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Pig Kidney (pig + kidney)
Selected AbstractsThermal deactivation and inhibition of D -Amino acid oxidase in permeabilized cells of the yeast Trigonopsis variabilisJOURNAL OF CHEMICAL TECHNOLOGY & BIOTECHNOLOGY, Issue 4 2004José A Moreno Abstract The inhibition of D -amino acid oxidase contained in permeabilized cells of the yeast Trigonopsis variabilis by ,-keto acids (pyruvic acid, phenylpyruvic acid and 4-methylthio-2-oxobutanoic acid), products of the transformation of the corresponding D -amino acids, was studied. In all cases, inhibition was of the mixed type and significant differences with respect to the inhibition shown by the enzyme from other sources such as pig kidney or the yeast Rhodotorula gracilis were observed. A study was also made of the thermal deactivation of the enzyme contained in permeabilized cells of T variabilis in the temperature range 30,50 °C in sodium phosphate and Tris hydroxylmethyl aminomethane + CaCl2 buffers. A deactivation mechanism with two steps in series is proposed to account for the variation in activity with time. The results suggest that the enzyme shows greater stability in phosphate buffer, with half-lives between 7.6 days at 30 °C and 8.6 h at 50 °C. Copyright © 2004 Society of Chemical Industry [source] Absence of 2,-deoxyguanosine-carbon 8-bound ochratoxin A adduct in rat kidney DNA monitored by isotope dilution LC-MS/MSMOLECULAR NUTRITION & FOOD RESEARCH (FORMERLY NAHRUNG/FOOD), Issue 4 2008Thierry Delatour Abstract The contribution of DNA adduct formation in the carcinogenic action of the mycotoxin ochratoxin A (OTA) has been subject to much debate. Recently, a carbon-bonded ochratoxin A-2,-deoxyguanosine adduct (dGuoOTA) formed by photochemical reaction in vitro has been shown by 32P-postlabeling/TLC to comigrate with a spot detected in DNA isolated from rat and pig kidney following exposure to OTA. Considering the large body of evidence arguing against covalent DNA binding of OTA and the poor resolution and specificity of postlabeling analysis, we developed a stable isotope dilution LC-MS/MS method to analyze dGuoOTA in kidney DNA isolated from rats treated with OTA. dGuoOTA and nitrogen-15-labeled dGuoOTA (15N5 -dGuoOTA) were prepared by photoirradiation of OTA in the presence of dGuo or nitrogen-15-labeled dGuo. Conditions for DNA hydrolysis were optimized using a synthetic oligonucleotide containing dGuoOTA to ensure complete release of dGuoOTA. The LOD of the method (S/N > 3) was 10 fmol dGuoOTA on-column. However, dGuoOTA was not detected in DNA samples isolated from male F344 rats treated with OTA for up to 90 days at doses known to cause renal tumor formation. Detection limits, calculated for each individual sample based on the absolute LOD and the amount of DNA injected, were as low as 3.5 dGuoOTA/109 nucleotides. These data are consistent with previous results showing lack of DNA adduct formation by OTA and demonstrate that dGuoOTA is not formed in biologically relevant amounts under physiological conditions in vivo. [source] Structural analysis of ,-Gal and new non-Gal carbohydrate epitopes from specific pathogen-free miniature pig kidneyPROTEINS: STRUCTURE, FUNCTION AND BIOINFORMATICS, Issue 13 2008Yun-Gon Kim Abstract The major barrier in transplantation of pig organs into humans is the presence of surface carbohydrate antigens (e.g., the Gal,1-3Gal,1-4GlcNAc-R (,-Gal) epitope) expressed on pig endothelial cells. In this study, total N -glycans from membrane glycoproteins derived from specific pathogen-free miniature pig kidney are identified by MALDI-TOF, negative ion ESI MS/MS and normal-phase HPLC (NP-HPLC) combined with exoglycosidase digestion. Over 100 N -glycans, including sialylated and neutral types, were identified. As well as the known ,-Gal antigens, some of these glycans contained novel non-Gal carbohydrate antigens such as (Neu5Gc-Gal-GlcNAc) and Gal,1-3Lewisx (Gal-Gal-(Fuc)GlcNAc) which have not been reported before in N -glycans from pig organs. The ability of MALDI, ESI, and HPLC to measure the relative proportions of the glycans was evaluated. The HPLC resolution was insufficient for accurate work and some minor differences were noted in the ionization efficiencies of different glycan groups when measured by the two mass spectrometric techniques. However, the results indicated that the relative quantity of ,-Gal epitope was in the region of 50% of the complex glycans. High-mannose type glycans were also abundant (35,43%) but appeared to be ionized more efficiently than the complex glycans by ESI than by MALDI. [source] Morphological changes induced in the pig kidney by extracorporeal shock wave lithotripsy: Nephron injuryTHE ANATOMICAL RECORD : ADVANCES IN INTEGRATIVE ANATOMY AND EVOLUTIONARY BIOLOGY, Issue 1 2003Youzhi Shao Abstract While shock wave lithotripsy (SWL) is known to cause significant damage to the kidney, little is known about the initial injury to cells along the nephron. In this study, one kidney in each of six juvenile pigs (6,7 weeks old) was treated with 1,000 shock waves (at 24 kV) directed at a lower pole calyx with an unmodified HM-3 lithotripter. Three pigs were utilized as sham-controls. Kidneys were fixed by vascular perfusion immediately after SWL or sham-SWL. Three of the treated kidneys were used to quantitate lesion size. Cortical and medullary samples for light (LM) and transmission electron microscopy (TEM) were taken from the focal zone for the shock waves (F2), the contralateral kidney, and the kidneys of sham-SWL pigs. Because preservation