Urinary excretion of cyclophosphamide in humans, determined by phosphorus-31 nuclear magnetic resonance spectroscopy

Phosphorus-31 NMR spectroscopy was used to analyze urine samples from patients treated with cyclophosphamide (CP) on 2 consecutive days. CP and all of its known phosphorylated metabolites except the tautomeric pair 4-hydroxycyclophosphamide/aldophosphamide, i.e. carboxycyclophosphamide (CXCP), dechloroethylcyclophosphamide (DCCP), alcophosphamide, ketophosphamide, and phosphoramide mustard (PM), were determined. Several other signals corresponding to unknown CP-related compounds were observed. Seven of them were identified; all were hydrolysis products of CP or its metabolites (one from CP, two from CXCP, three from DCCP, and one from PM). Twenty-four-hour urinary excretion of unmetabolized CP was not significantly different on the first (17% of the daily administered dose) and second (16%) days of treatment. The amounts of phosphorylated metabolites excreted in 24-hr urine samples were much higher after the second CP dose (37%) than after the first (20%), suggesting autoinduction of CP metabolism. CXCP and its two degradation products (accounting for 7-10% of CXCP) were by far the major metabolites (11.5 and 23% after the first and second doses, respectively). DCCP plus its degradation products and alcophosphamide each represented 2-3% on the first day of treatment and 5% on the second day of treatment. Levels of PM and its degradation products were extremely low (0.3 and 0.6% after the first and second CP doses, respectively), as were those of ketophosphamide (0.4 and 0.6% on the first and second days of treatment, respectively). We noted only modest interpatient variation in excreted levels of CP and all of its metabolites.     

The photolysis of dihalomethanes in surface seawater

Laboratory experiments were carried out with different types of natural and artificial seawaterto study the aqueous degradation kinetics of the photolabile compounds CH2I2, CH2Brl, and CH2ClI. Irradiation studies were carried out with a 1-kW Xe lamp, optically filtered to simulate the solar spectrum at the earth's surface. Halocarbon concentrations in the samples were analyzed by purge-and-trap gas chromatography/mass spectrometry. Generally, the compounds studied were found to follow first-order removal kinetics on irradiation. However, in the case of CH2I2, deviations from first-order removal occurred after a few minutes of irradiation, probably indicating radical recombination. Photolytic lifetimes varied from 12 min for CH2I2 to 13 h for CH2ClI in natural surface seawater at 15 degrees C and an irradiation intensity corresponding to overhead sun (solar zenith angle = 0 degrees). Photolysis of CH212 in artificial and natural seawater generated CH2CII with a yield of 25-30%, suggesting that this reaction is an important source of marine CH2ClI. Dark-incubations of CH2I2 for up to one week showed that this compound does not undergo nucleophilic attack by chloride, indicating that photolysis is the main abiotic degradation mechanism of CH2I2 in seawater.     

Effect of hypothyroidism on the lipid composition of rat plasma and erythrocyte membranes

The effect of hydrothyroidism on plasma and erythrocyte membrane lipid components has been investigated. This pathological state is accompanied by a) a cholesterol increase of about 60% in plasma, and at the same time a 22% reduction in erythroycte membranes; b) 44% and 30% phospholipid level decreases in both plasma and red cell membranes, respectively; and c) almost unaffected phospholipid and fatty acid compositions of both plasma and erythrocyte membranes. All changes were corrected by treatment of the hypothyroid rats with triiodothyronine for two days. These findings suggest that in hypothyroid rats a reduced transfer of cholesterol from plasma to erythrocyte membrane probably takes place. This could explain, at least in part, the increased hematic cholesterol level observed in hypothyroid animals. In red cell membranes, the simultaneous decrease in cholesterol and phospholipid levels does not alter the cholesterol/phospholipid molar ratio, thus avoiding their abnormal function.     

Observations of brittle failure around a circular test tunnel

In 1987, AECL proposed several experiments for its Underground Research Laboratory. Among these was the Mine-by Experiment, the main focus of which was to investigate the formation and characteristics of the excavation disturbed zone adjacent to an underground opening. The extent and nature of this zone are important concerns for both the operational safety and the sealing of a vault to control the migration of radionuclides.The excavation response phase of the Mine-by Experiment, which involved the excavation, without explosives, of a 46-m-long, 3.5-m-diameter test tunnel in brittle unfractured granite, was carried out between January and July 1992. State-of-the-art instrumentation comprising both geomechanical and geophysical instruments was used to monitor the progressive failure process around the circular tunnel, and the in situ experiment was supplemented by extensive laboratory tests. This paper provides a detailed record of the brittle failure process that was observed during excavation of the tunnel.     

