Cost effective, robust, and reliable coupled separation techniques for the identification and quantification of phospholipids in complex biological matrices: application to insects.

The quantification of phospholipid classes and the determination of their molecular structures are crucial in physiological and medical studies. This paper's target analytes are cell membrane phospholipids, which play an important role in the seasonal acclimation processes of poikilothermic organisms. We introduce a set of simple and cost-effective analytical methods that enable efficient characterization and quantification of particular phospholipid classes and the identification and relative distribution of the individual phospholipid species. The analytical approach involves solid-phase extraction and high-performance thin-layer chromatography, which facilitate the separation of particular lipid classes. The obtained fractions are further transesterified to fatty acid methyl esters and subjected to gas chromatography coupled to flame ionization detection, which enables the determination of the position of double bonds. Phospholipid species separation is achieved by high-performance liquid chromatography with mass spectrometry, which gives information about the headgroup moiety and attached fatty acids. The total content of each phospholipids class is assessed by phosphorus determination by UV spectrophotometry. The simultaneous analysis of phosphorus, fatty acid residues, and phospholipid species provides detailed information about phospholipid composition. Evaluation of these coupled methods was achieved by application to an insect model, Pyrrhocoris apterus. High correlation was observed between fatty acid compositions as determined by gas chromatography and high-performance liquid chromatography analysis.

Placental growth factor may predict increased left ventricular mass index in patients with mild to moderate chronic kidney disease--a prospective observational study.

BACKGROUND: Placental growth factor [PlGF) is a cardiovascular (CV) risk marker, which is related to left ventricle hypertrophy (LVH) in animal models. Currently there are no data available regarding the possible relationship of PlGF and the development of LVH or diastolic dysfunction in patients with chronic kidney disease (CKD) and the relationship of PlGF to other CV risk factors in CKD patients. The aim of our study was to determine the possible association of PlGF and several other CV risk markers to echocardiographic parameters in CKD population. METHODS: We prospectively examined selected laboratory (PlGF, fibroblast growth factor-23 -FGF23, vitamin D, parathyroid hormone, extracellular newly identified RAGE-binding protein - EN-RAGE, B-type natriuretic peptide - BNP) and echocardiographic parameters in 62 patients with CKD 2-4. Mean follow-up was 36 ±10 months. Laboratory and echocardiographic data were collected 2-3 times, at the shortest interval of 12 months apart. Multivariate regression analysis was used to detect independent correlations of variables. RESULTS: Increased left ventricular mass index (LVMI, g/m2.7) was found in 29% patients with CKD 2-4, left ventricular (LV) diastolic dysfunction was detected in 74.1% patients (impaired LV relaxation in 43.5% patients and pseudonormal pattern in 30.6% patients). After 36 ± 10 months increased LVMI was found in 37.1% patients with CKD 2-4, LV diastolic dysfunction was detected in 75.8% patients (impaired LV relaxation in 43.5% patients and pseudonormal pattern in 32.3% patients). Following independent correlations were found: LVMI was related to PlGF, cholesterol, BNP, systolic blood pressure and serum creatinine. EN-RAGE correlated positively with left atrial diameter and inversely with E/A ratio. During the follow-up we found a significant increase in LVMI and left atrial diameter, whereas a significant decrease in LVEF was noted. CONCLUSION: According to our data, PlGF is independently related to increased LV mass in CKD, whereas EN-RAGE is more likely related to diastolic dysfunction in this population.

Three kinds of regulatory signals for production of juvenile hormone in females of the linden bug, Pyrrhocoris apterus.

Three types of regulation of the corpus allatum (CA) activity were defined in females of the linden bug Pyrrhocoris apterus. First, short-term inhibition of the CA activity was found in starved or fed long-day females, or in short-day females. Inhibitory factor(s) are transmitted to the CA via nerves, but in vitro they might reach the CA via the incubation medium. Origin of the inhibition is the pars intercerebralis (PI). The inhibitory effect is reversible during short-term incubation in vitro. This short-term inhibition can be quickly restored by the presence of the brain-suboesophageal ganglion (BR-SG) with the PI or removed, by the presence of the BR-SG without the PI or by the absence of the BR-SG. Short-term inhibition is sufficient to inhibit the CA of starved long-day females, but it is not strong enough to inhibit the CA of diapausing bugs. Second, developmental stimulation of the CA activity by feeding in long-day females is associated with growth in size of the CA. Stimulation proceeds slowly (days) in vivo and reaches the CA from the PI via nerves. Activity of the CA is irreversible in vitro; it is maintained without any further stimulation by the PI, i.e. in the presence of the BR-SG without PI or in the absence of the BR-SG. In the intact BR-SG-CC-CA the developmental stimulation of the CA is compensated by short-term inhibition of similar strength. Therefore, the activity of large CA within the intact BR-SG-CC-CA (stimulated + inhibited) is similar to the activity of the small denervated CA (no stimulation + no inhibition). Third, long-term inhibition of the CA activity in short-day females, produced by the diapause inducing photoperiod in the PI, reaches the CA via nerves. However, in contrast to the short-term inhibition of the CA, it is irreversible during short-term incubation in vitro. The long-term inhibition can only be removed several days after disconnection of the CA from the brain in vivo.

