Drug concentration in saliva.

It is possible to predict plasma concentrations of drugs by measurement in saliva, obviating the need for venipuncture. Using a selection of weakly acidic and basic drugs, we have found this prediction reliable for drugs largely nonionized at normal plasma pH (phenytoin, phenobarbital, antipyrine) but unreliable for ionized drugs (chlorpropramide, tolbutamide, propranolol, meperidine). Deliberate alteration of saliva flow rate and pH using different stimuli have produced twofold changes in saliva drug concentrations. Wide interindividual variability of saliva pH is the likely explanation for the inconstancy of saliva to plasma concentration ratios for ionized drugs.

In vitro cytokine profiles and their relevance to rejection following renal transplantation.

Graft rejection remains an important cause of renal allograft failure, despite improvements in immunosuppression and HLA typing. Although HLA matching is beneficial, ensuring an exact match it is often impractical. Thus, a reliable in vitro method for quantitating and qualitating alloreactivity is an important goal. In this study, we measured in vitro the cytokine secretion profiles of mononuclear cells from patients prior to renal transplantation by stimulating with anti-CD3 monoclonal antibody and suppressing with cyclosporine. Mononuclear cells from patients who subsequently developed acute cellular rejection secreted higher mean levels of interleukin (IL)-2 and gamma-interferon (IFN-gamma) than those from patients who had no rejection episodes. IFN-gamma secretion was significantly associated with rejection (P = 0.002), whereas IL-2 secretion did not quite reach statistical significance. There was no significant correlation between IL-4 levels and rejection. Although cyclosporine suppressed the secretion of both IL-2 and IFN-gamma, there was no difference in sensitivity to suppression between rejectors and nonrejectors. These results further emphasize the importance of the TH1 lymphocyte subset in renal allograft rejection. The IFN-gamma secretory capacity of alloreactive T cells may influence the outcome of a renal allograft by (1) activating graft infiltrating macrophages and/or (2) up-regulating HLA molecules on the graft.

Caecal perforation in a renal transplant patient with disseminated histoplasmosis.

A renal transplant patient developed a fatal caecal perforation after Histoplasma capsulatum infection acquired abroad. Disseminated histoplasmosis is an uncommon fungal infection, usually seen in patients with impaired immunity. The diagnosis should be considered in immunosuppressed patients who develop prolonged fever or whose health deteriorates unexpectedly after travelling overseas. The infection is endemic in parts of the United States of America but occurs all over the world. Rapid diagnosis is often possible by histological examination of infected tissues. Treatment if started early may lead to recovery, but if it is not treated it is usually fatal.

Isolation of organisms in CAPD peritonitis: use of nutrient broth cultures and Bactec blood culture media.

Cultures of the first cloudy dialysates from 51 consecutive episodes of peritonitis in patients undergoing continuous ambulatory peritoneal dialysis were carried out using concentrated nutrient broth and Bactec blood culture media in addition to primary culture on solid media. Thirty-seven episodes (73%) were positive on solid media, and 42 episodes (82%) were positive by both nutrient broth and Bactec methods. The value of Gram-stained smears and WBC counts on dialysates is discussed.

Use of a nitrogen detector for GLC determination of fluorouracil in plasma during single- and combined-agent chemotherapy.

A GLC assay for fluorouracil was developed and used to monitor plasma drug levels in patients on both single- and combined-agent chemotherapy. Fluorouracil is extracted from plasma, derivatized by flash methylation, and estimated using a thermionic nitrogen-phosphorus detector. The GLC determination was accurate at concentrations as low as Q.1 microgram/ml of human plasma. Other drugs commonly used in combination with fluorouracil did not interfere with the assay.

Comparison of serum calcium fractions during haemodialysis and peritoneal dialysis.

Total, ultrafiltrable and ionised calcium concentrations were determined in anaerobic serum from healthy volunteers, patients immediately before and after haemodialysis and patients on continuous ambulatory peritoneal dialysis (CAPD). Protein-bound, complexed and albumin-corrected total calcium concentrations were calculated from the results. During haemodialysis, complexed calcium did not change, whereas the other fractions increased. For patients on CAPD, the total, ionised and protein-bound calcium results were frequently lower than the reference group, whereas the ultrafiltrable and albumin-corrected total calcium results were within or higher than the reference group. Albumin-corrected total calcium for all subjects correlated better with ultrafiltrable calcium than with ionised calcium. It was concluded that low ionised calcium concentrations found in CAPD patients may be related to low albumin concentrations, and the concentration of physiologically active calcium may be normal in these patients.

