scholarly journals Metabolism of [1−14C]glyoxylate, [1−14C]-glycollate, [1−14C]glycine and [2−14C]-glycine by homogenates of kidney and liver tissue from hyperoxaluric and control subjects

1967 ◽  
Vol 105 (2) ◽  
pp. 701-707 ◽  
Author(s):  
Betty M. Dean ◽  
R. W. E. Watts ◽  
Wendy J. Westwick
Keyword(s):  
Carbon Dioxide ◽  
Liver Tissue ◽  
Kidney Tissue ◽  
Primary Hyperoxaluria ◽  
Human Kidney ◽  
Control Subjects ◽  
Glyoxylate Metabolism ◽  
And Control ◽  
Kidney Liver

1. The metabolism of [1−14C]glyoxylate to carbon dioxide, glycine, oxalate, serine, formate and glycollate was investigated in hyperoxaluric and control subjects' kidney and liver tissue in vitro. 2. Only glycine and carbon dioxide became significantly labelled with 14C, and this was less in the hyperoxaluric patients' kidney tissue than in the control tissue. 3. Liver did not show this difference. 4. The metabolism of [1−14C]glycollate was also studied in the liver tissue; glyoxylate formation was demonstrated and the formation of 14CO2 from this substrate was likewise unimpaired in the hyperoxaluric patients' liver tissue in these experiments. 5. Glycine was not metabolized by human kidney, liver or blood cells under the conditions used. 6. These observations show that glyoxylate metabolism by the kidney is impaired in primary hyperoxaluria.

The Identification of the Enzymes That Catalyse the Oxidation of Glyoxylate to Oxalate in the 100 000 g Supernatant Fraction of Human Hyperoxaluric and Control Liver and Heart Tissue

1973 ◽  
Vol 44 (3) ◽  
pp. 227-241 ◽  
Author(s):  
Dorothy A. Gibbs ◽  
R. W. E. Watts
Keyword(s):  
Lactate Dehydrogenase ◽  
Nicotinamide Adenine Dinucleotide ◽  
Liver Tissue ◽  
Primary Hyperoxaluria ◽  
Heart Tissue ◽  
Supernatant Fraction ◽  
Glycollate Oxidase ◽  
And Control ◽  
Oxalate Synthesis ◽  
Lactate Dehydrogenase Isoenzymes

1. The enzymic oxidation of glyoxylate to oxalate in the soluble (100 000 g supernatant) fraction of liver and heart tissue from a patient with primary hyperoxaluria and from a non-hyperoxaluric subject have been studied. 2. An oxidized nicotinamide—adenine dinucleotide (NAD+)-dependent and a non-NAD+-dependent oxidation of glyoxylate to oxalate were observed in the liver tissue from both sources. 3. Evidence is presented that lactate dehydrogenase has a major role in catalysing the reaction in both of the tissues studied. The non-NAD+-dependent oxidations which are catalysed by xanthine oxidase and glycollate oxidase in the liver are relatively unimportant, and they were not detected in the heart. 4. An enzyme that catalyses the oxidation of glycollate was also demonstrated in liver tissue. This had a different electrophoretic mobility from the lactate dehydrogenase isoenzymes. 5. These findings are discussed with particular reference to human primary hyperoxaluria in which excessive oxalate synthesis occurs.

scholarly journals The Globoseries Glycosphingolipid Sialosyl Galactosyl Globoside Is Found in Urinary Tract Tissues and Is a Preferred Binding Receptor In Vitro for Uropathogenic Escherichia coli Expressing pap-Encoded Adhesins

1998 ◽  
Vol 66 (8) ◽  
pp. 3856-3861 ◽  
Author(s):  
A. E. Stapleton ◽  
M. R. Stroud ◽  
S. I. Hakomori ◽  
W. E. Stamm
Keyword(s):  
Escherichia Coli ◽  
Urinary Tract ◽  
Kidney Tissue ◽  
Human Kidney ◽  
Blood Group Antigens ◽  
E Coli ◽  
Vaginal Epithelial Cells ◽  
History Of ◽  
Tract Infections

