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PubMed1976

Dipeptidyl peptidase IV, a kidney brush-border serine peptidase.

Dipeptidyl peptidase IV, an enzyme that releases dipeptides from substrates with N-terminal sequences of the forms X-Pro-Y or X-Ala-Y, was purified 300-fold from pig kidney cortex. The kidney is the main source of the enzyme, where it is one of the major microvillus-membrane proteins. Several other tissues contained demonstrable activity against the usual assay substrate glycylproline 2-naphthylamide. In the small intestine this activity was greatly enriched in the microvillus fraction. In all tissues examined, the activity was extremely sensitive to inhibition by di-isopropyl phosphorofluoridate (Dip-F), but relatively resistant to inhibition by phenylmethylsulphonyl fluoride. It is a serine proteinase which may be covalently labelled with [32P]Dip-F, and is the only enzyme of this class in the microvillus membrane. The apparent subunit mol.wt. estimated by sodium dodecyl-sulphate/polyacrylamide-gel electrophoresis and by titration with [32P]Dip-F was 130 000. Gel-filtration and sedimentation-equilibrium methods gave values in the region of 280 000, which is consistent with a dimeric structure, a conclusion supported by electron micrographs of the purified enzyme. Among other well-characterized serine proteinases, this enzyme is unique in its membrane location and its large subunit size. Investigation of the mode of attack of the peptidase on oligopeptides revealed that it could hydrolyse certain N-blocked peptides, e.g. Z-Gly-Pro-Leu-Gly-Pro. In this respect it is acting as an endopeptidase and as such may merit reclassification and renaming as microvillus-membrane serine peptidase.

Animals↗
PubMed1997

In vivo analysis of cadmium in battery workers versus measurements of blood, urine, and workplace air.

OBJECTIVES: To measure in vivo the cadmium concentrations in kidney cortex (kidney-Cd) and in superficial liver tissue (liver-Cd) of nickel cadmium battery workers, and to compare the results with other commonly used estimates of cadmium exposure (current concentrations of cadmium in blood (B-Cd) and urine (U-Cd)) or repeated measurements of cadmium in workplace air (CumAir-Cd). METHODS: The study comprised 30 workers with a range of duration of exposure of 11-51 years. 13 subjects were currently employed, whereas the other 17 had a median period without occupational exposure of eight years before the measurements. The in vivo measurements were made with an x ray fluorescence technique permitting average detection limits of 30 and 3 micrograms cadmium per g tissue in kidney and liver, respectively. RESULTS: 19 of 30 (63%) people had kidney-Cd and 13 of 27 (48%) had liver-Cd above the detection limits. Kidney-Cd ranged from non-detectable to 350 micrograms/g and liver-Cd from non-detectable to 80 micrograms/g. The median kidney-Cd and liver-Cd were 55 micrograms/g and 3 micrograms/g, respectively. Kidney-Cd correlated significantly with B-Cd (r, 0.49) and U-Cd (r, 0.70), whereas liver-Cd correlated significantly with U-Cd (r, 0.58). Neither kidney-Cd nor liver-Cd correlated with the CumAir-Cd. The prevalence of beta 2-microglobulinurea increased with increased liver-Cd. CONCLUSIONS: Current U-Cd can be used to predict the kidney-Cd and liver-Cd measured in vivo. In vivo measurements of kidney-Cd and liver-Cd were not shown to correlate with the individual cadmium exposure estimates, obtained by integration of the cadmium concentration in workplace air. There may be several reasons for this, including uncertainties in the estimate of the individual cumulative exposures as well as in the in vivo measurements. There was a suggestion of a relation between liver-Cd and tubular proteinuria.

Aged↗
PubMed1976

Decreased uptake of orotate in kidney tumors.

The uptake of isotope by three transplanted kidney tumors after i.p. injection of 3H-labeled orotate was less than 5% of that in the host kidney cortex. A decrease of similar magnitude in the uptake into the acid-soluble tissue fractions from the kidney tumors was observed with very low incorporation into RNA relative to the host kidney cortex. Total uptake of orotate-3H and incorporation into RNA were several fold higher in the normal kidney cortex than in the kidney medulla and for the kidney cortex both parameters were less in young than in old rats. The uptake of uracil-3H by two kidney tumors was approximately 40% of that in the kidney cortex of host rats. Although the uptake of orotate-3H in kidney cortex was greater than that for uracil-3H the reverse situation was observed with kidney tumors, indicating that renal neoplasia in the rat is accompanied by an altered pattern of uptake of these metabolites.

