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Methods for Analysis of Protein Glutathionylation and their Application to Photosynthetic Organisms

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Molecular Plant 2009年第2期2卷 218-235页

作者： Xing-Huang Gao Mariette Bedhomme daniel Veyel mirko Zaffagnini Stephane d. Lemaire Institut de Biotechnologie des Plantes UMR 8618 CNRS/Universite Paris-Sud 11 Batiment 630 Orsay 91405 Cedex France

Protein S-glutathionylation, the reversible formation of a mixed-disulfide between glutathione and protein thiols, is involved in protection of protein cysteines from irreversible oxidation, but also in protein redox regulation. Recent studies have implicated S-glutathionylation as a cellular response to oxidative/nitrosative stress, likely playing an important role in signaling. Considering the potential importance of glutathionylation, a number of methods have been developed for identifying proteins undergoing glutathionylation. These methods, ranging from analysis of purified proteins in vitro to large-scale proteomic analyses in vivo, allowed identification of nearly 200 targets in mammals. By contrast, the number of known glutathionylated proteins is more limited in photosynthetic organisms, although they are severely exposed to oxidative stress. The aim of this review is to detail the methods available for identification and analysis of glutathionylated proteins in vivo and in vitro. The advantages and drawbacks of each technique will be discussed as well as their application to photosynthetic organisms. Furthermore, an overview of known glutathionylated proteins in photosynthetic organisms is provided and the physiological importance of this post-translational modification is discussed.

关键词： environmental signals oxidative and photo-oxidative stress proteomics signal transduction Glutathionylation Methods Glutaredoxin Glutathione.

Serous pancreatic neoplasia, data and review

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World Journal of Gastroenterology 2017年第30期23卷 5567-5578页

作者： Christoph F dietrich Yi dong Christian Jenssen Valentina Ciaravino Michael Hocke Wen-Ping Wang Eike Burmester Kathleen Moeller Nathan SS Atkinson Paola Capelli mirko d’Onofrio Medizinische Klinik 2 Caritas-Krankenhaus Bad Mergentheim Ultrasound department First Affiliated Hospital of Zhengzhou University department of Ultrasound Zhongshan HospitalFudan University department of Internal Medicine Krankenhaus Markisch Oderland Strausberg/Wriezen department of Radiology G.B Rossi University HospitalUniversity of Verona department of Internal Medicine 2 Helios Hospital Meiningen GmbH department of Internal Medicine I Sana Klinikum Lichtenberg Translational Gastroenterology Unit Oxford University Hospitals NHS Foundation Trust department of Pathology G.B Rossi University HospitalUniversity of Verona department of Radiology GB Rossi University HospitalUniversity of Verona

AIM To describe the imaging features of serous neoplasms of the pancreas using ultrasound, endoscopic ultrasound, computed tomography and magnetic resonance *** This multicenter international collaboration enhances a literature review to date, reporting features of 287 histologically confirmed cases of serous pancreatic cystic neoplasms(SPN). RESULTS Female predominance is seen with most SPN presenting asymptomatically in the 5th through 7th decade. Mean lesion size was 38.7 mm, 98% were single, 44.2% cystic, 46% mixed cystic and solid, and 94% hypoechoic on B-mode ultrasound. Vascular patterns and contrast-enhancement profiles are described as hypervascular and *** The described ultrasound features can aid differentiation of SPN from other neoplastic lesions under most circumstances.

关键词： Guideline Cancer Ultrasound Endoscopic ultrasound Elastography

Contrast-enhanced ultrasound of the pancreas

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World Journal of Radiology 2010年第3期2卷 97-102页

作者： mirko d'Onofrio Anna Gallotti Francesco Principe Roberto Pozzi Mucelli department of Radiology University Hospital G.B.RossiUniversity of Verona

The introduction of contrast-enhanced ultrasonography(CEUS) has led to major improvements in the diagnostic capabilities of ultrasound(US).The innovative use of CEUS for study of the pancreas has created the need for a definition of the most frequent dynamic features of solid and cystic *** is less expensive compared to computed tomography and magnetic resonance imaging and is able to significantly improve the accuracy of US,allowing better characterization and staging of pancreatic pathologies.