ABSTRACT Women with a history of recurrent Escherichia coliurinary tract infections (UTIs) are significantly more likely to be nonsecretors of blood group antigens than are women without such a history, and vaginal epithelial cells (VEC) from women who are nonsecretors show enhanced adherence of uropathogenic E. coli isolates compared with cells from secretors. We previously extracted glycosphingolipids (GSLs) from native VEC and determined that nonsecretors (but not secretors) selectively express two extended globoseries GSLs, sialosyl galactosyl globoside (SGG) and disialosyl galactosyl globoside (DSGG), which specifically bound uropathogenicE. coli R45 expressing a P adhesin. In this study, we demonstrated, by purifying the compounds from this source, that SGG and DSGG are expressed in human kidney tissue. We also demonstrated that SGG and DSGG isolated from human kidneys bind uropathogenic E. coli isolates expressing each of the three classes ofpap-encoded adhesins, including cloned isolates expressing PapG from J96, PrsG from J96, and PapG from IA2, and the wild-type isolates IA2 and R45. We metabolically 35S labeled these five E. coli isolates and measured their relative binding affinities to serial dilutions of SGG and DSGG as well as to globotriaosylceramide (Gb3) and globotetraosylceramide (Gb4), two other globoseries GSLs present in urogenital tissues. Each of the five E. coli isolates bound to SGG with the highest apparent avidity compared with their binding to DSGG, Gb3, and Gb4, and each isolate had a unique pattern of GSL binding affinity. These studies further suggest that SGG likely plays an important role in the pathogenesis of UTI and that its presence may account for the increased binding of E. colito uroepithelial cells from nonsecretors and for the increased susceptibility of nonsecretors to recurrent UTI.

scholarly journals Inhibition of TNF-αReverses the Pathological Resorption Pit Profile of Osteoclasts from Patients with Acute Charcot Osteoarthropathy

2015 ◽  
Vol 2015 ◽  
pp. 1-10 ◽  
Author(s):  
Nina L. Petrova ◽  
Peter K. Petrov ◽  
Michael E. Edmonds ◽  
Catherine M. Shanahan
Keyword(s):  
Osteoclast Formation ◽  
Diabetic Patients ◽  
Blood Monocytes ◽  
Control Subjects ◽  
Osteoclastic Resorption ◽  
Healthy Control ◽  
Macrophage Colony ◽  
And Control ◽  
Charcot Osteoarthropathy

We hypothesised that tumour necrosis factor-α(TNF-α) may enhance receptor activator of nuclear factor-κβligand- (RANKL-) mediated osteoclastogenesis in acute Charcot osteoarthropathy. Peripheral blood monocytes were isolated from 10 acute Charcot patients, 8 diabetic patients, and 9 healthy control subjects and culturedin vitroon plastic and bone discs. Osteoclast formation and resorption were assessed after treatment with (1) macrophage-colony stimulating factor (M-CSF) and RANKL and (2) M-CSF, RANKL, and neutralising antibody to TNF-α(anti-TNF-α). Resorption was measured on the surface of bone discs by image analysis and under the surface using surface profilometry. Although osteoclast formation was similar in M-CSF + RANKL-treated cultures between the groups (p>0.05), there was a significant increase in the area of resorption on the surface (p<0.01) and under the surface (p<0.01) in Charcot patients compared with diabetic patients and control subjects. The addition of anti-TNF-αresulted in a significant reduction in the area of resorption on the surface (p<0.05) and under the surface (p<0.05) only in Charcot patients as well as a normalisation of the aberrant erosion profile. We conclude that TNF-αmodulates RANKL-mediated osteoclastic resorptionin vitroin patients with acute Charcot osteoarthropathy.

scholarly journals Characterization of the Hoechst 33342 side population from normal and malignant human renal epithelial cells

2008 ◽  
Vol 295 (3) ◽  
pp. F680-F687 ◽  
Author(s):  
Sanjai K. Addla ◽  
Mick D. Brown ◽  
Claire A. Hart ◽  
Vijay A. C. Ramani ◽  
Noel W. Clarke
Keyword(s):  
Stem Cell ◽  
Epithelial Cells ◽  
Side Population ◽  
Kidney Tissue ◽  
Human Kidney ◽  
Proliferative Potential ◽  
Stem Cell Markers ◽  
Hoechst 33342 ◽  
Tissue Samples