Animals↗
PubMed1995

Detection of human 11 beta-hydroxysteroid dehydrogenase isoforms using reverse-transcriptase-polymerase chain reaction and localization of the type 2 isoform to renal collecting ducts.

11 beta-Hydroxysteroid dehydrogenase (11 beta-HSD), responsible for the interconversion of hormonally active cortisol to inactive cortisone, dictates specificity for the mineralocorticoid receptor (MR) in the distal nephron and colon. Two isoforms of human 11 beta-HSD have been cloned, an NADP(H)-dependent (type 1) dehydrogenase/oxo-reductase enzyme, and a high-affinity NAD-dependent (type 2) unidirectional dehydrogenase. Using the reverse-transcriptase polymerase chain reaction (RT-PCR) amplification of RNA extracted from human adult tissues, type 1 11 beta-HSD mRNA was found in decidua, placenta, liver, lung, spleen, kidney medulla, cerebellum and pituitary, but was absent in kidney cortex, sigmoid and rectal colon, salivary gland and thyroid. In contrast, type 2 11 beta-HSD mRNA was found only in placenta and in the classical mineralocorticoid target tissues, kidney cortex, kidney medulla, sigmoid and rectal colon, salivary gland, and colonic epithelial cell lines (AAC1 and RGC28). In situ hybridization studies of renal cortex, cortico-medullary junction and medulla using a 35S-labeled antisense cRNA probe for type 2 human 11 beta-HSD, revealed specific localization of type 2 11 beta-HSD mRNA expression exclusively to renal cortical and medullary collecting ducts. Type 1 and type 2 isoforms of human 11 beta-HSD are expressed in a distinct tissue-specific fashion, in keeping with the proposed differences in their physiological roles. Type 2 11 beta-HSD is found predominantly in mineralocorticoid target tissues where it serves to protect the MR in an autocrine fashion.

11-beta-Hydroxysteroid Dehydrogenases↗
PubMed1999

Binding of kidney nuclear proteins to the 5'-flanking region of the rat gene for Ca2+-binding protein regucalcin: involvement of Ca2+/calmodulin signaling.

Ca2+-binding protein regucalcin is expressed in the kidney cortex of rats, as assayed by Northern blot analysis. The existence of kidney nuclear factor which binds to the 5'-flanking region of the rat regucalcin gene was investigated. When nuclear extracts obtained from the kidney cortex of rats were used in gel mobility-shift assays, two protein-DNA complexes were uniquely formed with the DNA fragment containing the 5'-flanking region of the rat regucalcin gene. Competition gel shift experiments indicated the specific binding region of kidney cortex nuclear proteins in the 5'-flanking region of the rat regucalcin gene. The two nuclear protein-DNA complexes were formed with the same mobility in rat kidney cortex and liver, which possess detectable amounts of regucalcin mRNA in Northern blot analysis. The binding activities of nuclear factors from kidney cortex to the 5'-flanking region of the rat regucalcin gene were inhibited by a single intraperitoneal administration of trifluoperazine, an antagonist of calmodulin, to rats. The present study demonstrates that kidney cortex nuclear proteins specifically bind to the 5'-flanking region of the rat regucalcin gene, and that the binding activity may be partly mediated through the Ca2+/calmodulin-dependent process.

Animals↗
PubMed1981

[Comparative study of acid SH-dependent proteinases from the swine renal cortex].

The presence of porcine kidney cortex of the SH-dependent enzymes, lysosomal carboxypeptidase B and cathepsins B, H and L, was demonstrated. A method for isolation of these enzymes was described which involves gel filtration through Sephadex G-100, covalent chromatography on mercury-organic Sepharose 4B and rechromatography on Sephadex G-100. Specific activity of lysosomal carboxypeptidase B was increased 9000-fold by BAA splitting and that of cathepsin B 240-fold by BAPA splitting. The molecular weights of lysosomal carboxypeptidase B and cathepsin B were found to be 50,000 and 26,000, respectively. Both enzymes were identical with appropriate enzymes obtained from other sources and differed in their properties from high-molecular-weight SH-proteinase or porcine kidney cortex.

