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Article: Protein thiolation index in microvolumes of plasma

Daniela Giustarini / Dalle-Donne, Isabella / Colombo, Graziano / Milzani, Aldo / Santucci, Annalisa / Rossi, Ranieri

Analytical biochemistry. 2021 Apr. 01, v. 618

2021

Abstract: Protein Thiolation Index (PTI) has been recently proposed as a new biomarker of oxidative stress. It is calculated by measuring both free thiols and S-thiolated proteins in plasma with the assumption that this redox ratio is altered by a pro-oxidant ... ...

Abstract	Protein Thiolation Index (PTI) has been recently proposed as a new biomarker of oxidative stress. It is calculated by measuring both free thiols and S-thiolated proteins in plasma with the assumption that this redox ratio is altered by a pro-oxidant stimulus. Here the original protocol was modified and adapted to the use of microvolumes of blood collected by finger prick and down to 3 μl blood was shown to be the lowest volume suitable for this kind of analysis. The new procedure was used to evaluate both the circadian rhythm and the annual fluctuations of PTI in healthy humans.
Keywords	biomarkers ; circadian rhythm ; oxidative stress ; thiols
Language	English
Dates of publication	2021-0401
Publishing place	Elsevier Inc.
Document type	Article
ZDB-ID	1110-1
ISSN	1096-0309 ; 0003-2697
ISSN (online)	1096-0309
ISSN	0003-2697
DOI	10.1016/j.ab.2021.114125
Database	NAL-Catalogue (AGRICOLA)

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Article ; Online: Protein thiolation index in microvolumes of plasma.

Daniela Giustarini / Dalle-Donne, Isabella / Colombo, Graziano / Milzani, Aldo / Santucci, Annalisa / Rossi, Ranieri

Analytical biochemistry

2021 Volume 618, Page(s) 114125

Abstract	Protein Thiolation Index (PTI) has been recently proposed as a new biomarker of oxidative stress. It is calculated by measuring both free thiols and S-thiolated proteins in plasma with the assumption that this redox ratio is altered by a pro-oxidant stimulus. Here the original protocol was modified and adapted to the use of microvolumes of blood collected by finger prick and down to 3 μl blood was shown to be the lowest volume suitable for this kind of analysis. The new procedure was used to evaluate both the circadian rhythm and the annual fluctuations of PTI in healthy humans.
MeSH term(s)	Adult ; Aged ; Blood Proteins/chemistry ; Female ; Humans ; Male ; Middle Aged ; Plasma/chemistry ; Sulfhydryl Compounds/chemistry
Chemical Substances	Blood Proteins ; Sulfhydryl Compounds
Language	English
Publishing date	2021-01-30
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	1110-1
ISSN	1096-0309 ; 0003-2697
ISSN (online)	1096-0309
ISSN	0003-2697
DOI	10.1016/j.ab.2021.114125
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: The age-dependent decline of the extracellular thiol-disulfide balance and its role in SARS-CoV-2 infection

Daniela Giustarini / Annalisa Santucci / Desirée Bartolini / Francesco Galli / Ranieri Rossi

Redox Biology, Vol 41, Iss , Pp 101902- (2021)

2021

Abstract: SARS–CoV-2 (COVID-19) infection can cause a severe respiratory distress syndrome. The risk of severe manifestations and mortality characteristically increase in the elderly and in the presence of non-COVID-19 comorbidity. We and others previously ... ...

