Response of glutamine metabolism to glutamine-supplemented parenteral nutrition.

BACKGROUND: Increasing evidence suggests that glutamine is important for the function of many organ systems and supports the use of glutamine-enriched total parenteral nutrition (TPN) during severe illness. However, the effect of prolonged glutamine supplementation on glutamine kinetics has not been studied. OBJECTIVE: We investigated the effect of 8-10 d of TPN enriched with glutamine dipeptides on glutamine kinetics. DESIGN: Twenty-three preoperative patients were randomly allocated to receive either TPN enriched with glutamine dipeptides (60 micromol glutamine*kg body wt(-1)*h(-1)) or isonitrogenous, isoenergetic, glutamine-free TPN. A primed, continuous, 6-h intravenous infusion of L-[5-(15)N]glutamine and L-[1-(13)C]leucine was given before (baseline) and 8-10 d after the TPN solutions were administered. Baseline measurements were performed after a 40-h administration of a standard solution of glucose and amino acids (no glutamine). RESULTS: Glutamine-enriched TPN increased the total appearance rate of glutamine (P: < 0.05) but did not inhibit or increase the endogenous appearance rate. The standard TPN solution also increased the glutamine appearance rate (P: < 0.05), but the change was much smaller than in the glutamine-supplemented group (P: < 0.01). The plasma glutamine concentration did not rise significantly during either treatment, suggesting increased tissue glutamine utilization, especially in the glutamine-supplemented group. CONCLUSION: In view of the enhanced glutamine requirements in response to trauma and disease by tissues such as those of the gut, the immune system, and the liver, increased glutamine availability during glutamine-enriched TPN may be beneficial preoperatively in patients with gastrointestinal disease.

Energy, substrate and protein metabolism in morbid obesity before, during and after massive weight loss.

OBJECTIVE: To investigate the effect of surgically induced weight loss on energy, substrate and protein metabolism of morbidly obese patients. DESIGN: A prospective, clinical intervention study of morbidly obese patients before and after surgical treatment. SUBJECTS: Eight morbidly obese patients (BMI 47.88+/-7.03). METHODS: Total energy expenditure (TEE; doubly labeled water method), sleeping metabolic rate (SMR; respiration chamber), body composition (deuterium oxide component of doubly labeled water), substrate metabolism (48 h dietary records, 48 h urine collection and gaseous exchange in the respiration chamber) and whole body protein turnover (primed-continuous infusion of L-[1-13C]-leucine) were measured before, 3 and 12 months after vertical banded gastroplasty (VBG). RESULTS: The TEE decreased as a result of a decreased SMR (64%) and non-SMR (36%; P=0.001). SMR as a function of fat-free mass (FFM) decreased after weight loss (P<0.05). The physical activity index (PAI), defined as TEE/SMR, was low and was not influenced by weight loss. Protein and carbohydrate oxidation decreased significantly after VBG (P<0.05), although 3 months after VBG protein oxidation did not decrease enough to prevent loss of FFM. The energy used for protein turnover was approximately 24% of SMR and did not change after weight loss. CONCLUSIONS: Compensatory processes that oppose weight loss of morbidly obese patients exist, as demonstrated by the disproportional reduction of SMR, and a low PAI. Protein turnover is not a major contributor to the disproportional reduction of SMR.

Glutamine appearance rate in plasma is not increased after gastrointestinal surgery in humans.

The metabolic response to surgical stress is characterized by muscle protein breakdown and mobilization of amino acids and has been postulated to furnish glutamine and other amino acids to the immune system, gut and liver. The present study was undertaken to investigate whether the whole body appearance rate (R(a))(3) of glutamine in plasma is increased after major elective surgery. Fourteen patients (8 males, 6 females) were measured prior to laparotomy and on the second postoperative day. Patients received a primed continuous 6-h infusion of L-[5-(15) N]glutamine and L-[1-(13)C]leucine, and arterial blood samples and muscle biopsies were taken for concentration and enrichment measurements. As expected, the metabolic response to surgery was characterized by a rise in whole body protein breakdown (n = 14, P < 0.001) and a decreased concentration of glutamine in plasma (n = 14, P < 0.001) and muscle (n = 8, P < 0.01). However, these catabolic changes were not reflected by an increase in the plasma R(a) of glutamine: 246 +/- 8 micromol. kg(-1). h(-1) before surgery vs. 241 +/- 10 micromol. kg(-1). h(-1) on the second postoperative day. We conclude that the whole body R(a) of glutamine in plasma is not increased 2 d after elective gastrointestinal surgery. Further studies are warranted to establish whether the lack of an increase in plasma glutamine R(a) provides a rationale for glutamine supplementation.

