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Nitroarene concentrations and direct-acting mutagenicity of diesel exhaust particulates fractionated by silica-gel column chromatography.

Diesel exhaust particulates were extracted with benzene-ethanol (3:1, v/v) and separated into five fractions by silica-gel column chromatography. Direct-acting mutagenic activity was assayed by the Ames test using the Salmonella typhimurium YG1024 strain. The total activity of five fractions was about four times greater than that of the crude extract, suggesting that the activities in the fractions were suppressed in the crude extract. Strong activity was observed in fraction 4 which was eluted with dichloromethane (61.5% of the total activity) and fraction 5 which was eluted with ethanol (35.3%). Nitropolycyclic aromatic hydrocarbons (NPAHs) were determined by high-performance liquid chromatography with chemiluminescence detection. They were found mainly in fraction 4, although one NPAH was in fraction 3 which was eluted with n-hexane-dichloromethane (3:1, v/v). Based on these results, 53.1% of the activity in fraction 4 was attributed to NPAHs. The contribution of 1-nitropyrene and 1,3-, 1,6- and 1,8-dinitropyrenes was great and that of the other NPAHs was small. The mutagenic compound in fraction 5 was not identified. Fractions 1 and 2, which were eluted with n-hexane, and fraction 3 suppressed the activity of fraction 4. Polycyclic aromatic hydrocarbons in fractions 2 and 3 were considered as possible suppressors of NPAHs.

Animals↗

Effects of irradiation by single or multiple fractions per day on transplantable murine mammary carcinoma.

Effects of different fractionation schedules of irradiation were studied in an experimental mammary adenocarcinoma transplanted in the back of mice. The number of fractions per day varied from one to three, keeping the daily dose constant. It was found that an increase of the number of fractions per day did not necessarily lead to a decrease in the tumour response, as might be expected for the same total dose. At a twelve-day overall time, three fractions per day did not give such a good result as one fraction per day, but at a nineteen-day overall time three fractions per day were about as good as one fraction per day; and below 45 Gy total dose, the result with three fractions per day even seemed to be better. The effects of actinomycin D administered during the irradiation treatment show that schedules with three fractions per day possibly take better advantage of repair of sublethal and potentially lethal damage than schedules with one fraction per day. Administration of misonidazole during the irradiation treatment led to a dose-modifying effect of 1.2 in one fraction per day schedules but had no significant effect in three fractions per day schedules. The present results may provide guidelines for clinical application of irradiation schedules with more than one fraction per day.

Adenocarcinoma↗

Calculation of the biological effect of fractionated radiotherapy: the importance of radiation-induced apoptosis.

The total effect (TE) has been calculated for two different fractionation formalisms: the consecutive and repetitive fractionation mechanism, using a modified linear quadratic (LQ) model which includes the effect of apoptosis. For a given total dose, an increase in TE is seen when increasing the dose per fraction as well as the apoptotic fraction (Fa). Also, the TE increases with increasing alpha/beta ratio (of the modified LQ model). The ratio of TE for tumour tissue and TE for late reacting tissue is calculated assuming the absence of apoptosis in late reacting tissue and a common value of alpha/beta (of the modified LQ model). The biological effect ratio (BR) is higher for a large Fa and low doses per fraction, than for large doses per fraction and a small Fa. Assuming a consecutive fractionation mechanism, the TE formalism is unable to predict a log cell kill of more than 3 for beta values of 0.010-0.028. It is less dependent on dose per fraction and Fa than the repetitive fractionation mechanism. The biological effect ratio is only slightly higher than 1, and is less influenced by Fa, dose per fraction and alpha/beta ratio. A repetitive fractionation mechanism is also consistent with the preliminary results of published fractionation experiments. The calculations indicate that designing fractionation regimes for optimization of biological effect is a process where the role of apoptotic cell inactivation must be maximized, and where the influence of mitotic cell inactivation may be of less importance.

Apoptosis↗

Characteristics of DNA fractionated on benzoylated DEAE-cellulose.

