Publications by authors named "Giorgio Ricci"

84 Publications

Predictors of post-traumatic complication of mild brain injury in anticoagulated patients: DOACs are safer than VKAs-comment.

Intern Emerg Med 2021 Mar 2. Epub 2021 Mar 2.

Department of Emergency Medicine, University of Verona, Verona, Italy.

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http://dx.doi.org/10.1007/s11739-021-02687-yDOI Listing
March 2021

New Factors Enhancing the Reactivity of Cysteines in Molten Globule-Like Structures.

Int J Mol Sci 2020 Sep 22;21(18). Epub 2020 Sep 22.

Department of Chemical Sciences and Technologies, University of Rome "Tor Vergata", 00133 Rome, Italy.

Protein cysteines often play crucial functional and structural roles, so they are emerging targets to design covalent thiol ligands that are able to modulate enzyme or protein functions. Some of these residues, especially those involved in enzyme mechanisms-including nucleophilic and reductive catalysis and thiol-disulfide exchange-display unusual hyper-reactivity; such a property is expected to result from a low p and from a great accessibility to a given reagent. New findings and previous evidence clearly indicate that p perturbations can only produce two-four-times increased reactivity at physiological pH values, far from the hundred and even thousand-times kinetic enhancements observed for some protein cysteines. The data from the molten globule-like structures of ribonuclease, lysozyme, bovine serum albumin and chymotrypsinogen identified new speeding agents, i.e., hydrophobic/electrostatic interactions and productive complex formations involving the protein and thiol reagent, which were able to confer exceptional reactivity to structural cysteines which were only intended to form disulfides. This study, for the first time, evaluates quantitatively the different contributions of p and other factors to the overall reactivity. These findings may help to clarify the mechanisms that allow a rapid disulfide formation during the oxidative folding of many proteins.
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http://dx.doi.org/10.3390/ijms21186949DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7555924PMC
September 2020

Ultra-rapid glutathionylation of chymotrypsinogen in its molten globule-like conformation: A comparison to archaeal proteins.

Sci Rep 2020 06 2;10(1):8943. Epub 2020 Jun 2.

Dipartimento di Scienze e Tecnologie Chimiche, Università di Roma "Tor Vergata", Rome, Italy.

Chymotrypsinogen, when reduced and taken to its molten globule-like conformation, displays a single cysteine with an unusual kinetic propensity toward oxidized glutathione (GSSG) and other organic thiol reagents. A single residue, identified by mass spectrometry like Cys1, reacts with GSSG about 1400 times faster than an unperturbed protein cysteine. A reversible protein-GSSG complex and a low pK (8.1 ± 0.1) make possible such astonishing kinetic property which is absent toward other natural disulfides like cystine, homocystine and cystamine. An evident hyper-reactivity toward 5,5'-dithiobis-(2-nitrobenzoic acid) (DTNB) and 1-chloro-2,4-dinitrobenzene (CDNB) was also found for this specific residue. The extraordinary reactivity toward GSSG is absent in two proteins of the thermophilic archaeon Sulfolobus solfataricus, an organism lacking glutathione: the Protein Disulphide Oxidoreductase (SsPDO) and the Bacterioferritin Comigratory Protein 1 (Bcp1) that displays Cys residues with an even lower pK value (7.5 ± 0.1) compared to chymotrypsinogen. This study, which also uses single mutants in Cys residues for Bcp1, proposes that this hyper-reactivity of a single cysteine, similar to that found in serum albumin, lysozyme, ribonuclease, may have relevance to drive the "incipit" of the oxidative folding of proteins from organisms where the glutathione/oxidized glutathione (GSH/GSSG) system is present.
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http://dx.doi.org/10.1038/s41598-020-65696-5DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7265447PMC
June 2020

Molecular Investigation of SARS-CoV-2 Proteins and Their Interactions with Antiviral Drugs.

Viruses 2020 04 14;12(4). Epub 2020 Apr 14.

Department of Chemical Sciences and Technologies, University of Rome Tor Vergata, Via della Ricerca Scientifica 1, 00133 Rome, Italy.

A new Coronavirus strain, named SARS-CoV-2, suddenly emerged in early December 2019. SARS-CoV-2 resulted in being dramatically infectious, with thousands of people infected. In this scenario, and without effective vaccines available, the importance of an immediate tool to support patients and against viral diffusion becomes evident. In this study, we exploit the molecular docking approach to analyze the affinity between different viral proteins and several inhibitors, originally developed for other viral infections. Our data show that, in some cases, a relevant binding can be detected. These findings support the hypothesis to develop new antiviral agents against COVID-19, on the basis of already established therapies.
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http://dx.doi.org/10.3390/v12040445DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7232184PMC
April 2020

Risk factors associated with intracranial bleeding and neurosurgery in patients with mild traumatic brain injury who are receiving direct oral anticoagulants.

Am J Emerg Med 2020 Feb 24. Epub 2020 Feb 24.

Department of Emergency Medicine, University of Verona, Verona, Italy.

Background: The established clinical risk factors for post-traumatic intracranial bleeding have not been evaluated in patients receiving DOACs yet.

Aim: Evaluating the association between clinic and patient characteristics and post-traumatic intracranial bleeding (ICH) in patients with mild traumatic brain injury (MTBI) and DOACs.

Methods: This is a retrospective observational study conducted on three Emergency Departments. Multivariate analysis provided association in terms of OR with the risk of ICH. The performance of the multivariate model, described in a nomogram, has been tested with discrimination and decision curve analysis.

Results: Of 473 DOACs patients with MTBI, 8.5% had post-traumatic ICH. On multivariable analysis, major dynamics (odds ratio [OR] 6.255), post-traumatic amnesia (OR 3.961), post-traumatic loss of consciousness (LOC, OR 7.353), Glasgow Coma Scale (GCS) score < 15 (OR 3.315), post-traumatic headache (OR 4.168) and visible trauma above the clavicles (OR 3.378) were associated with a higher likelihood of ICH. The multivariate model, used for the nomogram construction, showed a good performance (AUC bias corrected with 5000 bootstraps resample 0.78). The DCAs showed a net clinical benefit of the prognostic model.

