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Quantification of Global Protein Disulfides and Thiol-Protein Mixed Disulfides to Study the Protein Dethiolation Mechanisms

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American Journal of Analytical Chemistry 2013年第10期4卷 9-19页

作者： Lucia Coppo Raffaella Priora Sonia Salzano pietro ghezzi Paolo Di Simplicio Brighton and Sussex Medical School Trafford Centre Falmer Brighton UK Department of Neuroscience Pharmacology Unit University of Siena Siena Italy

The redox state of cellular thiols is widely studied because it was recently linked to many different diseases and pathologies. In this work we quantified the concentrations of protein disulfides (PSSP) and thiol-protein mixed disulfides (XSSP) in rat tissues (liver, kidney and heart) and cells (Raw 264.7) by an improved method of XSSP and PSSP determination after oxidative stress induced by diamide. Under native and denaturing conditions, a thiol block by N-ethymaleimide was introduced to avoid thiol exchange reaction activations by protein SH groups (PSH) (PSH + XSSP ←→ PSSP + XSH) and alterations of original XSSP/PSSP levels. Low molecular weight thiols (XSH) and PSH were respectively measured by HPLC on supernatants and on corresponding pellets by DTNB (Ellman’s reagent) after dithiothreitol reduction. PSSP concentrations of liver, heart and kidney were respectively 0.304, 0.605 and 0.785 μmoles/g and after diamide exposure they were significantly augmented of about 65%-70% in liver and heart, but not in the kidney. Normal XSSP, that were -20 times lower than normal PSSP were induced by diamide in liver and heart of about 40 times, but not in kidney. Thermodynamic criteria regarding the pKa values of thiols engaged as PSSP and GSSP were used to interpret dethiolation mechanisms via thiol exchange reactions.

关键词： Protein Disulfides S-Glutathionylation Thiol pKa Diamide Oxidative Stress

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Dealing with robot-assisted surgery for rectal cancer:current status and perspectives

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World Journal of Gastroenterology 2016年第2期22卷 546-556页

作者： Roberto Biffi Fabrizio Luca Paolo pietro Bianchi Sabina Cenciarelli Wanda Petz Igor Monsellato Manuela Valvo Maria Laura Cossu Tiago Leal ghezzi Kassem Shmaissany Department of Gastrointestinal Surgery European Institute of Oncology20141 MilanItaly

The laparoscopic approach for treatment of rectal cancer has been proven feasible and oncologically safe, and is able to offer better short-term outcomes than traditional open procedures, mainly in terms of reduced length of hospital stay and time to return to working activity. In spite of this, the laparoscopic technique is usually practised only in high-volume experienced centres, mainly because it requires a prolonged and demanding learning curve. It has been estimated that over 50 operations are required for an experienced colorectal surgeon to achieve proficiency with this technique. Robotic surgery enables the surgeon to perform minimally invasive operations with better vision and more intuitive and precise control of the operating instruments, thus promising to overcome some of the technical difficulties associated with standard laparoscopy. It has high-definition threedimensional vision, it translates the surgeon's hand movements into precise movements of the instruments inside the patient, the camera is held and moved by the first surgeon, and a fourth robotic arm is available as a fixed retractor. The aim of this review is to summarise the current data on clinical and oncologic outcomes of robot-assisted surgery in rectal cancer, focusing on short- and long-term results, and providing original data from the authors' centre.

关键词： Rectal cancer Robotic total mesorectalexcision Robotic surgery Robotic resection Circumferentialresection margin Lymph node yield Distalresection margin positivity

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