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Kidney Sections (kidney + section)
Selected AbstractsSelective imaging of positively charged polar and nonpolar lipids by optimizing matrix solution compositionRAPID COMMUNICATIONS IN MASS SPECTROMETRY, Issue 20 2009Yuki Sugiura Previous studies have shown that matrix-assisted laser desorption/ionization,imaging mass spectrometry (MALDI-IMS) is useful for studying the distribution of various small metabolites, particularly lipids. However, in this technique, selective ionization of the target molecules is imperative, particularly when analyzing small molecules. Since the sample clean-up procedures available for the MALDI-IMS of small metabolites are limited, the tissue sample will contain numerous molecular species other than the target molecules. These molecules will compete for ionization resulting in severe ion suppression. Hence, it is necessary to develop and optimize a sample preparation protocol for the target molecules. In this study, through model experiments using reference compounds, we optimized the composition of the matrix solution used for positively charged lipids in terms of the concentration of the organic solvent and presence/absence of alkali metal salts. We demonstrated that a high concentration of organic solvent in the matrix solution favors the preferential detection of lipids over peptides. The presence of alkali metal salts in the matrix solution was favorable for the detection of polar lipids, while a salt-free matrix solution was suitable for the detection of nonpolar lipids. Furthermore, potassium salts added to the matrix solution caused merging of various lipid adducts (adducts with proton, sodium, and potassium) into one single potassiated species. Using the optimized protocols, we selectively analyzed phosphatidylcholine (PC) and triacylglycerol (TG) with different fatty acid compositions in a rat kidney section. Copyright © 2009 John Wiley & Sons, Ltd. [source] Improved diagnoses of autoimmune hepatitis using an anti-actin ELISAJOURNAL OF CLINICAL LABORATORY ANALYSIS, Issue 5 2008Vincent Aubert Abstract The presence of antismooth muscle antibodies is one of the diagnostic criteria of autoimmune hepatitis. We evaluated a new anti-F-actin ELISA test and compared it with indirect immunofluorescence assay (IIFA) for antismooth muscle antibodies (ASMA). Two hundred and nine serum samples (35 autoimmune hepatitis, 174 other hepatopathies and control sera) were tested by IIFA on mouse stomach kidney sections for ASMA and by the Quanta Lite Actin ELISA for anti-F-actin antibodies. ASMA were detected in 26 of 35 sera from autoimmune hepatitis (74%) as compared with 25 (71%) with anti-actin antibodies, as well as in 25 of 49 (51%) samples from viral hepatitis as compared with 7 (14%) with anti-actin antibodies. With regards to autoimmune hepatitis, though sensitivity (74.3 vs 71.4%) and negative predictive value (93.5 vs 93.9%) of ASMA and anti-actin ELISA were comparable, anti-actin ELISA was significantly better than ASMA IIFA in terms of specificity (89.7 vs 74.7%), and positive predictive value (58.1 vs 37.1%). Although frequently positive in HCV samples, a comparable sensitivity but better specificity makes the anti-actin ELISA a useful tool in combination with ASMA IIFA for the screening and diagnosis of autoimmune hepatitis. J. Clin. Lab. Anal. 22:340,345, 2008. © 2008 Wiley-Liss, Inc. [source] Characterization of an Antibody to the Human Melatonin mt1 ReceptorJOURNAL OF NEUROENDOCRINOLOGY, Issue 1 2001L. M. Williams Abstract Melatonin acts via high affinity, G-protein coupled, seven transmembrane domain receptors. To precisely localize these receptors, antibodies were raised in chickens against a 15 amino acid fragment at the intracellular C -terminal region of the human melatonin receptor subtype mt1 (DSSNDVADRVKWKPS, mt1338,352). A chimeric form of the receptor with a hydrophilic Flag peptide (DYKDDDDK) in sequence with the extracellular N -terminus (Flag-mt1) was generated by polymerase chain reaction and expressed in mammalian cell lines. An IgY antibody (Y31), which gave high antibody titres by enzyme-linked immunosorbent assay, was used to localize Flag-mt1 in stably transfected cells by immunofluoresence. Flag-mt1 localization with Y31 was identical to that obtained with the M5 antibody directed against the Flag epitope and was mainly localized to the Golgi apparatus with some staining at the cell surface. No staining was seen in untransfected cells with either antibody. Y31 staining was abolished using antibody preabsorbed with peptide antigen. Y31 immunofluorescence in fetal human kidney sections was restricted to nephrogenic regions and matched that of 2-(125I)iodomelatonin binding and mt1 gene expression by in situ hybridization. Y31 