The fundamental changes which predispose for renal cell carcinoma (RCC) are poorly characterized. It is hypothesized that “cancer stem cells” may be influential in carcinogenesis, and the epithelial side population (SP) is enriched for stemlike cells in other epithelial cancers. In this study, we have isolated and characterized the SP and non-SP (NSP) populations from normal (NK) and malignant (RCC) human kidney tissue. NK specimens were taken from patients undergoing non-renal cancer surgery and paired malignant and macroscopically normal tissue samples were taken from patients undergoing surgery for RCC. The Hoechst 33342 dye efflux technique was used to isolate epithelial SP and NSP from normal and malignant human renal tissue. Cellular subpopulations were phenotyped for lineage, cell cycle, and putative stem cell markers, and functionally characterized using in vitro colony-forming and proliferation assays. The SP constituted 3.8 ± 0.4 and 5.9 ± 0.9% of epithelial cells in NK and RCC, respectively, of which 14.1 ± 3.5 and 13.2 ± 3.6% were shown to be in G0. SP cells demonstrated greater proliferative potential in colony-forming efficiency, long-term culture, and spheroids assays and were shown to be maintained upon tissue culture passage. We have shown that the renal SP is enriched for quiescent cells, with a high proliferative capacity and stemlike properties. The population is, however, heterogeneous, confirming that the terms “SP cell” and “stem cell” cannot be used interchangeably.

Thyroxine monodeiodination in normal human kidney tissue in vitro

1986 ◽  
Vol 112 (4) ◽  
pp. 536-540 ◽  
Author(s):  
Niels Boye
Keyword(s):  
Competitive Inhibition ◽  
Kidney Tissue ◽  
Human Kidney ◽  
Cell Fraction ◽  
Type I ◽  
Ph Optimum ◽  
Normal Human Kidney ◽  
Normal Human ◽  
Rat Tissue

Abstract. The present study deals with thyroxine monodeiodination in normal human kidney. To allow for comparison with previous reports, the present methods are similar to those used by others in rat tissue studies. The microsomal cell fraction of normal human kidney tissue was obtained by differential ultracentrifugation. The microsomes were incubated under various conditions and the deiodination products assayed with radioimmunoassay. A type I 5'-monodeiodinase was demonstrated, pH optimum around 6.5. Competitive inhibition was observed of T3 generation from T4 by rT3 with a Km of 3.0 μm and a Ki of 4 μm. Vmax was 26.1 pmol/min/mg protein. Likewise rT3 was generated from added T4, but it was rapidly degraded, while T3 was relatively stable as is the case in rat tissue preparations. Propylthiouracil inhibited 5'-deiodination in a dose dependent fashion with complete abolishment of deiodination at propylthiouracil concentration of 10−4m. Ipodate inhibited the reaction with complete inhibition at 10−2 m. The data demonstrate that a human kidney particulate cell-fraction contained considerable amounts of T4 deiodinases, very similar to the type I deiodinase of various rat tissue, although the handling of rT3 and the inhibitory action of this iodothyronine on T4 to T3 conversion seem to be slightly different in the two species.

scholarly journals High throughput single cell RNA-seq of developing mouse kidney and human kidney organoids reveals a roadmap for recreating the kidney

2017 ◽  
Author(s):  
Alexander N. Combes ◽  
Belinda Phipson ◽  
Luke Zappia ◽  
Kynan T. Lawlor ◽  
Pei Xuan Er ◽  
...  
Keyword(s):  
Single Cell ◽  
Kidney Tissue ◽  
Human Kidney ◽  
Population Based ◽  
Mouse Kidney ◽  
Rna Seq ◽  
Kidney Morphogenesis ◽  
Human Ipsc ◽  
Kidney Organoids

