ABSTRACT
Acute renal failure (ARF) is a frequent complication of Gram-negative sepsis, with a high risk of mortality. Lipopolysaccharide (LPS)-induced ARF is associated with hemodynamic changes that are strongly influenced by the overproduction of nitric oxide (NO) through the cytokine-mediated up-regulation of inducible NO synthase. LPS-induced reductions in systemic vascular resistance paradoxically culminate in renal vasoconstriction. Collagen XVIII is an important component of the extracellular matrix expressed in basement membranes. Its degradation by matrix metalloproteases, cathepsins and elastases results in the formation of endostatin, claimed to have antiangiogenic activity and to be a prominent vasorelaxing agent. We evaluated the expression of endostatin/collagen XVIII in an endotoxemic ARF model. ARF was induced in C57BL/6 mice by intraperitoneal injection of LPS (10 mg/kg) followed by sacrifice 4 and 12 h later. Kidney tissue was the source of RNA and protein and the subject of histological analysis. As early as 4 h after LPS administration, blood urea, creatinine and NO levels were significantly increased compared to control. Endostatin/collagen XVIII mRNA levels were 0.71 times lower than sham-inoculated mice 4 h after LPS inoculation, returning to normal levels 12 h after LPS inoculation. Immunohistological examination revealed that acute injury caused by LPS leads to an increase of endostatin basement membrane staining in association with the decrease of CD31 endothelial basement membrane staining. These results indicate that in the early phase of endotoxemic ARF the endostatin levels were not regulated by gene expression, but by the metabolism of collagen XVIII.
Subject(s)
Animals , Mice , Acute Kidney Injury , Collagen Type XVIII/metabolism , Endostatins/metabolism , Endotoxemia/metabolism , Gene Expression , Blotting, Western , Collagen Type XVIII/genetics , Creatinine/blood , Endostatins/genetics , Immunohistochemistry , Lipopolysaccharides , Nitric Oxide/blood , Reverse Transcriptase Polymerase Chain Reaction , RNA, Messenger , Urea/bloodABSTRACT
Acute renal failure (ARF) is a frequent complication of Gram-negative sepsis, with a high risk of mortality. Lipopolysaccharide (LPS)-induced ARF is associated with hemodynamic changes that are strongly influenced by the overproduction of nitric oxide (NO) through the cytokine-mediated up-regulation of inducible NO synthase. LPS-induced reductions in systemic vascular resistance paradoxically culminate in renal vasoconstriction. Collagen XVIII is an important component of the extracellular matrix expressed in basement membranes. Its degradation by matrix metalloproteases, cathepsins and elastases results in the formation of endostatin, claimed to have antiangiogenic activity and to be a prominent vasorelaxing agent. We evaluated the expression of endostatin/collagen XVIII in an endotoxemic ARF model. ARF was induced in C57BL/6 mice by intraperitoneal injection of LPS (10 mg/kg) followed by sacrifice 4 and 12 h later. Kidney tissue was the source of RNA and protein and the subject of histological analysis. As early as 4 h after LPS administration, blood urea, creatinine and NO levels were significantly increased compared to control. Endostatin/collagen XVIII mRNA levels were 0.71 times lower than sham-inoculated mice 4 h after LPS inoculation, returning to normal levels 12 h after LPS inoculation. Immunohistological examination revealed that acute injury caused by LPS leads to an increase of endostatin basement membrane staining in association with the decrease of CD31 endothelial basement membrane staining. These results indicate that in the early phase of endotoxemic ARF the endostatin levels were not regulated by gene expression, but by the metabolism of collagen XVIII.
Subject(s)
Acute Kidney Injury/metabolism , Collagen Type XVIII/metabolism , Endostatins/metabolism , Endotoxemia/metabolism , Gene Expression , Animals , Blotting, Western , Collagen Type XVIII/genetics , Creatinine/blood , Endostatins/genetics , Immunohistochemistry , Lipopolysaccharides , Mice , Mice, Inbred C57BL , Nitric Oxide/blood , RNA, Messenger , Reverse Transcriptase Polymerase Chain Reaction , Urea/bloodABSTRACT
The application of a new fluorogenic pre-column derivatizing reagent, 2-(1-pyrenyl)ethyl chloroformate (PEOC), is reported for the separation and detection of biogenic polyamines using column liquid chromatography. The development of the method included the optimization of excitation and emission wavelengths, efficient gradient programming, derivatization temperature, time, and pH. Minimum detection limits, linear ranges, reproducibility, and recovery from analyzed samples were determined. The procedure was applied to hydrolyzed serum samples taken from healthy individuals and cancer patients. Separation of PEOC-derivatized polyamines from the serum hydrolysis by-products was successful and detection limits were more favorable than those previously reported for 9-fluorenyl-methyl chloroformate-derivatized polyamines.
Subject(s)
Biogenic Amines/blood , Chromatography, High Pressure Liquid/methods , Fluorescent Dyes , Formic Acid Esters , Indicators and Reagents , Pyrenes , Biogenic Amines/chemistry , Humans , Spectrometry, FluorescenceSubject(s)
Mastoiditis/etiology , Osteomyelitis/etiology , Temporal Bone/injuries , Child , Chronic Disease , Humans , Male , Time FactorsABSTRACT
The membrane electrode based on a synthetic chalcocite (Cu(2)S) single crystal responds primarily to the activity of copper(I) ions in solution. The experimental selectivity coefficient with respect to copper(II) ions is in good agreement with the value calculated on the basis of solubility products of both sulphides. The electrode has been calibrated with metal-ion buffers containing a strongly complexing ligand. TETREN, and can be used as an indicator in titrations of copper with EDTA and TETREN. Comparison of an experimental titration curve with one calculated with the aid of the program HALTAFALL showed good agreement in the case of TETREN, but there were discrepancies for the EDTA titration, which are attributed to the presence and complexation of copper(I) ions. The electrode has also been applied in metal titrations with Cu(2+) as indicator ion, though the potential changes observed were smaller than predicted. All titrations showed errors less than 1%.
ABSTRACT
The necessity of prolonged administration of the vaccine Polyvaccinum prompted this study on the effect of the preserving agent on the nasal mucosa. Buffered physiologic saline solution containing 0.1--0.4% phenol, and the vaccine containing 0.1--0.4% phenol or 0.01% merthiolate were applied for 12 weeks on the nasal mucosa of rabbits. The nasal mucosa, parenchymal organs and adrenals were studied macroscopically and histologically, and immunologic studies were made. On the basis of the results, preservation of Polyvaccinum mite with 0.4% phenol is suggested.