A decision support system to detect morphologic changes of chromatin arrangement in normal-appearing cells

Several studies have described malignancy-associated changes (MACs) of chromatin arrangement in the nuclei of apparently normal cells adjacent to and distant from an invasive cancer area. MAC assessment is a hard task, since it requires a deep knowledge of morphologic features of chromatin arrangement. The aim of this work is to verify the reproducibility of the subjective evaluation of the expert on the basis of a decision support system (DSS) that automatically and objectively reproduces MAC diagnosis. A set of 61 patients with suspected clinical diagnosis for lung cancer has been taken into account. The scientist who first described MAC defined each patient as MAC positive or negative on the basis of the MAC diagnosis performed on all cells of the related cytologic sample. A DSS based on an artificial neural network has been set up to learn the relation between 14 morphometric and texture parameters, computed on each nucleus by image processing techniques, with the MAC diagnosis of the expert on each cell. The results show that an objective automatic assessment on MAC by the DSS can effectively support the MAC diagnosis. The method adopted in this approach may be also appropriate for other problems, where an automatic classification of visually inspected patterns of biological micro- and submicrostructure is needed.     

The membrane-spanning proteoglycan NG2 binds to collagens V and VI through the central nonglobular domain of its core protein

NG2 is a membrane-spanning proteoglycan with a primary structure unique among cell surface or extracellular matrix proteins. To characterize the interaction between NG2 and extracellular matrix proteins, we have used a eukaryotic expression system to produce and purify several recombinant fragments covering not only the entire ectodomain of NG2 but also distinct subdomains of the molecule. Using a solid phase binding assay with various extracellular matrix proteins, we have identified two main ligands for NG2, namely, collagens V and VI. Consistent with previous models of glycosaminoglycan attachment, roughly 50% of the recombinant NG2 fragments containing the central domain have chondroitin sulfate chains attached to the protein core. These glycosaminoglycan chains are not directly involved in collagen binding, since chondroitinase-treated fragments exhibit an unimpaired ability to bind to both collagens. Using more restricted recombinant fragments of NG2, we mapped the binding site for both collagens to the central domain of NG2. Electron microscopy after rotary shadowing of native NG2 molecules indicates that this extended nonglobular domain provides a flexible connection joining the two N- and C-terminal globular regions of NG2. Rotary shadowing of mixtures of NG2 and collagen V or VI confirms a direct interaction between the molecules and indicates that the collagens align with the central region of NG2, giving the appearance of a rod between the N- and C-terminal globules.     

The effects of PEG-interleukin-2 and interleukin-2 on essential hypertension and cellular immune function in the spontaneously hypertensive rat

The effects of recombinant human interleukin-2 covalently linked to polyethylene glycol (PEG-IL-2) or interleukin-2 (IL-2) on hypertension and in vitro suppressor T cell function in the spontaneously hypertensive rats (SHR) were investigated. Male young prehypertensive (4 weeks old) SHRs and adult (10 weeks old) SHRs with established hypertension were injected with low (5,000 units (u)/kg) or high (50,000-100,000 u/kg) dose of PEG-IL-2 or IL-2 as a single bolus or repeated injections. Systolic blood pressure was measured twice weekly using the tail-cuff technique. Systolic blood pressure in the PEG-IL-2 or IL-2 treated animals, irrespective of age, dose, or route of injection, did not differ significantly from that measured in vehicle-treated controls over a 10 week period. Mean arterial pressure measured by intra-arterial catheter was 159 +/- 7 mm Hg 10 weeks after treatment with repeated injections of 5,000 u/kg of PEG-IL-2 and 158 +/- 9 mm Hg in vehicle-treated controls. All rats injected with IL-2 had IL-2-specific IgG antibody in their sera. None of the PEG-IL-2 treated rats had any detectable anti-IL-2 antibodies in their sera. Thus, PEG-IL-2 showed far less immunogenicity than IL-2. Suppressor T (Ts) cells generated from adult SHR spleen cells failed to suppress pokeweed mitogen (PWM)-driven immunoglobulin G (IgG) synthesis. PEG-IL-2 or IL-2 supplementation both in vitro and in vivo restored the ability of adult SHR to generate Ts cells able to inhibit IgG synthesis. Our data suggest that PEG-IL-2 or IL-2 administration does correct a prominent defective Ts cell activity found in adult SHR, but that correction of this immune abnormality is not attended by an attenuation of hypertension.     

Synthesis and pharmacological activities of some 2,3,4,5-tetrahydro[1,5]benzo[f]thiazepines

For the purpose of prophylactic treatment of splenic aneurysms, both examinations and treatments should be minimally invasive. Here, we report a case of a patient who underwent three-dimensional arterial computed tomography (CT) and laparoscope-assisted splenectomy with aneurysm resection as a combination of minimally invasive examination and treatment.