Carbamylated haemoglobin in normal, diabetic and uraemic patients.

Carbamylated haemoglobin arises from the non-enzymic modification of haemoglobin monomers by isocyanate derived from the spontaneous dissociation of urea. We measured carbamylated haemoglobin by high performance liquid chromatography in healthy subjects, non-uraemic hospital patients, diabetics, and different groups of uraemic patients. Carbamylated haemoglobin levels were found to be raised in uraemic subjects, but were independent of age, sex, glycaemic state and haemodialysis procedure. There was no significant difference in carbamylated haemoglobin levels between two groups of patients having different modes of dialysis treatment, probably indicating a similar degree of uraemic exposure in these patients.

Cellular versus vascular rejection in transplant kidneys. Correlation of radionuclide and Doppler studies with histology.

The presence of two distinct subtypes of renal allograft rejection are well documented by histological studies. The differentiation between vascular rejection (VR) and cellular rejection (CR) is essential for proper management by avoiding the need for unnecessary and potentially harmful immunosuppressive treatment of VR. A histological pattern with features that are similar and confusable with some cases of rejection may be seen in cyclosporin A toxicity (CyT). To evaluate the efficiency of Guy's perfusion index (GPI) and the Doppler pulsatility index (DPI) in differentiating these two histological subtypes, a prospective study was designed in which a total of 140 radionuclide tests and 133 ultrasounds scans performed on the same day on 58 patients during the first 3 months post-transplant were analysed, and the results correlated with the histological findings of 84 renal biopsies. Results show that the GPI had a sensitivity of 86.5% and a specificity of 94% in differentiating VR and CyT from CR, while the DPI had values of 83% and 69%, respectively. Chi-squared analysis showed a higher significant association between the GPI and histology (P < 0.0001) compared to that of the DPI and histology (P < 0.005), while Youden's index (J) showed a significant difference (P < 0.05) between GPI and DPI. It is concluded that GPI is more sensitive and specific than DPI in differentiating transplants that are well perfused from those with poor perfusion (VR and CyT).

Urinary protein excretion and the diagnosis of graft rejection or renal dysfunction in renal transplant patients.

Urinary proteins have been found to be a sensitive marker of renal damage caused by nephrotoxic agents. An electrophoretic method was used to investigate the potential value of the pattern of urinary protein excretion in 14 cyclosporin-treated renal transplant patients, to differentiate between graft rejection episodes and other causes of renal dysfunction. Urinary protein excretion consistent with renal damage was observed in all of the patients studied, with no marked differences between those with signs of graft rejection, those with renal dysfunction, or those with stable renal function.

Malaria: a laboratory risk.

A case is described of malaria contracted in a clinical laboratory by accidental self-inoculation with infected blood.

An automated enzymatic inulin assay, capable of full sinistrin hydrolysis.

Renal inulin clearance remains the standard by which other methods of measuring glomerular filtration rate are judged. A fully automated enzymatic assay capable of use with linear configuration inulin was recently published (Summerfield AL, et al. Clin Chem 1993; 39:2333-7). Sinistrin, a readily soluble preparation of polyfructan with side branching, is more suitable for clinical use and far more widely used in Europe. By modifying the incubation phase of samples with inulinase, incorporating a kinetic modification to the method of fructose analysis, and increasing the buffer strengths, we report a fully automated system, with minimal sample prehandling capable of complete sinistrin hydrolysis, and adapted for use on the Cobas Mira.

Peptiduria in experimental Fanconi syndrome in rats.

1. The plasma concentrations and urinary output of proline and hydroxyproline contained in peptides were measured in normal rats and in rats with Fanconi syndrome produced by injection of sodium maleate. All animals received a prior injection of 10 mg of the dipeptide L-prolyl-L-hydroxy-proline to increase plasma and urinary peptide content. There was a significant increase of urinary output of the two imino acids contained in peptides and a fall in their plasma concentrations. 2. It is concluded that increased output of peptides derived from collagen degradation in the experimental Fanconi syndrome in rats is at least in part due to diminished tubular reabsorption of these compounds from the glomerular filtrate. The results are claimed to be relevant to the increased output of urinary peptides in the Fanconi syndrome in man.

Plasma levels of 5-fluorouracil after oral and intravenous administration in cancer patients.