Chromatography on BD-cellulose columns with a salt gradient and formamide separates cellular DNA into two fractions (fraction I eluted within the salt gradient, fraction II with formamide), the proportions of these two fractions (ca. 2:1) being similar for DNA from a number of eucaryotic organisms. Yeast DNA was chosen for a detailed study of the mode of fractionation. Several physicochemical parameters, binding to nitrocellulose filters, sensitivity towards nuclease S1, labelling properties in vivo, and hybridization properties of the two DNA fractions were compared. It was shown that both fractions are native DNA and that the fractionation does not depend on the size or the (G + C) content of the DNA. Fraction I DNA contains only a small portion of molecules having single-stranded ends. Fraction II DNA is a heterogeneous population, containing molecules with peculiar structural characteristics: (a) It contains DNA molecules with single-stranded ends and/or gaps sensitive to nuclease S1; labeling experiments suggested that these are molecules undergoing repair and replication. (b) Another portion of fraction II is molecules sensitive to nuclease S1 in regions which are not single-stranded. (c) A third portion is DNA which, after treatment with nuclease S1, is still strongly bound to the resin. Indications that the segregation may be due to the presence of specific DNA sequences comes from the above experiments and from the finding that fraction I DNA is enriched in ribosomal genes and fraction II DNA in tRNA genes.

Centrifugation, Density Gradient↗

The effects of fractionated gamma irradiation on induction of mammary carcinoma in normal and estrogen-treated rats.

The effects of dose fractionation on induction of mammary carcinoma were studied in normal and estrogen-treated female rats of the inbred WAG/Rij strain. Groups of 40 animals received total-body doses of 1 or 2 Gy of (137)Cs gamma radiation, administered in fractions of 2.5, 10 or 40 mGy with intervals of 12 h, or in fractions of 10 mGy with intervals of 2, 5 or 24 h. The irradiations were started at the age of 8 weeks. Estrogen treatment was accomplished by implantation of a pellet containing estrogen at the age of 6 weeks. All mammary tumors were resected and classified histologically as carcinoma or fibroadenoma. The age-specific incidence of mammary carcinoma was compared with that in control groups of unirradiated normal or estrogen-treated rats and was expressed as excess normalized risk, using lifetime statistical analysis with both parametric and nonparametric methods. The data were also compared to the results of single-dose experiments reported in previous papers. Fractionated irradiation increased the risk of mammary cancer in both normal and estrogen-treated rats compared to the corresponding unirradiated control group. The excess normalized risk per unit of total dose was approximately equal with or without estrogen treatment. Without estrogen treatment, the effects of the single-dose and fractionated irradiations were approximately equal. In estrogen-treated animals, however, single-dose irradiation was up to 15 times more carcinogenic than the fractionated exposures. This fractionation effect appeared to vanish for total doses below approximately 0.3 Gy. With estrogen treatment, the excess normalized risk was significantly higher for dose fractions of 40 mGy than for fractions of 10 mGy. The risk was also markedly higher for fractionation intervals of 2 or 5 h than for intervals of 12 or 24 h. The results of these experiments show that the effects of dose fractionation on the induction of mammary carcinoma may depend on hormonal status, the total dose delivered, the dose per fraction, and the fractionation interval.

Animals↗

Water fraction of lumbar vertebral bone marrow estimated from chemical shift misregistration on MR imaging: normal variations with age and sex.

OBJECTIVE: Our objective was to use the chemical-shift misregistration effect of MR imaging to measure the variation in the water fraction in vertebral bone marrow with respect to patient age and sex. SUBJECTS AND METHODS: We examined prospectively 211 subjects (5-84 years old; 108 male and 103 female subjects) who had no history of hematologic disorders. We obtained midsagittal proton density-weighted spin-echo images (2500/22 [TR/TE]) with a narrow sampling bandwidth (6250 Hz). Misregistered signals from lipid protons of 6-10 mm (upward shift in the readout direction) were obtained with the chemical-shift effect. By measuring the mean signal intensities of the water voxels and the water-plus-lipid voxels, we calculated a region of interest in each vertebral body for L1 to L3. The water fraction of bone marrow was then calculated as 100 x (signal intensity of water voxels/signal intensity of water-plus-lipid voxels). Individual water fractions were calculated from the average values for the water fractions of the L1-L3 bodies. RESULTS: In the male subjects, the water fraction showed a rapid decrease in the older patients in the 5- to 34-year-old group. (Mean water fractions for male patients who were 5-14, 15-24, and 25-34 years old were 75.2%, 69.0%, and 53.7%, respectively.) For male patients more than 25 years old, the water fraction remained almost constant (approximately 50%). In female subjects who were 5-44 years old, the water fraction exceeded 69% and remained almost constant. However, the water fraction rapidly decreased in women more than 45 years old. (Mean water fractions for female patients who were 35-44, 45-54, and 55-64 years old were 70.9%, 61.1%, and 49.7%, respectively.) A comparison between male subjects and female subjects showed that the water fraction for women who were 25-54 years old exceeded the water fraction for men in the same age group (25-34 years old, p < .0001; 35-44 years old, p < .001; 45-54 years old, p < .05). CONCLUSION: MR imaging showed that male and female subjects convert hematopoietic marrow to fatty marrow in the lumbar vertebral bodies in significantly different ways. The data obtained for the normal water fraction of bone marrow may prove useful for evaluating diseases that affect marrow.