Animals↗
PubMed1993

The relationship between mercury concentration in human organs and different predictor variables.

Samples from different tissues were collected from autopsies of individuals of the general population of the Stockholm area, Sweden. The samples were analysed for total mercury content using radiochemical neutron activation analysis. Average concentrations of mercury in occipital cortex, abdominal muscle, pituitary gland and kidney cortex were, 10.6 (2.4-28.7), 3.3 (0.9-5.4), 25.0 (6.3-77) and 229 (21.1-810) micrograms/kg wet weight, respectively. Possible predictor variables for mercury concentrations were tested in multiple linear regression models. An effect of a number of tooth surfaces with amalgam was seen in occipital lobe cortex, abdominal muscle and pituitary gland, but not in kidney cortex. In occipital lobe cortex and abdominal muscle, concentrations of mercury increased with age. Explanations discussed include: that a significant fraction of the mercury retained from amalgam fillings has a very long biological half-life; a decreasing capacity of mercury excretion with age; or higher fish consumption in the older individuals. In kidney cortex mercury concentrations decreased with age. The reason for this remains unclear, but it might indicate a decreasing capacity of mercury excretion with age. Chronic alcohol abuse was associated with decreased concentrations of mercury in occipital cortex.

Adolescent↗
PubMed1978

[Effect of oxygen metabolism in kidney tissue on the development of plasma erythropoietic properties].

The oxygen metabolism (pO2, the constant of the oxygen rate consumption, the coefficient of the oxygen distribution), the activity of oxid-reduced and hydrolytic enzymes, the blood supply of the kidney cortex and medulla, were recorded. These data were compared with the erythropoietin activity of plasma in the rats with phenylhydrazine anemia and posttransfusion polycytemia. An obvious reverse interrelationship existed between plasma erythropoietin activity and value of pO2 and blood supply of the cortex. The activity of the above enzymes was practically unchanged. The data obtained suggest that erythropoietin is formed in the cortex of the kidney and its production is controlled by the degree of oxygen supply of the kidney tissue.

Anemia↗
PubMed1981

Selenium content and glutathione peroxidase activity in tissues of young cattle fed supplemented whole milk diets.

Three groups of 5 calves each were fed (from birth to 12 weeks of age) 3 vitamin-mineral supplemented whole milk diets differing only in their selenium (Se) content (0.03, 0.23, and 0.53 microgram/g of solids). Glutathione peroxidase (GSH-Px) activities and Se concentrations were determined in blood at weekly intervals and in selected tissues after 12 weeks. There were marked differences among the tissues in GSH-Px activity, most notably in the liver, lungs, and adrenal glands, depending on whether H2O2 or cumene hydroperoxide was used to assay the enzyme. Tissues having the highest GSH-Px activity on a weight basis, when assayed with the H2O2 substrate, were erythrocytes and testes. Enzyme activity in thymus, brain, striated muscle, adipose tissue, and blood plasma was lowest among the tissues analyzed. The Se concentration on a fresh tissue-weight basis was highest in kidney cortex, intermediate in kidney medulla, testes, liver, and spleen, and lowest in striated muscle, adipose tissue, and blood plasma. The Se concentration was increased in liver, kidney cortex, spleen, and heart, with increasing increments of dietary Se, whereas differences were not observed in testes and adipose tissue.

Animal Feed↗
PubMed1979

Metabolism in hypothermically perfused human kidney carcinoma: incorporation rate of leucine into proteins.

Protein synthesis in tumour-involved human kidneys was studied before, during and after hypothermic perfusion. Before and after perfusion the incorporation rate of leucine into tumour and kidney cortex proteins was determined by incubation of tissue slices at 37 degrees C. During perfusion the incorporation of leucine from the perfusate into tumour and kidney cortex proteins was determined. Before hypothermic perfusion the incorporation rate of leucine into proteins at 37 degrees C was almost the same in kidney cortex and tumour. Leucine was incorporated linearly with time into kidney cortex and tumour proteins during hypothermic perfusion but the incorporation rate was 3-4 times higher into kidney cortex proteins than into tumour proteins. After 6 days of hypothermic perfusion the leucine incorporation rate into proteins at 37 degrees C was depressed by 50% in kidney cortex and by 90% in tumour tissue. The specific activity of leucine in the perfusate decreased during the perfusion period indicating a release of leucine from degradation of proteins. It is concluded that the effect of hypothermic perfusion on protein synthesis was more pronounced in the tumour than in the normal renal parenchyma.

