Noninvasive in vivo 13C-NMR spectroscopy in the rat to study the pharmacokinetics of 13C-labeled xenobiotics.

Noninvasive NMR methodology has been developed to enable monitoring of 13C-labeled xenobiotics in the rat in vivo. 2,2-Dichloro-1-(2-chlorophenyl)-1-(4-chlorophenyl)-[3-13C]-propane can be detected in the liver of intact rats by in vivo 13C surface coil NMR spectroscopy after ip administration of the compound. The experiments were performed at 1.9 and 9.4 Tesla. The intrahepatic changes of the signal intensity of the labeled compound were followed as a function of time. In the days following administration, the concentration decreased and dropped to values below the detection limit after 12 days. The study demonstrates the feasibility of studies on pharmacokinetics of 13C-labeled compounds in the rat using noninvasive, in vivo surface coil NMR spectroscopy in animals. The sensitivity allows the detection of a single dose of the drug of 200 mg/kg, but can be improved.

Long term follow-up of multiple sclerosis by standardized, non-contrast-enhanced magnetic resonance imaging.

The usefulness of non-contrast-enhanced, standardized magnetic resonance imaging for the longterm follow-up of MS patients was evaluated in a retrospective study in 36 patients with clinically definite MS. All had remitting-relapsing diseases courses. Sixteen patients remained clinically stable during follow-up. Mean duration of follow-up was 22 months (SD: 11). A mean number of 3 MRI examinations was performed in each of the patients (SD: 1). Subclinical evolution was detected in 56% of the stable patients, indicating that clinical data alone are insufficient to assess disease activity. The relapsing patients showed significantly more and larger changes on MRI than stable patients (P less than 0.001), indicating that MRI is well suited as a follow-up parameter in conjunction with clinical data. The time courses of these quantitative changes and of the qualitative changes of putative MS lesions on MRI are discussed. It is concluded that MRI is a good indicator of global disease activity in multiple sclerosis patients, which makes MRI very useful for the evaluation of therapeutic trials.

Elimination of matrix and spectral interferences in the measurement of lead and cadmium in urine and blood by electrothermal atomic absorption spectrometry with deuterium background correction.

Direct measurement of lead and cadmium in blood and urine by electrothermal atomic absorption spectrometry with deuterium background correction (D2-AAS) is prone to severe matrix and spectral interferences. We overcame these effects by coating the L'vov platform with ammonium molybdate, reducing the atomization time, introducing a post-atomization cooling step, carefully selecting ashing and atomization temperatures, and using an appropriate procedure for matrix modification. To determine Pb and Cd in blood and urine, we used matrix-matched calibration curves. With the proposed procedure for sample preparation, both Pb and Cd in whole blood can be determined in the same diluted sample. Results obtained by D2-AAS correlate closely with those by Zeeman-corrected AAS. Detection limits (mean blank + 3 SDblank) for Pb in urine and blood were 4 micrograms/L. For cadmium, the detection limits were 0.4 and 0.1 micrograms/L for urine and blood analysis, respectively. Between-run CVs were less than 5.0%.

Determination of gadolinium in biological materials using graphite furnace atomic absorption spectrometry with a tantalum boat after solvent extraction.

A method was developed for the determination of gadolinium (Gd) in biological material using graphite furnace atomic absorption spectrometry (GFAAS). The element is first extracted into methyl isobutyl ketone and then reextracted into hydrochloric acid. Factors influencing the recovery of extraction such as pH, choice of chelating agents, and hydrochloric acid concentration have been investigated. The element is determined under STPF (stabilized temperature platform furnace) conditions with atomization from a tantalum boat. Under optimized furnace conditions, the use of the tantalum boat improved sensitivity substantially compared to the use of pyrolytically coated graphite tubes. Around 150 measurements could be performed with 1 boat. Memory effects, being a common problem in the GFAAS determination of lanthanoids, were no longer observed after insertion of the boat. The characteristic mass and detection limit (2SD; SD = standard deviation) of the Gd determination are 1000 and 2060 pg, respectively. The precision evaluated as the relative standard deviation (RSD) of six analyses was below 10% for tissue Gd concentrations ranging from 0.92 to 72.0 micrograms g-1. The recovery of added analyte ranged between 92.0% and 99.3%. The method was found to be suitable for studying the pharmacokinetics and biodistribution of Gd in rats.