Conclusions: Clinical risk factors can be used in DOACs patients to better define the risk of post-traumatic ICH.
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http://dx.doi.org/10.1016/j.ajem.2020.02.046DOI Listing
February 2020

Thirty-day mortality in atrial fibrillation patients with gastrointestinal bleeding in the emergency department: differences between direct oral anticoagulant and warfarin users.

Intern Emerg Med 2020 03 21;15(2):311-318. Epub 2019 Nov 21.

Department of Emergency Medicine, Girolamo Fracastoro Hospital of San Bonifacio, Azienda Ospedaliera Scaligera, San Bonifacio, Verona, Italy.

More clinical data are required on the safety of direct oral anticoagulants (DOACs). Although patients treated with warfarin and DOACs have a similar risk of bleeding, short-term mortality after a gastrointestinal bleeding (GIB) episode in DOAC-treated patients has not been clarified. The objective of this study was to assess differences in 30-day mortality in patients treated with DOACs or warfarin admitted to the emergency department (ED) for GIB. This was a multicentre retrospective study conducted over 2 years. The study included patients evaluated at three different EDs for GIB. The baseline characteristics were included. Subsequently, we assessed the differences in past medical history and clinical data between the two study groups (DOAC and warfarin users). Differences between the two groups were evaluated using Kaplan-Meier curves. Among the 284 patients presenting GIB enrolled in the study period, 39.4% (112/284) were treated with DOACs and 60.6% (172/284) were treated with warfarin. Overall, 8.1% (23/284) of patients died within 30 days. Among the 172 warfarin-treated patients, 8.7% (15/172) died within 30 days from ED evaluation. In the 112 DOAC-treated patients, the mortality rate was 7.1% (8/112). The Cox regression analysis, adjusted for possible clinical confounders, and the Kaplan-Meier curves did not outline differences between the two treatment groups. The present study shows no differences between DOACs and warfarin in short-term mortality after GIB.
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http://dx.doi.org/10.1007/s11739-019-02229-7DOI Listing
March 2020

Ultra-Rapid Glutathionylation of Ribonuclease: Is this the Real Incipit of its Oxidative Folding?

Int J Mol Sci 2019 Oct 31;20(21). Epub 2019 Oct 31.

Department of Chemical Sciences and Technologies, University of Rome "Tor Vergata", 00133 Rome, Italy.

Many details of oxidative folding of proteins remain obscure, in particular, the role of oxidized glutathione (GSSG). This study reveals some unknown aspects. When a reduced ribonuclease A refolds in the presence of GSSG, most of its eight cysteines accomplish a very fast glutathionylation. In particular, one single cysteine, identified as Cys95 by mass spectrometry, displays 3600 times higher reactivity when compared with an unperturbed protein cysteine. Furthermore, the other five cysteines show 40-50 times higher reactivity toward GSSG. This phenomenon is partially due to a low p value of most of these cysteines (average p = 7.9), but the occurrence of a reversible GSSG-ribonuclease complex ( = 0.12 mM) is reasonably responsible for the extraordinary hyper-reactivity of Cys95. Neither hyper-reactivity nor some protein-disulfide complexes have been found by reacting a reduced ribonuclease with other natural disulfides i.e., cystine, cystamine, and homocystine. Hyper-reactivity of all cysteines was observed toward 5,5'-dithiobis-(2-nitrobenzoic acid). Given that GSSG is present in high concentrations in the endoplasmic reticulum, this property may shed light on the early step of its oxidative folding. The ultra-rapid glutathionylation of cysteines, only devoted to form disulfides, is a novel property of the molten globule status of the ribonuclease.
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http://dx.doi.org/10.3390/ijms20215440DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6862303PMC
October 2019

Direct Oral Anticoagulant Treatment and Mild Traumatic Brain Injury: Risk of Early and Delayed Bleeding and the Severity of Injuries Compared with Vitamin K Antagonists.

J Emerg Med 2019 Dec 21;57(6):817-824. Epub 2019 Oct 21.

Department of Emergency Medicine, University of Verona, Verona, Italy.

Background: The risk of intracranial hemorrhage (ICH) in patients taking direct oral anticoagulants (DOACs) after mild traumatic brain injury (MTBI) is unclear.

Objectives: To assess the differences in the risk of developing early, delayed, and comprehensive bleeding after MTBI among patients treated with DOACs as compared with those treated with vitamin K antagonists (VKAs).

Methods: All MTBI patients taking oral anticoagulants in our emergency department between June 2017 and August 2018 were included. All patients on oral anticoagulants underwent immediate cerebral computed tomography (CT) and a second CT scan after 24 h of clinical observation.

Results: There were 451 patients enrolled: 268 were on VKAs and 183 on DOACs. Of the DOAC-treated patients, 7.7% (14/183) presented overall intracranial bleeding, compared with 14.9% (40/268) of VKA-treated patients (p = 0.026). Early bleeding was present in 5.5% (10/183) of DOAC-treated patients and in 11.6% (31/268) of VKA-treated patients (p = 0.030). Multivariable analysis showed that VKA therapy (odds ratio [OR] 2.327), high-energy impact (OR 11.229), amnesia (OR 2.814), loss of consciousness (OR 5.286), Glasgow Coma Scale score < 15 (OR 4.719), and the presence of lesion above the clavicles (OR 2.742) were associated with significantly higher risk of global ICH. A nomogram was constructed to predict ICH using these six variables. Discrimination of the nomogram revealed good predictive abilities (area under the receiver operating characteristic curve: 0.817).

Conclusions: DOAC-treated patients seem to have lower risk of posttraumatic intracranial bleeding compared with VKA-treated patients.
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http://dx.doi.org/10.1016/j.jemermed.2019.09.007DOI Listing
December 2019

Hemodialysis biomarkers: total advanced glycation end products (AGEs) against oxidized human serum albumin (HSAox).

Acta Diabetol 2019 Dec 7;56(12):1323-1331. Epub 2019 Sep 7.

Department of Chemical Sciences and Technologies, University of Rome Tor Vergata, 00133, Rome, Italy.