1. Plasma levels of 5-fluorouracil (5FU) have been determined in eleven cancer patients after 0.5 g and 1.0 g intravenous doses, and in one patient after paired 1.0 g oral and intravenous doses. 2. The plasma half-life after the 0.5 g intravenous dose was relatively constant, irrespective of the stage and spread of the disease. 3. Plasma kinetics of the drug were dose dependent. Doubling of the intravenous dose produced a 1.5-fold increase in plasma half life, a two-fold increase in initial plasma drug concentration, and a three-fold increase in area under the concentration/time curve. 4. In one patient receiving paired 1.0 g intravenous and oral doses nine weeks apart, an increase in the bioavailability of the drug coincided with a marked clinical regression in palpable intra-abdominal metastases. 5. The significance of measuring plasma drug kinetics and their relationship to drug efficacy and toxicity are discussed.

Peptide excretion in experimental Fanconi syndrome in the rat.

A study has been made of urinary peptide output in rats before and after production of a Fanconi syndrome induced by a single injection of sodium maleate. There was an unequivocal increase of urinary peptides on the first and second days after the injection, without any detectable change in the concentration of plasma peptides. 2. Similar results were obtained in osteolathyritic rats in which skeletal lesions had been produced by ingestion of beta-aminopropionitrile. 3. The fractional amino acid content of urinary peptides after maleate and beta-aminopropionitrile is shown to be significantly different from that in control animals. 4. Evidence is presented that the increased output of peptides is mainly due to increased renal clearance similar to that previously described for amino acids, glucose and several electrolytes in this type of experimental Fanconi syndrome.

Pharmacokinetics of lidocaine and its deethylated metabolite: dose and time dependency studies in man.

The concentrations of lidocaine and of its deethylated metabolite, MEGX, were measured in blood following the intravenous administration of 50 and 100 mg lidocaine hydrochloride, the oral administration of 100, 300, and 500 mg lidocaine hydrochloride monohydrate, and the oral administration of 300 mg lidocaine hydrochloride monohydrate every 8 h for seven doses, to three healthy volunteers. The range of values for the parameters defining the disposition kinetics of lidocaine were: terminal half-life, 50-231 min; total clearance, 13-17 ml/min/kg; initial dilution space, 0.13-2.5 liters/kg; and volume of distribution at steady state, 0.6-4.5 liters/kg. Lidocaine absorption from solution was rapid, but due to presystemic hepatic metabolism, the availability was low, the range of average values lying between 0.19 and 0.38. No dose or time dependency in lidocaine and monoethylglycinexylidide pharmacokinetics following the single dose studies of lidocaine were noted. Effective hepatic blood flow, based on total clearance and availability measurements, was estimated to be 18-27 ml/min/kg. The concentrations of MEGX were approximately one-third of those of lidocaine following intravenous lidocaine and were comparable following oral lidocaine, but as predicted, the dose normalized area under the MEGX concentration-time curve was constant and independent of the route of administration of lidocaine. In two subjects, the blood concentrations of lidocaine and MEGX following multiple doses of oral lidocaine were those predicted from the single dose studies. In the third subject, the degree of accumulation of lidocaine was greater than predicted. The reasons and mechanism for this difference between subjects on multiple dosing remains unclear.

Acute renal failure following accidental cutaneous absorption of phenol: application of NMR urinalysis to monitor the disease process.

An unusual case of acute renal failure is reported following accidental cutaneous absorption of phenol and exposure to dichloromethane. Renal function during the onset of the nephrotoxic episode and the subsequent recovery period was monitored using a combination of standard clinical biochemical techniques and high resolution 1H-NMR urinalysis. The initial urine biochemical patterns (up to 2 weeks following exposure) showed amino aciduria, glycosuria and lactic aciduria consistent with renal cortical necrosis. There followed a period of polyuria revealing a biochemical pattern (succinic aciduria, dimethylaminuria and N,N-dimethylglycinuria) consistent with renal papillary damage. Haemodialysis was required for a period of 3 weeks and the patient was discharged 42 days after admission to hospital when renal function was normal by standard clinical chemistry criteria (urea, potassium, sodium, creatinine, calcium, phosphate, urine glucose and protein). 1H-NMR spectroscopic urinalysis revealed residual renal biochemical abnormalities consistent with renal papillary damage that were not detected by conventional analytical techniques. One year after the incident the patient is still polyuric, passing up to 3 l of urine a day.