ABSTRACT
Formalin-fixed tissues are an important source of biological samples for biomedical research. However, proteomics analysis of formalin-fixed tissues has been set aside by formalin-induced protein modifications, which reduce protein extraction efficiency. In this study, a two level full factorial experimental design (2(4)) was used to determine the effects of the extracting conditions in the efficiency of protein recovery from formalin-fixed kidney samples. The following variables were assessed: temperature of extraction, pH of extraction, composition of the extracting buffer and the use ultrasonic energy applied with probe. It is clearly demonstrated that when hating and ultrasonic energy are used in conjunction, a 7-fold increase (p < 0.05) in protein extraction is obtained if compared to extracting conditions for which neither heating nor ultrasonic energy are used. The optimization study was done following the amount of protein extracted by UV (Nanodrop(®) technology, protein ABS at 280 nm) and by 1D SDS-PAGE. Extracts obtained with the optimized conditions were subjected to LC-MALDI MS/MS. A total of 112 proteins were identified.
Subject(s)
Formaldehyde , Kidney/chemistry , Proteins/isolation & purification , Chromatography, Liquid , Electrophoresis, Polyacrylamide Gel , Humans , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization , Tandem Mass SpectrometrySubject(s)
Calciphylaxis/therapy , Combined Modality Therapy , Debridement , Thiosulfates/therapeutic use , Anti-Bacterial Agents/therapeutic use , Antiphospholipid Syndrome/complications , Carcinoma, Renal Cell/complications , Carcinoma, Renal Cell/surgery , Cardiovascular Diseases/complications , Cardiovascular Diseases/drug therapy , Cinacalcet , Clostridium Infections/complications , Clostridium Infections/drug therapy , Clostridium Infections/surgery , Clostridium perfringens , Foot Ulcer/drug therapy , Foot Ulcer/etiology , Foot Ulcer/microbiology , Foot Ulcer/surgery , Humans , Hyperparathyroidism, Secondary/drug therapy , Hyperparathyroidism, Secondary/etiology , Hyperparathyroidism, Secondary/surgery , Kidney Failure, Chronic/complications , Kidney Failure, Chronic/surgery , Kidney Failure, Chronic/therapy , Kidney Neoplasms/complications , Kidney Neoplasms/surgery , Kidney Transplantation , Male , Middle Aged , Naphthalenes/therapeutic use , Postoperative Complications/etiology , Postoperative Complications/therapy , Renal Replacement Therapy , Staphylococcal Infections/complications , Staphylococcal Infections/drug therapy , Staphylococcal Infections/surgeryABSTRACT
No disponible
Subject(s)
Humans , Gentamicins/pharmacokinetics , Drug Resistance, Microbial , Catheter-Related Infections/prevention & control , Peritoneal Dialysis/adverse effects , Peritonitis/prevention & controlABSTRACT
No disponible
Subject(s)
Humans , Male , Middle Aged , Calciphylaxis/drug therapy , Renal Insufficiency, Chronic/complications , Renal Replacement Therapy/adverse effects , Vascular Calcification/physiopathologySubject(s)
Antibiotic Prophylaxis , Catheter-Related Infections/prevention & control , Peritoneal Dialysis/adverse effects , Peritonitis/prevention & control , Administration, Topical , Candidiasis/etiology , Candidiasis/prevention & control , Catheter-Related Infections/epidemiology , Catheter-Related Infections/etiology , Catheter-Related Infections/microbiology , Catheters, Indwelling , Device Removal , Gentamicins/administration & dosage , Gentamicins/adverse effects , Gentamicins/therapeutic use , Gram-Negative Bacterial Infections/epidemiology , Gram-Negative Bacterial Infections/etiology , Gram-Negative Bacterial Infections/prevention & control , Humans , Peritoneal Dialysis/instrumentation , Peritonitis/epidemiology , Peritonitis/etiology , Peritonitis/microbiology , Staphylococcal Infections/epidemiology , Staphylococcal Infections/etiology , Staphylococcal Infections/prevention & controlSubject(s)
Hemolytic-Uremic Syndrome/therapy , Hypertension/drug therapy , Pre-Eclampsia/therapy , Puerperal Disorders/therapy , Adult , Angiotensin II Type 1 Receptor Blockers/therapeutic use , Anti-Inflammatory Agents/therapeutic use , Antihypertensive Agents/therapeutic use , Cesarean Section , Combined Modality Therapy , Emergencies , Female , Hemolytic-Uremic Syndrome/drug therapy , Humans , Labetalol/therapeutic use , Methyldopa/therapeutic use , Plasmapheresis , Pre-Eclampsia/drug therapy , Prednisone/therapeutic use , Pregnancy , Puerperal Disorders/drug therapy , Remission InductionABSTRACT
No disponible
Subject(s)
Humans , Catheter-Related Infections/prevention & control , Peritoneal Dialysis/methods , Peritonitis/prevention & control , Risk FactorsABSTRACT
No disponible
Subject(s)
Humans , Female , Pregnancy , Adult , Pre-Eclampsia , Hypertension/complications , Hemolytic-Uremic Syndrome/complications , Pulmonary Edema/complications , Acute Kidney Injury/physiopathology , Pregnancy ComplicationsSubject(s)
Edema/etiology , Genital Diseases, Male/etiology , Peritoneal Dialysis/adverse effects , Aged , Humans , MaleABSTRACT
No disponible
Subject(s)
Humans , Male , Aged, 80 and over , Edema/etiology , Genital Diseases, Male/etiology , Peritoneal Dialysis/adverse effects , Tomography, X-Ray ComputedABSTRACT
No disponible
Subject(s)
Humans , Male , Aged , Peritonitis/etiology , Peritoneal Dialysis/adverse effects , Renal Insufficiency, Chronic/complicationsABSTRACT
No disponible