Abstract	SARS–CoV-2 (COVID-19) infection can cause a severe respiratory distress syndrome. The risk of severe manifestations and mortality characteristically increase in the elderly and in the presence of non-COVID-19 comorbidity. We and others previously demonstrated that the low molecular weight (LMW) and protein thiol/disulfide ratio declines in human plasma with age and such decline is even more rapid in the case of inflammatory and premature aging diseases, which are also associated with the most severe complications of COVID-19 infection. The same decline with age of the LMW thiol/disulfide ratio observed in plasma appears to occur in the extracellular fluids of the respiratory tract and in association with many pulmonary diseases that characteristically reduce the concentrations and adaptive stress response of the lung glutathione. Early evidence in literature suggests that the thiol to disulfide balance of critical Cys residues of the COVID-19 spike protein and the ACE-2 receptor may influence the risk of infection and the severity of the disease, with a more oxidizing environment producing the worst prognosis.With this hypothesis paper we propose that the age-dependent decline of LMW thiol/disulfide ratio of the extracellular fluids, could play a role in promoting the physical (protein-protein) interaction of CoV-2 and the host cell in the airways. Therefore, this redox-dependent interaction is expected to affect the risk of severe infection in an age-dependent manner. The hypothesis can be verified in experimental models of in vitro CoV-2 infection and at the clinical level in that LMW thiols and protein thiolation can now be investigated with standardized, reliable and versatile laboratory protocols. Presenting the verification strategy of our hypothesis, we also discuss available nutritional and ancillary pharmacological strategies to intervene on the thiol/disulfide ratio of extracellular fluids of subjects at risk of infection and COVID-19 patients.
Keywords	SARS-CoV-2 ; COVID-19 ; Spike S protein ; ACE-2 receptor ; Aging ; Oxidative stress ; Medicine (General) ; R5-920 ; Biology (General) ; QH301-705.5
Subject code	610
Language	English
Publishing date	2021-05-01T00:00:00Z
Publisher	Elsevier
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Blood Thiol Redox State in Chronic Kidney Disease

Maria Lisa Garavaglia / Daniela Giustarini / Graziano Colombo / Francesco Reggiani / Silvia Finazzi / Marta Calatroni / Lucia Landoni / Nicola Marcello Portinaro / Aldo Milzani / Salvatore Badalamenti / Ranieri Rossi / Isabella Dalle-Donne

International Journal of Molecular Sciences, Vol 23, Iss 2853, p

2022 Volume 2853

Abstract: Thiols (sulfhydryl groups) are effective antioxidants that can preserve the correct structure of proteins, and can protect cells and tissues from damage induced by oxidative stress. Abnormal levels of thiols have been measured in the blood of patients ... ...

Abstract	Thiols (sulfhydryl groups) are effective antioxidants that can preserve the correct structure of proteins, and can protect cells and tissues from damage induced by oxidative stress. Abnormal levels of thiols have been measured in the blood of patients with moderate-to-severe chronic kidney disease (CKD) compared to healthy subjects, as well as in end-stage renal disease (ESRD) patients on haemodialysis or peritoneal dialysis. The levels of protein thiols (a measure of the endogenous antioxidant capacity inversely related to protein oxidation) and S -thiolated proteins (mixed disulphides of protein thiols and low molecular mass thiols), and the protein thiolation index (the molar ratio of the S -thiolated proteins to free protein thiols in plasma) have been investigated in the plasma or red blood cells of CKD and ESRD patients as possible biomarkers of oxidative stress. This type of minimally invasive analysis provides valuable information on the redox status of the less-easily accessible tissues and organs, and of the whole organism. This review provides an overview of reversible modifications in protein thiols in the setting of CKD and renal replacement therapy. The evidence suggests that protein thiols, S -thiolated proteins, and the protein thiolation index are promising biomarkers of reversible oxidative stress that could be included in the routine monitoring of CKD and ESRD patients.
Keywords	chronic kidney disease ; end-stage renal disease ; haemodialysis ; peritoneal dialysis ; protein thiols ; S -thiolated proteins ; Biology (General) ; QH301-705.5 ; Chemistry ; QD1-999
Language	English
Publishing date	2022-03-01T00:00:00Z
Publisher	MDPI AG
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Effects of Physiological and Pathological Urea Concentrations on Human Microvascular Endothelial Cells

Graziano Colombo / Alessandra Altomare / Emanuela Astori / Lucia Landoni / Maria Lisa Garavaglia / Ranieri Rossi / Daniela Giustarini / Maria Chiara Lionetti / Nicoletta Gagliano / Aldo Milzani / Isabella Dalle-Donne

International Journal of Molecular Sciences, Vol 24, Iss 1, p

2022 Volume 691

Abstract: Urea is the uremic toxin accumulating with the highest concentration in the plasma of chronic kidney disease (CKD) patients, not being completely cleared by dialysis. Urea accumulation is reported to exert direct and indirect side effects on the ... ...