Aims: Nephropathic patients show higher levels of advanced glycation end products (AGEs) and oxidized human serum albumin (HSAox) compared to healthy subjects. These two classes of compounds are formed as the result of oxidative insults; for this reason, they can be useful oxidative stress biomarkers. The present study examines the variation of AGEs and HSAox in hemodialysis (HD) patients before and after dialysis session, evaluating the impact of different dialytic techniques and filters on their removal.

Methods: A total of 50 healthy subjects (control group) and 130 HD patients were enrolled in the study. Hemodialysis patients were subdivided based on dialytic techniques: 109 in diffusive technique and 22 in convective technique. We monitored HSAox, AGEs and other laboratory parameters at early morning in healthy subjects and in HD patients before and after the dialysis procedures.

Results: The level of HSAox decreases after a single dialytic session (from 58.5 ± 8.8% to 41.5 ± 11.1%), but the concentration of total AGEs increases regardless of adopted dialytic techniques (from 6.8 ± 5.2 µg/ml to 9.2 ± 4.4 µg/ml). In our study, levels of HSAox and total AGEs are similar in diabetic and non-diabetic HD patients. The increase in total AGEs after dialysis was only observed using polysulfone filters but was absent with polymethacrylate filters.

Conclusions: HSAox is a simple and immediate method to verify the beneficial effect of a single dialysis session on the redox imbalance, always present in HD patients. Total AGEs assayed by ELISA procedure seem to be a less reliable biomarker in this population.
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http://dx.doi.org/10.1007/s00592-019-01413-7DOI Listing
December 2019

Glutathione Transferase P1-1 an Enzyme Useful in Biomedicine and as Biomarker in Clinical Practice and in Environmental Pollution.

Nutrients 2019 Jul 27;11(8). Epub 2019 Jul 27.

Department of Chemical Sciences and Technologies, University of Rome Tor Vergata, Via della Ricerca Scientifica 1, 00133 Rome, Italy.

Glutathione transferase P1-1 (GSTP1-1) is expressed in some human tissues and is abundant in mammalian erythrocytes (here termed e-GST). This enzyme is able to detoxify the cell from endogenous and exogenous toxic compounds by using glutathione (GSH) or by acting as a ligandin. This review collects studies that propose GSTP1-1 as a useful biomarker in different fields of application. The most relevant studies are focused on GSTP1-1 as a biosensor to detect blood toxicity in patients affected by kidney diseases. In fact, this detoxifying enzyme is over-expressed in erythrocytes when unusual amounts of toxins are present in the body. Here we review articles concerning the level of GST in chronic kidney disease patients, in maintenance hemodialysis patients and to assess dialysis adequacy. GST is also over-expressed in autoimmune disease like scleroderma, and in kidney transplant patients and it may be used to check the efficiency of transplanted kidneys. The involvement of GSTP in the oxidative stress and in other human pathologies like cancer, liver and neurodegenerative diseases, and psychiatric disorders is also reported. Promising applications of e-GST discussed in the present review are its use for monitoring human subjects living in polluted areas and mammals for veterinary purpose.
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http://dx.doi.org/10.3390/nu11081741DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6723968PMC
July 2019

Trends and determinants of Emergency Room admissions for asthma: A retrospective evaluation in Northeast Italy.

World Allergy Organ J 2019 3;12(7):100046. Epub 2019 Jul 3.

Asthma Center and Allergy Unit, Verona University and General Hospital, Verona, Italy.

Background: Asthma still represents a cause of death and hospital admissions worldwide. Our study aimed at analyzing the trend of Emergency Room (ER) asthma admissions in Northeast Italy in order to investigate the relevance of specific patient-related determinants and environmental triggers (pollens, mold spores, and pollutants).

Methods: Retrospective data from admissions for asthma exacerbations registered between the years 2013 and 2015 in two main ERs in Northeast Italy were collected. Data about patients' age, sex and nationality were recorded. Classification of disease severity followed the current Italian ER triage scoring system (white: no need for emergency treatment; green: need for fast treatment; yellow: severe condition; red: life-threatening condition). Data on pollen/mold spore counts and pollutants were analyzed.

Results: Overall, 1745 ​ER admissions for asthma were registered, with a persistent and significant increase year by year. A slight prevalence of females and patients over 50 years old was observed. Immigrants accounted for 32%, 36% and 26% of admissions respectively in 2013, 2014 and 2015. The prevalence of immigrants' admissions was significantly higher when comparing the relative ratio of immigrant populations/Italian nationals (p ​< ​0.05). The admissions were coded as follows: white, 6.30%; green, 35.36%; yellow, 39.37%; red, 18.97%. People aged ≥50 years were more frequently admitted with a red code, but the trend was not statistically significant (p ​= ​0,0815). By contrast, amongst immigrants there was a higher prevalence of white and green codes observed in comparison with Italian nationals. Grass pollen peak and PM high levels represented environmental determinants of ER admissions increase.

Conclusions: The increasing rate of asthma-related ER admissions highlights the need for implementing asthma control strategies. Investigating the traits of patients referring to ER for asthma exacerbations, as well as environmental-related determinants, may help in identifying at-risk individuals and in orienting preventive strategies accordingly. Immigrants represent the most vulnerable sub-population, and their potential difficulties in accessing treatments and health services should be specifically addressed. Overall, implementing patient education in order to improve treatment adherence, as well as providing an asthma action plan to every asthmatic patient, continue to be the most urgent needs.
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http://dx.doi.org/10.1016/j.waojou.2019.100046DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6612754PMC
July 2019

The impact of ionizing irradiation on liver detoxifying enzymes. A re-investigation.

Cell Death Discov 2019 8;5:66. Epub 2019 Feb 8.

1Department of Chemical Sciences and Technologies, University of Rome "Tor Vergata", Rome, Italy.