AbstractRecent advances in our capacity to differentiate human pluripotent stem cells to human kidney tissue are moving the field closer to novel approaches for renal replacement. Such protocols have relied upon our current understanding of the molecular basis of mammalian kidney morphogenesis. To date this has depended upon population based-profiling of non-homogenous cellular compartments. In order to improve our resolution of individual cell transcriptional profiles during kidney morphogenesis, we have performed 10x Chromium single cell RNA-seq on over 6000 cells from the E18.5 developing mouse kidney, as well as more than 7000 cells from human iPSC-derived kidney organoids. We identified 16 clusters of cells representing all major cell lineages in the E18.5 mouse kidney. The differentially expressed genes from individual murine clusters were then used to guide the classification of 16 cell clusters within human kidney organoids, revealing the presence of distinguishable stromal, endothelial, nephron, podocyte and nephron progenitor populations. Despite the congruence between developing mouse and human organoid, our analysis suggested limited nephron maturation and the presence of ‘off target’ populations in human kidney organoids, including unidentified stromal populations and evidence of neural clusters. This may reflect unique human kidney populations, mixed cultures or aberrant differentiation in vitro. Analysis of clusters within the mouse data revealed novel insights into progenitor maintenance and cellular maturation in the major renal lineages and will serve as a roadmap to refine directed differentiation approaches in human iPSC-derived kidney organoids.

scholarly journals Tissue distribution of a menthyl-conjugated oligodeoxyribonucleotide antisense to PAI-1 mRNA.

2005 ◽  
Vol 52 (4) ◽  
pp. 849-856
Author(s):  
Janusz Szemraj ◽  
Khalid N I Al-Nedawi ◽  
Ewa Chabielska ◽  
Wlodzimierz Buczko ◽  
Zofia Pawlowska
Keyword(s):  
Rat Kidney ◽  
Inhibitory Effect ◽  
Organ Distribution ◽  
Ribonuclease Protection Assay ◽  
Ribonuclease Protection ◽  
And Control ◽  
Kidney Liver ◽  
Pai 1

The inhibitory effect of numerous analogues of PO-16, an hexadecadeoxyribonucleotide antisense to sequences -22 to -17 of PAI-1 mRNA coding for a fragment of the signal peptide, on the expression of PAI-1 in endothelial cells, and physiological consequences of the subsequently reduced PAI-1 activity tested in vitro and in vivo, were described in our previous studies. Of particular interest was PO-16 5'-O-conjugated with menthyl phosphorothioate (MPO-16R). In this work, tissue localisation of MPO-16R labelled with [(35)S] phosphorothioate at the 3'-end, was determined. [(35)S]MPO-16R and control [(35)S]MPO-16R-SENSE oligonucleotides were administered intravenously into 22 rats and organ distribution of the labelled bioconjugates was assessed after 24 and 48 h. For this purpose, tissue sections were subjected to autoradiography, and quantitated by liquid scintillation after solubilisation. Overall clearance of radioactivity was already seen after 24 h, with the radioactivity recovered mainly in the kidney and liver. A smaller fraction of radioactivity was also retained in the spleen and heart. The kidney concentration of the labelled probe was higher than that of liver by 50%. The distribution of PAI-1 mRNA in untreated rat kidney, liver, spleen and heart established by two independent techniques: Ribonuclease Protection Assay and Real-Time PCR, shows the same pattern as that observed for [(35)S]MPO-16R antisense.

scholarly journals Graft immaturity and safety concerns in transplanted human kidney organoids

2019 ◽  
Vol 51 (11) ◽  
pp. 1-13 ◽  
Author(s):  
Sun Ah Nam ◽  
Eunjeong Seo ◽  
Jin Won Kim ◽  
Hyung Wook Kim ◽  
Hong Lim Kim ◽  
...  
Keyword(s):  
Therapeutic Option ◽  
Kidney Tissue ◽  
Human Kidney ◽  
Ips Cells ◽  
Attractive Potential ◽  
Transplanted Kidney ◽  
Filtration Barrier ◽  
Kidney Organoids