Automated isolation of high-purity plasma albumin for isotope ratio measurements.

Measurement of the incorporation of labeled amino acids in plasma albumin, isolated from plasma sampled at different time points after infusion start is a well-known technique to study human albumin synthesis. Unfortunately, no chromatographic method has been described yet, enabling the automated isolation of high-purity albumin from large numbers of plasma samples as is required to study the kinetics of this process. Therefore, we developed a fast protein liquid chromatographic method, capable of processing 200 microliters amounts of plasma in 74 min (injection to injection). The system can run unattended as the FPLC system is connected to a sample processor equipped with a polyether ether ketone (PEEK) sample loop and a cooled sample tray. Albumin isolation was divided into three steps. First, plasma samples were injected onto a 1-ml Blue Sepharose HiTrap affinity column, equilibrated with 50 mmol/l phosphate buffer (pH 7.0). After elution of non-binding protein, switching the solvent to phosphate buffer with 1.5 mol/l sodium chloride eluted albumin. The resulting albumin fraction was desalted on-line by directing it through two consecutive HiTrap 5-ml desalting columns, whereafter it was retained in the system within a 5-ml PTFE loop, connected to a motor value. After switching this valve, thus bypassing the sample loop, the phosphate buffers were changed automatically to Tris buffers. Final purification involved elution of the captured fraction over a 1-ml ion-exchange Resource Q column, using a sodium chloride gradient, ranging from 0 to 0.5 mol/l in Tris buffer (20 mmol/l, pH 7.5). A more than 99% purity of the final albumin fraction was confirmed by capillary electrophoresis.

[Glutamine as a key ingredient in protein metabolism]. / Glutamine als spil van de eiwitbenutting.

Glutamine has a number of unique properties which suggest that this amino acid plays an important role in health and disease. Glutamine is considered a conditionally essential amino acid, because during periods of severe metabolic stress the body's requirements of glutamine may exceed the individual's ability to produce sufficient amounts of the amino acid. Studies with glutamine-enriched nutrition show beneficial effects on nitrogen balance, muscle protein metabolism, gastrointestinal mucosa, and immune status. In certain patient categories addition of glutamine reduces the number of infectious complications, improves long-term survival, and shortens hospital stay, e.g. bone marrow transplantation patients, neonates with severely subnormal weight, patients with multiple organ failure, multi-trauma patients. More studies are needed to document the dose-response and to identify the patients that are likely to benefit from glutamine supplementation.

Glutamine: the pivot of our nitrogen economy?

Glutamine serves as a shuttle of useful nontoxic nitrogen, supplying nitrogen from glutamine-producing (eg, muscle) to glutamine-consuming tissues. True production rates of glutamine are difficult to measure, but probably are less than 60 to 100 g/d for a 70-kg man. During catabolic stress increased amounts of glutamine are released from muscle, consisting of protein derived glutamine, newly synthesized glutamine, and glutamine losses from the intramuscular free pool. The large and rapid losses of free muscle glutamine are difficult to restore, presumably as a result of disturbances in the Na+ electrochemical gradient across the cell membrane. Whereas increased amounts of glutamine are released from muscle, glutamine consumption by the immune system (liver, spleen) also is enhanced. Thus, during catabolic stress changes occur in the flow of glutamine between organs. These changes are not necessarily reflected by alterations in the whole-body appearance rate of glutamine. In contrast with the gut, where glutamine is taken up in a concentration dependent manner, the immune system actively takes up glutamine despite decreased plasma concentrations. Supplementation with glutamine influences uptake by both the gut and the immune system, as evidenced by increased mucosal glutamine concentrations and gut glutathione production. There is evidence suggesting that this improves gut barrier function. Although the benefit of glutamine supplementation is most evident from experimental studies, clinical studies on the effect of glutamine do exist and suggest that glutamine supplementation has beneficial effects with regard to patient outcome.