By looking at many studies describing the impact of ionizing irradiations in living mice on a few key detoxifying enzymes like catalase, superoxide dismutase, glutathione peroxidase, glutathione reductase and glutathione transferase, we noted conflicting evidences: almost all papers finalized to demonstrate the protective effects of natural or synthetic drugs against the damage by irradiations, described also a relevant inactivation of these enzymes in the absence of these compounds. Conversely, no inactivation and even enhanced activity has been noted under similar irradiation modality in all studies supporting the "adaptive response". Motivated by these curious discrepancies, we performed irradiation experiments on living mice, explanted mouse livers and liver homogenates observing that, in all conditions the activity of all these enzymes remained almost unchanged except for a slight increase found in explanted livers. Our results put a question about many previous scientific reports in this field.
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http://dx.doi.org/10.1038/s41420-019-0148-8DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6368569PMC
February 2019

Red Blood Cell Distribution Width Improves Reclassification of Patients Admitted to the Emergency Department with Acute Decompensated Heart Failure.

J Med Biochem 2018 Jul 1;37(3):299-306. Epub 2018 Jul 1.

Department of Cardiology and Intensive Care Cardiology, University of Verona, Verona, Italy.

Background: The usual history of chronic heart failure (HF) is characterized by frequent episodes of acute decompensation (ADHF), needing urgent management in the emergency department (ED). Since the diagnostic accuracy of routine laboratory tests remains quite limited for predicting short-term mortality in ADHF, this retrospective study investigated the potential significance of combining red blood cell distribution width (RDW) with other conventional tests for prognosticating ADHF upon ED admission.

Methods: We conducted a retrospective study including visits for episodes of ADHF recorded in the ED of the Uni versity Hospital of Verona throughout a 4-year period. Demo - graphic and clinical features were recorded upon patient presentation. All patients were subjected to standard Chest X-ray, electrocardiogram (ECG) and laboratory testing in - cluding creatinine, blood urea nitrogen, B-type natriuretic peptide (BNP), complete blood cell count (CBC), sodium, chloride, potassium and RDW. The 30-day overall mortality after ED presentation was defined as primary endpoint.

Results: The values of sodium, creatinine, BNP and RDW were higher in patients who died than in those who survived, whilst hypochloremia was more frequent in patients who died than in those who survived. The multivariate model, incorporating these parameters, displayed a modest efficiency for predicting 30-day mortality after ED admission (AUC, 0.701; 95% CI, 0.662-0.738; p=0.001). Notably, the inclusion of RDW in the model significantly enhanced prediction efficiency, with an AUC of 0.723 (95% CI, 0.693-0.763; p<0.001). These results were confirmed with net reclassification improvement (NRI) analysis, showing that combination of RDW with conventional laboratory tests resulted in a much better prediction performance (net reclassification index, 0.222; p=0.001).

Conclusions: The results of our study show that prognostic assessment of ADHF patients in the ED can be significantly improved by combining RDW with other conventional laboratory tests.
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http://dx.doi.org/10.1515/jomb-2017-0054DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6298466PMC
July 2018

The extreme hyper-reactivity of Cys94 in lysozyme avoids its amorphous aggregation.

Sci Rep 2018 10 30;8(1):16050. Epub 2018 Oct 30.

Department of Chemical Sciences and Technologies, University of Rome "Tor Vergata", Rome, Italy.

Many proteins provided with disulfide bridges in the native state undergo amorphous irreversible aggregation when these bonds are not formed. Here we show that egg lysozyme displays a clever strategy to prevent this deleterious aggregation during the nascent phase when disulfides are still absent. In fact, when the reduced protein assembles into a molten globule state, its cysteines acquire strong hyper-reactivity towards natural disulfides. The most reactive residue, Cys94, reacts with oxidized glutathione (GSSG) 3000 times faster than an unperturbed protein cysteine. A low pK of its sulfhydryl group (6.6/7.1) and a productive complex with GSSG (K = 0.3 mM), causes a fast glutathionylation of this residue (t = 3 s) and a complete inhibition of the protein aggregation. Other six cysteines display 70 times higher reactivity toward GSSG. The discovery of extreme hyper-reactivity in cysteines only devoted to structural roles opens new research fields for Alzheimer's and Parkinson diseases.
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http://dx.doi.org/10.1038/s41598-018-34439-yDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6207692PMC
October 2018

Thiol disulfide exchange reactions in human serum albumin: the apparent paradox of the redox transitions of Cys.

FEBS J 2018 09 2;285(17):3225-3237. Epub 2018 Aug 2.

Department of Chemical Sciences and Technologies, University of Rome Tor Vergata, Italy.

Human serum albumin (HSA) is characterized by 17 disulfides and by only one unpaired cysteine (Cys ), which can be free in the reduced albumin or linked as a mixed disulfide with cysteine, or in minor amount with other natural thiols, in the oxidized albumin. In healthy subjects, the level of the oxidized form is about 35%, but it rises up to 70% after oxidative insults or in patients with kidney diseases. Oxidized albumin is therefore considered a short-term biomarker of oxidative stress as its level may increase or decrease under appropriate redox inputs in discrete temporal spans. This paper defines, for the first time, the kinetic properties of reduced and oxidized Cys of HSA in their reactions with natural disulfides and thiols. Kinetic constants support the evidence that the Cys redox oscillations observed in vivo are mainly due to the interaction with cysteine and cystine without the involvement of any enzymatic support. This study gives also a plausible explanation for the absence of involvement of the 17 disulfides naturally present in HSA in these redox transitions. This inert behavior toward cysteine is marginally due to solvent accessibility or flexibility factors of these bonds but mainly to their strong thermodynamic stability, which is caused essentially by a proximity effect. A similar mechanism is likely at play in the many proteins that maintain disulfide bridges in a reducing medium like the cytosol.
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http://dx.doi.org/10.1111/febs.14609DOI Listing
September 2018

Association of Short- and Medium-Term Particulate Matter Exposure with Risk of Mortality after Spontaneous Intracerebral Hemorrhage.

J Stroke Cerebrovasc Dis 2018 Sep 24;27(9):2519-2523. Epub 2018 May 24.

Department of Neuroscience, Azienda Ospedaliera Universitaria Integrata Verona, Verona, Italy.

Objective: We investigated the association of short- and medium-term particulate matter (PM) exposure with risk of mortality in patients with spontaneous intracerebral hemorrhage (ICH) identified according to strict etiologic criteria.