Aged↗
PubMed1988

Concentrations of some trace elements in hair, liver and kidney from autopsy subjects--relationship between hair and internal organs.

Autopsy samples of hair, liver and kidney cortex from 30 Swedish subjects (Caucasian) were analysed for Ag, Co, Cr, Hg, Sb, Se and Zn by neutron activation analysis (NAA). The following elements were observed to have higher concentrations in hair than in liver and kidney cortex: Ag, (Co), Cr and Hg (on a dry weight basis). Selenium was highly concentrated in kidney cortex, and Fe in liver. The observed concentrations of most of the elements were very variable for each tissue. In particular, Co values for hair showed more than a 5000-fold difference. However, Se values for hair were relatively constant. A highly significant positive correlation was observed between Hg concentrations in hair and kidney cortex. Somewhat weaker correlations were found between Hg in kidney cortex and liver and Se in hair and kidney cortex. The concentration of Co in liver correlated with that in kidney cortex. It was concluded that, with the exception of mercury, and to a lesser degree selenium, hair analysis did not provide a useful measure of the trace element status of the subjects included in this study.

Autopsy↗
PubMed2005

Evaluation of absolute volume of human fetal kidney's cortex and medulla during gestation.

BACKGROUND: Human fetal kidney is quite different from the mature kidney, both macroscopically and hystologically. Lobulated surface of the human fetal kidney reflects its inner organisation. AIM: To determine the fetal kidneys' volume according to the gestational age, to establish periods of their maximal and minimal growth and to compare these values for various gestational ages. METHODS: Forty five human fetal kidneys aged from IV to X lunar months were analyzed. Kidneys were divided into nine groups according to their gestational age. The volumes of cortex and medulla were determined using stereological methods. The results were statistically analyzed and the periods of significant growth of these structures were marked. RESULTS: Fetal kidney's cortex and medulla grew continually with a very high coefficient of linear correlation with crown-rump length. The cortex/medulla ratio was minimal in the first half of V lunar month, when medulla grew most rapidly and it was maximal immediately before birth, when cortex achieved its maximum. CONCLUSION: This study was an effort to provide some parameters which would help in the future investigations of the development of human fetal kidney.

Crown-Rump Length↗
PubMed1999

Expression and localization of the neuronal glycine receptor beta-subunit in human, rabbit and rat kidneys.

The glycine receptor (GlyR) is a ligand-gated Cl- channel composed of two transmembrane subunits, alpha and beta, and gephyrin. The goal of this study was to determine whether the alpha- and/or beta-subunits of the GlyR are expressed in human, rabbit and/or rat kidneys. Screening of human and rat kidney cortex cDNA libraries identified polymerase chain reaction products that were identical to the neuronal GlyR beta-subunit. Sequencing revealed that rat kidney cortex and neuronal GlyR beta-subunits were identical. RNA isolated from the S2 segment of rabbit renal proximal tubules (RPT) and rat and rabbit kidney cortex was amplified following reverse transcription and gave similar results to that of human and rat kidney cDNA libraries. Degenerate primers against all GlyR alpha-subunits did not yield a product from rat and rabbit kidney cortex RNA, or from human and rat kidney cortex cDNA libraries. Immunofluorescence studies localized the beta-subunit and gephyrin to the basolateral membrane of rabbit RPT. These results provide compelling evidence for the GlyR beta-subunit, but not the alpha-subunit, in human, rabbit and rat kidney cortex.

Animals↗
PubMed2002

Application of quantitative competitive polymerase chain reaction for measurements of mRNA from antioxidative enzymes in the diabetic rat retina and kidney.