Nutritional depletion and dietary manipulation: effects on the immune response.

The association between nutritional depletion and the increased susceptibility for infectious diseases has been recognized for a long time. The complexity of the immune system, however, makes it difficult to unravel the underlying mechanisms. It appears that depletion adversely affects virtually all components of the immune system. This review provides an overview over the specific requirements of substrates by immune cells and the effects of nutritional depletion on various components of the immune response, with special attention to gut-associated lymphoid tissue. The literature concerning effects of dietary interventions with specific nutrients on the immune response is also discussed. Finally, we offer a hypothesis with regard to the improvement of composition of "trauma" nutrition solutions.

Absence of glutamine isotopic steady state: implications for the assessment of whole-body glutamine production rate.

1. During infusion of [5-15N]glutamine in patients with gastrointestinal cancer we unexpectedly observed a gradual decrease in time of the appearance rate (Ra) of glutamine in plasma. Here we investigate whether the failure to achieve a plateau isotopic enrichment in plasma is, among other factors, due to incomplete equilibration of the glutamine tracer with the large intramuscular free glutamine pool.2. Plasma and intramuscular glutamine enrichment were measured during 6-11 h infusions of L-[5-15N]glutamine and L-[1-13C]glutamine in post-absorptive patients admitted to hospital for elective abdominal surgery. L-[1-13C]Leucine and L-[ring-2H5]phenylalanine were infused to measure the proportion of glutamine appearing in plasma directly due to its release from protein.3. The glutamine tracer entered muscle, but the rise in intramuscular glutamine enrichment was small, presumably as a result of the enormous size of the intramuscular glutamine pool and the limited speed of entry of glutamine into muscle. In each patient the intramuscular glutamine enrichment was lower than that in plasma (P<0.001), and both increased with tracer infusion time (P<0.001), indicating incomplete equilibration of the glutamine tracer.4.A comparison of the results obtained by the two glutamine tracers indicated that recycling of the nitrogen label contributed to about 15% of the decrease in Ra.5. There was a gradual reduction in the glutamine release from proteolysis, which contributed to 16-21% of the decline in Ra.6. We conclude that slow equilibration of the glutamine tracer with the large muscle glutamine pool significantly contributes to the absence of isotopic steady state. Consequently, the appearance rate of glutamine in plasma measured during short tracer infusion periods (hours) considerably overestimates the whole-body glutamine flux.

Contribution of tubular anion and cation secretion to residual renal function in chronic dialysis patients.

The clearance of organic ions by the tubules may contribute to the removal of uremic waste products in dialysis patients. The renal excretion of an exogenous anion p-aminohippurate (PAH) was investigated in 10 peritoneal dialysis patients and 10 hemodialysis patients during one clearance period and compared with the clearance of creatinine (Ccr) and inulin (CIn). The clearance period was 24 hours in the peritoneal dialysis patients and one interdialytic interval of 3 days divided in 4 parts [CPA-D] in hemodialysis patients. In peritoneal dialysis patients the renal clearance of total PAH (median 14.3 ml/min, range 3.8-33.0) exceeded the CIN (median 3.2 ml/min, range 1.6-11.2, p < 0.005) and Ccr (median 4.0 ml/min, range 1.7-15.0, p < 0.005). A positive correlation was found between the tubular clearances of creatinine (cationic pathway) and of total PAH (anionic pathway, r: 0.72, p <0.02). In hemodialysis patients the clearance of total PAH (CPA: median 2.0, range 0.8-9.6; CPD: median 3.8, range 1.7-15.4) also exceeded the clearance of inulin (CPA: median 1.5, range 0.2-3.4; CPD: median 2.7, range 0.9-4.4) in the beginning and the end of the interdialytic interval (p < 0.005). The CIN and the clearance of total PAH increased during the interdialytic interval, but the Ccr (CPA: median 2.2, range 0.4-8.9, CPD: median 2.9, range 1.2-4.6) remained stable. Thus, the change in tubular clearance of creatinine and PAH was opposite during the interdialytic interval: it increased for total PAH and decreased for creatinine. The CTPAH/CIN ratio in hemodialysis patients was lower than in peritoneal dialysis patients. In CPA it was median 1.6 (range 1.1-5.6, p < 0.05) and in CPD it was median 1.7 (range 1.1-5.0, p < 0.02) and in the peritoneal dialysis patients it was median 3.6 (range 1.5-9.1). We conclude that tubular clearances contribute to the residual renal function in dialysis patients, but the tubular handling of anions and cations in relation to the residual GFR is different between peritoneal and hemodialysis patients. A difference in clearance of organic acids caused by the dialysis techniques may be an explanation for the differences in clinical outcome between the two dialysis modalities.