Adolescent↗

Pharmacological activity of three fractions obtained by smoking cannabis through a water pipe.

In a collaborative work, six chemical fractions obtained from smoke of cannabis, prepared by the Department of Biological Chemistry, University of Athens, were assayed pharmacologically at the Department of Psychobiology, Escola Paulista de Medicina, Brazil. The chemical fractions, obtained through burning cannabis mixed with a special tobacco through a water pipe similar to that used by Greek smokers, were: fraction IS composed of water-soluble compounds, fraction IIS made up of water-insoluble substances that condensed at the top of the water phase due to their low solubility; and fraction IIIS, of water-insoluble highly volatile substances, corresponding to the part that reaches the human lungs. Fractions IB, IIB and IIIB were placebo fractions obtained from burning only the special tobacco, called "tubeki", which is mixed with cannabis by the Greek smokers. The six fractions were assayed pharmacologically, in a blind fashion, using the following methods: barbiturate sleeping time in mice, catatonia in mice, corneal reflex in rabbits (Gayer test), and spontaneous motor activity in mice. Fraction IS, and its placebo fraction IB, were inactive in all methods employed, revealing no water-soluble active substances in cannabis. Fraction IIS revealed to be the most active as it induced significant effects in all 4 methods used; its IIB placebo fraction was inactive. Fraction IIIS, composed of the only substances that reach the human lungs, was moderately active; it produced significant effects on barbiturate sleeping time of mice and on the corneal reflex of rabbits, but was inactive in inducing catatonia and in decreasing spontaneous motor activity of mice. These results indicate that devices such as the water pipe used by Greek smokers retain active principles from cannabis.

Animals↗

Fractionation of human blood lymphocytes on Helix pomatia A haemagglutinin coupled to sepharose beads.

Treatment of human blood lymphocytes with neuraminidase has previously been shown to uncover receptors for the A haemagglutinin of the snail Helix pomatia (HP). Neuraminidase-treated lymphocytes were now fractionated on columns charged with large Sepharose particles to which HP had been coupled covalently. HP-receptor negative (HP-) lymphocytes passed the columns while HP-receptor positive (HP+) lymphocytes were retained. The latter cells were eluted by addition of the competitive hapten N-acetyl-D-galactosamine (D-GalNAc). The total yield of cells recovered after fractional was 60-80% Surface marker studies indicated that there was no selective loss of any of the major lymphocyte subpopulations. The fraction that passed the columns (fraction I) consisted of approximately 10% of all lymphocytes. It contained approximately 1% HP+cells and approximately 3% of all lymphocytes forming rosettes which sheep erythrocytes (E+ cells) present before fractionation. 50-55% of the lymphocytes in this fraction had surface-bound immunoglobulin (SIg+ cells) and complement receptors (EAC+ cells). Of the SIg+ cells, approximately 60% were true B cells while the remaining 40% had IgG adsorbed to their surface. The majority of the B cells were recovered in this fraction. The lymphocytes of this fraction responded poorly to T-cell mitogen but had an enhanced K-cell activity to chicken erythrocytes. Elution of the cells retained on the column with 0.1 mg/ml D-GalNAc gave a fraction II, consisting of approximately 15% of all lymphocytes. This fraction had a mixed composition. The majority of the cells (approximately 45%) were recovered by subsequent elution with 1.0 mg/ml D-GalNAc. This fraction III was strongly enriched with HP+ and E+ cells (T cells). About 10% of the HP+ cells in this fraction were SIg+. However, on the majority of these cells this surface-bound immunoglobulin was probably externally absorbed IgG. These HP+-SIg" cells were also EAC+ and had Fc receptors, as shown by rosette formation with IgG-coated bovine erythrocytes. The lymphocytes of fraction III responded most strongly to T-cell mitogen while their K-cell activity was weak.

