Classification of lung cancer patients and controls by chromatography of modified nucleosides in serum.

A wide spectrum of modified nucleosides has been quantified by high-performance liquid chromatography in serum of 49 male lung cancer patients, 35 patients with other cancers, and 48 patients hospitalized for nonneoplastic diseases. Data for 29 modified nucleoside peaks were normalized to an internal standard and analyzed by discriminant analysis and stepwise discriminant analysis. A model based on peaks selected by a stepwise discriminant procedure correctly classified 79% of the cancer and 75% of the noncancer subjects. It also demonstrated 84% sensitivity and 79% specificity when comparing lung cancer to noncancer subjects, and 80% sensitivity and 55% specificity in comparing lung cancer to other cancers. The nucleoside peaks having the greatest influence on the models varied dependent on the subgroups compared, confirming the importance of quantifying a wide array of nucleosides. These data support and expand previous studies which reported the utility of measuring modified nucleoside levels in serum and show that precise measurement of an array of 29 modified nucleosides in serum by high-performance liquid chromatography with UV scanning with subsequent data modeling may provide a clinically useful approach to patient classification in diagnosis and subsequent therapeutic monitoring.

Clinical evaluation and verification of the hyperthermia treatment planning system hyperplan.

PURPOSE: A prototype of the hyperthermia treatment planning system (HTPS) HyperPlan for the SIGMA-60 applicator (BSD Medical Corp., Salt Lake City, Utah, USA) has been evaluated with respect to clinical practicability and correctness. MATERIALS AND METHODS: HyperPlan modules extract tissue boundaries from computed tomography (CT) images to generate regular and tetrahedral grids as patient models, to calculate electric field (E-field) distributions, and to visualize three-dimensional data sets. The finite difference time-domain (FDTD) method is applied to calculate the specific absorption rate (SAR) inside the patient. Temperature distributions are calculated by a finite-element code and can be optimized. HyperPlan was tested on 6 patients with pelvic tumors. For verification, measured SAR values were compared with calculated SAR values. Furthermore, intracorporeal E-field scans were performed and compared with calculated profiles. RESULTS: The HTPS can be applied under clinical conditions. Measured absolute SAR (in W/kg), as well as relative E-field scans, correlated well with calculated values (+/-20%) using the contour-based FDTD method. Values calculated by applying the FDTD method directly on the voxel (CT) grid, were less well correlated with measured data. CONCLUSION: The HyperPlan system proved to be clinically feasible, and the results were quantitatively and qualitatively verified for the contour-based FDTD method.

Negative chemical ionization studied of human and food chain contamination with xenobiotic chemicals.

Negative chemical ionization mass spectrometry with a mixture of isobutane, methylene chloride, and oxygen as the reagent gas has been used to explore contamination of environmental substrates with xenobiotic chemicals. The substrates in question, fish tissue, human seminal plasma, and human adipose tissue, were cleaned up by one of the following three cleanup procedures: (1) continuous liquid-liquid extraction steam distillation; (2) gel-permeation chromatography; and (3) adsorption on activated carbon followed by elution with toluene. The third procedure was used only for the examination of planar polychlorinated aromatic hydrocarbons in environmental samples. Using these techniques, we have found evidence for contamination of fish samples with polychloronaphthalenes, polychlorostyrenes, polychlorobiphenyls, polychlorodibenzofurans, and polychlorodibenzodioxins among other chemicals. The polychlorodibenzodioxins appeared only in the spectra of extracts of fish obtained from the Tittabawassee River at Midland Michigan. The polychlorodibenzofuran ions appeared in NCI mass spectra of fish that were significantly contaminated (above 2 ppm) with polychlorobiphenyls. Toxic substances occurring in human seminal plasma included pentachlorophenol, hexachlorobenzene, DDT metabolites, and polychlorobiphenyls. We have investigated toxic substances in human seminal plasma because of the apparent decrease in sperm density in U.S. males over the last 30 years. Results of screening human adipose tissue for contamination with xenobiotic chemicals have been largely coincident with result of the EPA human monitoring program. Polychlorobiphenyls, DDT metabolites, nonachlor, and chlordane have appeared in most samples examined. Detection limits for all of these chemicals were of the order of 1 ppb.