Cimetidine improves GFR-estimation by the Cockcroft and Gault formula.

In some patients with renal disease 24-hour cimetidine aided creatinine clearances cannot equal GFR even after administration of the maximum daily dose of cimetidine. Short duration cimetidine aided creatinine clearances can equal GFR but are inconvenient for clinical use and can be inaccurate due to incomplete urine collection. We studied how accurately GFR can be estimated without the need to collect urine, by applying the Cockcroft and Gault formula (CCock) on a single plasma creatinine concentration, after oral administration of 3 x 800 mg cimetidine during the preceding 24 hours. GFR was measured as standard clearance, using continuous infusion of 125I-iothalamate. Nineteen patients with various renal diseases, plasma creatinine < 180 mumol/l and body mass index between 15 and 30 kg/m2 were included. After cimetidine administration, plasma creatinine values remained stable for 6 hours, despite rapidly decreasing plasma cimetidine values during the same period, in all 15 patients with GFR > 40 ml/min/1.73 m2. Tubular creatinine secretion was blocked completely in 14 of them. With cimetidine both accuracy and precision of the Cockcroft clearance improved: the mean (+/- SD) ratio of CCock to GFR decreased from 1.28 (+/- 0.21) to 0.98 (+/- 0.11) (p < 0.001) and the standard deviation of the difference (CCock-GFR) decreased from 9.23 to 7.07 ml/min/1.73 m2 (p < 0.05). With cimetidine the Cockcroft clearance correlated well with GFR (r = 0.974, p < 0.001) and this was as good as the correlation, between GFR and a 4-hour standard creatinine clearance (r = 0.972, p < 0.001). In conclusion, with a minimum of inconvenience, this method provides the clinician with accurate information on GFR for the outpatient follow-up of patients with a mild-to-moderate decrease in renal function, provided that no gross discrepancy between total bodyweight and muscle mass is present.

Clearance ratios of amylase isoenzymes and IgG subclasses: do they reflect glomerular charge selectivity?

The clearance ratios of endogenous plasma proteins with the same size but a different charge, such as the amylase isoenzymes and the immunoglobulin (Ig) G subclasses, have been used to assess glomerular charge selectivity in man. These proteins are, however, subject to tubular reabsorption. In this study we measured the IgG subclass/IgG clearance ratios for IgG1 (pI 8.0-9.5), IgG2 (pI 7.0-7.5) and IgG4 (pI < 6) in 6 healthy volunteers. Our results suggested a selective influence of tubular reabsorption: the IgG1/IgG clearance ratio was 0.68 +/- 0.14 (mean +/- SD) and lower than IgG2/IgG (2.02 +/- 1.06, p < or = 0.01). IgG4/IgG was 0.89 +/- 0.39. In addition, we studied the clearance ratios of pancreatic (PA, pI 7.0) and salivary amylase (SA, pI 5.9-6.4) and of IgG1 and IgG2 in 8 patients with minimal change nephrotic syndrome (MCNS), 11 patients recovering from acute tubular necrosis (ATN) and 9 healthy volunteers (controls). In MCNS glomerular charge selectivity is lost, while in recovering ATN tubular function is severely disturbed. The PA/SA clearance ratio was 3.25 +/- 0.89 in controls, reflecting intact glomerular charge selectivity. In MCNS patients the PA/SA clearance ratio had decreased to 1.21 +/- 0.23 (p < or = 0.001). In ATN patients the PA/SA clearance ratio was reduced as well: 1.55 +/- 0.41 (p < or = 0.001), although the aselective nature of the proteinuria and the modest albuminuria indicated intact glomerular charge selectivity. The IgG1/ IgG2 clearance ratio was 0.54 +/- 0.15 in controls, again suggesting preferential tubular reabsorption of IgG1. In MCNS patients the IgG1/IgG2 clearance ratio was 0.16 +/- 0.10 (p < or = 0.001); this probably reflects the relatively increased glomerular sieving of IgG2 when glomerular charge selectivity is lost. In ATN patients the IgG1/IgG2 clearance ratio was 1.07 +/- 0.47 (p < or = 0.001), which suggests a partial loss of preferential reabsorption of IgG1. It was concluded that the PA/SA clearance ratio is influenced by loss of tubular function and therefore does not reflect glomerular charge selectivity specifically. The IgG1/IgG2 ratio cannot be used to assess glomerular charge selectivity either because of the interference of selective tubular reabsorption of the subclasses. These findings put the assessment of glomerular charge using endogenous proteins in a new light and bring forward the necessity to interpret these ratios with the utmost cautiousness.

