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A Dawnay

Publications and source records attributed to A Dawnay.

At least 19 recordsLinked to original sources
PubMed2005

Advanced glycation end product free adducts are cleared by dialysis.

Plasma advanced glycation end product (AGE) free adducts are increased up to 50-fold among patients on dialysis. We examined the ability of hemodialysis (HD) and continuous ambulatory peritoneal dialysis (CAPD) to clear these compounds. The AGE free adducts Nepsilon-carboxymethyl-lysine (CML) and Nepsilon-(1-carboxyethyl)lysine (CEL) and the hydroimidazolones derived from glyoxal (G-H1), methylglyoxal (MG-H1), and 3-deoxyglucosone (3DG-H) were determined by LC-MS/MS and pentosidine by HPLC with fluorimetric detection in ultrafiltrates of plasma, urine, or PD effluent as appropriate from patients on HD (n = 8) or PD (n = 8), and from healthy controls (n = 8). Among patients on HD, all free AGEs predialysis were significantly higher than in controls and were decreased with dialysis. The removal of MG-H1 and 3DG-H was comparable to that of urea, whereas that of CML and pentosidine was some 20% higher; in contrast, the removal of CEL and G-H1 was 25% lower. Among patients on CAPD, free AGEs in PD effluent increased with increasing dwell time. The combined renal and peritoneal 24-h excretion rates of CML (4.7 micromol), CEL (6.5 micromol), 3DG-H (16.6 micromol), and pentosidine (0.08 micromol) were twofold higher than the amount excreted in healthy controls, whereas MG-H1 was ninefold higher (59 micromol); the combined clearances of all free AGEs except pentosidine were lower than in healthy controls. Impaired renal clearance contributes to increased plasma free AGEs in uremia, but the increased excretion rate among patients on PD demonstrates that there was also an increased synthesis of free AGEs. Both HD and PD are able to remove free AGEs.

Adult↗
PubMed2003

Renal clearance of glycation adducts: anti-glycation defence in uraemia and dialysis.

Increased plasma concentrations of free AGEs (advanced glycation end products) in uraemia are due to impaired renal clearance exacerbated by dietary load and breakdown of the elevated protein-bound AGEs. Increased protein-bound AGEs most probably reflect increased synthesis due to the increased oxidative and carbonyl stress of uraemia, which is exacerbated by the dialysis procedure. Use of biocompatible peritoneal dialysis fluids and haemodialysis membranes will be beneficial, but strategies to further reduce AGE formation are required. These should focus on the hydroimidazalone AGEs, which are quantitatively the most important, and not only on the conventionally monitored AGEs N (epsilon)-carboxymethyl-lysine and pentosidine.

Enzyme-Linked Immunosorbent Assay↗
PubMed2003

Removal of advanced glycation end products in clinical renal failure by peritoneal dialysis and haemodialysis.

AGEs (advanced glycation end products) accumulate markedly in the plasma of human subjects with renal failure. We investigated the efficiency of removal of AGEs from the circulation by PD (peritoneal dialysis) and HD (haemodialysis) therapy. Free AGEs were measured by LC-MS/MS in blood plasma before dialysis, in dialysis fluid effusate after a 2-12 h dwell time in the peritoneal cavity of PD subjects, and in the HD dialysate before and after HD therapy. In clinical uraemia, the concentrations of free AGEs in blood plasma were increased up to 50-fold. For example, levels of MG-H1 (methylglyoxal-derived hydroimidazolone) were: normal controls, 110+/-46 nM; PD subjects, 1876+/-676 ( P <0.01); HD subjects, 5496+/-1138 nM ( P <0.001). In PD subjects, the AGE concentration in the effusate increased with increasing dwell time, reaching a maximum at a concentration higher than that in plasma for some AGEs at 4-12 h. This may reflect AGE formation in the peritoneal cavity. In HD, AGE concentrations in HD fluid were decreased markedly from the start to the end of a dialysis session, except that levels of the methylglyoxal-derived AGEs N (epsilon)-(1-carboxyethyl)lysine and MG-H1, and of pentosidine, remained 5-fold higher than control levels. Inadequate clearance of free AGEs may be linked to the increased risk of cardiovascular disease in patients with renal failure.