Abstract	Urea is the uremic toxin accumulating with the highest concentration in the plasma of chronic kidney disease (CKD) patients, not being completely cleared by dialysis. Urea accumulation is reported to exert direct and indirect side effects on the gastrointestinal tract, kidneys, adipocytes, and cardiovascular system (CVS), although its pathogenicity is still questioned since studies evaluating its side effects lack homogeneity. Here, we investigated the effects of physiological and pathological urea concentrations on a human endothelial cell line from the microcirculation (Human Microvascular Endothelial Cells-1, HMEC-1). Urea (5 g/L) caused a reduction in the proliferation rate after 72 h of exposure and appeared to be a potential endothelial-to-mesenchymal transition (EndMT) stimulus. Moreover, urea induced actin filament rearrangement, a significant increase in matrix metalloproteinases 2 (MMP-2) expression in the medium, and a significant up- or down-regulation of other EndMT biomarkers (keratin, fibrillin-2, and collagen IV), as highlighted by differential proteomic analysis. Among proteins whose expression was found to be significantly dysregulated following exposure of HMEC-1 to urea, dimethylarginine dimethylaminohydrolase (DDAH) and vasorin turned out to be down-regulated. Both proteins have been directly linked to cardiovascular diseases (CVD) by in vitro and in vivo studies. Future experiments will be needed to deepen their role and investigate the signaling pathways in which they are involved to clarify the possible link between CKD and CVD.
Keywords	urea ; HMEC-1 ; CVD ; CKD ; vasorin ; differential proteomics ; Biology (General) ; QH301-705.5 ; Chemistry ; QD1-999
Subject code	570 ; 610
Language	English
Publishing date	2022-12-01T00:00:00Z
Publisher	MDPI AG
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Nitric Oxide-Related Oxidative Stress and Redox Status in Health and Disease

Darko Modun / Daniela Giustarini / Dimitrios Tsikas

Oxidative Medicine and Cellular Longevity, Vol

2014 Volume 2014

Keywords	Cytology ; QH573-671 ; Biology (General) ; QH301-705.5 ; Science ; Q
Language	English
Publishing date	2014-01-01T00:00:00Z
Publisher	Hindawi Publishing Corporation
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Superior Properties of N-Acetylcysteine Ethyl Ester over N-Acetyl Cysteine to Prevent Retinal Pigment Epithelial Cells Oxidative Damage

Gian Marco Tosi / Daniela Giustarini / Lorenzo Franci / Alberto Minetti / Francesco Imperatore / Elena Caldi / Paolo Fiorenzani / Anna Maria Aloisi / Anna Sparatore / Ranieri Rossi / Mario Chiariello / Maurizio Orlandini / Federico Galvagni

International Journal of Molecular Sciences, Vol 22, Iss 2, p

2021 Volume 600

Abstract: Oxidative stress plays a key role in the pathophysiology of retinal diseases, including age-related macular degeneration (AMD) and diabetic retinopathy, which are the major causes of irreversible blindness in developed countries. An excess of reactive ... ...