Tissue signaling pathways in the regulation of life-span and reproduction in females of the linden bug, Pyrrhocoris apterus.

Molecular studies on Drosophila melanogaster do not provide consistent results with regard to the hormonal regulation of a trade-off between life-span and fecundity. To unravel the physiological basis of the cost of reproduction without affecting animal's genotype, a new insect model, Pyrrhocoris apterus, was employed. Reproduction was manipulated by surgical ablations of tissues implicated in reproductive endocrinology, namely the pars intercerebralis (PI) of the brain, the corpus allatum (CA) and the ovary, and the response of life-span to these interventions under diapause-promoting short days and reproduction-promoting long days was measured. Life-span of long-day females increased in the following order: control (high fecundity)=ovary-ablation (no egg production)

Photoperiodic and food signals control expression pattern of the clock gene, period, in the linden bug, Pyrrhocoris apterus.

The temporal expression pattern of the circadian clock gene period was compared between heads of the linden bug, Pyrrhocoris apterus , kept under diapause-promoting short days (SD) and diapause-preventing long days (LD) using a real-time PCR quantification. Diapause or reproduction was programmed by photoperiod during the larval stage, but the first difference in per mRNA abundance between SD and LD insects was observed only after adult ecdysis. The expression level of per mRNA was markedly higher, up to more than 10-fold, in the destined-to diapause animals compared with those scheduled for reproduction. Up-regulation of per transcript was restricted to an early diapause peak, with the maximum expression on days 3 to 5 after adult ecdysis. Starvation reduced the peak level of per mRNA to about 50% of the value found in feeding females in the SD conditions, but per mRNA abundance was similarly low in fasting and feeding females in LD. Photoperiodic refractoriness in either wild-type postdiapause adults or in a selected nondiapause variant of P. apterus was associated with reproduction and low, LD-like levels of per mRNA under both SD and LD. Overall, the data suggest that the photoperiodic programming itself has no direct effect on per mRNA abundance, but it does determine the response of per transcript to food signals during subsequent expression of diapause/reproduction physiology.

Soluble receptor for advanced glycation end products in patients with decreased renal function.

BACKGROUND: Advanced glycation end products (AGEs) accumulate in patients with decreased renal function and exert various toxic effects through the receptor for AGEs (RAGE). Soluble RAGE (sRAGE) is a naturally occurring inhibitor of AGE-RAGE action. The aim of the study is to describe the relationship of sRAGE to renal function and dialysis modalities. METHODS: The studied group consisted of 81 patients: 25 patients with various degrees of decreased renal function, 20 long-term hemodialysis (HD) patients, 15 peritoneal dialysis (PD) patients, and 21 healthy age-matched subjects. sRAGE was assessed immunochemically (enzyme-linked immunosorbent assay), and routine biochemical parameters were measured by means of certified methods. RESULTS: sRAGE level correlates positively with serum creatinine concentration (r = 0.50; P < 0.05), and its relationship to creatinine clearance is hyperbolic. sRAGE levels are elevated significantly, mainly in patients with end-stage renal disease (3,119.0 +/- 968.4 pg/mL in HD patients and 3,652.7 +/- 1,677.7 pg/mL in PD patients versus 1,405.1 +/- 426.1 pg/mL in controls; both P < 0.001 versus controls). In PD patients, sRAGE is detectable in spent dialysate (median, 75.8 pg/mL), correlates with its serum levels (r = 0.67; P < 0.05), and is related to protein losses in dialysate. In HD patients, sRAGE levels increase by 50% (P < 0.001) from 0 to 15 minutes during both HD and hemodiafiltration, and then decrease until the end of the session. CONCLUSION: Serum sRAGE levels increase in patients with decreased renal function, mainly patients with end-stage renal disease. It remains to be elucidated whether the increase is caused just by decreased renal function or whether sRAGE is upregulated to protect against toxic effects of AGEs.