Glycation End Products, Advanced↗
PubMed2001

Plasma OLC is elevated in mild experimental uremia but is not associated with hypertension.

BACKGROUND: Little is known about the renal handling of endogenous ouabain-like compound (OLC). The aim of this study was to determine the normal renal clearance of OLC and the effect of mild experimental uremia on plasma OLC and its clearance. METHODS: Male Wistar rats were studied 8 weeks after subtotal (5/6th) nephrectomy (n = 8) and compared with a control sham-operated group (n = 8). RESULTS: Plasma creatinine and OLC were higher in uremic animals compared with controls (creatinine 76+/-5.6 micromol/L v 45+/-9.6 micromol/L, respectively, P < .00005; OLC 195+/-62 pmol/L v 121+/-62 pmol/L, P < .02). Creatinine clearance and OLC clearance were lower in uremic animals compared with controls (creatinine 1.06+/-0.12 mL/min v 1.58+/-0.32 mL/min, respectively, P < .002; OLC 23.6+/-10.4 microL/min v 33.2+/-11.4 microL/min, P < .05). There were no significant differences (all P > .05) between the uremic and control groups in the fractional clearance of OLC (uremic 2.3%+/-1.0% v control 2.2%+/-1.0%), OLC excretion rate (uremic 6.2+/-2.4 pmol/24 h v control 5.0+/-1.1 pmol/24 h) or in the mean systolic blood pressure (BP) (uremic 132+/-13 mm Hg v control 126+/-3 mm Hg). The amount of OLC excreted per unit of functioning nephron mass was 78% higher in uremic animals than in controls. The rate of tubular absorption varied linearly with filtered load, did not differ between groups, and showed no evidence of saturation. CONCLUSIONS: The kidneys are an important excretion route for plasma OLC and moderate but significant increases may occur without inducing hypertension in the short term. The low fractional clearance of OLC is most likely due to tubular absorption and/or catabolism.

Animals↗
PubMed2000

Serum inhibin, activin and follistatin in postmenopausal women with epithelial ovarian carcinoma.

OBJECTIVE: To investigate the role of serum inhibin A, inhibin pro-alphaC immunoreactivity, activin A, and follistatin in postmenopausal women with epithelial ovarian cancer. DESIGN: Case-control study. SAMPLE: e Serum samples from 27 postmenopausal women with epithelial ovarian cancer and 54 controls from the general population participating in an ovarian cancer screening trial. RESULTS: Women with epithelial ovarian cancer had significantly higher serum levels of pro-alphaC immunoreactivity (P = 0.03), activin A (P = 0.004) and follistatin (P = 0.04), but not inhibin A (P = 0.13). Using the 90th centile in the control group as the cut off, pro-alphaC levels were elevated in 41% of women with epithelial ovarian cancer, while inhibin A was elevated in only 15%. Using the 95th centile as the cut off, serum pro-alphaC was elevated in only 11% of women with epithelial ovarian cancer (3/27), while activin A was elevated in 48% (11/23). Follicle stimulating hormone levels were significantly lower in women with epithelial ovarian cancer (P = 0.01). Although, inhibin-related peptides can modulate follicle stimulating hormone levels, there was no correlation between inhibin A, pro-alphaC immunoreactivity, activin A or follistatin and follicle stimulating hormone. CONCLUSION: These data demonstrate that though there is preferential secretion of precursor forms of the alpha subunit rather than dimeric inhibin A by epithelial ovarian cancer, pro-alphaC is unlikely to be a useful tumour marker. Activin A is more commonly elevated in postmenopausal women with epithelial ovarian cancer and its role as a tumour marker in the diagnosis and screening of epithelial ovarian cancer warrants further evaluation.

Activins↗
PubMed2000

Mass spectrometric monitoring of albumin in uremia.

