Improved methods of estimating root length using a photocopier,a light box and a bar code reader

Photocopying was found to be a rapid method of making a permanent record of a root sample. The method used produced a copy with white roots against a black background. Manual estimates of root length were made from photocopies using a light box. The number of intersections visible when laid over a copy of a white on black regular square grid was counted. Automated estimates of root length were made by scanning a photocopy with a bar code reader in place of a pen in a computer-driven graph plotter. Roots >0.2 mm diameter were resolved with precision and speed.     

Bypassing prefertilization barriers to hybridization in Nicotiana using in vitro pollination and fertilization

Summary In vitro pollination of placenta attached ovules was useful in bypassing unilateral incongruity barriers for several Nicotiana interspecific hybrid combinations (N. tabacum cv. Ky 17 X N. amplexicaulis, Ky 17 X N. benthamiana, and Ky 17 X N. repanda). By measuring the pollen tube growth over time, prefertilization barriers were determined to be the cause of the incongruity. Seedling necrosis was a problem in the development of the N. amplexicaulis hybrid and it prevented maturation of the N. repanda hybrid. Callus produced from cotyledons of the N. amplexicaulis hybrid eventually resulted in plants that survived to maturity. This procedure was not successful for the N. repanda materials. The N. amplexicaulis and N. benthamiana hybrids were sterile but following chromosome doubling by midrib culture, male and female fertile plants were produced.Conventional hybridization, fertilized ovule culture, and in vitro pollination were unsuccessful in obtaining hybrids of Ky 17 crossed with N. arentsii or N. bonariensis. Apparently, strong postfertilization barriers prevent the production of viable seed of these hybrids. Each of the N. repanda — N. tabacum reciprocal hybrids could not be rescued using callus culture; this adds support to the existence of strong sexual postfertilization barriers. A recent report, however, showed that it was possible to obtain this hybrid using the technique of somatic hybridization. Thus, it appears that it may also be possible to obtain asexual hybrids of N. arentsii and N. bonariensis with N. tabacum.The investigations reported herein were supported by USDA/SEA/CRGO Project 59-2213-1-1-613-0 and the paper (No. 86-3-137) is published with approval of the Director of the Kentucky Agricultural Experiment StationThe research reported in this paper is in partial fulfillment of the Ph.D. requirements for the senior author     

M241 (CD1) expression on B lymphocytes

The human thymus leukemia-like antigens (CD1a-c) consist of three similar glycoproteins found on subpopulations of normal thymocytes, T cell acute leukemias, and cutaneous dendritic cells. The CD1c antigen recognized by the M241 monoclonal antibody was detected on the circulating mononuclear cells of three children with severe combined immunodeficiency disease (SCID). Two-color immunofluorescence analysis demonstrated that M241 expression (43 to 95%) was limited to cells expressing the B cell-restricted antigens B4 (CD19), B1 (CD20), and surface immunoglobulin. To confirm M241 expression on normal cells of the B lineage rather than aberrant expression limited to SCID B cells, its expression was demonstrated serologically and biochemically on purified B cells from spleen, tonsil, and peripheral blood. Parallel analyses with monoclonal antibodies NA1/34 and 4A76 demonstrated that the CD1a and CD1b molecules were negative on all B cells that were studied. It has been hypothesized that the CD1 molecules represent the human counterpart of the murine thymus leukemia antigens due to their similar size, limited tissue distribution, and association with beta 2-microglobulin. This study suggests that a subset of CD1 antigens detected by M241 (CD1c) may represent a human analog of a murine Qa antigen due to its extended distribution on normal peripheral B cells.     

Formation of organic products in self-radiolyzed calcium carbonate

Summary Calcium carbonate labeled with carbon-14 was self-irradiated by means of the -decay of its carbon-14. A number of products containing one or two carbon atoms were identified by high performance liquid chromatography. Formic and oxalic acids were produced in relatively high yields, while glyoxylic, glycolic, and acetic acids, as well as formaldehyde and methanol, were formed in lower yields. These results support the suggestion that carbonates subjected to ionizing radiation could have been a source of carbon for organic synthesis on the primitive earth.     