Evaluation of segmentation algorithms for generation of patient models in radiofrequency hyperthermia.

Time-efficient and easy-to-use segmentation algorithms (contour generation) are a precondition for various applications in radiation oncology, especially for planning purposes in hyperthermia. We have developed the three following algorithms for contour generation and implemented them in an editor of the HyperPlan hyperthermia planning system. Firstly, a manual contour input with numerous correction and editing options. Secondly, a volume growing algorithm with adjustable threshold range and minimal region size. Thirdly, a watershed transformation in two and three dimensions. In addition, the region input function of the Helax commercial radiation therapy planning system was available for comparison. All four approaches were applied under routine conditions to two-dimensional computed tomographic slices of the superior thoracic aperture, mid-chest, upper abdomen, mid-abdomen, pelvis and thigh; they were also applied to a 3D CT sequence of 72 slices using the three-dimensional extension of the algorithms. Time to generate the contours and their quality with respect to a reference model were determined. Manual input for a complete patient model required approximately 5 to 6 h for 72 CT slices (4.5 min/slice). If slight irregularities at object boundaries are accepted, this time can be reduced to 3.5 min/slice using the volume growing algorithm. However, generating a tetrahedron mesh from such a contour sequence for hyperthermia planning (the basis for finite-element algorithms) requires a significant amount of postediting. With the watershed algorithm extended to three dimensions, processing time can be further reduced to 3 min/slice while achieving satisfactory contour quality. Therefore, this method is currently regarded as offering some potential for efficient automated model generation in hyperthermia. In summary, the 3D volume growing algorithm and watershed transformation are both suitable for segmentation of even low-contrast objects. However, they are not always superior to user-friendly manual programs for contour generation. When the volume growing algorithm is used, the contours have to be postprocessed with suitable filters. The watershed transformation has a large potential if appropriately developed to 3D sequences and 3D interaction features. After all, the practicality and feasibility of every segmentation method critically depend on various details of the user software as pointed out in this article.

Structure-activity relationships of chlorinated benzenes as inducers of different forms of cytochrome P-450 in rat liver.

Hexachlorobenzene (HCB) produced increases in ethoxyresorufin (ERR) O-deethylase, aryl hydrocarbon hydroxylase (AHH) and aminopyrine N-demethylase activities in rat liver phenobarbital which were intermediate between those produced by phenobarbital and 3,4-benzpyrene (BP), alpha-Naphthoflavone (ANF) selectivity inhibited ERR activity in BP and HCB-induced microsomes (94% and 88%). Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of liver microsomes indicated that HCB did not produce a detectable increase in a polypeptide with electrophoretic properties similar to those of purified cytochrome P-448 (Mr = 56,000). However, HCB did induce a polypeptide with Mr = 53,000 corresponding to one of two polypeptide bands induced by BP. This polypeptide may represent a second form of cytochrome P-448. Purification of HCB to remove possible dibenzo-p-dioxin impurities did not alter the 'mixed-type' induction produced by HCB. In contrast to HCB, all other chlorinated benzenes tested resembled phenobarbital as inducers.

Silicic acid chromatographic separation of polychlorinated biphenyls and pesticides: some contaminants and limitations.

The Armour and Burke method for separating polychlorinated biphenyls (PCBs) from pesticides involves the use of silicic acid. However, we detected di-2-ethylhexyl phthalate, PCBs, and other impurities as interfering contaminants in several batches of silicic acid. The presence of H2SO4 in some batches of silicic acid is inferred. The acid may be responsible for the production of contaminants which interfere in gas-liquid chromatographic analyses. Contaminants in silicic acid are reduced by extracting the adsorbent with solvent, and/or partitioning the concentrated pesticide fraction with 1N NaOH. These purification procedures provide separations relatively free of impurities, but result in reduced adsorbent activity. PCB-pesticide separations are reproducible only within a given batch lot of silicic acid because of the varying adsorbent characteristics of each lot. Alternative adsorbents should be explored for most PCB-pesticide separations.