BACKGROUND: Advanced glycation end products (AGEs) are a novel class of uremic toxins. In plasma, they are present in proteins and also in low molecular mass peptides. AGE-modified peptides are thought to bind and modify plasma proteins. Monitoring of the consequent increase in molecular mass of serum albumin may be used in surveillance of the clinical management of uremia. METHODS: We investigated molecular mass changes of human serum albumin (HSA) glycated by glucose and methylglyoxal in vitro and of subjects with moderate renal impairment, end-stage renal disease (ESRD), ESRD on hemodialysis, and normal healthy controls by matrix-assisted laser desorption ionization mass spectrometry. RESULTS: Fatty acid-free HSA had a molecular mass of 66,446 +/- 114 D. Mean (+/-SD) molecular mass increases were HSA minimally glycated by glucose 399 +/- 88 D (N = 5, P < 0.001), HSA highly glycated by glucose 6780 +/- 122 D (N = 5, P < 0.001), HSA minimally glycated by methylglyoxal 73 +/- 121 D (N = 5, P > 0.05), and HSA without fatty acid removal 535 +/- 90 D (N = 5, P < 0.001). For HSA of human subjects, mean (+/- SD) molecular mass increases were normal healthy controls 243 +/- 97 D (N = 5), moderate renal impairment 350 +/- 83 D (P > 0.05 with respect to controls, N = 5), ESRD 498 +/- 128 (P < 0.02 with respect to controls, N = 3), and ESRD on hemodialysis 438 +/- 85 D (P < 0.02 with respect to controls, N = 5). The mean molecular mass of albumin of all groups was increased significantly with respect to that of fatty acid free albumin (P < 0.001). CONCLUSIONS: Only ESRD was associated with a significant increase in the molecular mass of HSA in vivo. Since this mass increase was very low and much lower than reported for AGE-modified peptides, it may reflect AGE formation on HSA by alpha-oxoaldehydes that accumulate in uremia, rather than modification of albumin by AGE-modified peptides. The molecular mass of HSA in vivo was indicative of a minimal and not a high extent of glycation.

Adult↗
PubMed2000

Analysis of human ouabainlike compound by micellar electrokinetic chromatography.

In this preliminary study we have optimised micellar electrokinetic chromatography (MEKC, a form of capillary electrophoresis) to enable the chromatographic and spectral characteristics of human ouabainlike compound (OLC) to be investigated. Sera from fifty patients were combined to form a pool (100 ml) whilst urine (92.5 ml) was obtained from a normal healthy volunteer. Both samples were initially concentrated and partially purified by solid phase extraction before further purification by sequential HPLC separations. Final volumes for both extracts were 100 microl. MEKC was performed on a HP(3D) CE instrument with voltage set at 20 KV, capillary temperature at 20 degrees C, injection time 4 s, sample volume 10 nl, with detection by photodiode array. A compound was found in both serum and urine that had similar elution and spectral characteristics to authentic ouabain. We conclude that MEKC is potentially a useful tool for the analysis of human OLC.

Cardenolides↗
PubMed1998

Comparison of in vitro AGE formation between standard PD fluid and a novel bicarbonate/lactate formulation.

Peritoneal advanced glycation end-product (AGE) formation may be accelerated by glucose degradation products produced as a consequence of heat sterilization of peritoneal dialysis (PD) fluid. The formation of these degradation products is reduced if the glucose is separated from the buffers during heat sterilization. This pilot study compared in vitro AGE formation in PD fluid (1.36% and 3.86% glucose) heat sterilized in a two-compartment bag (bicarbonate/lactate buffer) with that in a standard, single-compartment bag (lactate buffer, Dianeal). Peritoneal dialysis fluids were incubated with human serum albumin (HSA, 1 g/L), as a model protein, at 37 degrees C for 0, 5, and 20 days. Formation of AGEs was assessed by measuring fluorescence at each time point. Advanced glycation end-product formation was greater in lactate PD fluid compared with bicarbonate/lactate PD fluid of equivalent glucose strength. Advanced glycation end-product formation in the lactate PD fluid containing 1.36% glucose was comparable to that in the bicarbonate/lactate PD fluid containing 3.86% glucose. The rate of increase in fluorescence per day was greater in the first 5 days of incubation than in the subsequent 15 days. These results are compatible with the presence of greater amounts of glucose-degradation products in the standard single-compartment bag resulting in enhanced AGE formation.

Bicarbonates↗
PubMed1998

The pathogenesis and consequences of AGE formation in uraemia and its treatment.