Decrease in Methoprene tolerant and Taiman expression reduces juvenile hormone effects and enhances the levels of juvenile hormone circulating in males of the linden bug Pyrrhocoris apterus.

Juvenile hormone (JH) produced by the corpus allatum (CA) stimulates vitellogenesis and reduces the synthesis of hexamerin proteins in adult females of Pyrrhocoris apterus. At present it is unknown whether the signaling pathway involving the JH receptor gene Methoprene tolerant (Met) and its binding partner Taiman (Tai), regulates the synthesis of accessory gland proteins (ACPs) and hexamerin proteins or effects male survival. Knockdown of genes by injecting Met dsRNA or Tai dsRNA, reduced the amount of ACPs whilst enhancing the amount of hexamerin mRNA in the fat body and the release of hexamerin proteins into haemolymph, as occurs after the ablation of CA. Lifespan was enhanced by injecting Met but not Tai dsRNA. Diapause associated with the natural absence of JH had a stronger effect on all these parameters than the ablation of CA or the knockdown of genes. This indicates there is an additional regulating agent. Both Met and Tai dsRNA induced a several fold increase in JH (JH III skiped bisepoxide) but a concurrent loss of Met or Tai disabled its function. This supports the view that the Met/Tai complex functions as a JH receptor in the regulation of ACPs and hexamerins.

Why is the number of days required for induction of adult diapause in the linden bug Pyrrhocoris apterus fewer in the larval than in the adult stage?

Adult females of Pyrrhocoris apterus, programmed for diapause by short-day (SD) photoperiod and those programmed for reproduction by long-day (LD) retain photoperiodic information in continuous darkness (DD) until death. However, if the interruption of SD by DD is made in the course of diapause programming in adults, then the incidence of diapause depends on the number of SD cycles received before DD, with no evidence that the photoperiodic clock is free-running at DD to complete diapause induction. These results indicate that the photoperiodic clock is stopped after transfer to DD and the information accumulated before transfer to DD is maintained. Diapause programming in the adult stage requires 9-10 SD cycles to induce diapause in 80% of individuals. However, if the diapause programming starts after ecdysis of LD-larvae to the last instar, only 3 SD cycles before transfer to DD are required for diapause in 80% of individuals. Surprisingly, if the newly ecdysed last instar LD-larvae, sensitive to photoperiod, are transferred to DD (thus they did not experience any SD), diapause occurs in 40% of the individuals. Thus, diapause 'information' is present in LD-larvae and is responsible for a lower number of SD required for diapause induction in the larval than in the adult stage.

Brain norepinephrine identified by mass spectrometry is associated with reproductive status of females of the linden bug Pyrrhocoris apterus.

Several biogenic amines, including controversial presence of norepinephrine (NE), were identified by the high performance liquid chromatography equipped with electrospray ionisation mass spectrometry in brain complexes of adult females of Pyrrhocoris apterus. Quantitative analysis was performed by the high performance liquid chromatography coupled to electrochemical detector. Levels of NE, dopamine (DA), octopamine (OA) and 5-hydroxytryptamine (5-HT) in brain complexes were measured in reproductive vs. diapause females. In field collected samples, levels of NE and DA were significantly higher in reproductive (May) than in non-reproductive (Sep, Oct, Feb) females. In laboratory females, NE is higher in long day photoperiod (reproduction) than in short day photoperiod (diapause) already from the first week of the adult age, while DA shows differences between the two contrasting photoperiods only from the second week of the adult age. No association between reproductive status and levels of OA and 5-HT was found.

Juvenile hormone signaling during reproduction and development of the linden bug, Pyrrhocoris apterus.