In vivo inter-organ protein metabolism of the splanchnic region and muscle during trauma, cancer and enteral nutrition.

The study of protein kinetics has entered a new era by the recognition that whole body protein turnover only poorly reflects the true events occurring in several organs and with regard to the multitude of proteins present in the body. It is also increasingly recognized that the simultaneous synthesis and degradation of proteins is important in regulation and adaptation during several metabolic conditions like starvation, feeding, after trauma, and during exercise. Especially important is the recognition that the kinetics of individual proteins may change in opposite directions, thereby leading to fluxes of alpha-amino-nitrogen that serve to adapt to and survive a changing environment. At present, much emphasis is put upon molecular biological regulation. However, it is important that the metabolic processes that occur in the intact organism are still poorly defined. New technology allows the exploration of these processes, which should therefore prompt the initiation of further research in this area.

Circadian rhythm in glomerular transport of macromolecules through large pores and shunt pathway.

In previous studies we have demonstrated that the circadian rhythm in renal clearance of serum proteins is more pronounced than the variability in glomerular filtration rate, and that the highest day-night fluctuations are found for the largest proteins. To analyze whether additional circadian rhythmicity in size-selective glomerular transport could explain these phenomena, we measured renal clearances of inulin and dextrans in a range of 30 to 90 A over a period of one day and compared these data with renal clearance in proteins. Eight patients with nephrotic syndrome and a GFR > 60 ml/min and 6 healthy volunteers were studied during a protocol of bed rest and spaced protein and fluid intake. After administration of a loading dose, inulin and dextran were continuously infused. Blood and urine were sampled every three hours. In patients, but not in normals, fractional clearances of dextrans larger than 45 A showed a circadian rhythm with a peak in daytime and a close phase-relationship with the rhythm in GFR. The day-night differences were the most pronounced for the largest dextrans. Analysis of the day-night differences in a computer model showed circadian variability in transport through the shunt pathway and through large pores. These results can, to an important degree, explain our previous observations on circadian variability in renal clearance of proteins in patients with nephrotic syndrome.

Time course of inulin and creatinine clearance in the interval between two haemodialysis treatments.

BACKGROUND: Urinary volume of haemodialysis patients with residual renal function increases during the interdialytic interval. The contribution of GFR to this change in water and solute excretion has not been quantified in detail. The creatinine clearance (Clc) as a determinant of the GFR may overestimate GFR caused by the tubular secretion of creatinine. Cimetidine has been used to inhibit the secretion of creatinine in non-dialysed patients. No data are available on its usefulness in haemodialysis patients. METHODS: Two identical interdialytic intervals (DI) of 3 days (DI-1, DI-2) were investigated in 11 patients. The interval between DI-1 and DI-2 was 1 week. During DI-2 cimetidine 800 mg daily was administered. Each DI was divided in four urine-collection periods. RESULTS: The water and solute excretion in DI-1 and DI-2 were similar. Urinary production increased from 0.37 +/- 0.30 ml/min to 0.66 +/- 0.33 ml/min (P < 0.05), inulin clearance (Cli) increased from 1.8 +/- 1.1 ml/min to 2.7 +/- 1.2 ml/min (P < 0.05), fractional sodium excretion from 9.0 +/- 5.7% to 14.5 +/- 9.0% (P < 0.05). In contrast to Cli the Clc showed no increase during the interdialytic interval both in DI-1 and DI-2. The overestimation of GFR by creatinine (Clc-Cli) decreased during DI-1 from 1.35 +/- 1.69 ml/min to 0.26 +/- 0.60 (P < 0.05) and during DI-2 from 1.01 +/- 1.33 ml/min to 0.10 +/- 0.67 (P < 0.01). The ratio Clc/Cli decreased during DI-1 from 1.78 +/- 0.53 to 1.09 +/- 0.19 (P < 0.01) and during DI-2 from 2.02 +/- 1.13 to 1.05 +/- 0.30 (P < 0.01). All parameters were not different between the comparable days of DI-1 and DI-2. CONCLUSION: We conclude that the urinary volume in the interdialytic interval is directly related to changes in GFR. During the interdialytic interval GFR increased and tubular secretion of creatinine decreased. The administration of cimetidine did not improve the accuracy of Clc as a measurement of GFR in end-stage renal failure.