Urinary proteins in four rodent species.

1. Mean urinary protein concentration levels are significantly higher in male Peromyscus leucopus than females (98.4 and 72.4 mg/dl). 2. Only females showed a significant correlation between weight and urinary protein concentration (r = 0.75 vs r = 0.03). 3. In intraspecific sexual electrophoretic comparisons of P. leucopus and P. maniculatus non-denatured urinary protein, four and two common bands were identified, respectively. Males of both species showed an extra protein band. 4. Four common electrophoretically separable denatured urinary protein bands were observed between 14,200 and 116,000 mol. wt in male and female P. leucopus and female P. gossypinus. Three of the four major protein bands were also found in P. maniculatus. Male Reithrodontomys megalotis pattern showed none of the major bands.

[Laser-based endoscopic measurement of airway dimensions]. / Lasergestützte endoskopische Vermessung der grossen Atemwege.

Bronchoscopy is the technique of choice for the evaluation of a stenosis in the large airways. However, no system has been successfully employed for the bronchoscopic measurement of airway stenosis. The purpose of these study was the development and validation of a method for measuring the cross-sectional areas in the large airways. Furthermore, this application should be used for the 3D-reconstruction and visualisation of airway stenosis. A laser probe inserted into the operating channel of the bronchoscope enabled assessment of the distance between the images and the tip of the bronchoscope by means of projecting a ring of light on to the endoluminal wall. Image distortion due to the wide-angle lens was corrected by a computer program developed by us. Plastic tubes with known diameters were used for validation. Additionally, distortion-corrected bronchoscopic images were compared with distortion-free videoscopic image analysis of tracheal slices taken from pigs. When plastic tubes were used, the correlation coefficient (r) was slightly higher (r = 0,99, p < 0,01) than the correlation of cross-sectional areas between bronchoscopic and videoscopic images of tracheal slices (r = 0,88, p < 0,01). Furthermore the system could be used in a few patients for 2D and 3D measurement and visualisation of airway stenosis. Application of the present method offer quantitative assessment of airway stenosis located in the large airways.

Influence of patient models and numerical methods on predicted power deposition patterns.

BACKGROUND: A hyperthermia planning system has been developed for generating patient and applicator models as well as calculating and visualizing E-field and temperature distributions. Significant dependencies on models and algorithms have been found. METHODS: Computerized tomography (CT) data sets are first transformed into so called 'labelled CT-volume'-data sets of equal resolution, which are used for segmentation. The first type of patient model obtained subsequently is based on regions with specified electrical properties representing tissues or organs (so called 'region-based model'). The second patient model renders a direct transformation of Hounsfield Units (HU) to electrical constants (so called 'HU-based model'). The FDTD-method (finite difference time domain) is then applied on a cubic lattice employing either an auxiliary 'sub-cubic lattice' (for HU-based segmentation) or a tetrahedron grid (for region-based segmentation) to assign the electrical properties, both representing the anatomy of the patient. E-field distributions are corrected by a post-processing procedure with respect to the geometry of interfaces defined by the tetrahedron grid. For comparison, the VSIE method (volume surface integral equation) is performed on the same tetrahedron grid. The applicator model assumes eight half-wavelength dipole antennas fed with constant voltages with water as background medium. RESULTS: For both numerical methods (FDTD, VSIE) the resulting antenna input impedances as well as the current distributions along the antennas were quite similar and almost insensitive to the particular geometry model (region-based, HU-based). In contrast to that, the power deposition patterns in the interior of the patient depended strongly on those models. Major differences can be related to different labels of the tissue type bone in the HU-based model in comparison to the definition via regions. Conversely, comparable results were obtained using the VSIE method and the FDTD method on the region-based patient model with a posteriori correction at the tetrahedron grid points. SAR (specific absorption rate) elevations up to a factor of 10 were predicted when employing region-based models. Those peaks might correspond to specific toxicity of electromagnetic radiation clinically known as hot spot phenomena or musculo-skeletal syndromes. Conversely, HU-based models generated quite homogeneous power deposition patterns with fluctuations of at most factor 2. CONCLUSION: The methods employing region-based geometry models such as the VSIE method and FDTD method in conjunction with a posteriori correction at tissue interfaces result in comparable E-field distributions for regional hyperthermia. Due to its shorter calculation time, the FDTD method is currently used in the clinic. Predictions derived from HU-based models without prior corrections of tissue specifications are not always supported by clinical experience.

