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1.
Methods Mol Biol ; 2602: 107-122, 2023.
Article in English | MEDLINE | ID: mdl-36446970

ABSTRACT

The family of ubiquitin C-terminal hydrolases (UCHs(releases ε-linked amide bonds positioned at the C-terminus of ubiquitin. UCHL3 is a highly conserved and dual functional member of this family, recognizing C-terminal extensions of two paralogous modifiers: ubiquitin and NEDD8. The Saccharomyces cerevisiae orthologue of UCHL3, namely, Yuh1, is the only UCH family member in this organism. Like UCHL3, Yuh1 recognizes ubiquitin as well as Rub1, the direct orthologue of NEDD8 in S. cerevisiae. We describe here a method for examining the activity of bacteria and yeast expressed Yuh1 by monitoring the C-terminal trimming of UBB + 1 and Rub1 + 1 through immunoblotting and the increased AMC fluorescence readout detected through a plate reader.


Subject(s)
Saccharomyces cerevisiae Proteins , Ubiquitin Thiolesterase , Saccharomyces cerevisiae/genetics , Proteolysis , Ubiquitin , Amides , Ubiquitins
2.
Eur J Clin Invest ; 40(12): 1074-80, 2010 Dec.
Article in English | MEDLINE | ID: mdl-20678121

ABSTRACT

BACKGROUND: Prolonged surgical procedures involving stress, extended general anaesthesia and a long pre-surgical fasting period may have systemic effects such as alterations in saliva flow rate and composition. These may compromise the patient's electrolytes and fluid balance and cause dehydration, systemic stress and oxidative changes. PATIENTS AND METHODS: Saliva was collected prior and following surgery from 20 patients and 20 control subjects. The saliva samples were analysed for flow rates and levels of the following: calcium (Ca), magnesium (Mg), total protein, albumin and lactate dehydrogenase (LDH), total antioxidant status (TAS), uric acid (UA), superoxide dismutase (SOD), carbonyls, matrix metalloproteinases (MMPs) -2, -3 and -9 and heat shock proteins (HSPs) 70 and 90. RESULTS: Salivary levels of Ca, Mg, protein, albumin and LDH were higher in post-surgical patients by 70% (P = 0·002), 88% (P = 0·0001), 120% (P = 0·13), 111% (P = 0·039) and 492% (P = 0·006) respectively than that in healthy controls. Salivary antioxidants in the surgical patients were higher while salivary carbonyls remained unchanged. Salivary TAS levels in pre- and post-surgical patients were higher by 63% (P = 0·001) and 85% (P = 0·0001) respectively, UA concentrations by 92% (P = 0·014) and 81% (P = 0·036) respectively and SOD values by 47% (P = 0·61) and 112% (P = 0·049) respectively. Salivary concentrations of MMP3 were higher in pre- and post-surgical patients by 23% (P = 0·067) and 30% (P = 0·044) respectively. CONCLUSIONS: Local salivary, oral and systemic-induced alterations should be prevented. Moreover, salivary collection and analysis may be a new, efficient tool in the monitoring of patients undergoing major surgery. Further related research is necessary.


Subject(s)
Elective Surgical Procedures , Saliva/chemistry , Salivation/physiology , Adult , Albumins/analysis , Antioxidants/analysis , Calcium/analysis , Female , HSP70 Heat-Shock Proteins/analysis , HSP90 Heat-Shock Proteins/analysis , Humans , Lactate Dehydrogenases/analysis , Magnesium/analysis , Male , Matrix Metalloproteinases/analysis , Middle Aged , Postoperative Period , Preoperative Period , Superoxide Dismutase/analysis , Uric Acid/analysis
3.
Ann Plast Surg ; 62(1): 70-4, 2009 Jan.
Article in English | MEDLINE | ID: mdl-19131724

ABSTRACT

During the 2006 war between Israel and Lebanon, 282 Israeli soldiers were evacuated to Rambam Health Care Campus. Of these, 210 were admitted for observation or treatment, and 15 of these were admitted to the Department of Plastic and Reconstructive Surgery. Thirty-five other soldiers, hospitalized in other departments, required the care of Plastic Surgeons, either for conservative or surgical treatment. The injury profile observed was consistent with data from previous low-intensity warfare, which demonstrated that over 80% of injuries were produced by fragmentation weapons, such as artillery, mortarshells, rockets, and missiles. It differs, however, from our experience in previous wars and our expectations regarding burn wounds, both in incidence and severity, which were significantly lower as compared with the past. This article presents our management of extensive soft tissue injuries, and details 3 representative cases. It highlights the role of the Plastic Surgeon as part of the whole treatment in this type of injury and helps to predict the needs of the medical system in preparation for the future.


Subject(s)
Military Personnel , Plastic Surgery Procedures/methods , Soft Tissue Injuries/surgery , Adult , Humans , Israel , Lebanon , Male , Surgery, Plastic , Warfare , Young Adult
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