Drawbacks of the constant-infusion technique for measurement of renal function.

We investigated the validity of the steady-state constant infusion method (CIM), in which quantitative urinary recovery and constant plasma concentrations of the solute infused are required. Successive 3-h clearances of inulin and p-aminohippuric acid (PAH) were determined for 27 h in 25 patients with renal disease. Results were compared with the standard method of bladder clearance (StM) and with a modified CIM (ModCIM). The 24-h urinary recovery was incomplete for both inulin and PAH. Mean 24-h ModCIM inulin clearance overestimated StM by 4.5 ml.min-1 x 1.73 m-2 (range 0-9, P < 0.001) independent of the extent of renal impairment and pointed to slow distribution and/or extrarenal clearance of inulin. For PAH, the difference between ModCIM and StM clearance was related to the average PAH clearance by ModCIM and StM (r = 0.78). Furthermore, neither plasma inulin nor PAH became completely constant, because of the circadian rhythm in renal function. In conclusion, the conditions of the steady-state CIM technique are not fulfilled, and the method is not suitable for accurate measurement of inulin and PAH clearance, especially when the clearance is low.

Circadian variation of urinary albumin excretion in pregnancy.

OBJECTIVE: The hypothesis was tested that circadian variations in urinary albumin excretion of pregnant women in the third trimester of normal pregnancy are different from nonpregnant individuals. DESIGN: Circadian variability in urinary albumin excretion was studied both in pregnant women and in nonpregnant controls either during normal daily activities or in contrast during continuous bedrest with standardised fluid and food intake to study endogenously generated rhythm. SETTING: Outpatient department and metabolic ward of a university hospital. MAIN OUTCOME MEASURES: Urinary albumin excretion during fixed time periods over the day and the night. RESULTS: Both ambulant pregnant and nonpregnant women have circadian rhythm in urinary albumin excretion but pregnant women have, firstly, a higher 24 h urinary albumin excretion, secondly, smaller relative day-night differences than nonpregnant controls and thirdly, in the metabolic ward some pregnant women demonstrate absence of rhythm. CONCLUSIONS: The higher albumin excretion during pregnancy in 24 h urine collections can largely be explained by a higher excretion during the night, compared with that of nonpregnant women. The day-night difference in urinary albumin excretion of ambulant pregnant women is exogenously determined.

Phase I and pharmacologic study of the novel indoloquinone bioreductive alkylating cytotoxic drug E09.