Cell Membrane↗

[Partial purification and use of Trypanosoma cruzi glycosidic fractions for Chagas disease diagnosis].

Protein and glycoprotein fractions are used for Chagas' disease diagnosis, but they are degraded by parasite endogenous proteases. Therefore protease inhibitors are used for their conservation increasing the antigen cost. The possibility of using protease-resistant glycosidic fractions could solve this problem allowing to obtain stable antigens, with a high sensitivity at a lower cost. This proposal is reinforced by the existence of anti-galactosyl antibodies against T. cruzi oligosaccharic fractions in sera from chagasic patients. The aim of this work was to obtain T. cruzi glycosidic fractions and to evaluate their possible use for Chagas' disease serologic diagnosis. Total protein and glycoproteins from the four stages of T. cruzi were obtained. The glycosidic fractions were obtained by exhaustive proteolysis with Proteinase K. Protein, glycoprotein and glycosidic profiles were analysed by SDS-PAGE followed by staining proteins (Coomassie/silver) or glycoproteins and glycosidic fractions (APABGP). Antigenicity of the different fractions was determined by Western Blot and luminography, using control hyperimmune serum anti-epimastigote and a "pool" of chagasic human sera. Finally, the capacity of the glycosidic fractions to discriminate chagasic and non-chagasic human sera and the sensitivity of the fractions respect to control serum, were determined by ELISA. Main findings were: a) there are peptides, glycopeptides and glycosidic fractions that are common and specific to parasite stages; b) there are more antigenic glycoproteins in epimastigotes and metacyclics than in trypomastigotes and amastigotes; c) there is a correlation between the glycosidic fraction-pattern and the host type; d) glycosidic fractions can be used as antigens to discriminate, by ELISA, chagasic and non chagasic patients, but show lower titers with respect to total proteic and glycoprotein antigens. It is concluded that it is possible to develop a diagnostic kit for Chagas' disease employing glycosidic fractions, eliminating the disadvantages of the current kits.

Animals↗

Coronary artery bypass grafting in patients with ejection fractions below forty percent. Early and late results in 466 patients.

The outcome of patients undergoing coronary artery bypass grafting with preoperative ejection fractions below 40% was evaluated to determine if a specific level of ventricular dysfunction resulted in unacceptably poor short-term or long-term survival rates. Left ventricular ejection fractions were segregated into groups of five percentage points each starting from 35% to 39% and progressing down to 10% to 14%. In evaluating the six ejection fraction groups between 10% and 39%, we found no significant differences among them with regard to previous myocardial infarctions, left ventricular end-diastolic pressure (LVEDP), age, preoperative New York Heart Association (NYHA) class, or number of vessels bypassed. Eighty-four percent were men and 16% women. From 1976 through 1982, 466 patients were distributed among these groups, all having ejection fractions below 40% (mean 30% +/- 3% SEM). There were significant differences (p = 0.001) in both the hospital and long-term survival (36 months) of patients with preoperative ejection fractions from 20% to 39% (425 patients) as compared to those with preoperative ejection fractions from 10% to 19% (41 patients). Hospital survival rate was 89% for patients with ejection fractions from 20% to 39% but only 63% for patients with ejection fractions below 20%. Similarly, at 3 years, patients with ejection fractions of 20% to 39% had an average survival rate of 60% as compared to an average survival rate of 15% for those with ejection fractions below 20%. Neither the preoperative LVEDP nor the intraoperative ischemic arrest time significantly predicted survival. In all survivors, NYHA class decreased from an average of 3.00 to 1.25 in surviving patients following bypass at a mean follow-up of 29 +/- 5 months. It is concluded that ejection fraction is an excellent predictor of short-term and long-term survival following coronary artery bypass grafting. Patients with ejection fractions of 10% to 19% have a significantly reduced short-term and long-term survival rate as compared to patients with ejection fractions of 20% or more.

Angina Pectoris↗

Fractionation and characterization of rat hepatocytes isolated from ethanol-induced fatty liver.