Abstract	Oxidative stress plays a key role in the pathophysiology of retinal diseases, including age-related macular degeneration (AMD) and diabetic retinopathy, which are the major causes of irreversible blindness in developed countries. An excess of reactive oxygen species (ROS) can directly cause functional and morphological impairments in retinal pigment epithelium (RPE), endothelial cells, and retinal ganglion cells. Antioxidants may represent a preventive/therapeutic strategy and reduce the risk of progression of AMD. Among antioxidants, N-acetyl-L-cysteine (NAC) is widely studied and has been proposed to have therapeutic benefit in treating AMD by mitigating oxidative damage in RPE. Here, we demonstrate that N-acetyl-L-cysteine ethyl ester (NACET), a lipophilic cell-permeable cysteine derivative, increases the viability in oxidative stressed RPE cells more efficiently than NAC by reacting directly and more rapidly with oxidizing agents, and that NACET, but not NAC, pretreatment predisposes RPE cells to oxidative stress resistance and increases the intracellular reduced glutathione (GSH) pool available to act as natural antioxidant defense. Moreover, we demonstrate the ability of NACET to increase GSH levels in rats’ eyes after oral administration. In conclusion, even if experiments in AMD animal models are still needed, our data suggest that NACET may play an important role in preventing and treating retinal diseases associated with oxidative stress, and may represent a valid and more efficient alternative to NAC in therapeutic protocols in which NAC has already shown promising results.
Keywords	oxidative stress ; retinopathy ; retinal pigment epithelium ; age-related macular degeneration ; N-acetyl-L-cysteine ; N-acetyl-L-cysteine ethyl ester ; Biology (General) ; QH301-705.5 ; Chemistry ; QD1-999
Subject code	610
Language	English
Publishing date	2021-01-01T00:00:00Z
Publisher	MDPI AG
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Antihypertensive, cardio- and neuro-protective effects of Tenebrio molitor (Coleoptera

Federica Pessina / Maria Frosini / Paola Marcolongo / Fabio Fusi / Simona Saponara / Alessandra Gamberucci / Massimo Valoti / Daniela Giustarini / Paolo Fiorenzani / Beatrice Gorelli / Valeria Francardi / Maurizio Botta / Elena Dreassi

PLoS ONE, Vol 15, Iss 5, p e

Tenebrionidae) defatted larvae in spontaneously hypertensive rats.

2020 Volume 0233788

Abstract: In pre-hypertension, moderate control of blood pressure (BP) can be obtained by a nutritional approach. The effects of a diet enriched with defatted larvae of the mealworm Tenebrio molitor (Coleoptera: Tenebrionidae) (TM) endowed with ACE inhibitory ... ...

Abstract	In pre-hypertension, moderate control of blood pressure (BP) can be obtained by a nutritional approach. The effects of a diet enriched with defatted larvae of the mealworm Tenebrio molitor (Coleoptera: Tenebrionidae) (TM) endowed with ACE inhibitory activity was studied in both spontaneously hypertensive rats (SHR) and in the age-matched normotensive Wistar Kyoto strain. These were fed for 4 weeks with standard laboratory rodent chow supplemented with or without TM or captopril. In SHR, the TM diet caused a significant reduction in BP, heart rate and coronary perfusion pressure, as well as an increase in red blood cell glutathione/glutathione disulphide ratio. Rat brain slices of SHR were more resistant to oxidative stress and contained lower levels of inflammatory cytokines, while vascular and liver enzyme-activities were not affected. These results suggest that TM can be considered a new functional food that can lower BP in vivo and thus control cardiovascular-associated risk factors such as hypertension.
Keywords	Medicine ; R ; Science ; Q
Subject code	610
Language	English
Publishing date	2020-01-01T00:00:00Z
Publisher	Public Library of Science (PLoS)
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: SARS-CoV2 infection impairs the metabolism and redox function of cellular glutathione

Desirée Bartolini / Anna Maria Stabile / Sabrina Bastianelli / Daniela Giustarini / Sara Pierucci / Chiara Busti / Carmine Vacca / Anna Gidari / Daniela Francisci / Roberto Castronari / Antonella Mencacci / Manlio Di Cristina / Riccardo Focaia / Samuele Sabbatini / Mario Rende / Antimo Gioiello / Gabriele Cruciani / Ranieri Rossi / Francesco Galli

Redox Biology, Vol 45, Iss , Pp 102041- (2021)

2021

Abstract: Viral infections sustain their replication cycle promoting a pro-oxidant environment in the host cell. In this context, specific alterations of the levels and homeostatic function of the tripeptide glutathione have been reported to play a causal role in ... ...