Does lead play a role in the development of chronic renal disease?

For many decades lead has been considered to be one of the causes of chronic renal failure. A critical analysis of the available epidemiological studies, however, indicates that the relationship between lead exposure and the development of chronic renal failure is largely circumstantial. Indeed, several aspects remain obscure: relative and absolute risk, risk factors, duration of exposure, pathology and diagnostic criteria. Moreover, methodological problems related to cohort selection (i.e. the 'Healthy worker effect'), study design and definition of renal dysfunction may limit the value of the epidemiological results. Although the available literature suggests that lead may play a direct or contributory role in the development of chronic renal failure it is concluded that additional detailed epidemiological studies are required.

Acromegaly, multinodular goiter and silent polyostotic fibrous dysplasia. A variant of the McCune-Albright syndrome.

A 36-year-old woman is reported with a possible variant of the McCune-Albright syndrome. The triad was incomplete because of the absence of skin pigmentation and since the sexual precocity was not evident. The presence of a pituitary mass and the secretory dynamics of growth hormone and prolactin were suggestive of a mammosomatotroph cell adenoma. A toxic multinodular goiter was also associated, but unique was the spontaneous normalization of the thyroid function. Unusual was the silent evolution of the polyostotic fibrous dysplasia, which was only fortuitously discovered during magnetic resonance imaging of the pituitary region. Treatment of the acromegaly with the long-acting somatostatin analogue octreotide resulted in an important inhibition of the GH secretion and in a reduction of the volume of the pituitary adenoma.

Improved correlation of magnetic resonance imaging (MRI) with clinical status in multiple sclerosis (MS) by use of an extensive standardized imaging-protocol.

In a previous study we have shown that the sensitivity of magnetic resonance imaging (MRI) for the detection of multiple sclerosis (MS) lesions was improved significantly, especially in the infratentorial region, by use of an extensive standardized MRI-protocol consisting of sagittal T1, axial protondensity and axial T2, and sagittal protondensity and sagittal moderately T2-weighted images. The goal of the present study was to assess whether the clinical correlation of the visualized lesions had improved accordingly. Using a scoring system based on lesion dimensions, we compared 70 MRI examinations performed in 25 patients with definite MS, with the relevant clinical data as given by the Expanded Disability Status Scale (EDSS) and Functional System scale (FS). We found a significant correlation (r = 0.66, P = 0.0001) between the MRI score and the EDSS. Significant correlations also existed between MRI scores and cerebellar and brainstem FS scores. These correlations were consistently higher than those reported by other authors. We conclude that a standardized MRI examination, including sagittal protondensity and moderately T2-weighted images, should be performed in every MS patient. The improved clinical correlation could be of importance in follow-up studies when assessing the efficacity of therapy.

Experimental cerebral infarction in the rat: utility of Gd-DOTA for MR imaging.

Nineteen rats presenting a very small (about 2-mm diameter), superficial cerebral infarction were studied with MR-imaging (1.89 Tesla) before and after injection of Gd-DOTA. Fifteen rats examined 1 or 2 hr, and 2, 3, 4, 5, 7, or 10 days after lesion induction, received 0.5 mmol Gd-DOTA/kg body weight. Four other rats presenting a 4-day-old lesion, received 0.1, 0.2, or 0.6 mmol Gd-DOTA/kg. Each rat underwent one imaging study comprising T2-weighted spin-echo (SE) images (3000/100) with subsequent injection of Gd-DOTA followed by 12 consecutive series of T1-weighted SE images (320/40), each taking 6 min. Using 0.5 mmol Gd-DOTA/kg, early (immediate) and long-lasting (more than 1 hour) visualization of lesions of varying age (1 hr to up to 10 days) was possible and at appropriate time intervals after injection, the visualization of the lesion was clearer and more complete than with T2-weighted images. Even in the rats studied with smaller doses of the contrast agent (0.1 or 0.2 mmol/kg), postcontrast T1-weighted images provided superior delineation of the lesions as compared to T2-weighted images.

Iron overload and bone disease in chronic dialysis patients.

