces DNA strand breaks in many cells, including keratinocytes. Benzoyl peroxide toxicity is presumably mediated by the formation of reactive free radicals and by the consumption of intracellular antioxidants. In this work we investigated the effect of both the lipophilic antioxidant et-tocopherol and the hydrophilic thiol donor N-acetylcysteine (NAC) in human keratinocyte line HaCaT exposed to benzoyl peroxide. A protective effect against benzoyl peroxide cytotoxicity was achieved when cells were grown on a alpha-tocopherol layer. On the contrary, the addition of alpha-tocopherol dissolved in ethanol had a pro-oxidant effect, leading to an enhancement of benzoyl peroxide toxicity. Cytotoxicity was also reduced adding NAC to the culture medium; the presence of both NAC and alpha-tocopherol exerts a synergistic cytoprotection.
2004
- Effetti della N-acetilcisteina in condizioni di stress ossidativo
[Abstract in Atti di Convegno]
Iannone, A; Bellei, E; Bergamini, S; Rota, C; Tomasi, A; Albertazzi, A.
abstract
Effetti della N-acetilcisteina in condizioni di stress ossidativo
2004
- Markers of inflammation in end stage renal disease patients. A role for antioxidants
[Abstract in Atti di Convegno]
Iannone, A; Bellei, E; Bergamini, S; Rota, C; Tomasi, A
abstract
Markers of inflammation in end stage renal disease patients. A role for antioxidants
2004
- Markers of oxidative stress and antioxidant status in haemodialysis patients
[Relazione in Atti di Convegno]
Iannone, A; Bergamini, S; Bellei, E; Rota, C; Lucchi, L; Albertazzi, A; Tomasi, A
abstract
Study of oxidative stress markers and antioxidant status in haemodialysis patients
2004
- Redox regulation and antioxidant in shock
[Abstract in Atti di Convegno]
Iannone, A; Bellei, E; Bergamini, S; Rota, C; Tomasi, A
abstract
Redox regulation and antioxidant in shock
2004
- Relationship of asymmetric dimethylarginine to haemodialysis hypotension
[Articolo su rivista]
Bergamini, Stefania; L., Vandelli; Bellei, Elisa; Rota, Cristina; P., Manfredini; Tomasi, Aldo; Albertazzi, Alberto; Iannone, Anna
abstract
Hypotension is one of the major complications in patients undergoing haemodialysis (HD), that is well evident in patients defined as hypotension-prone. The mechanisms underlying the hypotensive episodes are not known. We carried out a clinical study on hypotension-prone HD patients to test the existence of a dysregulation in the nitric oxide (NO) generating pathway. Since asymmetric dimethylarginine (ADMA) is an endogenous compound which regulates NO synthesis, we measured its variation in plasma of stable-HD and hypotension-prone patients before, during, and at the end of HD. Before HD, the hypotension-prone patients have higher ADMA levels than stable-HD patients. The HD procedure significantly removes ADMA from plasma of stable-HD patients, while in the hypotension-prone ADMA levels are unchanged at the end of the HD. Moreover, in the hypotension-prone patients, during the hypotensive episode, a dramatic drop of ADMA levels is observed, followed by a rapid increase at the end of the HD. The symmetric dimethylarginine (SDMA), which has no effect on NO synthesis, is also high in plasma of both groups of HD patients compared to normal subjects, and in both groups its levels at the end of HD are significantly reduced. The hypotension-prone patients have basal TNF-alpha levels lower than the stable-HD groups, that significantly increase during the hypotensive episode. On the basis of these findings, we suggest that the hypotensive syndrome could be related to a dysregulation between ADMA metabolism and clearance due both to cytokines release and to an extremely fast ADMA clearance during HD, leading to an increase in NO blood levels.
2004
- Role of nitric oxide generating pathway in haemodialysis hypotension
[Poster]
Bergamini, S; Rota, C; Bellei, E; Albertazzi, A; Vandelli, L; Tomasi, A; Iannone, A
abstract
Role of nitric oxide in haemodialysis hypotension
2003
- Measuring oxidative stress and redox state in patients: meaning and perspectives
[Poster]
Tomasi, A; Bergamini, S; Bellei, E; Rota, C; Iannone, A.
abstract
Measuring oxidative stress and redox state in patients with ESRD.