Abstract	Viral infections sustain their replication cycle promoting a pro-oxidant environment in the host cell. In this context, specific alterations of the levels and homeostatic function of the tripeptide glutathione have been reported to play a causal role in the pro-oxidant and cytopathic effects (CPE) of the virus. In this study, these aspects were investigated for the first time in SARS-CoV2-infected Vero E6 cells, a reliable and well-characterized in vitro model of this infection. SARS-CoV2 markedly decreased the levels of cellular thiols, essentially lowering the reduced form of glutathione (GSH). Such an important defect occurred early in the CPE process (in the first 24 hpi). Thiol analysis in N-acetyl-Cys (NAC)-treated cells and membrane transporter expression data demonstrated that both a lowered uptake of the GSH biosynthesis precursor Cys and an increased efflux of cellular thiols, could play a role in this context. Increased levels of oxidized glutathione (GSSG) and protein glutathionylation were also observed along with upregulation of the ER stress marker PERK. The antiviral drugs Remdesivir (Rem) and Nelfinavir (Nel) influenced these changes at different levels, essentially confirming the importance or blocking viral replication to prevent GSH depletion in the host cell. Accordingly, Nel, the most potent antiviral in our in vitro study, produced a timely activation of Nrf2 transcription factor and a GSH enhancing response that synergized with NAC to restore GSH levels in the infected cells. Despite poor in vitro antiviral potency and GSH enhancing function, Rem treatment was found to prevent the SARS-CoV2-induced glutathionylation of cellular proteins. In conclusion, SARS-CoV2 infection impairs the metabolism of cellular glutathione. NAC and the antiviral Nel can prevent such defect in vitro.
Keywords	COVID-19 ; SARS-CoV-2 ; Glutathione ; Thiols ; Nrf2 ; Protein glutathionylation ; Medicine (General) ; R5-920 ; Biology (General) ; QH301-705.5
Subject code	570
Language	English
Publishing date	2021-09-01T00:00:00Z
Publisher	Elsevier
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: A seleno-hormetine protects bone marrow hematopoietic cells against ionizing radiation-induced toxicities.

Desirée Bartolini / Yanzhong Wang / Jie Zhang / Daniela Giustarini / Ranieri Rossi / Gavin Y Wang / Pierangelo Torquato / Danyelle M Townsend / Kenneth D Tew / Francesco Galli

PLoS ONE, Vol 14, Iss 4, p e

2019 Volume 0205626

Abstract: 2,2'-diselenyldibenzoic acid (DSBA) is a chemical probe produced to explore the pharmacological properties of diphenyldiselenide-derived agents with seleno-hormetic activity undergoing preclinical development. The present study was designed to verify in ... ...

Abstract	2,2'-diselenyldibenzoic acid (DSBA) is a chemical probe produced to explore the pharmacological properties of diphenyldiselenide-derived agents with seleno-hormetic activity undergoing preclinical development. The present study was designed to verify in vivo the drug's properties and to determine mechanistically how these may mediate the protection of tissues against stress conditions, exemplified by ionizing radiation induced damage in mouse bone marrow. In murine bone marrow hematopoietic cells, the drug initiated the activation of the Nrf2 transcription factor resulting in enhanced expression of downstream stress response genes. This type of response was confirmed in human liver cells and included enhanced expression of glutathione S-transferases (GST), important in the metabolism and pharmacological function of seleno-compounds. In C57 BL/6 mice, DSBA prevented the suppression of bone marrow hematopoietic cells caused by ionizing radiation exposure. Such in vivo prevention effects were associated with Nrf2 pathway activation in both bone marrow cells and liver tissue. These findings demonstrated for the first time the pharmacological properties of DSBA in vivo, suggesting a practical application for this type of Se-hormetic molecules as a radioprotective and/or prevention agents in cancer treatments.
Keywords	Medicine ; R ; Science ; Q
Subject code	616
Language	English
Publishing date	2019-01-01T00:00:00Z
Publisher	Public Library of Science (PLoS)
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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