A procedure for fractionation of hepatocytes according to density is presented. Hepatocytes were prepared from rats fed an ethanol-containing liquid diet, a liquid diet without ethanol, or a chow diet. The hepatocytes were fractionated in a Metrizamide density gradient and the cell fractions obtained were characterized with respect to protein synthesis, fatty acid oxidation, and esterification and ATP content. From ethanol-fed rats, two fractions of hepatocytes were obtained, one with a density of approximately 1.04 gm. per ml. and one with a density of ca. 1.08 gm. per ml. Cells from the lighter fraction contained 3 times more triacylglycerol than cells from the heavier fraction. Fifty per cent of the cells from the lighter fraction contained large (greater than 5 micron) lipid droplets as compared to 6% in the heavier fraction. The average yield from a 170-gm. rat of lipid-rich and lipid-poor cells was about 65 x 10(6) and 170 x 10(6) cells, respectively. Electron micrographs of the liver before perfusion and of the isolated and washed cell fraction revealed intact ultrastructure. Fractionation of hepatocyte suspensions from rats fed a high fat liquid diet without ethanol or a chow diet normally yielded two cell fractions with densities of ca. 1.08 and 1.10 gm. per ml. and 1.10 and 1.12 gm. per ml., respectively. The metabolic capacity of the cell fractions was unaltered by the fractionation procedure as judged by the ATP content, the rate of palmitate oxidation and esterification, and the rate of protein synthesis.

Adenosine Triphosphate↗

Acute effects of external and internal pacing on ejection fraction in patients with severe cardiac failure.

The present study aims at evaluating the acute hemodynamic effects of external and internal (3 and 15 mA) pacing on 30 patients with severe cardiac failure. The global and regional ejection fraction were calculated by repeated radionuclide ventriculography at rest (control) and during increasing pacing rates. Blood pressure was measured at the end of each scintigraphy data acquisition period. The pacing rate was increased by 10 beats/min in every step and varied between +10 to +60 beats/min above the resting heart rate. The ejection fraction during pacing was lower than the resting ejection fraction in every pacing mode (resting ejection fraction = 23.30 +/- 7.67%, external pacing = 19.36 +/- 9.30%, p < 0.05, internal pacing 3 mA = 22.15 +/- 7.00%, p = N.S., internal pacing 15 mA = 19.92 +/- 6.95%, p < 0.05). The resting ejection fraction was higher than the ejection fraction in every pacing rate. In 4 out of 30 patients the ejection fraction was higher in every pacing mode and rate than the resting ejection fraction. The regional ejection fraction of the interventricular septum during internal pacing was lower (p < 0.001) than resting and/or external pacing. In conclusion, pacing in patients with severe cardiac failure reduces the control ejection fraction. This reduction increases as the pacing rate increases. The reduction is greater with internal pacing of 15 mA while the hemodynamic effect in external pacing did not differ significantly from internal pacing. Overall, the resting ejection fraction was higher than the paced ejection fraction, although in some patients pacing improved their resting ejection fraction.

Adult↗

Coupling gravitational and flow field-flow fractionation, and size-distribution analysis of whole yeast cells.

This work continues the project on field-flow fractionation characterisation of whole wine-making yeast cells reported in previous papers. When yeast cells are fractionated by gravitational field-flow fractionation and cell sizing of the collected fractions is achieved by the electrosensing zone technique (Coulter counter), it is shown that yeast cell retention depends on differences between physical indexes of yeast cells other than size. Scanning electron microscopy on collected fractions actually shows co-elution of yeast cells of different size and shape. Otherwise, the observed agreement between the particle size distribution analysis obtained by means of the Coulter counter and by flow field-flow fractionation, which employs a second mobile phase flow as applied field instead of Earth's gravity, indicates that yeast cell density can play a major role in the gravitational field-flow fractionation retention mechanism of yeast cells, in which flow field-flow fractionation retention is independent of particle density. Flow field-flow fractionation is then coupled off-line to gravitational field-flow fractionation for more accurate characterisation of the doubly-fractionated cells. Coupling gravitational and flow field-flow fractionation eventually furnishes more information on the multipolydispersity indexes of yeast cells, in particular on their shape and density polydispersity.

Cell Fractionation↗

Carcinogenic oil fractionation and nuclear enlargement in mouse skin.