This present study applied quantitative competitive polymerase chain reaction (QC-PCR) in the analyses of mRNA expression of the endogenous antioxidative enzymes CuZn superoxide dismutase (SOD), MnSOD, catalase, and glutathione peroxidase in tissue samples from the retina and kidney cortex of diabetic rats. RNA was extracted from snap-frozen retinas and pieces of the kidney cortex of male Wistar rats with streptozotocin (STZ)-induced diabetes and control rats. The mRNA levels were analyzed using QC-PCR. The animals were kept in the laboratory for 1 and 6 months, respectively, and fed a normal or probucol- (1% wt/wt) enriched diet. By using QC-PCR, relative mRNA levels of all antioxidative enzymes could be estimated in the retina and kidney cortex. In the retina, the relative catalase mRNA concentration was about 1/10 that of the other enzymes. After 6 months of diabetes, there was a 100% increase of the catalase (median, 0.012 [range, 0.008 to 0.017] v 0.006 [0.005 to 0.010]; P =.011) and a 50% increase of the glutathione peroxidase mRNA levels (0.88 [0.44 to 1.12] v 0.52 [0.31 to 0.79]; P =.044). In the kidney cortex, the relative glutathione peroxidase mRNA level was 10 to 15 times higher, and catalase mRNA level about half of those of CuZnSOD and MnSOD. After 1 month of diabetes, there was an increase only of the glutathione peroxidase mRNA levels, by 170% (17.59 [6.19 to 29.49] v 6.96 [2.34 to 9.04]; P =.047). We conclude that quantification of mRNA can provide difficulties when the amount of sample RNA is limited and/or the gene expression is low. The present study shows QC-PCR to be useful as a tool for measuring expression of mRNA not only in the kidney cortex but also in small tissue samples like the retina. Our results indicate modestly increased mRNA expression of catalase and glutathione peroxidase in the retina and likewise modestly increased mRNA expression of glutathione peroxidase in the kidney cortex of rats with STZ-induced diabetes. Extended studies, also including enzyme activities, are needed before any effect of hyperglycemia on the overall enzyme activity can be established.

Animals↗
PubMed1996

[Study on the significance of mercury accumulation in the brain from dental amalgam fillings through direct mouth-nose-brain transport].

The transport of mercury (Hg) from the oro-nasal to the cranial cavity via a direct route was investigated. In 55 deceased persons, Hg concentrations were measured in the olfactory bulb and the trigeminal ganglion, and the number of dental amalgam fillings was assessed. For the purpose of comparison, Hg concentrations were also determined in the occipital lobe cortex, the pituitary gland and the kidney cortex. Quantitative Hg analysis was performed by cold vapor atomic absorption spectroscopy after acid digestion using high pressure microwave treatment. In the olfactory bulb (geom. mean 17.4 micrograms/kg w. w.), the Hg concentration was significantly higher than in the occipital lobe cortex (geom. mean 9.2 micrograms/kg w. w.) (p < 0.0001). No significant difference was found between the Hg concentration in the trigeminal ganglion (geom. mean 12 micrograms/kg w. w.) and the occipital lobe cortex (alpha = 0.005; p = 0.0342). Regression analysis did not reveal a statistically significant correlation between the number of dental amalgam fillings and the Hg content in the olfactory bulb and the trigeminal ganglion, respectively (alpha = 0.01). Therefore, these results do not support the hypothesis of a significant flow o Hg from dental amalgam fillings to the cranial cavity by a direct oro-nasal route. In contrast, a statistically significant correlation exists between the number of dental amalgam fillings and the Hg concentration in the kidney cortex (r2 = 0.317; p < 0.0001), and, to a lesser extent, the Hg concentration in the occipital lobe cortex (r2 = 0.17; p = 0.0016). The highest Hg concentrations (geom. mean 93.1 micrograms/kg w. w.) were detected in the kidney cortex, followed by the pituitary gland (geom. mean 30.0 micrograms/kg w. w.). In this study, Hg concentration in the pituitary gland did not correlate with the number of dental amalgam fillings.

Adolescent↗
PubMed2001

Changes in zinc and copper homeostasis in human livers and kidneys associated with exposure to environmental cadmium.