Juvenile hormone (JH), a sesquiterpenoid produced by the insect corpus allatum gland (CA), prevents metamorphosis in larvae and stimulates vitellogenesis in adult females. Whether the same JH signaling pathway regulates both processes is presently unknown. Here, we employ the robust JH response during reproduction and development of the linden bug, Pyrrhocoris apterus, to compare the function of key JH-signaling genes encoding the JH receptor, Methoprene-tolerant (Met), its binding partner Taiman (Tai), and a JH-inducible protein, Krüppel-homolog 1 (Kr-h1). RNA interference (RNAi) with Met or Tai, but not Kr-h1, blocked ovarian development and suppressed vitellogenin gene expression in the fat body of females raised under reproduction-inducing conditions. Loss of Met and Tai matched the effects of CA ablation or the natural absence of JH during reproductive diapause. Stimulation of vitellogenesis by treatment of diapausing females with a JH mimic methoprene also required both Met and Tai in the fat body, whereas Kr-h1 RNAi had no effect. Therefore, the Met-Tai complex likely functions as a JH receptor during vitellogenesis. In contrast to Met and Kr-h1 that are both required for JH to prevent precocious metamorphosis in P. apterus larvae, removal of Tai disrupted larval ecdysis without causing premature adult development. Our results show that while Met operates during metamorphosis in larvae and reproduction in adult females, its partner Tai is only required for the latter. The diverse functions of JH thus likely rely on a common receptor whose actions are modulated by distinct components.

Endocrine regulation of non-circadian behavior of circadian genes in insect gut.

The linden bug Pyrrhocoris apterus exhibits a robust diapause response to photoperiod. Photoperiod strongly affected basal levels of circadian gene transcripts in the gut, via the neuroendocrine system. Cryptochrome 2 (cry2) mRNA level was much higher in diapause promoting short days (SD) than in reproduction promoting long days (LD), while Par Domain Protein 1 (Pdp1) mRNA level was higher in LD than in SD. The effect of photoperiod on gene expression was mediated by the neurosecretory cells of the pars intercerebralis (PI) and the juvenile hormone (JH) producing corpus allatum (CA). In LD-females, CA ablation resulted in SD-like levels of gene transcripts, while PI ablation had little effect. Conversely, in SD-females, CA ablation had only a little effect, while PI ablation resulted in LD-like levels of gene transcripts. Thus, the CA is responsible for LD-like characteristics of gene expression in reproducing females and the PI is responsible for SD-like characteristics of gene expression in diapausing females. A simultaneous ablation of both PI and CA revealed two roles of PI in SD-females: (1) inhibition of CA, and (2) weak CA-independent stimulation of cry2 mRNA. Overall, our results indicate that peripheral circadian gene expression in the gut reflects the physiological state of females (with respect to diapause or reproduction) rather than the external light-dark cycle.

Novel roles for the corpus allatum hormone in the cost of sexual interactions in the linden bug Pyrrhocoris apterus.

The cost of sexual interactions, usually expressed as a reduction of life-span, is a fundamental but poorly understood aspect of life. According to a widely accepted view, a rise in the "pro-aging" juvenile hormone (JH) might contribute to the decrease of life span caused by sexual interactions. We tested this hypothesis using the linden bug Pyrrhocoris apterus by removing the corpus allatum (CA), the source of JH. If JH is causally involved in the cost of sexual interactions, then the absence of CA (JH) should decrease the negative effect of sexual interactions on survival. As expected, ablating the CA significantly prolonged life-span of both virgin females and virgin males. Mated insects of both sexes lived significantly shorter than virgins. However, contrary to prediction, the decrease of life span by sexual interactions was similar in control and CA-ablated males, and was even enhanced in CA-ablated females. Another unexpected finding was that males paired with CA-ablated females lived almost as long as virgin males and significantly longer than did males paired with control females, although ablating the female CA did not cause any decrease in mating activity. On the other hand, females paired with CA-ablated males lived only slightly longer than did females paired with control males. These results highlight several important points. (1) In both genders, the negative effect of sexual interactions on insect's survival is not mediated by the insect's own CA. (2) The male CA has only minor effect on female survival, while (3) the female CA (JH) is principally responsible for the sex-induced reduction in the male survival.

Receptor for advanced glycation end products--soluble form and gene polymorphisms in chronic haemodialysis patients.