Flow cytometry, morphometry and histopathology as biomarkers of benzo[a]pyrene exposure in brown bullheads (Ameiurus nebulosus).

Brown bullheads were given a single intraperitoneal dose of 0, 5, 25 or 125 mg kg-1 benzo[a]pyrene (BaP), a carcinogenic polycyclic aromatic hydrocarbon, and evaluated over 18 months. Flow cytometric analyses of hepatocyte DNA content indicated an increase in DNA synthesis in BaP-exposed fish prior to day 14 post-exposure. Thereafter, all flow cytometric variables returned to initial levels. Histopathological evaluation of livers from fish sampled at 18 months revealed significant differences among treatments in the amount of hepatic macrophage ceroid pigmentation and basophilic staining intensity. No neoplasms or changes in blood cell DNA content were detected. Significant morphometric variations existed among fish, but differences between sexes overshadowed differences attributable to dose. Flow cytometry yielded no evidence of long-term DNA alterations from a single exposure to BaP; however, the differences detected by DNA analysis shortly after the toxic event suggest that flow cytometric cell cycle analysis may be useful for documenting continuing exposures.

Microcontaminants and reproductive impairment of the Forster's tern on Green Bay, Lake Michigan--1983.

For the 1983 nesting season, Forster's tern (Sterna forsteri) reproductive success was significantly impaired on organochlorine contaminated Green Bay, Lake Michigan compared to a relatively uncontaminated inland location at Lake Poygan, Wisconsin. Compared with tern eggs from Lake Poygan, eggs from Green Bay had significantly higher median concentrations of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), other polychlorinated dibenzo-p-dioxins (PCDDs), total polychlorinated biphenyls (PCBs), total (three congeners) non-ortho, ortho' PCBs, five individual PCB congeners known to induce aryl hydrocarbon hydroxylase (AHH) and several other organochlorine contaminants. Conversions of analytical concentrations of TCDD and PCB congeners based on relative AHH induction potencies allowed for estimation of total 2,3,7,8-TCDD equivalents. Two PCB congeners, 2,3,3',4,4'- and 3,3',4,4',5-pentachlorobiphenyl (PeCB) accounted for more than 90% of the median estimated TCDD equivalents at both Green Bay and Lake Poygan. The median estimated TCDD equivalents were almost 11-fold higher in tern eggs from Green Bay than in eggs from Lake Poygan (2175 and 201 pg/g). The hatching success of Green Bay sibling eggs from nests where eggs were collected for contaminant analyses was 75% lower at Green Bay than at Lake Poygan. Hatchability of eggs taken from other nests and artificially incubated was about 50% lower for Green Bay than for Lake Poygan. Among hatchlings from laboratory incubation, those from Green Bay weighed approximately 20% less and had a mean liver weight to body weight ratio 26% greater than those from Lake Poygan. In both field and laboratory, mean minimum incubation periods were significantly longer for eggs from Green Bay compared to Lake Poygan (8.25 and 4.58 days, respectively). Mean minimum incubation time for Green Bay eggs in the field was 4.37 days longer than in the laboratory. Hatchability was greatly improved when Green Bay eggs were incubated by Lake Poygan adults in an egg-exchange experiment, but was sharply decreased in Lake Poygan eggs incubated in Green Bay nests. Nest abandonment and egg disappearance were substantial at Green Bay but nil at Lake Poygan. Thus, not only factors intrinsic to the egg, but also extrinsic factors (parental attentiveness), impaired reproductive outcome at Green Bay. The epidemiological evidence from this study strongly suggested that contaminants were a causal factor. AHH-active PCB congeners (intrinsic effects) and PCBs in general (extrinsic effects) appeared to be the only contaminants at the concentrations measured in eggs, capable of producing the effects that were observed at Green Bay.