Methods: We conducted a retrospective analysis of prospectively collected data from consecutive patients with spontaneous ICH admitted to the emergency department of the University Hospital of Verona from March 2011 to December 2014. Outcome measures were mortality within 1 month after ICH and significant hematoma expansion (HE) defined as an absolute growth of more than 12.5 mL or a relative increase of more than 50% from baseline to follow-up computed tomography scan.

Results: A final number of 308 patients were included. In the adjusted model, higher PM and PM values in the last 3 days (odds ratio [OR] 1.827, 95% confidence interval [CI] 1.057-3.159, P = .031 and OR 1.949, 95% CI 1.025-3.704, P = .042, respectively) and in the last 4 weeks (OR 4.975, 95% CI 2.174-11.381, P < .001 and OR 9.781, 95% CI 3.425-27.932, P < .001, respectively) before ICH were associated with higher mortality rate. No association was found between PM exposure and significant HE.

Conclusions: PM exposure in the short- and medium-term before spontaneous ICH was associated with risk of 1-month mortality, independent of predictors such as age, sex, stroke severity, intraventricular hemorrhage, ICH volume, ICH location, ICH etiologic subtype, significant HE, antithrombotic therapy, atrial fibrillation, and blood glucose levels.
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http://dx.doi.org/10.1016/j.jstrokecerebrovasdis.2018.05.007DOI Listing
September 2018

Erythrocyte glutathione transferase in kidney transplantation: a probe for kidney detoxification efficiency.

Cell Death Dis 2018 02 19;9(3):288. Epub 2018 Feb 19.

Department of Chemical Sciences and Technologies, University of Rome, Tor Vergata, Via della Ricerca Scientifica, 00133, Rome, Italy.

Erythrocyte glutathione transferase (e-GST) is overexpressed in case of increased blood toxicity and its level correlates with the kidney disease progression. Thus, it represents a probe of kidney efficiency against circulating toxins. We measured the activity of e-GST in patients with transplant kidney from living and cadaver donors, correlated its level to biochemical parameters of kidney function, and measured the level of oxidized albumin as a probe of oxidative stress using a new simple procedure. Interestingly, the activity of e-GST in transplant patients from cadaver donors (N = 153) is very high (11.7 U/g) compared to healthy subjects (N = 80) ( 5.6 U/g). Lower values were observed in transplant patients with kidney from living donors (N = 16) (9.8 U/g). Except for steroids, no correlation has been found with the immunosuppressive therapies and routine clinical and laboratory parameters. Also serum oxidized albumin, which reveals oxidative stress, is significantly higher in transplant patients from cadaver donors (53%) compared to that from living donors (36%). Overall, these data indicate that most of transplant kidneys from cadavers lost part of the detoxifying power against circulating toxins and suffer a relevant oxidative stress compared to those coming from living donors. A case report suggests that e-GST could represent a very early marker of incipient graft rejection. In conclusion, e-GST may be used to check the decline or maintenance of the kidney detoxification competence during post-transplantation course.
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http://dx.doi.org/10.1038/s41419-018-0289-3DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5833821PMC
February 2018

Association between short- and medium-term air pollution exposure and risk of mortality after intravenous thrombolysis for stroke.

J Thromb Thrombolysis 2018 Feb;45(2):293-299

Section of Clinical Biochemistry, University of Verona, Verona, Italy.

The exposure to air pollutants may increase both incidence and mortality of stroke. We aimed to investigate the association of short- and medium-term exposure to particulate matter (PM) and nitrogen dioxide (NO) with the outcome of intravenous thrombolysis (IVT) for stroke. We conducted a retrospective analysis based on data prospectively collected from 944 consecutive IVT-treated stroke patients. The main outcome measure was 3-month mortality. The secondary outcome measures were causes of neurological deterioration (≥ 1 NIHSS point from baseline or death < 7 days), including intracerebral hemorrhage, cerebral edema (CED), and persistence or new appearance of hyperdense cerebral artery sign. In the adjusted model, higher PM and PM values in the last 3 days and 4 weeks before stroke were independently associated with higher mortality rate [hazard ratio (HR) 1.014, 95% confidence intervals (CI) 1.005-1.024, p = 0.003; HR 1.079, 95% CI 1.055-1.103, p = 0.001; HR 1.019, 95% CI 1.005-1.032, p = 0.008; and HR 1.015, 95% CI 1.004-1.027, p = 0.007; respectively]. Higher PM and PM values in the last 4 weeks were associated with higher CED rate [odd ratio (OR) 1.023, 95% CI 1.007-1.040, p = 0.006; and OR 1.017, 95% CI 1.003-1.032, p = 0.021; respectively]. No significant association between PM or NO and other causes of neurological deterioration was observed. Higher exposure to PM in the last 3 days and 4 weeks before stroke may be independently associated with 3-month mortality after IVT. Higher exposure to PM in the last 4 weeks before stroke may also be independently associated with CED after IVT.
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http://dx.doi.org/10.1007/s11239-017-1589-7DOI Listing
February 2018

Paraoxonase, arylesterase and lactonase activities of paraoxonase-1 (PON1) in obese and severely obese women.

Scand J Clin Lab Invest 2018 Feb - Apr;78(1-2):18-24. Epub 2017 Nov 23.

g Department of Medical Sciences, Postgraduate School of Digestive Diseases , University of Ferrara , Ferrara , Italy.