Bone pathology was studied in 27 patients showing either iron or aluminium accumulation in bone. These patients belonged to a group of 120 unselected chronic haemodialysis patients in whom transiliac bone biopsies had been obtained. Group A consisted of 12 patients with bone iron deposits (positive Perls' staining at the calcified bone boundary, CBB) and only minimal aluminium accumulation (bone aluminium below 20 micrograms/g wet weight). Group B included 15 patients with pronounced aluminium accumulation (positive aluminium staining at the CBB and bone aluminium of 20 micrograms/g wet weight or more) and without significant bone iron deposition (negative Perls' staining at the CBB). Bone diseases were classified as early hyperparathyroidism, osteitis fibrosa, mixed disease, osteomalacia, adynamic bone disease or other bone condition using osteoid volume, relative osteoblastic activity (ROBA%), and the presence of fibrosis, as criteria. In group A, 5 of 12 patients showed adynamic bone disease, a fairly uncommon condition in the general population of non-parathyroidectomised dialysis patients. In fact, in a control group of 80 patients without iron and without aluminium overload, only five patients showed adynamic bone disease. In group B, 8 of 15 patients showed osteomalacia, and 2 of 15 presented with mixed disease, which is in agreement with the established relationship between bone aluminium accumulation and the occurrence of defective bone mineralisation. It is concluded that iron overload in dialysis patients is associated with an increased frequency of adynamic bone disease.

Value of serum aluminium monitoring in dialysis patients: a multicentre study.

The clinical relevance of regular serum aluminium monitoring in dialysis patients was investigated in a multicentre study by 6-monthly determination of the serum aluminium during 4 consecutive years. In a group totalling 1193 patients, a striking decrease of mean serum aluminium was observed the last 2 years of the study. This phenomenon was accompanied by a substantial reduction of the prescribed dose of aluminium hydroxide (Al(OH)3) and its partial replacement by calcium carbonate (CaCO3) and/or magnesium hydroxide (Mg(OH)2). Under this policy serum phosphate control remained satisfactory. In all the centres, water treatment was found to be adequate, yielding dialysate aluminium around 2 micrograms/l. Dialysis patients with clinically overt liver disease showed a significantly greater median serum aluminium concentration than that observed in a control dialysis population. Compared to the latter group, the median serum aluminium concentration of dialysis patients with diabetes mellitus did not differ significantly. Results further indicated that patients with biopsy-proven osteomalacia presented a significantly greater median serum aluminium compared to that of patients without osteomalacia. We demonstrated that a serum aluminium of 60 micrograms/l provides a relatively sensitive (82%) and specific (86%) index for the detection of aluminium-related bone disease (ARBD). Provided the aluminium determinations are performed by a qualified laboratory, serum monitoring in dialysis patients (a) allows the safer use of aluminium-containing phosphate binders, and (b) is of value in the diagnosis of overload/toxicity.

Improved sensitivity of MRI in multiple sclerosis by use of extensive standardized procedures.

The relative value of two different MRI procedures for the assessment of infratentorial extension in multiple sclerosis (MS) was studied. Multislice spin-echo techniques were used overall. Procedure A consisted of parasagittal T1-weighted images (500/30) and axial T2-weighted images (2500/30, 2500/120). Procedure B consisted of parasagittal T2-weighted images (1600/35, 1600/90). In the parasagittal T2-weighted images clear visualization of MS lesions is achieved because signal intensities of CSF and normal nervous tissue are nearly identical. All images were performed with a 0.5 Tesla MR system. Data were obtained in 98 patients with definite (N = 30) or probable MS (N = 68). Areas with abnormal signal intensity in the infratentorial regions (brainstem, cerebellum, and/or cervical spinal cord) were identified in 44% of the patients with procedure A and in 64% with procedure B. The standard application of the combination of both procedures improves the sensitivity of the MR examination for the diagnosis of MS, the delineation of infratentorial lesions and the correlation between clinical and MR data without excessively increasing imaging time.

Risk of aluminium intoxication in long-term acetate Redy dialysis.