关键词： Contrast-enhanced ultrasonography Ultrasound Pancreatic diseases

Pancreatic neuroendocrine neoplasms: Magnetic resonance imaging features according to grade and stage

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World Journal of Gastroenterology 2017年第2期23卷 275-285页

作者： Riccardo de Robertis Sara Cingarlini Paolo Tinazzi Martini Silvia Ortolani Giovanni Butturini Luca Landoni Paolo Regi Roberto Girelli Paola Capelli Stefano Gobbo Giampaolo Tortora Aldo Scarpa Paolo Pederzoli mirko d'Onofrio department of Radiology Casa di Cura Pederzoli Course in Inflammation Immunity and CancerUniversity of Verona Verona Comprehensive Cancer Network Department of Medical OncologyG.B.Rossi HospitalUniversity of Verona department of Oncology Casa di Cura Pederzoli department of Pancreatic Surgery Casa di Cura Pederzoli department of Pathology Casa di Cura Pederzoli

AIMTo describe magnetic resonance (MR) imaging features of pancreatic neuroendocrine neoplasms (PanNENs) according to their grade and tumor-nodes-metastases stage by comparing them to histopathology and to determine the accuracy of MR imaging features in predicting their biological *** study was approved by our institutional review board; requirement for informed patient consent was waived due to the retrospective nature of the study. Preoperative MR examinations of 55 PanNEN patients (29 men, 26 women; mean age of 57.6 years, range 21-83 years) performed between June 2013 and december 2015 were reviewed. Qualitative and quantitative features were compared between tumor grades and stages determined by histopathological *** defined margins were more common in G2-3 and stage III-IV PanNENs than in G1 and low-stage tumors (P d high specificity in the identification of G2-3 and stage III-IV tumors (90.3% and 96%, 95%CI: 73.1-97.5 and 77.7-99.8). The mean apparent diffusion coefficient value was significantly lower in G2-3 and stage III-IV lesions compared to well differentiated and low-stage tumors (1.09 × 10-3 mm2/s vs 1.45 × 10-3 mm2/s and 1.10 × 10-3 mm2/s vs 1.53 × 10-3 mm2/s, P = 0.003 and 0.001). Receiving operator characteristic analysis determined optimal cut-offs of 1.21 and 1.28 × 10-3 mm2/s for the identification of G2-3 and stage III-IV tumors, with sensitivity and specificity values of 70.8/80.7% and 64.5/64% (95%CI: 48.7-86.6/60-92.7 and 45.4-80.2/42.6-81.3).CONCLUSIONMR features of PanNENs vary according to their grade of differentiation and their stage at diagnosis and could predict the biological behavior of these tumors.

关键词： Pancreatic neuroendocrine tumor World Health Organization classification 2010 diffusion-weighted imaging European Neuroendocrine Tumor Society staging system Magnetic resonance imaging

Mass-forming pancreatitis: Value of contrast-enhanced ultrasonography

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World Journal of Gastroenterology 2006年第26期12卷 4181-4184页

作者： mirko d'Onofrio Giulia Zamboni Alessia Tognolini Roberto Malagò Niccolò Faccioli Luca Frulloni Roberto Pozzi Mucelli department of Radiology University of Verona VeronaItaly department of Gastroenterology University of Verona VeronaItaly

AIM： To assess the utility of contrast-enhanced ultrasonography （CEUS） with a second-generation contrast medium in the differential diagnosis between mass-forming pancreatitis and pancreatic carcinoma. METHOdS： From our radio-pathology database, we retrieved all the patients affected by mass-forming pancreatitis or pancreatic carcinoma who underwent CEUS. We evaluated the results of CEUS in the study of the 173 pancreatic masses considering the possibilities of a differential diagnosis between mass-forming pancreatitis and pancreatic tumor by identifying the ＂parenchymographic＂ enhancement during the dynamic phase of CEUS, which was considered diagnostic for mass-forming pancreatitis. RESULTS： At CEUS, 94% of the mass-forming pancreatitis showed intralesional parenchymography. CEUS allowed diagnosis of mass-forming pancreatitis with sensitivity of 88.6%, specificity of 97.8%, positive predictive value of 91.2%, negative predictive value of 97.1%, and overall accuracy of 96%. CEUS significantly increased the diagnostic confidence in the differential diagnosis between mass-forming pancreatitis and pancreatic carcinoma, with receiver operating characteristic curve areas from 0.557 （P = 0.1608） for baseline US to 0.956 （P 〈 0.0001） for CEUS. CONCLUSION： CEUS allowed diagnosis of massforming pancreatitis with diagnostic accuracy of 96%. CEUS significantly increases the diagnostic confidence with respect to basal US in discerning mass-forming pancreatitis from pancreatic neoplasm.