Obesity is independently associated with disturbances in lipid and lipoprotein metabolism, oxidative stress, and is a well-established independent risk factor for cardiovascular diseases (CVD). Human paraoxonase 1 (PON1) is a pleotropic high-density lipoprotein (HDL)-associated enzyme with antioxidant and anti-inflammatory proprieties that have been suggested to contribute to the athero-protective function of the lipoprotein. The aim of this study was to investigate whether obesity is associated with PON1 activity and whether this association is influenced by oxidative stress, inflammation and HDL cholesterol (HDL-C) concentration. The promiscuous activities, arylesterase and paraoxonase, and the putative physiological activity, lactonase, of PON1 were assessed in the serum of 214 obese and severely obese, 101 overweight and 129 normal-weight women. Levels of high-sensitivity C-reactive protein (hs-CRP), hydroperoxides (by-products of lipid oxidative damage) and lipid profiles were also evaluated. Arylesterase activity was the only activity that significantly differed across the groups (ANOVA, p < .01), with the greatest decrease observed in individuals with body mass index (BMI) > 40 kg/m compared to controls (p < .001). This activity was also inversely, although weakly (r = -0.160, p < .001) correlated with the BMI, and the association was independent of age and levels of oxidative stress and inflammation, but not of HDL-C concentration. In conclusion, our results suggest that the apparent obesity-associated decrement of PON1 activity might simply reflect the decrease in concentration of its plasmatic carrier.
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http://dx.doi.org/10.1080/00365513.2017.1405274DOI Listing
September 2018

Red blood cell distribution width independently predicts 1-month mortality in acute decompensation of cirrhotic patients admitted to emergency department.

Eur J Gastroenterol Hepatol 2018 Jan;30(1):33-38

Section of Clinical Biochemistry, University of Verona, Verona.

Aim: The aim of this study was to explore whether red blood cell distribution width (RDW) can help predict the risk of short-term mortality in patients with acute decompensation of cirrhosis.

Patients And Methods: We carried out a retrospective analysis of all patients consecutively admitted to the emergency department (ED) of the University Hospital of Verona (Italy) for acute decompensation of liver cirrhosis, between 1 June 2013 and 31 December 2016. The RDW value was measured at ED admission, along with collection of clinical features and other laboratory data, and was then correlated with severity of disease (Chronic Liver Failure Consortium Acute Decompensation score; CLIF-C AD score) and 1-month mortality.

Results: The final study population consisted of 542 patients, 80 (14.8%) of whom died within 30 days after ED admission. The median RDW of patients who died was significantly higher than the median RDW of those who survived (17.4 vs. 15.5%; P<0.001). The percentage of patients who died significantly increased across different RDW quartiles (6.8, 9.7, 11.5 and 32.1%, P<0.001). In univariate analysis, significant correlation was observed between RDW and clinical severity of acute decompensate cirrhosis (Child-Pugh score: r=0.198, P<0.001; Model for End-Stage Liver Disease score: r=0.311, P=0.001; CLIF-C AD: 0.127, P=0.005). The combination of RDW and CLIF-C AD score exhibited better performance for predicting 1-month mortality than the CLIF-C AD score alone (area under the curve=0.769 vs. 0.720; P=0.006). In multivariate analysis, RDW was independently associated with a 1.2-2.3 higher risk of 1-month mortality.

Conclusion: The assessment of RDW at ED admission may improve risk stratification of patients with acute decompensation of cirrhosis.
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http://dx.doi.org/10.1097/MEG.0000000000000993DOI Listing
January 2018

The Role of Red Blood Cell Distribution Width for Predicting 1-year Mortality in Patients Admitted to the Emergency Department with Severe Dyspnoea.

J Med Biochem 2017 Jan 25;36(1):32-38. Epub 2017 Jan 25.

Section of Clinical Biochemistry, University of Verona, Verona, Piazzale LA Scuro, 37100 - Verona, Italy.

Background: Universally accepted and validated instruments for predicting the outcome of patients presenting to the emergency department (ED) with severe dyspnoea do not exist so far, nor are they regularly used by the emergency physicians. This study hence aimed to establish whether red blood cell distribution width (RDW) may be a predictive parameter of 1-year mortality in a population of patients admitted to the ED with severe dyspnoea attributable to different underlying disorders.

Methods: We retrospectively evaluated all the patients undergoing arterial blood gas analysis for severe dyspnoea (irrespective of the cause) during admission to ED of University Hospital of Verona from September 1, 2014 to November 31, 2014.

Results: The final study population consisted of 287 patients for whom complete clinical and laboratory information was available. Overall, 36 patients (12.5%) died after a 1-year follow-up. The RDW value was found to be considerably increased in patients who deceased during the follow-up compared to those who survived (17.2% versus 14.8%; p<0.001). In both univariate and multivariate analyses, the RDW value was found to be a significant predictor of 1-year mortality. In particular, patients with RDW ≥ 15.0% displayed a 72% increased risk of 1-year mortality after multiple adjustments.

Conclusions: The measurement of RDW, a very simple and inexpensive laboratory parameter, may represent an important factor for predicting medium-term mortality in patients presenting to the ED with severe dyspnoea.
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http://dx.doi.org/10.1515/jomb-2016-0026DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5471657PMC
January 2017

Early in-hospital variation of red blood cell distribution width predicts mortality in patients with acute heart failure.

Int J Cardiol 2017 Sep 6;243:306-310. Epub 2017 May 6.

Section of Clinical Biochemistry, University of Verona, Verona, Italy.

Background: Some studies showed that the value of red blood cell distribution width (RDW) at admission may predict clinical outcomes in patients with acutely decompensated heart failure (ADHF). Therefore, this study was planned to investigate whether in-hospital variations of RDW may also predict mortality in this condition.

Methods: The final study population consisted of 588 patients admitted to the local Emergency Department (ED), who were hospitalized for ADHF. The RDW was measured at ED admission and after 48h and 96h of hospital stay. In-hospital variations from admission value, expressed as absolute variation (DeltaRDW) or percent variation (Delta%RDW), were then correlated with 30- and 60-day mortality.

Results: Overall, 87 (14.8%) and 118 (20.1%) patients with ADHF died at 30 or 60days of follow-up. Delta%RDW after 96h of hospital stay independently predicted 30-day mortality (odds ratio, 1.12; 95% CI, 1.07-1.18). An increase >1% of Delta%RDW after 96h of hospital stay independently predicted both 30-day (odds ratio, 2.86; 95% CI, 1.67-4.97) and 60-day (odds ratio, 3.06; 95% CI, 1.89-4.96) mortality. A similar trend was observed for DeltaRDW, since an increase after 96h of hospital stay was associated with a nearly 4-fold higher 30-day mortality (odds ratio, 3.65; 95% CI, 2.02-6.15).