Advanced glycation endproducts (AGEs) accumulate in uraemia as a consequence of diminished clearance of low molecular weight forms which retain their reactivity and may subsequently combine with circulating and tissue macromolecules. Successful renal transplantation is the only form of renal replacement therapy which effectively clears these circulating AGEs; both haemodialysis and peritoneal dialysis are comparatively ineffective although high-flux haemodialysis confers some benefits. De novo AGE formation may be accelerated in uraemia due to carbonyl and oxidative stress leading to further accumulation. The consequences for the patient with chronic renal failure may be acceleration of vascular disease, renal failure progression and dialysis-related amyloidosis. Accelerated peritoneal AGE formation as a consequence of treatment with peritoneal dialysis fluids may be detrimental to peritoneal membrane function but does not appear to contribute to systemic elevation of AGEs.

Cardiovascular Diseases↗
PubMed1997

Development of enzyme immunoassay for endogenous ouabain-like compound in human plasma.

Widespread evidence supports the existence of an endogenous digitalis-like compound in mammals. We report here the development of a novel enzyme immunoassay for ouabain that, in conjunction with a detailed HPLC study, identifies a ouabain-like compound (OLC) in extracted human plasma. The assay is sensitive-minimum detection limit for OLC 37 pmol/L (11 pmol/L in plasma)-and has a working range (between-assay CV < 10%) of 180-10000 pmol/L (54-3000 pmol/L in plasma). Mean recoveries of ouabain added to plasma ranged from 90% to 100%, and plasma extracts diluted in parallel to the standard curve. Plasma OLC concentrations in 10 healthy volunteers averaged 92 pmol/L (range 55-168), assuming 100% cross-reactivity of OLC in the ouabain assay. HPLC analysis with two distinct chromatographic conditions demonstrated that endogenous human plasma OLC co-eluted with authentic ouabain. The enzyme immunoassay is rapid and easy to perform and will support further investigation of the nature of this controversial endogenous steroid.

Chromatography, High Pressure Liquid↗
PubMed1995

Identification of auto-transplanted parathyroid tissue by Tc-99m methoxy isobutyl isonitrile scintigraphy.

Parathyroid tissue is sometimes auto-transplanted into the forearm after 'total' parathyroidectomy in patients with renal failure. Recurrent hyperparathyroidism demands identification of the source of PTH secretion which cannot be assumed to be the forearm. To this end, Tc-99m methoxy isobutyl isonitrile (MIBI) scintigraphy was used to identify functioning auto-transplanted parathyroid tissue in seven patients undergoing renal replacement therapy (five with functioning renal transplants and two on haemodialysis). Serum PTH was estimated in venous blood taken proximal and distal to the forearm graft and from the contralateral arm, and subsequent Tc-99m MIBI scanning was carried out without knowledge of the PTH results. Five patients had a significant gradient in PTH levels between sites proximal and distal to the graft, and between the proximal site and the contralateral arm, suggesting functioning parathyroid tissue in the graft. Subsequent Tc-99m MIBI scintigraphy confirmed the activity of the auto-transplanted parathyroid tissue in these five patients. In the remaining two patients without a significant PTH gradient between the sampling sites, Tc-99m MIBI scintigraphy did not identify any functioning forearm parathyroid tissue. The scan results therefore correlated well with the gradients in PTH levels, suggesting that MIBI scintigraphy can be used to identify functioning auto-transplanted parathyroid tissue. The results also indicate that any patient who has undergone auto-transplantation of parathyroid tissue must have blood samples taken from veins proximal to the graft and either distal to it, or from the contralateral arm when parathyroid status is re-assessed, particularly when surgery is being considered for recurrent hyperparathyroidism.

Adult↗
PubMed1994

Dramatic increase in levels of placental protein 14 in amniotic fluid at 10-15 weeks' pregnancy.

Concentrations of placental protein 14 (PP14) were measured by radioimmunoassay in 103 amniotic fluid samples from 10 to 20 weeks of pregnancy. Low levels of PP14 were present at 10-12 weeks (median values 82-280 micrograms/l). After 12 weeks, the levels increased by two orders of magnitude to reach a peak at 18 weeks (median 11 mg/l). This pattern may reflect the obliteration of the extraembryonic coelom and direct apposition of the decidua with the chorio-amnion.

Amniotic Fluid↗
PubMed1994

Cyclosporin as initial treatment for Crohn's disease.