关键词： Contrast-enhanced ultrasonography Pancreatitis Pancreatic neoplasm Mass-forming pancreatitis

Contrast enhanced ultrasound with quantitative perfusion analysis for objective characterization of pancreatic ductal adenocarcinoma: A feasibility study

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World Journal of Radiology 2014年第3期6卷 31-35页

作者： mirko d'Onofrio Stefano Canestrini Stefano Crosara Riccardo de Robertis Roberto Pozzi Mucelli department of Radiology University Hospital G.B. RossiUniversity of Verona

The aim of this study was to determine whether contrast enhanced ultrasound(CEUS) quantitative perfusion analysis allows an objective characterization of ductal adenocarcinoma(AdK) of the pancreas. Patients with pancreatic AdK underwent CEUS. All examinations were performed on an Acuson S2000 system(Siemens, Erlangen, Germany) after the iv administration of 2.4 mL contrast agent(SonoVue, Bracco, Milan, Italy). All lesions were pathologically proved. An operator manu-ally drew different regions of interest within the tumor and the adjacent parenchyma to allow the quantita-tive perfusion analysis. The mean values of peak of enhancement, time to peak and ascending curve were calculated and compared using the Student's t test. The quantitative perfusion analysis was possible in all lesions. The mean values of the peak of enhancement, time to peak and ascending curve were 17.19%, 7.97 s and 159.52% s within the tumor and 33.57%, 8.89 s and 355.29% s within the adjacent parenchyma. The peak of enhancement and the ascending curve values were significantly different within the tumor and the ad-jacent parenchyma. Thus, CEUS allows the quantitative perfusion analysis of pancreatic ductal adenocarcinoma.

关键词： Contrast enhanced ultrasound Pancreatic adenocarcinoma Perfusion analysis Quantitative imaging

Liver volumetry:Is imaging reliable?Personal experience and review of the literature

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World Journal of Radiology 2014年第4期6卷 62-71页

作者： mirko d'Onofrio Riccardo de Robertis Emanuele demozzi Stefano Crosara Stefano Canestrini Roberto Pozzi Mucelli department of Radiology G.B. Rossi HospitalUniversity of Verona

The amount of the future liver remnant volume is fun-damental for hepato-biliary surgery, representing animportant potential risk-factor for the development ofpost-hepatectomy liver failure. despite this, there isno uniform consensus about the amount of hepaticparenchyma that can be safely resected, nor about themodality that should be chosen for this evaluation. Thepre-operative evaluation of hepatic volume, along witha precise identification of vascular and biliar anatomyand variants, are therefore necessary to reduce surgi-cal complications, especially for extensive *** studies have tried to validate imaging methods[ultrasound, computed tomography(CT), magneticresonance imaging] for the assessment of liver volume,but there is no clear evidence about the most accuratemethod for this evaluation. Furthermore, this volumet-ric evaluation seems to have a certain degree of error,tending to overestimate the actual hepatic volume,therefore some conversion factors, which should givea more reliable evaluation of liver volume, have been proposed. It is widespread among non-radiologists the use of independent software for an off-site volumetric analysis, performed on digital imaging and communica-tions in medicine images with their own personal com-puter, but very few studies have provided a validation of these methods. Moreover, while the pre-transplanta-tion volumetric assessment is fundamental, it remains unclear whether it should be routinely performed in all patients undergoing liver resection. In this editorial the role of imaging in the estimation of liver volume is dis-cussed, providing a review of the most recent literature and a brief personal series of correlations between liver volumes and resection specimens' weight, in order to assess the precision of the volumetric CT evaluation.

关键词： Liver Hepatectomy Ultrasound Computed tomography Magnetic resonance imaging

Left atrium veno-arterial extra corporeal membrane oxygenation as temporary mechanical support for cardiogenic shock:A case report

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World Journal of Clinical Cases 2023年第27期11卷 6531-6536页

作者： Rossana Lamastra david Michael Abbott Antonella degani Carlo Pellegrini Roberto Veronesi Stefano Pelenghi Chiara dezza Giulia Gazzaniga mirko Belliato department of Surgical Pediatric and Diagnostic SciencesUniversity of PaviaPavia 27100PVItaly department of Cardiothoracic Surgery Fondazione IRCCS Policlinico San MatteoPavia 27100PVItaly Clinical SurgicalDiagnostic and Pediatric Sciences DepartmentUniversity of PaviaItaly-Department of Cardiac SurgeryFondazione IRCCS Policlinico San Matteo FoundationPavia 27100PVItaly SC-AR2 Anestesia e Terapia Intensiva Cardiotoracica Fondazione IRCCS Policlinico San Matteo Pavia 27100PVItaly department of Cardiac Surgery 1 IRCCS University Hospital Foundation"San Matteo"Pavia 27100Italy

