Molecular analysis of the CDR3 encoding region of the immunoglobulin heavy chain locus in cerebrospinal fluid cells as a diagnostic tool in lymphomatous meningitis.

The diagnosis of leptomeningeal B-cell lymphoma is based on the identification of malignant B cells in the cerebrospinal fluid (CSF). Frequently, cytology does not allow clear distinction between neoplastic lymphoid cells and reactively transformed mononuclear cells. Individual B-cell clones can be identified on the basis of DNA sequences that encode the highly diverse complementarity-determining region (CDR3) of the immunoglobulin heavy chain locus (IgH). We studied CSF samples from 5 patients with B-cell malignancies and cytological evidence of leptomeningeal involvement, using polymerase chain reaction (PCR)-based high-resolution capillary electrophoresis and automated fluorescence analysis to detect PCR fragments. As controls, we assessed CSF specimens from 7 patients with inflammatory neurological diseases and three samples without pathological findings. In all patients with B-cell malignancies, a single PCR product was generated, indicating that CDR3-specific fragments were derived from monoclonal cell populations. CSF samples from patients with inflammatory diseases yielded multiple CDR3 amplicons, suggesting the presence of a polyclonal B-cell activation. No PCR product could be amplified in normal CSF samples. Automated fluorescence detection of CDR3 fragments is a highly sensitive and rapid method to distinguish neoplastic monoclonal and reactive polyclonal B-cell populations in the CSF.

The distribution of 14C-chloramphenicol in the Japanese quail (Coturnix coturnix japonica).

The distribution of 14C-labelled chloramphenicol after oral and intravenous administration to egg laying Japanese quail was studied by whole-body autoradiography. In the liver, kidneys, gizzard, intestinal contents (bile) and oviduct, the 14C-concentration was higher than that of the blood short time after injection and remained higher than the blood up to 4 days. From 4 hrs, the concentration of 14C in the egg yolks was higher than that of the blood and from 24 hrs the radioactivity in the albumen of the eggs in the oviduct was also higher than that of the blood. The peak concentration in the egg yolk was found in the second egg laid 2-4 days after administration of 14C-chloramphenicol. In the albumen the maximum concentration was found in the first laid egg 24-48 hrs after administration. In the egg yolks, about 30% of the radioactivity represented unchanged chloramphenicol up to 5 days after administration. It was also shown that about 5% of the injected 14C-chloramphenicol was exhaled as 14CO2 during the first 12 hrs and about 37% of the dose was excreted in the combined faeces and urine during the same period of time.

Fate and specific tissue retention of toxaphene in mice.

Series of female virgin and pregnant albino mice were i.v. injected with 14C-labelled- or unlabelled toxaphene (16 mg/kg b.w.). After survival times ranging from 1 min to 32 days the toxaphene distribution in the body was studied using whole-body autoradiography and capillary gas-chromatography. Autoradiographic studies have shown that after an initial accumulation in the liver, brown fat, lung, brain, kidney, and ovaria (corpora lutea) there was a gradual redistribution of radioactivity to the white fat within 4 h postinjection. The labelling was then decreasing rapidly and only negligible amounts of the radioactivity were present in the adipose tissue after 32 days. In the fetus only the liver and adrenals showed a distinct labelling. A specific and persistent accumulation of the label was detected in some zones of the adrenal cortex suggesting a possible direct interference of toxaphene with adrenal steroid hormone synthesis. The gas chromatographic pattern of toxaphene-derived residues in the tissue samples resembled that of the technical toxaphene, but was changing in different tissues with the time. The liver chromatograms indicated more extensive formation of metabolites.

Studies on the tissue disposition and fate of [14C]toxaphene in Japanese quail.

The disposition and the fate of a [14C]toxaphene preparation was studied in adult and juvenile female Japanese quail. The distribution of [14C]toxaphene in the body is dominated by high concentrations of radioactivity in the adipose tissue and the egg yolk, and, especially in juvenile birds, in the bone marrow. The [14C]toxaphene compounds present in the fat tend to be less polar than the parent [14C]toxaphene. More than half of the radioactive dose is excreted via the droppings, eggs, and preen gland within a few days, with biliary excretion playing a major role. The main part of the radioactivity in feces and urine consists of water soluble compounds, a part of which is indicated to be glucuronide and sulfate conjugates, but other metabolites are probably also present. A possible connection between the toxaphene residues in the bone and the known toxaphene induced osteomalacia in birds is discussed.