A non-solvent-refined oil, previously shown to produce tumours in a long-term skin painting assay and a positive result in the mouse skin nuclear enlargement test, was separated using silica gel elution techniques into eight fractions. Fraction 1, which contained saturated and 1-2 ring aromatic hydrocarbons, was used to dilute the other fractions to produce solutions equivalent to and twice their original concentration in the intact oil. Fraction 1, diluted fractions 2-8, the reconstituted oil, the original oil and a negative control oil were examined for their ability to produce nuclear enlargement. The degree of nuclear enlargement observed with fraction 1 was considered to indicate marginal or no carcinogenic activity. With the fraction that contained 2-3 ring aromatic compounds, weak carcinogenic activity could not be ruled out but it is unlikely that these components played a major role in the carcinogenicity of the oil. The greatest activity was identified in the fraction containing the total 3/4-6 ring polycyclic aromatic hydrocarbons (PAHs) and further fractionation of this suggested that the parent 4-6 ring PAHs were responsible for more than half the activity of the intact oil, despite being present at only 0.06%. The fraction that contained polar compounds showed no activity. The reconstituted oil showed slightly less activity than the original oil. Other fractions, which contained alkylated PAHs, did not seem to have much activity. These findings, which are in close agreement with published evidence on tumour induction by fractions of oils and coal liquids, suggest that the components responsible for inducing nuclear enlargement are the same as those responsible for skin tumour induction.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Differential in vitro degradation of particular Fos family members expressed by kainic acid in nuclear and cytosolic fractions of murine hippocampus.

Mice were injected with kainic acid (KA) at a convulsive dose, followed by homogenization of the hippocampus in the presence of different protease and phosphatase inhibitors, and subsequent preparation of nuclear and cytosolic fractions. An intraperitoneal injection of KA resulted in marked expression of particular Fos family members, including c-Fos, Fra-2, and Fos-B, but not Fra-1 proteins, in both fractions 2 to 18 h after administration. These fractions were individually incubated at 30 degrees C for 1 to 18 h for determination of in vitro degradation. Similarly rapid degradation was seen with c-Fos protein between nuclear fractions obtained 2 and 18 h after administration, while no significant degradation was found for c-Fos protein in cytosolic fractions obtained 2 h after administration during incubation. By contrast, in vitro incubation led to rapid degradation of c-Fos protein in cytosolic fractions obtained 18 h after administration. Degradation profiles were peculiar to each member protein in nuclear and cytosolic fractions obtained 2 and 18 h after administration. Dialysis prevented degradation of c-Fos protein in nuclear fractions without markedly affecting that in cytosolic fractions in a manner independent of the time after administration. The addition of inhibitors for phosphatases, but not for proteases, accelerated the degradation of c-Fos protein in nuclear fractions previously dialyzed. These results suggest that in vivo KA signals may modulate heterologous machineries responsible for breakdown of each Fos family member in a unique manner in nuclear fractions, rather than cytosolic fractions, of murine hippocampus.

Animals↗

Kinetics of radioiodinated species in subcellular fractions from rat hearts following administration of iodine-123-labelled 15-(p-iodophenyl)-3-(R,S)-methylpentadecanoic acid (123I-BMIPP).

It is recognized that iodine-123-labelled 15-(p-iodophenyl)-3-(R,S)-methylpentadecanoic acid (123I-BMIPP) slowly washes out of the myocardium. The mechanism for the washout was investigated in normal rat hearts by analyses of the subcellular distribution and lipid classes based on the BMIPP metabolism. Rat hearts were excised at 1-120 min after intravenous injection of 123I-BMIPP. After counting the radioactivity, the hearts were digested with Nagarse and homogenized, and then fractionated into the cytosolic, mitochondrial, microsomal and crude nuclear fractions by centrifugations. The radioactivity of each fraction was counted, and the lipid classes were analysed by radio-thin-layer chromatographic and high-performance liquid chromatographic methods. The heart uptake of 123I-BMIPP was maximal at 5 min (6.81%+/-0.36% ID/g), and 41% of the radioactivity disappeared within 120 min. The myocardial radioactivity was immediately distributed into the cytosolic, mitochondrial, microsomal and crude nuclear fractions. The distribution (%) of each fraction was almost identical from 5 min through 120 min. The cytosolic fraction was always the major site of radioactivity deposition (60%), and the time-activity curve of the cytosolic fraction paralleled that of the whole heart throughout the 120-min study period. In the cytosolic fraction, most of the radioactivity was incorporated into the triglyceride class, and the rest was present in the free fatty acid, phospholipid (phosphatidylcholine) and diglyceride classes. In the mitochondrial fraction, the radioactivity was mostly incorporated into the phospholipid class (phosphatidylethanolamine), followed by free fatty acids. The final metabolite of 123I-BMIPP, 123I-p-iodophenylacetic acid (123I-PIPA), initially appeared in the mitochondrial fraction as early as 1 min, and subsequently in the cytosolic fraction at 5 min. Another intermediary metabolite, 123I-p-iodophenyldodecanoic acid (123I-PIPC12), was found only in the mitochondrial fraction after 5 min. In conclusion, the slow washout kinetics of 123I-BMIPP from the myocardium mainly reflects the turnover rate of the triglyceride pool in the cytosol. The BMIPP metabolism, i.e. initial alpha-oxidation followed by subsequent cycles of beta-oxidation, was confirmed in vivo. The participation of the mitochondria in the metabolism was also proven.