was used to immunoprecipitate biotinylated membrane proteins from Flag-mt1 stably transfected and untransfected CHO cells. Western blotting of immunoprecipitated proteins revealed two major bands specific to stably transfected cells, one at 63 kDa and one at 86 kDa. The first band almost certainly corresponds to the glycosylated form of Flag-mt1 and the second band to receptor dimers. Thus, Y31 antibody is suitable for use in detecting the human mt1 receptor subtype in tissues and in transfected cells. [source] Assessing the antifungal activity and toxicity profile of amphotericin B lipid complex (ABLC; Abelcet®) in combination with caspofungin in experimental systemic aspergillosisJOURNAL OF PHARMACEUTICAL SCIENCES, Issue 6 2004Olena Sivak Abstract The purpose of this study was to assess the antifungal activity and renal and hepatic toxicity of amphotericin B lipid complex (ABLC; Abelcet®) following co-administration of Caspofungin to rats infected with Aspergillus fumigatus. Aspergillus fumigatus inoculum (1.3,2.3,×,107 colony forming units [CFU]) was injected via the jugular vein; 48 h later male albino Sprague,Dawley rats (350,400 g) were administered either a single intravenous (IV) dose of Fungizone® (1 mg AmpB/kg), ABLC (1 or 5 mg AmpB/kg), or an equivalent volume of normal saline (NS) (vehicle control) once daily for 4 days. Rats were further randomized into groups to receive 3 mg/kg Caspofungin or physiologic saline IV once daily for 4 days. To assess antifungal activity, brain, lung, heart, liver, spleen, and kidney sections were homogenized with NS (2 mL; 1 g of each tissue/mL) and a 0.1-mL aliquot was spread plated onto a Sabouraud dextrose agar plate. The plates were incubated for 48 h at 37°C, at which time the numbers of CFU were determined and corrected for tissue weight. To assess renal and hepatic toxicity, serum creatinine and aspartate aminotransferase levels were determined. Fungizone and ABLC at a dosing regimen of 1 mg/kg i.v. once daily for four consecutive days and Caspofungin at a dosing regimen of 3 mg/kg i.v. once daily for four consecutive days had similar effectiveness in decreasing the total number of Aspergillus fumigatus CFUs found in all organs analyzed compared to non-treated controls. A combination of ABLC (1 mg/kg i.v.,×,4 days) and Caspofungin (3 mg/kg i.v.,×,4 days) significantly decreased the total number of Aspergillus fumigatus CFUs found in all organs analyzed compared to Caspofungin alone and non-treated controls. ABLC at a dosing regiment of 5 mg/kg i.v. once daily for four consecutive days was more effective in decreasing the total number of Aspergillus fumigatus CFUs found in all organs analyzed compared to Fungizone or ABLC alone at 1 mg/kg and Caspofungin alone at 3 mg/kg. However, a combination of ABLC (5 mg/kg i.v.,×,4 days) and Caspofungin (3 mg/kg i.v.,×,4 days) was not more effective than ABLC at 5 mg/kg or the combination of ABLC at 1 mg/kg and Caspofungin 3 mg/kg in reducing the total number of Aspergillus fumigatus CFUs compared to controls. Except for non-treated infected control rats, none of the treatment groups tested displayed a greater than 50% increase in serum creatinine concentrations from baseline. In addition, only ABLC at a dosing regimen of 1 mg/kg i.v. once daily for four consecutive days displayed a greater than 50% increase in AST concentration from baseline. Taken together, these findings suggest that ABLC at 5 mg/kg once daily,×,4 days appears to be the best therapeutic choice in this animal model. © 2004 Wiley-Liss, Inc. and the American Pharmacists Association J Pharm Sci 93:1382,1389, 2004 [source] Bioimaging TOF-SIMS of tissues by gold ion bombardment of a silver-coated thin sectionMICROSCOPY RESEARCH AND TECHNIQUE, Issue 6 2004Håkan Nygren Abstract The imaging time-of-flight secondary-ion-mass-spectrometry (TOF-SIMS) method was utilized to address the problem of cholesterol localization in rat tissues. Rat kidneys were fixed, cryoprotected by sucrose, frozen, sectioned by cryoultramicrotomy, and dried at room temperature. The samples were either covered with a thin silver layer or analyzed uncovered in an imaging TOF-SIMS instrument equipped with an Au -source. The yield of desorbed secondary ions for some species was up to 600-fold higher after silver coating of the samples. Reference samples of cholesterol were silver-coated and analyzed by TOF-SIMS to define significant peaks, specific for cholesterol. Such peaks were found at m/z = 386 (C27H46O+), m/z = 493 (C27H46O107Ag+), m/z = 495 (C27H46O109Ag+), m/z = 879 (C54H92O2107Ag+), and m/z = 881 (C54H92O2109Ag+). The silver-cationized cholesterol (493 , m/z , 495) signal was localized by imaging TOF-SIMS in the kidney sections and showed a high cholesterol content in the kidney glomeruli. A more