BACKGROUND: A novel bioreductive alkylating indoloquinone compound, E09 [3-hydroxy-5-aziridinyl-1-methyl-2-(1H-indole-4,7-indione)- prop-F128b-en-alpha-ol], has been shown to have distinct antitumor activity against solid tumors, excellent activity under hypoxic conditions, but no notable bone marrow toxicity in preclinical models. PURPOSE: A phase I study was carried out to determine the toxicity, maximum tolerated dose (MTD), pharmacology, and antitumor response of E09. METHODS: E09 was administered as a 5-minute intravenous infusion once every 3 weeks to 32 patients with solid tumors. The starting dose of 2.7 mg/m2 was one tenth of the mouse equivalent of lethal dose to 10% of animals (MELD10). Dose was escalated by 100% until the area under the curve (AUC) at the MELD10 was reached, following a Fibonacci-like schedule. The pharmacokinetics of E09 and its metabolite E05A with an open aziridine ring was determined using a new high-pressure liquid chromatographic method and noncompartmental calculation of kinetic parameters. The sigmoid Emax model was used to fit pharmacokinetic parameters to toxicity. The renal function and proteinuria were quantitated and were further evaluated by determining renal clearance ratios of immunoglobulin G (IgG) to albumin and pancreatic amylase to salivary amylase. RESULTS: The 32 patients were treated with a total of 85 assessable courses of E09. The dose-limiting toxicity was proteinuria, which was accompanied by sodium and water retention. All symptoms were reversible on day 15 except in two patients, who developed acute renal failure. The ratios of IgG to albumin and pancreatic amylase to salivary amylase suggested a loss of glomerular negative charge consistent with a minimal change glomerulopathy. The pharmacokinetics of E09 showed its rapid elimination from the central compartment but with wide interpatient variation in the overall disposition of the drug. Total plasma clearance of E09 ranged from 3.2 to 24 L/min. The AUC of E09 was linearly related to the administered dose. The relationship between the AUC and proteinuria was best fitted by the sigmoid Emax model (r = .98). In two patients with adenocarcinoma of unknown primary site and in a third patient with bile duct cancer, a partial response was observed. CONCLUSIONS: The MTD of E09 was determined to be 27 mg/m2. The standard approach of drug administration is considered unsuitable because of potential renal toxicity and wide variability in the pharmacokinetics of E09. Individual dose adjustments based on plasma concentration measurements are recommended to combine maximally achievable exposure with tolerable toxicity.

Urinary sodium excretion in patients with nephrotic syndrome, and its circadian variation.

We analysed sodium excretion and its circadian variation in 70 patients with nephrotic syndrome and 19 healthy controls over 1-3 days, with a regimen of bed rest and constant sodium intake around the clock. We sampled urine and blood and took their blood pressure every 3 h. We also scored 60 renal biopsies for presence of interstitial fibrosis and tubular atrophy. Peripheral oedema was estimated in 37 patients. Fifty-nine patients excreted > 10 mmol sodium per 24 h, in equilibrium with dietary intake. In group A (n = 24), sodium excretion followed a normal circadian rhythm, with a daytime peak. In group B (n = 35), 29 had reversed circadian rhythm with a night-time peak, and 6 had no apparent rhythm. Nephrotic syndrome was more severe in group B than in A (serum albumin 19.5 vs. 24.1 g/l, p < 0.05; oedema 7.0 vs. 3.8 kg, p < 0.01). Group B also had signs of more advanced renal disease (GFR 49 vs. 99 ml/min; number of biopsies with tubulo-interstitial damage: 20/28 vs. 4/23; p < 0.001). Reversed sodium rhythm was associated with reversed circadian rhythms for GFR, effective renal plasma flow and urine flow, and blunting or reversal of the day-night differences in blood pressure and plasma renin activity. Eleven patients had urinary sodium excretion < 1 mmol/24 h. With respect to severity of nephrosis, they resembled group B, but GFR and incidence of tubulointerstitial lesions were like group A. Half of the patients with nephrotic syndrome had reversed circadian rhythm for sodium excretion.(ABSTRACT TRUNCATED AT 250 WORDS)

Circadian rhythm of glomerular filtration rate in patients after kidney transplantation.

Within 6 months of a kidney transplantation the graft can be regarded as an organ deprived of its innervation. We analysed whether the transplanted kidney has a diurnal rhythm of its glomerular filtration rate (GFR) similar to the GFR rhythm that has been demonstrated in normal individuals and in patients with nephrotic syndrome. GFR was measured by inulin clearances every 3 h during 1 day of bed-rest and identical food and fluid intake per 3 h in seven patients, 4-7 months after a successful kidney transplantation, and in 10 healthy volunteers. Similar to these healthy subjects, a normal circadian rhythm of GFR was detected in all but one patient with a maximum of 57 (range 45-82) ml/min in daytime, a minimum of 47 (range 36-70) ml/min during the night and a relative amplitude of 21 (range 10-41)%. The circadian rhythm of GFR was absent in the patient with the lowest value of GFR (39 ml/min). In conclusion, GFR has a circadian rhythm in patients studied within 6 months of a kidney transplantation, despite the absence of renal innervation.