2003
- Nitric oxide and hypotension in haemodialysis patients.
[Abstract in Rivista]
Iannone, Anna; Bellei, Elisa; Bergamini, Stefania; Rota, C.; Vandelli, L.; Albertazzi, Alberto; Tomasi, Aldo
abstract
proceedings
2003
- Plasma antioxidants modifications during haemodialysis
[Poster]
Bellei, E; Manfredini, P; Lucchi, L; Bergamini, S; Rota, C; Tomasi, A; Vandelli, L; Albertazzi, A; Banni, S; Iannone, A.
abstract
The high incidence of atherosclerosis (stroke and ischemic heart disease) in patients with end-stage renal failure (ESRF), associated with clinical and/or laboratory signs of systemic inflammation, has been considered to be in part due to the long-term haemodialysis (HD).
The contact with the artificial surface of the extra-corporeal circuit may promote free radicals formation, which is responsible for the increased lipid peroxidation observed in these patients.
It has been also demonstrated that they have a decreased intracellular antioxidant capability.
To test the presence of oxidative stress and evaluate also the antioxidant status during the dialytic session, we performed a clinical study in a group of functionally anephric patients on maintenance HD treatment with three weekly HD sessions, using a low-flux cellulose diacetate membrane and a freshly prepared bicarbonate buffer.
2003
- Redox regulation, cytokines, and nitric oxide in inflammation
[Capitolo/Saggio]
Tomasi, Aldo; Bergamini, Stefania; Rota, C.; Iannone, Anna
abstract
non disponibile
2003
- “Role of NO in hypotension prone dialytic patients”
[Abstract in Rivista]
Iannone, Anna; Bellei, Elisa; Bergamini, Stefania; Rota, Cristina; Vandelli, Lorenza; Albertazzi, Alberto; Tomasi, Aldo
abstract
not available
2002
- Antioxidants modulates benzoyl-peroxide toxicity in human keratinocytes
[Abstract in Rivista]
Manfredini, P; Bellei, E; Bergamini, S; Rota, C; Iannone, A; Tomasi, A; Albertazzi, A.
abstract
Benzoyl peroxide (BP) is a free-radical generating compound, widely used in the polymer industry and also in pharmaceuticals as antimicrobial agent to treat acne. However, BP causes irritation and contact dermatitis in about 1% of patients. Concern over the use of this compound is motivated by the demonstration that it can also act as skin tumor promoter in mice. In addition, BP induces DNA strand breaks in many cells, included keratinocytes. In vitro, the tossicity associated with BP is presumably mediated by formation of benzoyloxyl radical, responsable for membrane damage.
In this work we investigated the effect of both the lipophilic antioxidant a-tocopherol and the thiol donor N-acetylcysteine (NAC) in human keratinocyte line HaCaT exposed to BP.
2002
- N-Acetylcysteine negatively modulates nitric oxide production in endotoxin-treated rats through inhibition of NF-ĸB activation
[Articolo su rivista]
C., Rota; Bergamini, Stefania; F., Daneri; Tomasi, Aldo; F., Virgili; Iannone, Anna
abstract
N-Acetylcysteine (NAC) has a wide spectrum of biological activities, either related to the ability to increase intracellular thiols or directly acting as an antioxidant. We used an in vivo animal model to study NAC modulation of nitric oxide (NO) production in response to lipopolysaccharide treatment. A comparison was made between NAC and the N-[3-(aminomethyl)benzyl] acetamidine (1400W), an inhibitor of the inducible NO synthase (iNOS). Both inhibit NO production, although NAC lacks any effect if given when iNOS is already induced; this indicates that the decrease of NO generation is not due to an effect on iNOS activity. We found that the DNA binding activity of nuclear transcription factor-kappaB in peripheral blood cells was inhibited by NAC given before lipopolysaccharide, whereas tumor necrosis factor-a secretion was not affected.
