Oncogenic transformation by fractionated doses of neutrons

Oncogenic transformation was assayed after C3H 10T1/2 cells were irradiated with monoenergetic neutrons; cells were exposed to 0.23-, 0.35-, 0.45-, 5.9-, and 13.7-MeV neutrons given singly or in five equal fractions over 8 h. At the biologically effective neutron energy of 0.45 MeV, enhancement of transformation was evident with some small fractionated doses (below 1 Gy). When transformation was examined as a function of neutron energy at 0.5 Gy, enhancement was seen for cells exposed to three of the five energies (0.35, 0.45, and 5.9 MeV). Enhancement was greatest for cells irradiated with 5.9-MeV neutrons. Of the neutron energies examined, 5.9-MeV neutrons had the lowest dose-averaged lineal energy and linear energy transfer. This suggests that enhancement of transformation by fractionated low doses of neutrons may be radiation-quality dependent.     

Relationship of endogenous abscisic Acid to sucrose level and seed growth rate of soybeans

It has been proposed that abscisic acid (ABA) may stimulate sucrose transport into filling seeds of legumes, potentially regulating seed growth rate. The objective of this study was to determine whether the rate of dry matter accumulation in seeds of soybeans (Glycine max L.) is correlated with the endogenous levels of ABA and sucrose in those sinks. The levels of ABA and sucrose in seed tissues were compared in nine diverse Plant Introduction lines having seed growth rates ranging from 2.5 to 10.0 milligrams dry weight per seed per day. At 14 days after anthesis (DAA), seeds of all genotypes contained less than 2 micrograms of ABA per gram fresh weight. Levels of ABA increased rapidly, however, reaching maxima at 20 to 30 DAA, depending upon tissue type and genotype. ABA accumulated first in seed coats and then in embryos, and ABA maxima were higher in seed coats (8 to 20 micrograms per gram fresh weight) than in embryos (4 to 9 micrograms per gram fresh weight. From 30 to 50 DAA, ABA levels in both tissues decreased to less than 2 micrograms per gram fresh weight. Levels of sucrose were also low early in development, less than 10 milligrams per gram fresh weight at 14 DAA. However, by 30 DAA, sucrose levels in seed coats had increased to 20 milligrams per gram fresh weight and remained fairly constant for the remainder of the filling period. In contrast, sucrose accumulated in embryos throughout the filling period, reaching levels greater than 40 milligrams per gram fresh weight by 50 DAA. Correlation analyses indicated that the level of ABA in seed coats and embryos was not directly correlated to the level of sucrose measured in those tissues or to the rate of seed dry matter accumulation during the linear filling period. Rather, the ubiquitous pattern of ABA accumulation early in development appeared to coincide with water uptake and the rapid expansion of cotyledons occurring at that time. Whole tissue sucrose levels in embryos and seed coats, as well as sucrose levels in the embryo apoplast, were generally not correlated with the rate of dry matter accumulation. Thus, it appears that, in this set of diverse soybean genotypes, seed growth rate was not limited by endogenous concentrations of ABA or sucrose in reproductive tissues.     

Sedimentary records of accelerated nutrient loading in Florida lakes

Transfer functions relating trophic state (Carlson's TSI_{chlorophyll-a}) to present day accumulation rate of (1) nutrients, (2) cations, and (3) organic sediment, are computed using Binford's ²¹⁰Pb-dilution method. As computed from surficial sediments of 27 lakes, former trophic states are reconstructed for recent (²¹⁰Pb-dated) sedimentary histories of 14 lakes. Of the three kinds of models potentially available, model (3) (TSI vs. organic accumulation) is the weakest statistically, and may be unduly influenced by exceptional deposition and/or preservation of allochthonous organic matter. At present, however, it is the only model applicable to all 14 of the histories tested. Results are encouraging in that model accurately predicts observed TSI's in several mesotrophic and eutrophic lakes. Clearly significant increases (accelerations) are inferred only for a of the most eutrophic lakes of the set, while the model consistently overpredicts TSI's of the 7 most oligotrophic lakes. As Whitmore's diatom-assemblage index is a better predictor of TSI than is model (3) in the one eutrophic lake in which it has been tested, we expect more persuasive results when models (1) and (2) can be tested within a more complete set of analytical data. We were surprised to find 3 severely disturbed lakes among the 12 that show little or no acceleration in rate of eutrophication in recent decades, but we defer attempts at explanation until former nutrient loading can be tested by model (1).     