Animals↗

Comparison of the response of human FaDu squamous cell carcinoma in nude mice after hypofractionated-accelerated regimens and "curative" fractionation schedules.

BACKGROUND: It has been suggested that tumors respond differently after irradiation with 10 or more fractions than with less fractionated regimens and that extrapolation from experiments with only a few fractions to "curative" regimens may be invalid. To test this hypothesis, we compared hypofractionated-accelerated treatments with "curative" fractionation schedules in human squamous cell carcinoma in nude mice. MATERIAL AND METHODS: FaDu tumors were transplanted subcutaneously into the hindleg of NMRI nu/nu mice. TCD50 values, i.e., the dose necessary to control 50% of the tumors locally, were determined after irradiation under ambient blood flow conditions with 5 and 10 fractions in 5 days, 10 fractions in 10 days, and 30 fractions in 15 days, 6 weeks or 10 weeks. RESULTS: TCD50 values of the hypofractionated regimens were not significantly different and varied from 41 to 46 Gy. The number of fractions given in the same overall time had no measurable effect on local tumor control. The TCD50 after 30 fractions in 6 weeks was 30 Gy higher than after the hypofractionated regimens. This effect was caused by a substantial increase of TCD50 with overall treatment time, the dose recovered per day was 0.82 Gy (95% CI 0.66; 0.96). alpha/beta eff determined from all data was 58 Gy (13; infinite). CONCLUSIONS: The results of the present study compare well with our previous findings after different "curative" fractionation schedules in the same tumor. Thus, our study does not support that tumors respond differently after application of 10 or more fractions compared to less fractionated regimens. The biological mechanisms that govern the radiation response of FaDu tumors appear to be the same for hypofractionated-accelerated and "curative" regimens. Since only one tumor was investigated, these results cannot be generalized at the present time.

Animals↗

Association of Chernobyl-derived 239+240Pu, 241Am, 90Sr and 137Cs with different molecular size fractions of organic matter in the soil solution of two grassland soils.

Radiocesium is normally bound only rather weakly and unspecifically by humic substances, in contrast to the actinides Pu and Am. Recently, however, it was observed that fallout 137Cs in the soil solution from an Of-horizon of a podzol forest soil (slightly decomposed plant material) was associated essentially only with one single size fraction of the humic substances. In deeper soil layers with well humified material (AOh-horizon), radiocesium was associated with all size fractions of the dissolved organic matter (DOM). To examine whether this unexpected behaviour is also observable for DOM isolated from other soils, we determined the association of fallout 137Cs, 90Sr, 238Pu, 239+240Pu and 241Am with various size fractions of DOM from in situ soil solutions isolated from two layers (0-2 cm and 2-5 cm) of two grassland soils (a soddy podzolic soil and a peat soil) within the 10 km zone of the nuclear reactor at Chernobyl (Ukraine). The four size fractions of DOM as obtained by gel filtration of the soil solution were (mean nominal molecular weight in daltons): fraction I: > or = 2000, fraction II: 1300; fraction III: 560, fraction IV: inorganic compounds. The results for the well humified DOM (humus accumulation horizon of podzol, deeper layer of peat soil) showed that Pu and Am are essentially associated with the high molecular weight fractions, while Sr is present only in the 'inorganic' fraction. Radiocesium is found in all the size fractions separated. A quite similar pattern was also found for Pu, Am, and Sr in the soil solution from only slightly decomposed plant material (0-2 cm of peat soil), but not for radiocesium. This radionuclide was again essentially only observable in one single low molecular weight fraction of DOM. The above results thus support our recent observations in the different horizons of a forest podzol mentioned above, even though no reason for the different binding of radiocesium by well humified soil organic matter and by only slightly decomposed plant material can be given at present. The data demonstrate, however, that information on only the total amount of a radionuclide in the soil solution will not be sufficient to interpret or predict its fate adequately in the soil.

Americium↗