2002
- “VITAMIN E MODULATES BENZOYL PEROXIDE TOXICITY IN HUMAN KERATINOCYTES”
[Relazione in Atti di Convegno]
Manfredini, P; Bellei, Elisa; Bergamini, Stefania; Rota, Cristina; Tomasi, Aldo; Albertazzi, Alberto; Iannone, Anna
abstract
not available
2001
- Cytokine and nitric oxide redox regulation in vivo
[Abstract in Atti di Convegno]
Tomasi, A; Bellei, E; Bergamini, S; Rota, C; Iannone, A.
abstract
Inflammation and oxidative stress, at the molecular level, induce peroxidation and modify redox equlibrium.
2001
- N-Acetylcysteine inhibits in vivo nitric oxide production by inducible nitric oxide synthase
[Articolo su rivista]
Bergamini, Stefania; C., Rota; R., Canali; Mg, Staffieri; F., Daneri; Bini, Anna; F., Giovannini; Tomasi, Aldo; Iannone, Anna
abstract
non disponibile
2000
- Influence of different hemodialysis membranes on red blood cell susceptibility to oxidative stress
[Articolo su rivista]
L., Lucchi; Bergamini, Stefania; B., Botti; R., Rapana; A., Ciuffreda; P., Ruggiero; M., Ballestri; Tomasi, Aldo; Albertazzi, Alberto
abstract
Oxidative stress is crucial in red blood cell (RBC) damage induced by activated neutrophils in in vitro experiments. The aim of the study was to evaluate whether the bioincompatibility phenomena occurring during hemodialysis (HD) (where neutrophil activation with increased free radical production is well documented) may have detrimental effects on RBC. We evaluated RBC susceptibility to oxidative stress before and after HD in 15 patients using Cuprophan, cellulose triacetate, and polysulfone membrane. RBC were incubated with t-butyl hydroperoxide as an oxidizing agent both in the presence and in the absence of the catalase inhibitor sodium azide. The level of malonaldehyde (MDA), a product of lipid peroxidation, was measured at 0, 5, 10, 15, and 30 min of incubation. When Cuprophan membrane was used, the MDA production was significantly higher after HD, indicating an increased susceptibility to oxidative stress in comparison to pre-I-ID. The addition of sodium azide enhanced this phenomenon. Both cellulose triacetate and polysulfone membranes did not significantly influence RBC susceptibility to oxidative stress. Neither the level of RBC reduced glutathione nor the RBC glutathione redox ratio changed significantly during HD with any of the membranes used. The RBC susceptibility to oxidative stress was influenced in different ways according to the dialysis membrane used, being increased only when using the more bioincompatible membrane Cuprophan, where neutrophil activation with increased free radical production is well documented. The alterations found in this study might contribute to the reduced RBC longevity of HD patients where a bioincompatible membrane is used.
1999
- Prooxidant activity of ferrioxamine in isolated rat hepatocytes and linoleic acid micelles
[Articolo su rivista]
Bergamini, Stefania; C., Rota; M., Staffieri; Tomasi, Aldo; Iannone, Anna
abstract
The complex iron-desferrioxamine (ferrioxamine) is considered chemically unreactive, and not able to participate in redox cycle reactions. Desferrioxamine-dependent toxicity is, however, described in both human and animal studies. The aim of this work was to test the possibility that chelated iron, under certain circumstances, could enter redox reactions, giving an explanation of desferrioxamine side effects, Carefully prepared ferrioxamine, to obtain a 1:1 desferrioxamine:iron ratio, was added to isolated rat hepatocytes and to linoleic acid micelles. A strong prooxidant and cytotoxic effect was observed in the cells, also potentiating tert-butyl hydroperoxide-induced lipid peroxidation. In micelles, the prooxidant effect was observed only in the presence of ascorbate, which is oxidized during the process, giving rise to ascorbyl radical. Ferrioxamine, under the experimental conditions used, did not release iron, indicating that the prooxidant effect was due to iron redox cycling. The addition of desferrioxamine prevented both ferrioxamine- and tert-butyl hydroperoxide-induced lipid peroxidation and cytotoxicity. Concurrently, a nitroxide radical was detected, an indication of the radical scavenger activity of the hydroxamic moiety. No radical species was observed when ferrioxamine was added to the same system. The prooxidant effect of ferrioxamine gives a possible explanation of the reported human and animal desferrioxamine toxicity. When, in compartmentalized regions, the ratio of desferrioxamine:metal reaches 1:1, ferrioxamine is formed. In the absence of metal-free desferrioxamine, ferrioxamine can participate in redox cycling reactions, initiating lipid peroxidation and cytotoxicity.