Conclusion: Despite it remains unclear whether RDW is a real risk factor or an epiphenomenon in ADHF, these results suggest that more aggressive management may be advisable in ADHF patients with increasing anisocytosis during the first days of hospitalization.
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http://dx.doi.org/10.1016/j.ijcard.2017.05.023DOI Listing
September 2017

Proteomic and functional analyses reveal pleiotropic action of the anti-tumoral compound NBDHEX in Giardia duodenalis.

Int J Parasitol Drugs Drug Resist 2017 08 29;7(2):147-158. Epub 2017 Mar 29.

Department of Infectious Diseases, Istituto Superiore di Sanità, viale Regina Elena 299, 00161 Rome, Italy. Electronic address:

Giardiasis, a parasitic diarrheal disease caused by Giardia duodenalis, affects one billion people worldwide. Treatment relies only on a restricted armamentarium of drugs. The disease burden and the increase in treatment failure highlight the need for novel, safe and well characterized drug options. The antitumoral compound NBDHEX is effective in vitro against Giardia trophozoites and inhibits glycerol-3-phosphate dehydrogenase. Aim of this work was to search for additional NBDHEX protein targets. The intrinsic NBDHEX fluorescence was exploited in a proteomic analysis to select and detect modified proteins in drug treated Giardia. In silico structural analysis, intracellular localization and functional assays were further performed to evaluate drug effects on the identified targets. A small subset of Giardia proteins was covalently bound to the drug at specific cysteine residues. These proteins include metabolic enzymes, e.g. thioredoxin reductase (gTrxR), as well as elongation factor 1B-γ (gEF1Bγ), and structural proteins, e.g. α-tubulin. We showed that NBDHEX in vitro binds to recombinant gEF1Bγ and gTrxR, but only the last one could nitroreduce NBDHEX leading to drug modification of gTrxR catalytic cysteines, with concomitant disulphide reductase activity inhibition and NADPH oxidase activity upsurge. Our results indicate that NBDHEX reacts with multiple targets whose roles and/or functions are specifically hampered. In addition, NBDHEX is in turn converted to reactive intermediates extending its toxicity. The described NBDHEX pleiotropic action accounts for its antigiardial activity and encourages the use of this drug as a promising alternative for the future treatment of giardiasis.
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http://dx.doi.org/10.1016/j.ijpddr.2017.03.006DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5377010PMC
August 2017

Rapid and well tolerated action of idarucizumab for antagonizing dabigatran in a patient needing urgent thrombolysis: a case report.

Blood Coagul Fibrinolysis 2017 Oct;28(7):576-579

aLaboratory of Clinical Chemistry and Hematology, General Hospital of Verona bEmergency Department, University Hospital of Verona cSection of Clinical Biochemistry, University of Verona, Verona, Italy.

: Dabigatran is a direct oral anticoagulant drug exhibiting clinical benefits over vitamin K antagonists. A procedure for reversing the anticoagulant effect of direct oral anticoagulants may be needed in emergency clinical settings, and is traditionally accomplished by using plasma products or hemostatic physical interventions. Idarucizumab, a specific antidote for dabigatran, has recently become available. This compound can be rapidly administered by intravenous injection and is effective in reversing anticoagulation in few minutes. We describe here the case of a 78-year-old woman taking dabigatran for atrial fibrillation, who was admitted to the emergency department with a diagnosis of acute cerebral ischemia. Dabigatran plasma levels on admission (74 ng/ml) were measured with diluted thrombin time. Idarucizumab was immediately administered and dabigatran plasma concentration suddenly decreased to less than 2 ng/ml. Successful systemic thrombolysis could hence be performed with full recovery.
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http://dx.doi.org/10.1097/MBC.0000000000000634DOI Listing
October 2017

Emergency Department Readmission in Elderly Patients After Acute Rhythm or Rate Control Treatment for Atrial Fibrillation.

J Atr Fibrillation 2016 Aug-Sep;9(2):1387. Epub 2016 Aug 31.

Professor of Internal Medicine, Department of Internal Medicine, Azienda Ospedaliera Universitaria Integrata,Verona, Italy, Chief of Postgraduate School of Emergency Medicine, University of Verona, Italy.

Atrial fibrillation (AF) is an age-related increasing disease, characterized by a high number of relapses frequently leading the patients to Emergency Department (ED). Despite AF relapses may be clinically heterogeneous, a proper management requires either a fast and effective restore of the sinus rhythm or a satisfactory control of the ventricular rate. Whether the strategy adopted in the ED could affect the course of disease is still debated. Therefore, the aim of our study was to evaluate the number of ED readmission for AF related symptoms and the event-free period in patients older than 70 years previously treated in ED for an AF recurrence, in order to assess a possible relationship with the acute strategy. An overall number of 302 recurrences of AF were drawn randomly, regarding 102 patients (mean age 78 years). We found that 206 cases (68.2%) were treated with rhythm restoration strategy (RR) whereas 96 (31.8%) with rate control strategy (RC). The median following event-free period was 118.6 +/- 15.5 and 212.9 +/- 37.1 days (p < 0.05) for RR and RC strategy, respectively. Within 6 months, 124 (60.1%) out of RR group patients and only 44 (45.8%) out of RC group patients had to be readmitted to ED for AF related symptoms (whether a recurrence or inefficient rate control symptoms) (p< 0.05). This advantage was substantially confirmed (79.1% vs 65.6% respectively, p < 0.05) after a 12 months follow-up. Our results indicate that acute treatment of AF may affect the long-term outcome of the disease and the ED readmission rate of the patient. Ventricular rate control seems to be associated with a longer event-free period if compared to the rhythm control strategy in the elderly patients. This suggests an age-based work-up of patients admitted to the ED, preferentially using ventricular rate control in elderly subjects.
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http://dx.doi.org/10.4022/jafib.1387DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5129682PMC
August 2016

Glutathione transferase P1-1 as an arsenic drug-sequestering enzyme.

Protein Sci 2017 02 14;26(2):317-326. Epub 2016 Dec 14.

ACRF Rational Drug Discovery Centre, St. Vincent's Institute of Medical Research, Fitzroy, Victoria, 3065, Australia.