Childhood Crohn's disease may cause significant morbidity. T cell activation is considered to be central to Crohn's disease pathology, and as cyclosporin is a powerful inhibitor of T cell activation, and has been used in adult Crohn's disease with encouraging results, it may offer the prospect of remission if given early in the course of disease. Children with newly diagnosed Crohn's disease or those relapsing off treatment were therefore given cyclosporin or conventional treatment (enteral nutrition or corticosteroids) by random allocation. Evaluation was performed initially and at two months. Twenty four children were studied (10 on cyclosporin and 14 on conventional treatment; one child on cyclosporin withdrew). Significant clinical improvement occurred in the group on conventional treatment, but not in the cyclosporin group. Colonoscopic improvement was noted in 5/9 on cyclosporin and 8/14 on conventional treatment, but neither group produced a significant fall in median colonoscopic index. Histological improvement was seen in 7/8 on cyclosporin and 8/13 on conventional treatment, but cyclosporin was not significantly better. Cyclosporin produced improved clinical and histological appearance without matched improvement in blood disease indices. It was not better than conventional treatment, and simple oral administration is probably not suitable for newly diagnosed patients with Crohn's disease.

Adolescent↗
PubMed1993

Glycated albumin in serum and dialysate of patients on continuous ambulatory peritoneal dialysis.

1. Chronic use of hyperosmolar glucose solutions in continuous ambulatory peritoneal dialysis may cause glycation of peritoneal structural proteins which could contribute to membrane dysfunction and ultrafiltration failure. To determine whether glycation can occur in the environment of the dialysate, we have carried out studies using albumin as a model protein. 2. Glycated albumin was measured in the serum and dialysate of 46 patients on continuous ambulatory peritoneal dialysis (31 non-diabetic patients, 15 diabetic patients). Dialysate and serum glycated albumin (ranges 1.0-12.7% and 0.9-10.2%, respectively) were related to each other (r = 0.988, P < 0.001), but dialysate glycated albumin was significantly higher than serum glycated albumin (P < 0.0001), with the dialysate to serum glycated albumin ratio being greater than unity in 76% of patients (mean ratio 1.14). This implies either preferential transfer of glycated albumin across the peritoneal membrane or intraperitoneal glycation during the dwell period. 3. In vitro, significant glycation occurred in dialysate during a 6 h incubation period (P < 0.01) at a rate related to the glucose concentration in the dialysate (rs = 0.63, P < 0.05). The glycation rate was not significantly affected (P = 0.05) by factors other than the glucose concentration. 4. Our results demonstrate that protein glycation occurs within the peritoneum during continuous ambulatory peritoneal dialysis. Further studies are required to establish the relationship of glycation of structural proteins in the peritoneal membrane to membrane function.

Adolescent↗
PubMed1993

Serum glycated albumin and fructosamine in renal dialysis patients.

The effect of uraemia on protein glycation was studied by measuring glycated albumin and fructosamine in 50 non-diabetic dialysis patients (31 continuous ambulatory peritoneal dialysis, CAPD, 19 haemodialysis). After correction for serum albumin concentration, glycated albumin (g/100 g) was increased in the haemodialysis group (1.94 +/- 0.40) compared with both CAPD patients (1.46 +/- 0.37; p < 0.001) and controls (1.52 +/- 0.29; p < 0.001), but did not differ between CAPD patients and controls (p > 0.05). Serum fructosamine, corrected for either serum albumin or total protein concentration (mumol/100 g), was raised in CAPD (828 +/- 90, 386 +/- 41, respectively) and haemodialysis patients (802 +/- 123, 391 +/- 42, respectively) compared with controls (609 +/- 69, 332 +/- 27, respectively; p < 0.0001 in all cases), but did not differ between the two dialysis groups (p > 0.05). A single haemodialysis cycle had no effect on the measurement of glycated albumin or fructosamine (p > 0.05). The results confirm that glycated protein levels are generally raised in dialysis patients. In CAPD patients, altered albumin metabolism resulting from large peritoneal losses is likely to have caused a decrease in the amount of albumin glycated, an effect less apparent on the concentration of fructosamine because of the additional contribution of glycated globulins.

Adolescent↗