diffuse distribution of cholesterol was also found over areas representing the cytoplasm or plasma membrane of the epithelial cells in the proximal tubules of rat kidney. Microsc. Res. Tech. 65:282,286, 2004. © 2005 Wiley-Liss, Inc. [source] Matrix-assisted laser desorption/ionization imaging mass spectrometry of oxaliplatin derivatives in heated intraoperative chemotherapy (HIPEC)-like treated rat kidneyRAPID COMMUNICATIONS IN MASS SPECTROMETRY, Issue 4 2010Amina Bouslimani Oxaliplatin [1,2-diaminocyclohexane (dach)-Pt complex] is a platinum anticancer drug which is mainly used in the treatment of advanced colorectal cancer, particularly in Heated Intraoperative Chemotherapy (HIPEC) for the treatment of colorectal peritoneal carcinomatosis. In order to better understand the penetration of oxaliplatin in treated tissues we performed a direct imaging of tissue sections from HIPEC-like treated rat kidney using matrix-assisted laser desorption/ionization (MALDI) mass spectrometry. This procedure allowed the detection and localization of oxaliplatin and its metabolites, the monocysteine and monomethionine complexes, in kidney sections. Specifically, oxaliplatin and its metabolites were localized exclusively in the kidney cortex, suggesting that it did not penetrate deeply into the organ. Based on these results, an imaging analysis of human tumors collected after HIPEC is currently in progress to assess the distribution of oxaliplatin and/or metabolites with the aim of defining clinical conditions to improve drug penetration. Copyright © 2010 John Wiley & Sons, Ltd. [source] Developing Ovarian Follicles Inhibit the Endotoxin-Induced Glomerular Inflammatory Reaction in Pseudopregnant RatsAMERICAN JOURNAL OF REPRODUCTIVE IMMUNOLOGY, Issue 5 2004Marijke M. Faas Problem:, We tested the hypothesis that developing ovarian follicles produce factors inhibiting the endotoxin induced inflammatory response. Method of study:, Pseudopregnant rats were treated with FSH to induced follicular development (FSH-rats). For control we used untreated pseudopregnant rats (PSP-rats) and rats in the follicular phase of the ovarian cycle (C-rats). All rats were infused with either saline or endotoxin. Three days after the infusion rats were sacrificed and kidney specimens were snapfrozen. Cryostat kidney sections were stained for the presence of monocytes, granulocytes, CD11a- and CD11b-positive cells and for ICAM-1 expression. Results:, The results show that induction of follicular development in pseudopregnant rats inhibited glomerular infiltration of monocytes and CD11b+ cells, while it did not affect the other parameters, i.e. glomerular granulocyte number, CD11a+ cells and glomerular ICAM-1 expression. Conclusion:, Developing ovarian follicles produce factors inhibiting monocyte responses to endotoxin. [source] CXCR3+CD4+ T cells are enriched in inflamed kidneys and urine and provide a new biomarker for acute nephritis flares in systemic lupus erythematosus patientsARTHRITIS & RHEUMATISM, Issue 1 2009P. Enghard Objective The high frequency of CD4+ T cells in interstitial infiltrates of patients with lupus nephritis suggests a contribution of these cells to local pathogenesis. The aim of this study was to examine the role of CXCR3 and the chemokine CXCL10 in recruiting these cells into the kidney and to determine whether the infiltrating T cells could be monitored in the urine to provide a reliable biomarker for acute lupus nephritis. Methods The frequencies of CD3+ T cells, CXCR3+ cells, and CXCL10+ cells were determined by immunohistochemical and immunofluorescence analyses of kidney sections from 18 patients with lupus nephritis. The frequency of CXCR3+CD4+ T cells was determined by flow cytometry of peripheral blood and urine from 38 patients with systemic lupus erythematosus (SLE), and the values were compared with disease activity as determined by the Systemic Lupus Erythematosus Disease Activity Index. Results In renal biopsy tissues from patients with lupus nephritis, a mean of 63% of the infiltrating cells expressed CXCR3, ,60% of them were T cells, and the CXCR3+ cells colocalized with CXCL10-producing cells. In biopsy tissues from SLE patients with acute nephritis, ,50% of the urinary CD4+ T cells were CXCR3+, as compared with 22% in the peripheral blood, and the frequency of urinary CXCR3+CD4+ T cells correlated with disease activity. Moreover, the number of urinary CD4+ T cells reflected nephritis activity, and elevation above 800 CD4+ T cells per 100 ml of urine sharply delineated active from inactive nephritis. Conclusion CXCR3+ T cells are recruited into the inflamed kidneys, are enriched in the urine, and are a valuable marker of nephritis activity in SLE. They also present a potential target for future therapies. [source] | |||||||||||||