Sarcomeric domain organization within single skinned rabbit psoas fibers and its effects on laser light diffraction patterns.

Total intensity and fine structure of first-order laser light diffraction maxima from single skinned rabbit psoas fibers were studied. Total intensity of the diffraction maxima was measured as a function of the incidence angle (omega-scan). In the most homogeneous fibers, most of the intensity in the diffraction maxima is confined to a rather narrow range of incidence angles. Fibers with less homogeneous striation patterns, apparently composed of several regions of distinct sarcomere length and tilt of striation (domains), give rise to several narrow intensity peaks in their omega-scans. Left and right first-order diffraction lines produce omega-scans of almost identical shape, composed of one or more intensity peaks, with each pair of corresponding peaks separated by about the same angle. The data indicated that in single skinned rabbit psoas fibers, light diffraction is dominated by Bragg diffraction and that the peaks within omega-scans can be directly correlated with domains within the illuminated fiber segment. In the most homogeneous fiber segments the diameter of domains, estimated from the width of the corresponding maxima in the omega-scans, could almost be as large as the fiber diameter. On average, from the number of peaks in the omega-scans two to three domains with an average length of approximately 250-350 microns can be identified in a fiber cross-section. Therefore, on average only a small number of domains (8 per mm) are found within skinned rabbit psoas fiber segments. In contrast, the number of substructural lines within the diffraction maxima is large even for microscopically homogeneous fibers. Substructural lines appear to be present only when several domains are illuminated simultaneously. Separation and width of these substructural lines are approximately inversely proportional to the length of the illuminated region of the fiber. These data suggest that the substructural lines are due to interference between domains, illuminated simultaneously by a light source with a high degree of spatial coherence (laser). The relevance of these findings for measurements of sarcomere length by laser light diffraction is discussed.     

Effect of insertions, deletions, and double-strand breaks on homologous recombination in mouse L cells.

We have used DNA-mediated gene transfer to study homologous recombination in cultured mammalian cells. A family of plasmids with insertion and deletion mutations in the coding region of the herpes simplex type 1 thymidine kinase (tk) gene served as substrates for DNA-mediated gene transfer into mouse Ltk- cells by the calcium phosphate technique. Intermolecular recombination events were scored by the number of colonies in hypoxanthine-aminopterin-thymidine selective medium. We used supercoiled plasmids containing tk gene fragments to demonstrate that an overlap of 62 base pairs (bp) of homologous DNA was sufficient for intermolecular recombination. Addition of 598 bp of flanking homology separated from the region of recombination by a double-strand gap, deletion, or insertion of heterologous DNA increased the frequency of recombination by 300-, 20-, or 40-fold, respectively. Linearizing one of the mutant plasmids in a pair before cotransfer by cutting in the area of homology flanking a deletion of 104 bp or an insertion of less than 24 bp increased the frequency of recombination relative to that with uncut plasmids. However, cutting an insertion mutant of greater than or equal to 24 bp in the same manner did not increase the frequency. We show how our data are consistent with models that postulate at least two phases in the recombination process: homologous pairing and heteroduplex formation.     

X-ray diffraction evidence for cross-bridge formation in relaxed muscle fibers at various ionic strengths

Equatorial x-ray diffraction patterns from single skinned rabbit psoas fibers were studied at various ionic strengths to obtain structural information regarding cross-bridge formation in relaxed muscle fibers. At ionic strengths between 20 and 50 mM, the intensity of the 11 reflection, I11, of the relaxed state was close to that of the rigor state, whereas the intensity of the 10 reflection, I10, was approximately twice that of rigor reflection. Calculations by two-dimensional Fourier synthesis indicated that substantial extra mass was associated with the thin filaments under these conditions. With increasing ionic strength between 20 and 100 mM, I10 increased and I11 decreased in an approximately linear way, indicating net transfer of mass away from the thin filaments towards the thick filaments. These results provided evidence that cross-bridges were formed in a relaxed fiber at low ionic strengths, and that the number of cross-bridges decreased as ionic strength was raised. Above mu = 100 mM, I10 and I11 both decreased, indicating the onset of increasing disorder within the filament lattice.