Chlorinated paraffins: disposition of a highly chlorinated polychlorohexadecane in mice and quail.

Uniformly 14C-labelled 1-chloro-polychlorohexadecane (PCHD) of high chlorine content (69% w/w) was given to Japanese quail and to C57Bl mice perorally (p.o.) and intravenously (i.v.). The degradation of PCHD to 14CO2, measured during 8 h, was found to be minute (about 1% of dose) in both species after either route of administration. In mice 66 and 43% of dose was eliminated in the feces during 96 h following p.o. and i.v. administration, respectively; the urinary excretion was about 3% in both cases. In quail, the combined fecal and urinary excretion during 96 h after p.o. administration was 58% of dose. The autoradiographic distribution following p.o. administration showed some general similarities between mice and quail; high radioactivities were present in bile, liver, kidney, and intestinal contents up to 24 h after administration. In addition, in quail high radioactivity was present in the hypophysis, retina, blood, and egg yolk. In mice strong accumulation and retention was observed in the corpora lutea up to 30 days after administration. A long time retention in fat occurred in both species.

Metabolic fate of chlorinated paraffins: degree of chlorination of [1-14C]-chlorododecanes in relation to degradation and excretion in mice.

The disposition of three [1-14C]-chlorododecanes (MCDD, PCDD I and PCDD II; 17.4%, 55.9%, and 68.5% chlorination) was studied in C57Bl mice. [1-14C]-lauric acid (LA) was studied as reference compound. Fifty-two percent (MCDD), 32% (PCDD I), and 8% (PCDD II) of the radioactive doses were exhaled as 14CO2 during 12 h after i.v. injection. Similar results were obtained after p.o. administration. In addition to a marked labelling of the liver and fat, the distribution patterns observed at 24 h after administration revealed an uptake of radioactivity in tissues with high cell turnover/high metabolic activity, e.g., intestinal mucosa, bone marrow, salivary glands and thymus. The concentration of radioactivity in these sites and the exhalation of 14CO2, which were inverse to the degree of chlorination, indicate that the chloroalkanes are degraded to metabolites which can be utilized in the intermediary metabolism. A similar, although more pronounced, distribution pattern and 14CO-2-exhalation (70% of i.v. dose) was observed after LA administration. The long time retention of heptane-soluble radioactivity in liver and fat (indicating unmetabolized substance) increased with degree of chlorination. On the contrary, the administration of LA and the chlorododecanes MCDD and PCDD I, but not of PCDD II, resulted in a selective labelling of the central nervous system 30-60 days after injection.

Metabolism of 2,4',5-trichlorobiphenyl: role of the intestinal microflora in the formation of bronchial-seeking methyl sulphone metabolites in mice.

Groups of germ-free and conventional mice were treated with 2,4',5-trichlorobiphenyl (triCB) and [35S]cysteine or [35S]methionine, respectively. Control animals received the labelled amino acids only. Conventional mice accumulated significantly more extractable radioactivity both in lung and kidney tissues when compared to germ-free mice. The extracted radioactivity in lung and kidney tissues was shown to be due to the accumulation of methyl-[35S]sulphonyl-triCB. The low radioactivity in lungs of the germ-free mice was also shown to be due to the accumulation of small amounts of the sulphones. The results indicate an involvement of the intestinal flora in the formation of methyl sulphone metabolites of triCB.

Accumulation of polychlorinated biphenyls in steroidgenic tissue of gonads and adrenals in Japanese quail.

Female quail exhibited a high uptake of PCB in the yolk of growing follicles, in the thecal gland cells and the medullary interstitial tissue of the ovary and in the postovulatory and atretic follicles. Uptake in the shell gland was low. An elimination study of four days indicated that the major part of PCB is excreted via the eggs. Male quails showed a very low uptake of PCB in the testes and no accumulation was in the interstitium. The uptake in the fat, liver, and adrenals was similar to that in the females.