BACKGROUNd Veno-arterial extra corporeal membrane oxygenation(VA-ECMO)support is commonly complicated with left ventricle(LV)distension in patients with cardiogenic *** resolved this problem by transeptally converting VAECMO to left atrium veno-arterial(LAVA)-ECMO that functioned as a temporary paracorporeal left ventricular assist device to resolve LV *** our case LAVA-ECMO was also functioning as a bridge-to-transplant device,a technique that has been scarcely reported in the *** SUMMARY A 65 year-old man suffered from acute myocardial injury that required percutaneous *** than two weeks later,noncompliance to antiplatelet therapy led to stent thrombosis,cardiogenic shock,and cardiac *** VA-ECMO support was started,and the patient underwent a second coronary angiography with re-stenting and intra-aortic balloon pump *** VA-ECMO support was complicated by left ventricular distension which we resolved via ***,episodes of bleeding and sepsis complicated the clinical picture and the patient passed away 27 d after initiating *** This clinical case demonstrates that LAVA-ECMO is a viable strategy to unload the LV without another invasive percutaneous or surgical *** also demonstrate that LAVA-ECMO can also be weaned to a left ventricular assist device system.A benefit of this technique is that the procedure is potentially reversible,should the patient require VA-ECMO support again.A transeptal LV venting approach like LAVA-ECMO may be indicated over ImpellaTM in cases where less LV unloading is required and where a restrictive myocardium could cause LV *** ventricular over-distention is a well-known complication of peripheral VA-ECMO in cardiogenic shock and LAVA ECMO through transeptal cannulation offers a novel and safe approach for treating LV overloading,without the need of an additional percutaneous access.

关键词： Left atrium venoarterial extra corporeal membrane oxygenation Shock Case report

Prompt and Easy Activation by Specific Thioredoxins of Calvin Cycle Enzymes of Arabidopsis thaliana Associated in the GAPdH/CP12/PRK Supramolecular Complex

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Molecular Plant 2009年第2期2卷 259-269页

作者： Lucia Marri mirko Zaffagnini Valerie Collin Emmanuelle Issakidis-Bourguet Stephane d. Lemaire Paolo Pupillo Francesca Sparla Myroslawa Miginiac-Maslow Paolo Trost Laboratory of Molecular Plant Physiology Department of Experimental Evolutionary Biology University of Bologna Via Irnerio 42 1-40126 Bologna Italy Institut de Biotechnologie des Plantes UMR 8618 CNRS/Univ. Paris-Sud 1 1 91405 Orsay Cedex France

The Calvin cycle enzymes glyceraldehyde-3-phosphate dehydrogenase （GAPdH） and phosphoribulokinase （PRK） can form under oxidizing conditions a supramolecular complex with the regulatory protein CP12. Both GAPdH and PRK activities are inhibited within the complex, but they can be fully restored by reduced thioredoxins （TRXs）. We have investigated the interactions of eight different chloroplast thioredoxin isoforms （TRX f1, m1, m2, m3, m4, y1, y2, x） with GAPdH （A4, B4, and B8 isoforms）, PRK and CP12 （isoform 2）, all from Arabidopsis thaliana. In the complex, both A4-GAPdH and PRK were promptly activated by TRX f1, or more slowly by TRXs ml and m2, but all other TRXs were ineffective. Free PRK was regulated by TRX f1, m1, or m2, while B4- and Bs-GAPdH were absolutely specific for TRX fl. Interestingly, reductive activation of PRK caged in the complex was much faster than reductive activation of free oxidized PRK, and activation of A4-GAPdH in the complex was much faster （and less demanding in terms of reducing potential） than activation of free oxidized B4- or BB-GAPdH. It is proposed that CP12-assembled supramolecular complex may represent a reservoir of inhibited enzymes ready to be released in fully active conformation following reduction and dissociation of the complex by TRXs upon the shift from dark to low light. On the contrary, autonomous redox-modulation of GAPdH （B- containing isoforms） would be more suited to conditions of very active photosynthesis.

关键词： carbon metabolism enzymology light regulation metabolic regulation photosynthesis Arabidopsis.