BACKGROUND: The receptor for advanced glycation end products (RAGE) is involved in the pathogenesis of vascular and inflammatory diseases. The pathological effects mediated via RAGE are physiologically inhibited by soluble RAGE (sRAGE). Our aim was to study sRAGE and RAGE gene polymorphisms in haemodialysis (HD) patients. METHODS: A total of 261 stable HD patients were enrolled in the study and prospectively followed up for 30 months. At the begining of the study, sRAGE inflammatory and nutritional parameters were determined. RAGE polymorphisms were determined in a subgroup of 214 HD patients. A group of 100 healthy controls was used for comparison. RESULTS: In HD patients, sRAGE is elevated in comparison with healthy controls (3427+/-1508 vs 1758+/-637 pg/ml, P<0.001). It correlates negatively with residual diuresis (r=-0.193, P<0.05), with the acute phase reactants fibrinogen (r=-0.174, P<0.05) and orosomucoid (r=-0.135, P<0.05) and with the leucocyte count (r=-0.158, P<0.05). On the other hand, it is not related to the presence of diabetes mellitus, cardiovascular disease, nutritional status and mortality. The highest sRAGE levels are found in -429 CC and 2184 GG polymorphisms of the RAGE gene. The same results as for sRAGE were obtained for endogenous secretory RAGE (esRAGE), which correlated significantly with sRAGE (r=0.88, P<0.001). CONCLUSION: We conclude that in HD patients, sRAGE is increased due to decreased renal function, which is a very strong determinant of sRAGE levels, and is inversely related to inflammation. The highest sRAGE levels are influenced genetically. In our study, sRAGE levels were not related to mortality of HD patients.

Effect of hemodiafiltration on pregnancy-associated plasma protein A (PAPP-A) and related parameters.

BACKGROUND: Dialysis patients are at high risk of vascular/cardiovascular complications with multifactorial pathogenesis, and pregnancy-associated plasma protein A (PAPP-A) is one of the new markers related to cardiovascular risk. Because hemodiafiltration (HDF) is supposed to be better for cardiovascular status, the aim of this study was to describe whether it has any advantage concerning changes of PAPP-A and related molecules during the session in comparison with hemodialysis (HD). METHODS: The studied group consisted of 20 chronic hemodialysis patients. In each patient, PAPP-A and related parameters-IGFBP-4 (insulin like growth factor binding protein), IGF-I (insulin like growth factor), and two MMPs (matrix metalloproteinases)-2 and 9-were determined both during a single online HDF session (high-flux polysulfone membrane HF80, postdilution) and during a single HD session (low-flux polysulfone membrane F6, F7) at time 0 (start), 15 min, 120 min, and 240 min (end) of the session. RESULTS: PAPP-A, elevated at baseline in dialysis patients, changes significantly both during HDF and HD without significant differences between these two procedures (mean levels during HDF were 24.3, 53.9, 24.3, and 27.3 mIU/L). It increases more than two-fold from 0 to 15 min of the session (p < 0.001) and then decreases until the end of the session (p < 0.001). MMP-2 decreased slightly during both sessions (p < 0.001), and changes of other molecules were only minimal. CONCLUSION: A single HDF session compared to HD has no advantage in the decrease of PAPP-A and other tested molecules, all of them related to cardiovascular risk. Studies aimed at a long-term effect of both procedures on these parameters would be needed to further evaluate these therapeutical strategies.

Influence of oral vitamin E therapy on micro-inflammation and cardiovascular disease markers in chronic hemodialysis patients.

BACKGROUND: The aim of this study was to evaluate the influence of oral vitamin E therapy on serum concentrations of several markers of micro-inflammation and cardiovascular disease in chronic hemodialysis (HD) patients. METHODS: 29 HD patients were randomized into two groups: 15 patients were treated orally with 400 mg of vitamin E daily for a period of five weeks, and 14 patients received no antioxidant supplementation. Before and after vitamin E therapy, serum concentrations of vitamin E (high-performance liquid chromatography), pregnancy-associated plasma protein-A (immunochemical--TRACE assay), C-reactive protein (nephelometry), intercellular adhesion molecule-1 (ELISA), and E-selectin (ELISA) were measured. HD patients were compared with 16 healthy controls. RESULTS: Baseline serum concentrations of PAPP-A and CRP were significantly higher in HD patients than in healthy controls (PAPP-A: 26.23+/-11.94 vs. 11.41+/-1.94 mIU/L, p<0.001; CRP: 5.20+/-3.50 vs. 3.40+/-3.80 mg/L, p<0.05). After five weeks of oral vitamin E intake, serum PAPP-A, CRP, ICAM-1, and E-selectin concentrations remained unchanged in both groups of HD patients. CONCLUSION: Chronic micro-inflammation in HD patients is documented by the elevation of CRP and PAPP-A. A daily oral dose of 400 mg of vitamin E does not seem to be able to reduce enhanced oxidative stress and micro-inflammation in chronic HD patients.