Closed-circuit dialysis using the Redy sorbent cartridge to regenerate the dialysate has been incriminated in previous reports as a cause of severe fracturing osteomalacia and fatal encephalopathy in several patients treated with this procedure for 15-36 months. In a retrospective study, we compared 15 unselected patients who had received Redy dialysis for 66 +/- 14 months with 15 control patients dialysed with single passage of dialysate. Redy and control patients were matched for age, sex, and duration of dialysis. They belonged to two dialysis centres, situated in the same geographical area and having a common water supply. Mean serum and bone aluminium concentrations were slightly greater in the Redy group but the differences were not significant. Pathological fractures had occurred in two Redy patients and in one control, but could not be attributed to aluminium-induced bone disease. Although the histochemical staining for aluminium in bone was positive in six patients, diagnosis of aluminium-induced bone disease was made in one case only. The results of bone histomorphometry did not differ significantly between the two groups. Our findings may be explained by the strict application of the measures required to avoid aluminium contamination of the Redy dialysate, i.e. sufficient rinsing before dialysis, use of almost aluminium-free water, and of acetate-buffered dialysate.

Defect in succinate oxidation by isolated muscle mitochondria in a patient with symmetrical lesions in the basal ganglia.

A 3-year-old boy was referred for evaluation of psychomotor retardation. He had a waddling gait with proximal hypotonia and paresis. Computed tomography (CT scan) and magnetic resonance imaging (MRI) of the brain demonstrated symmetrical lesions in the basal ganglia suggesting bilateral necrosis. Lactate and pyruvate levels in blood and cerebrospinal fluid were persistently elevated. A biopsy of the quadriceps muscle showed normal light microscopic findings except for a slightly raised number of lipid droplets. Electron microscopy confirmed this and also showed a rather large number of subsarcolemmal mitochondria without crystalline inclusions. Biochemical studies showed a normal carnitine level and normal mitochondrial enzyme activities in muscle homogenate, including succinate-cytochrome c reductase. However, intact isolated mitochondria failed to oxidize succinate. An explanation for this paradoxical finding is a deficiency in that part of the coenzyme Q (CoQ) that is reduced by the succinate dehydrogenase complex. The differential diagnosis between Leigh's syndrome and infantile bilateral striatal necrosis (IBSN) is discussed. The role of neuroradiology in prompting complementary investigations is stressed.

Bone lead in dialysis patients.

We measured lead and calcium in multiple bone biopsies from 11 cadavers without known excessive past exposure to lead. Paired iliac crest, transiliac and tibial bone biopsies from these cadavers indicated that in bone biopsy specimens the lead/calcium ratio is more reproducible than the absolute lead concentration. There were no significant differences between the lead/calcium ratios from the iliac crest, transiliac, or tibial specimens. Transiliac bone biopsies from 35 patients (13 patients showing symptoms of slight or moderate degree of renal failure, medical history of gout and/or arterial hypertension and 22 lead workers with chelatable lead in excess of 1000 micrograms) indicated that the lead and the lead/calcium ratio in bone biopsies reflect body lead stores as estimated by the EDT A test (r = 0.87 and 0.83, respectively). Chemical and histological studies of transiliac biopsies previously obtained from 153 dialysis patients (from 8 dialysis centers from Belgium, France and Germany) for studies of aluminum-induced bone disease showed that chronic renal failure and dialysis do not cause accumulation of lead in bone and elevated bone lead does not appear to alter trabecular bone histomorphometry. We found that in 5% of the hemodialysis population studied, bone lead concentrations approximated levels found in active lead workers.

High resolution nuclear magnetic resonance imaging of the spinal cord in experimental demyelinating disease.

Chronic recurrent experimental allergic encephalomyelitis was induced in a strain 13 guinea pig by inoculation of isologous spinal cord homogenate. The spinal cord was obtained after perfusion with 4% paraformaldehyde and examined with nuclear magnetic resonance (NMR) imaging. Proton NMR spin echo images (repetition time: 3 s; echo times: 20 and 60 ms) were obtained from intact, isolated spinal cord in a 4.7 Tesla, 50 mm bore magnet. The slice thickness of the images was 380 microns and the inplane resolution was 40 X 40 microns. The images showed superficial areas of low signal intensity in the lateroventral regions of the white matter, in some instances with a seam of higher signal intensity. Neuropathologically, these abnormalities corresponded exactly to areas of demyelination. Control images did not show these abnormalities. The present high resolution imaging allowed a correlation between demyelination and abnormal NMR signals in a small laboratory animal with an inflammatory demyelinating disease.