Arsenic-based compounds are paradoxically both poisons and drugs. Glutathione transferase (GSTP1-1) is a major factor in resistance to such drugs. Here we describe using crystallography, X-ray absorption spectroscopy, mutagenesis, mass spectrometry, and kinetic studies how GSTP1-1 recognizes the drug phenylarsine oxide (PAO). In conditions of cellular stress where glutathione (GSH) levels are low, PAO crosslinks C47 to C101 of the opposing monomer, a distance of 19.9 Å, and causes a dramatic widening of the dimer interface by approximately 10 Å. The GSH conjugate of PAO, which forms rapidly in cancerous cells, is a potent inhibitor (K  = 90 nM) and binds as a di-GSH complex in the active site forming part of a continuous network of interactions from one active site to the other. In summary, GSTP1-1 can detoxify arsenic-based drugs by sequestration at the active site and at the dimer interface, in situations where there is a plentiful supply of GSH, and at the reactive cysteines in conditions of low GSH.
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http://dx.doi.org/10.1002/pro.3084DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5275733PMC
February 2017

Early function decline after ischemic stroke can be predicted by a nomogram based on age, use of thrombolysis, RDW and NIHSS score at admission.

J Thromb Thrombolysis 2017 Apr;43(3):394-400

Section of Clinical Biochemistry, University of Verona, Piazzale LA Scuro, 37100, Verona, Italy.

The availability of prediction tools for risk stratification after acute stroke is seen as a valuable perspective for tailored clinical management. This retrospective study was aimed to identify significant predictors of poor outcome in patients presenting with acute ischemic stroke, which could then be used for constructing a prediction model. The study population consisted of 837 patients admitted to the Stoke Unit of University Hospital of Verona (Italy) for acute ischemic stroke within 12 h of symptoms onset. In multivariate analysis, age, use of thrombolysis, red blood cell distribution width (RDW) and NIHSS score at admission were found to be significant predictors of 3-month functional decline. A nomogram constructed by integrating these four variables exhibited an area under the curve of 0.832 for predicting functional impairment. The >80% risk cut-off derived from the nomogram was associated with 0.91 positive predictive value, whereas a risk probability <10% displayed 0.93 negative predictive value for predicting functional impairment. These results demonstrate that a prediction tool integrating some important clinical, laboratory and demographic variables may enable an efficient risk stratification of poor outcome after acute stroke.
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http://dx.doi.org/10.1007/s11239-016-1456-yDOI Listing
April 2017

Evolution of Negative Cooperativity in Glutathione Transferase Enabled Preservation of Enzyme Function.

J Biol Chem 2016 Dec 4;291(52):26739-26749. Epub 2016 Nov 4.

From the Department of Chemical Sciences and Technologies,

Negative cooperativity in enzyme reactions, in which the first event makes subsequent events less favorable, is sometimes well understood at the molecular level, but its physiological role has often been obscure. Negative cooperativity occurs in human glutathione transferase (GST) GSTP1-1 when it binds and neutralizes a toxic nitric oxide adduct, the dinitrosyl-diglutathionyl iron complex (DNDGIC). However, the generality of this behavior across the divergent GST family and its evolutionary significance were unclear. To investigate, we studied 16 different GSTs, revealing that negative cooperativity is present only in more recently evolved GSTs, indicating evolutionary drift in this direction. In some variants, Hill coefficients were close to 0.5, the highest degree of negative cooperativity commonly observed (although smaller values of n are theoretically possible). As DNDGIC is also a strong inhibitor of GSTs, we suggest negative cooperativity might have evolved to maintain a residual conjugating activity of GST against toxins even in the presence of high DNDGIC concentrations. Interestingly, two human isoenzymes that play a special protective role, safeguarding DNA from DNDGIC, display a classical half-of-the-sites interaction. Analysis of GST structures identified elements that could play a role in negative cooperativity in GSTs. Beside the well known lock-and-key and clasp motifs, other alternative structural interactions between subunits may be proposed for a few GSTs. Taken together, our findings suggest the evolution of self-preservation of enzyme function as a novel facility emerging from negative cooperativity.
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http://dx.doi.org/10.1074/jbc.M116.749507DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5207182PMC
December 2016

Red Blood Cell Distribution Width Is an Independent Predictor of Outcome in Patients Undergoing Thrombolysis for Ischemic Stroke.

Semin Thromb Hemost 2017 Feb 3;43(1):30-35. Epub 2016 Nov 3.

Section of Clinical Biochemistry, University of Verona, Verona, Italy.

An appropriate and timely management, including early diagnosis and accurate prognostication, is the mainstay for managed care of patients with acute ischemic stroke. Since red blood cell distribution width (RDW) was found to be an independent predictor of clinical outcomes in patients with thrombotic disorders, we designed a retrospective observational study to investigate whether the RDW value may also retain predictive significance in stoke patients undergoing thrombolytic therapy. This retrospective study was based on all patients admitted to the Emergency Department (ED) of the University Hospital of Verona (Italy) with a diagnosis of ischemic stroke, who underwent systemic thrombolysis between January 2013 and June 2015. The RDW value along with basal clinical characteristics was recorded at ED admission. The final study population consisted of 316 patients. A significant association was found between stroke severity (NIHSS score) and RDW ( = 0.322;  < 0.001). The median RDW value in patients with clinical improvement after thrombolysis was significantly lower than in patients without (13.4 vs. 14.1%;  < 0.001). The diagnostic accuracy (area under the curve) of RDW for predicting the lack of neurological improvement was 0.667. In univariate analysis, RDW >14.5% was associated with increased rate of no neurological improvement (odds ratio [OR], 2.38; 95% confidence interval [CI], 1.37-4.13), an association remaining significant also in multivariate analysis (OR, 1.85; 95% CI, 1.13-3.32). Survivor curve analysis showed that patients with RDW values ≥14.5% had a higher risk of 1-year mortality and shorter survival. These results suggest that RDW assessment at ED admission may provide valuable diagnostic and prognostic information in patients with acute ischemic stroke.
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http://dx.doi.org/10.1055/s-0036-1592165DOI Listing
February 2017