1998
- Antioxidant activity of carotenoids: an electron spin resonance study of beta-carotene and lutein interaction with free radicals generated in a chemical system
[Articolo su rivista]
Iannone, Anna; C., Rota; Bergamini, Stefania; Tomasi, Aldo; L. M., Canfield
abstract
b-Carotene is thought to be a chainbreaking
antioxidant, even though we have no information
about the mechanism of its antioxidant activity.
Using electron-spin resonance (ESR) spectroscopy coupled
to the spin-trapping technique, we have studied
the effect of b-carotene and lutein on the radical adducts
of the spin-trap PBN (N-t-butyl-a-phenylnitrone)
generated by the metal-ion breakdown of
different tert-butyl hydroperoxide (tBOOH) concentrations
in methylene chloride. The peroxyl radical,
along with an oxidation product of PBN (the PBNOx),
trapped at room temperature from the breakdown of
high concentration of tBOOH (1 M), were quenched
by b-carotene or lutein, in competition with the spintrapping
agent. However, carotenoids were not able to
quench the alkoxyl and methyl radicals generated in
the reaction carried out in the presence of low tBOOH
concentration (1 mM). The reaction between carotenoids
and the peroxyl radical was also carried out in
the absence of the spin trap, at 77 K: Under these different
experimental conditions, we did not detect any
radical species deriving from carotenoids. In the same
system, a further evidence of the peroxyl radical
quenching by b-carotene and lutein was obtained. The
antioxidant activity of vitamin E was also tested, for
comparison with the carotenoids. In the presence ofatocopherol,
peroxyl and alkoxyl radicals were
quenched, and the tocopheroxyl radical was detected.
Our data provide the first direct evidence that carotenoids
quench peroxyl radicals. Under our experimental
conditions, we did not detect any carotenoid radical
species that could derive from the interaction with the
peroxyl radical. The radical-trapping activity of b-carotene
and lutein demonstrated in this chemical reaction
contributes to our understanding carotenoid antioxidant
action in biological systems.
1998
- “The highly reducing sugar 2-deoxy-d-ribose induces apoptosis in human fibroblasts by reduced glutathione depletion and cytoskeletal disruption”
[Articolo su rivista]
D., Kletsas; D., Barbieri; D., Stathakos; B., Botti; Bergamini, Stefania; Tomasi, Aldo; D., Monti; W., Malorni; C., Franceschi
abstract
2-deoxy-D-Ribose (dRib), the most reducing sugar, induces apoptosis in normal human fibroblasts, as judged by cytoplasmic shrinkage, chromatin condensation, DNA fragmentation and mitochondrial depolarization. This effect is independent from culture conditions, such as cell density and the presence or absence of serum in the culture milieu, suggesting that dRib-induced apoptosis is cell cycle-independent. dRib was found also to provoke disruption of the actin filament network and detachment from the substratum, while at the same time, interestingly, it increases the expression of several integrins and cell adhesion molecules. Furthermore, dRib was found to reduce the intracellular levels of reduced glutathione (GSH). The apoptotic process was not affected by the macromolecular-synthesis inhibitors cycloheximide and actinomycin D. On the contrary, the antioxidant N-acetyl-L-cysteine (NAG) fully blocks the dRib-induced apoptosis by preventing GSH depletion, while it also inhibits actin-filament-network disruption and mitochondrial depolarization. The above indicate that dRib induces apoptosis in human fibroblasts by a mechanism involving glutathione metabolism and oxidative stress, as well as disturbance of cytoskeletal integrity and cell adhesion.