AbstractFor chronic kidney disease, regeneration of lost nephrons with human kidney organoids derived from induced pluripotent stem (iPS) cells is proposed to be an attractive potential therapeutic option. It remains unclear, however, whether organoids transplanted into kidneys in vivo would be safe or functional. Here, we purified kidney organoids and transplanted them beneath the kidney capsules of immunodeficient mice to test their safety and maturity. Kidney organoid grafts survived for months after transplantation and became vascularized from host mouse endothelial cells. Nephron-like structures in grafts appeared more mature than kidney organoids in vitro, but remained immature compared with the neighboring mouse kidney tissue. Ultrastructural analysis revealed filtration barrier-like structures, capillary lumens, and tubules with brush border in the transplanted kidney organoids, which were more mature than those of the kidney organoids in vitro but not as organized as adult mammalian kidneys. Immaturity was a common feature of three separate differentiation protocols by immunofluorescence analysis and single cell RNA sequencing. Stroma of transplanted kidney organoid grafts were filled with vimentin-positive mesenchymal cells, and chondrogenesis, cystogenesis, and stromal expansion were observed in the long term. Transcription profiles showed that long-term maintenance after kidney organoid transplantation induced transcriptomic reprogramming with prominent suppression of cell-cycle-related genes and upregulation of extracellular matrix organization. Our data suggest that kidney organoids derived from iPS cells may be transplantable but strategies to improve nephron differentiation and purity are required before they can be applied in humans as a therapeutic option.

scholarly journals Ketogenesis from butyrate and acetate by the caecum and the colon of rabbits

1972 ◽  
Vol 130 (3) ◽  
pp. 785-790 ◽  
Author(s):  
S. J. Henning ◽  
F. J. R. Hird
Keyword(s):  
Carbon Dioxide ◽  
Liver Tissue ◽  
Ketone Bodies ◽  
Distal Colon ◽  
Proximal Colon ◽  
Enzyme Assays ◽  
Liver Slices

1. When studied in vitro, tissue from the caecum and the proximal colon of rabbits converted butyrate into ketone bodies. The conversion was similar to that observed with liver slices. The ketogenic activity was associated with the mucosa rather than the muscle of the gut wall and, in the colon, diminished as the distance from the caecal–colonic junction increased. 2. Tissue from the wall of the ileum, caecum, proximal colon and distal colon was also shown to metabolize [1-14C]butyrate to carbon dioxide. 3. Enzyme assays showed that in both liver tissue and caecal mucosa the activity of hydroxymethylglutaryl-CoA synthase was more than ten times that of acetoacetyl-CoA deacylase. Labelling experiments in vitro gave confirmation of the hydroxymethylglutaryl-CoA pathway. 4. The significance of the conversion of butyrate into ketone bodies is discussed.

Blood fuel metabolites in asthma during and after progressive submaximal exercise

1987 ◽  
Vol 73 (1) ◽  
pp. 81-86 ◽  
Author(s):  
G. E. Packe ◽  
J. Wiggins ◽  
B. M. Singh ◽  
M. Nattrass ◽  
A. D. Wright ◽  
...  
Keyword(s):  
Carbon Dioxide ◽  
Oxygen Consumption ◽  
Oxygen Saturation ◽  
Ketone Body ◽  
Respiratory Exchange Ratio ◽  
Carbon Dioxide Production ◽  
Hydrogen Ion ◽  
Control Subjects ◽  
Progressive Exercise ◽  
And Control

1. Ten male stable asthmatic subjects and 10 matched control subjects performed a progressive exercise test on a treadmill to 85% of their predicted maximum heart rate. 2. Blood lactate, pyruvate, hydrogen ion, glucose, alanine, glycerol and total ketone body concentrations were measured at frequent intervals during and up to 60 min after exercise. Carbon dioxide production, oxygen consumption, ventilation, respiratory exchange ratio and oxygen saturation were also measured during and up to 10 min after exercise. 3. There were no significant differences between the asthmatic and control subjects in levels of carbon dioxide production, oxygen consumption and ventilation. The respiratory exchange ratio was greater in the asthmatic subjects during recovery from exercise (P < 0.05). No changes in oxygen saturation were observed during exercise in either group. 4. In both asthmatic and control subjects, lactate, pyruvate, hydrogen ion, alanine and glycerol concentrations showed an increase from baseline levels, reaching maximum levels up to 10 min after exercise and returning to baseline within 1 h after exercise. Total ketone body concentrations decreased during exercise. 5. There were no significant differences between the asthmatic and control subjects in the concentration of any metabolite over the study period. 6. These data indicate that fuel metabolism during and after short-term progressive exercise is similar in stable asthmatic and normal subjects.

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