No benefit of hemodiafiltration over hemodialysis in lowering elevated levels of asymmetric dimethylarginine in ESRD patients.

BACKGROUND: It has been suggested that hemodiafiltration (HDF) is more efficient than hemodialysis (HD) in lowering plasma levels of the endogenous nitric oxide synthase inhibitor asymmetric dimethylarginine (ADMA), which is a strong and independent predictor of overall mortality in ESRD patients. METHODS/PATIENTS: Twenty ESRD patients (11 women) were studied during both a single online HDF session and a single HD session. In each patient, ADMA, L-arginine, SDMA, beta(2)-microglobulin and urea were measured at several time points. RESULTS: Although HDF was clearly superior to HD in decreasing plasma beta(2)-microglobulin, there was no difference in the elimination characteristics of ADMA. However, HDF but not HD eliminated the nitric oxide synthase substrate L-arginine, making HD superior in increasing the L-arginine/ADMA ratio. CONCLUSION: Neither HD nor HDF sufficiently removes the putative uremic toxin ADMA. The clinical significance of HD better improving the L-arginine/ADMA ratio (parameter of NO production) as compared to HDF needs to be determined.

Intravenous iron gluconate administration increases circulating PAPP-A in hemodialysis patients.

BACKGROUND: Pregnancy-associated plasma protein-A (PAPP-A) is a proatherosclerotic molecule, interrelated with oxidative stress in hemodialysis (HD) patients. As intravenous (IV) iron might enhance oxidative stress in HD patients, this study investigates circulating PAPP-A during HD session and after IV iron administration. METHODS: In 20 HD patients, plasma PAPP-A concentration was assessed immunochemically during 2 HD sessions (prior to HD and at 60, 130, and 240 min of HD session). Sodium ferric gluconate (62.5 mg) was given IV to all patients 65 min after the start of the second HD. RESULTS: Sixty-five min after IV iron application, there was a significant increase in plasma PAPP-A (from 36.0+/-9.9 to 79.6+/-28.9 mU/L, p<0.0001). At the end of this HD session, PAPP-A decreased significantly (p<0.0001), but still remained 1.5-fold greater compared with predialysis levels (p<0.0005). CONCLUSION: IV iron increases circulating PAPP-A, and in this way, it might contribute to more pronounced cardiovascular complications in HD patients.

Influence of parenteral iron therapy and oral vitamin E supplementation on neutrophil respiratory burst in chronic hemodialysis patients.

BACKGROUND: Bioincompatibility of hemodialysis (HD) membranes is responsible for neutrophil activation leading to oxidative stress, which can be further increased by intravenous (IV) iron (Fe) administration. The aim of our study was to monitor neutrophil respiratory burst during HD and to find out whether this process is influenced by IV Fe and oral vitamin E administration. METHODS: Within four HD sessions, blood samples were taken from seven chronic HD patients at time 0 (before HD), 60, 70, and 130 min of HD session. Neutrophil respiratory burst was assessed by luminol-enhanced chemiluminescence (CL). Plasma advanced oxidation protein products (AOPP) concentration was measured spectrophotometrically. During the second and the fourth HD, 62.5 mg of sodium ferric gluconate was applied IV in the 65th minute of HD. Before the last two HD, the patients were given orally 200 mg of vitamin E daily for 7 days. Patient's results were compared with healthy controls. RESULTS: Predialysis CL is higher in patients than in controls (1,926 +/- 436 vs. 1,083 +/- 325 RLU, p<.01). CL decreases in the 60th min of HD (1,926 +/- 436 vs. 1,220 +/- 599 RLU, p<.05); thereafter, it remains stable. After Fe application, CL increases at time 130 compared with CL at time 60 (1,303 +/- 269 vs. 877 +/- 292 RLU, p<.05). AOPP concentration is higher in patients than in controls (137.5 +/- 42.7 vs. 88.9 +/- 24.8 micromol/L, p<.01) and remains unaffected by vitamin E supplementation. After vitamin E intake, predialysis CL remains significantly higher than in controls, and changes in CL during HD are minimal despite Fe administration. CONCLUSION: HD patients' neutrophils generate more oxygen radicals than in healthy individuals. This production decreases during HD and then increases after IV Fe administration. Short-term vitamin E administration attenuates this fluctuation of neutrophil oxidative metabolism, without affecting the total degree of oxidative stress.