of the tissue occurred within minutes of SWL, the initial injury caused by the shock waves could be separated from secondary changes. No tissue damage was observed in contralateral sham-SWL kidneys, but treated kidneys showed signs of injury, with a lesion of 0.2% ± 0.1% of renal volume. Intraparenchymal hemorrhage and injury to tubules was found at F2 in both the cortex and medulla of SWL-treated kidneys. Tubular injury was always associated with intraparenchymal bleeding, and the range of tissue injury included total destruction of tubules, focal cellular fragmentation, necrosis, cell vacuolization, and membrane blebbing. The initial injury caused by SWL was cellular fragmentation and necrosis. Cellular vacuolization, membrane blebbing, and disorganization of apical brush borders appear to be secondary changes related to hypoxia. Anat Rec Part A 275A:979,989, 2003. © 2003 Wiley-Liss, Inc. [source] Renal and Cardiac Endothelial Heterogeneity Impact Acute Vascular Rejection in Pig-to-Baboon XenotransplantationAMERICAN JOURNAL OF TRANSPLANTATION, Issue 5 2009C. Knosalla Xenograft outcomes are dictated by xenoantigen expression, for example, Gal ,1, 3Gal (Gal), but might also depend on differing vascular responses. We investigated whether differential vascular gene expression in kidney and cardiac xenografts correlate with development of thrombotic microangiopathy (TM) and consumptive coagulation (CC). Immunosuppressed baboons underwent miniswine or hDAF pig kidney (n = 6) or heart (n = 7), or Gal-transferase gene-knockout (GalT-KO) (thymo)kidney transplantation (n = 14). Porcine cDNA miniarrays determined donor proinflammatory, apoptosis-related and vascular coagulant/fibrinolytic gene expression at defined time points; validated by mRNA, protein levels and immunopathology. hDAF-transgenic and GalT-KO xenografts, (particularly thymokidneys) exhibited prolonged survival. CC was seen with Gal-expressing porcine kidneys (3 of 6), only 1 of 7 baboons postcardiac xenotransplantation and was infrequent following GalT-KO grafts (1 of 14). Protective-type genes (heme oxygenase-I, superoxide dismutases and CD39) together with von Willebrand factor and P-selectin were upregulated in all renal grafts. Transcriptional responses in Gal-expressing xenografts were comparable to those seen in the infrequent GalT-KO rejection. In cardiac xenografts, fibrin deposition was associated with increased plasminogen activator inhibitor-1 expression establishing that gene expression profiles in renal and cardiac xenografts differ in a quantitative manner. These findings suggest that therapeutic targets may differ for renal and cardiac xenotransplants. [source] Preparation of active enzyme samples for IR studies of Na+/K+ -ATPaseBIOPOLYMERS, Issue 4-5 2002Detlef Thoenges Abstract In the case of the integral membrane protein Na+/K+ -ATPase, preparation of highly concentrated samples for IR difference spectroscopy often leads to inactivation of the enzyme. Therefore, we compared the activity of Na+/K+ -ATPase using different techniques of sample preparation. The loss of activity can be minimized by cooling the sample to 10°C and by the addition of glycerol and dithiothreitol. The activity of Na+/K+ -ATPase isolated from pig kidney is independent of the protein concentration whereas the enzyme from shark rectal gland is inactivated at concentrations above 1 ,g/,L and is thus unsuitable for IR experiments. © 2002 Wiley Periodicals, Inc. Biopolymers (Biospectroscopy) 67: 271,274, 2002 [source] Kinetics of PME/Pi in pig kidneys during cold ischemiaNMR IN BIOMEDICINE, Issue 7 2007Dominik von Elverfeldt Abstract Quality assessment of renal grafts via 31P magnetic resonance spectroscopy (MRS) has been investigated since 1986. As ATP concentrations decay rapidly during cold ischemia, the ratio of phosphomonoesters (PME) to inorganic phosphate (PiO) within the organ (PME/PiO) is commonly used as a quality marker and is considered to be the most reliable parameter. MRS did not lead to any delay in the transplantation procedure since it was performed during the time necessary for immunological matching (cross-match). Differences in the time period until transplantation call for extrapolation of the measured ratio to the end of cold ischemia before correlating with graft performance after transplantation. Therefore, quantitative determination of PME/PiO kinetics is essential. As a model for metabolite decay in human renal grafts, pig kidneys obtained from a slaughterhouse were monitored for up to 80,h via 31P MRS at 2,T. By employing chemical shift imaging (CSI) with a spatial resolution of approximately 1,×,1,×,4,cm3, it was possible to reduce partial volume effects significantly. The improved spectral resolution gained through CSI enabled reliable PME/PiO ratios to be determined only from those voxels containing renal tissue. Spectra were fitted automatically using the magnetic resonance user interface (MRUI), with prior knowledge obtained from unlocalized spectra when necessary. A monoexponential time dependence of PME/PiO for histidine,tryptophane,alpha-ketoglutarate (HTK)-perfused kidneys during cold ischemia was observed, and the determined value of the decay constant , was 0.0099,±,0.0012,h,1. In University of Wisconsin solution (UW)-perfused kidneys, an , of 0.0183,±,0.0053,h,1 was determined. Determination of the decay constant enables a usable extrapolation of PME/PiO for quality assessment of UW perfusion and a reliable extrapolation for HTK-perfused human renal grafts. Copyright © 2007 John Wiley & Sons, Ltd. [source] | ||||||||||||||||