This study was undertaken to assess changes in zinc and copper homeostasis in human tissues that could be attributed to human exposure to environmental cadmium, using samples of lung, liver and kidney cortex of 61 Queensland residents, aged 2 to 89 years, who had died of accidental causes. None of the subjects were exposed to cadmium in the workplace. Levels of zinc in liver and kidney cortex samples showed inverse associations with donor age whereas zinc in lung only showed inverse association with gender. Lung zinc levels in females were 14% lower than in males. Zinc in liver and kidney cortex samples were found to exist in at least two pools; one was associated with cadmium that bound to metallothionein (MT) and the other was associated with non-MT bound copper. In liver, the amounts of zinc in the MT pool were smaller compared to those in non-MT pool given that only 7% of zinc variations were explained by cadmium whereas 22% of the liver zinc variations were accounted for by non-MT bound copper. In sharp contrast, larger amounts of zinc in kidney cortex samples were in the MT pool, compared to those in the non-MT pool given that cadmium was found to explain 69% of total zinc variation whereas copper explained only 17% of kidney zinc variations. The levels of copper in liver were found to be increased by 45-50% in subjects with high cadmium exposure level, compared to subjects of similar ages with medium exposure level. The levels of zinc and copper in kidney cortex samples in the subjects with high cadmium exposure were both found to be significantly elevated compared to those found in the medium-exposure group whereas copper contents were about 19-23% greater than in medium- as well as low-exposure groups. Taken together these results indicate increased sequestration of zinc and copper in liver and kidney cortex samples. The increases in metal sequestrations were observed in liver samples having cadmium contents of greater than 1 microg/g wet weight and in kidney cortex having cadmium contents of greater than 26 microg/g wet weight. Zinc and copper contents in lung of this sample group, however, were not associated with cadmium due probably to lower exposure levels compared to those of liver and kidney.

Adolescent↗
PubMed1992

The human renal 25-hydroxyvitamin D3-1 alpha-hydroxylase: properties studied by isotope-dilution mass spectrometry.

The renal 25-hydroxyvitamin D3-1 alpha-hydroxylase activity has been measured in normal human kidney cortex, using a highly specific assay based on isotope-dilution mass spectrometry. The cortex was obtained from kidneys removed due to renal tumours. The subcellular distribution of 25-hydroxyvitamin D3-1 alpha-hydroxylase activity was studied. Enzyme activity was only observed in the mitochondrial fraction. Mitochondria from non-tumourous kidney cortex had a Vmax of 0.17 +/- 0.02 pmol min-1 mg-1 protein and the apparent Km was in the range of 14 mumol l-1. There was a tendency to a higher 25-hydroxyvitamin D3-1 alpha-hydroxylase activity in preparations from male kidney (0.21 +/- 0.03 pmol min-1 mg-1 protein) than female (0.12 +/- 0.02, P less than 0.05). A significant inverse correlation between serum phosphate and 25-hydroxyvitamin D3-1 alpha-hydroxylase activity was found. No correlation was observed between enzyme activity and serum levels of 1,25-dihydroxyvitamin D (total and free index), PTH, total calcium or ionized calcium. The results indicate that there is a sex difference in human 25-hydroxyvitamin D3-1 alpha-hydroxylase activity similar to the one observed in laboratory animals. Furthermore, the data support the hypothesis that serum phosphate is a major regulator of 1,25-dihydroxyvitamin D3 production in man.

25-Hydroxyvitamin D3 1-alpha-Hydroxylase↗
PubMed1993

Expression cloning of human and rat renal cortex Na/Pi cotransport.

We have isolated two cDNA clones, NaPi-2 and NaPi-3, by screening rat kidney cortex and human kidney cortex cDNA libraries, respectively, for expression of sodium-dependent phosphate transport in Xenopus laevis oocytes. Substrate specificity and a detailed kinetic analysis (Na, Pi, H+ concentrations) suggested that expressed uptake activities relate to proximal tubular brush border membrane Na/Pi cotransport. NaPi-2 cDNA contains 2464 bp encoding a protein of 637 aa; NaPi-3 cDNA contains 2573 bp encoding a protein of 639 aa. NaPi-2- and NaPi-3-deduced protein sequences show high homology to each other but are different from the protein sequence deduced from the previously cloned NaPi-1 cDNA (from rabbit proximal tubules). Hydropathy profile predictions suggest at least eight membrane-spanning regions in NaPi-2/3-related proteins. In vitro translation results in proteins of the expected size and suggests glycosylation. Northern blot analysis shows corresponding mRNA species (approximately 2.7 kb) in kidney cortex of various species but no hybridization with RNAs isolated from a variety of other tissues (including intestinal segments); a hybridization signal (approximately 4.8 kb) was observed only in the lung (human). We conclude that we have structurally identified two closely related proteins most likely involved in human and rat renal brush border Na/Pi cotransport.

Amino Acid Sequence↗