Calcium and contractions of isolated smooth muscle cells from rat myometrium

Smooth muscle cells were isolated from estrogenized rat myometrium by collagenase digestion. Electron microscopic examination and measurement of cell lengths by image-splitting micrometry were carried out after fixation with acrolein. Mean lengths of cells before and after isolation were 81.7 and 66.9 micron, respectively. Responses of cells were compared with contractions of isolated strips recorded isometrically. Effects of carbachol and KCl were examined in 2 mM Ca, 2 mM Ca + 4 mM EGTA, and 2 mM Ca + 10(-8) M nitrendipine solution. Carbachol and KCl produced concentration-dependent shortening of isolated cells maximal at 30 s after addition. The concentrations of carbachol required to produce shortenings were about 100-fold less than those required to produce isometric contractions; but no major difference was observed in the concentration dependence of cell shortening and isometric contraction produced by potassium-induced depolarization. In 2 mM Ca solution, there was a phasic response, followed by a tonic response such that more than 50% of maximum cell shortening was maintained for 10 min. However, in 2 mM Ca + 4 mM EGTA or 10(-8) M nitrendipine, the tonic contraction was abolished and cells rapidly relaxed after 30 s. If carbachol was added to cells after varying times in the EGTA-containing solution, the ability to initiate a contraction declined exponentially with a half-time of 160 s. Effects of depolarization by KCl were examined in 2 mM Ca plus nitrendipine and 2 mM Ca + 4 mM EGTA solution. Shortening occurred in 2 mM Ca solution by depolarization but not if nitrendipine was added. Though shortening was not observed in 2 mM Ca + 4 mM EGTA solution by KCl, subsequent addition of carbachol induced shortening. These results suggested that there was an intracellular Ca store site from which Ca was released by carbachol and which was not affected by depolarization in the absence of external Ca. No evidence was obtained that the contraction persists in Ca2+-free medium in isolated cells, which is in agreement with previous findings in small muscle strips in which only a similar transient response was obtained.     

Cellular responses to ionizing radiation: effects of interrupting DNA repair with chemical agents

This review is concerned with the influence of different classes of chemical agents on cellular repair of DNA damage induced by ionizing radiation. Single-strand break rejoining is little affected by inhibitors of DNA synthesis; however, such inhibitors do lead to a persistence of double-strand breaks in the DNA, and this correlates with an enhancement of chromosome aberrations and cell killing. Experiments with antagonists of topoisomerase II suggest an intriguing role for this DNA unwinding enzyme in double-strand break repair. Interference with poly(ADP-ribose) synthesis, by means of the inhibitor 3-aminobenzamide, does not have a clear-cut effect on recovery from ionizing radiation damage. Various substances (for example, caffeine and trypsin) affect DNA repair via a modulation of the cell cycle, altering the time available to the cell for repairing potentially lethal DNA damage before such damage is 'fixed' by the process of DNA replication. Finally, disturbing cellular energy metabolism, and depressing the level of ATP, can inhibit the repair of radiation damage.     

The human glutathione S-transferase. Studies on the kinetic, stability and inhibition characteristics of the erythrocyte enzyme

Bromosulphophthalein and N-ethylmaleimide, inhibitors of glutathione S-transferase (EC 2.5.1.18 RX: glutathione R. transferase), have been used to identify variant forms of the erythrocyte enzyme. One 'atypical' sample was detected and was shown to have appreciably different kinetic and stability properties. These inhibitors may be useful in surveys of variation in this group of enzymes.     

A ten-month study of endogenous testosterone levels and behaviour in outdoor-living female rhesus monkeys (Macaca mulatta)

Endogenous testosterone levels were measured in association with sexual, aggressive, and social/affiliative behaviors in 11 outdoor-housed female rhesus monkeys over a ten-month period. Several behaviors (sex directed toward the male, sex received from the male, aggression directed toward the male, submission directed toward the male, submission directed toward the female, and groom another female) were significantly (p<0.05) positively correlated with testosterone in from one to five females. No trends were strong enough across all females to suggest that any of these correlations have species-wide significance. Factor analysis revealed clearcut clusters of behaviors, but elevations in testosterone were not strongly associated with any of these clusters. It is concluded that endogenous testosterone levels have little measurable effect on overt behavior in female rhesus monkeys.     

Sequence similarities between chicken intestinal 110-kDa ATPase and myosin I-like enzymes

We report the partial amino acid sequence of chicken intestinal microvillar 110-kDa protein that, as a complex with calmodulin, has previously been shown to exhibit myosin-like ATPase and actin-binding activities. The sequence shows a high degree of similarity to the sequence of a novel vertebrate myosin I-like heavy chain encoded by a cDNA isolated from bovine intestine. This confirms that the bovine and chicken proteins are the first examples of Acanthamoeba myosin I-like proteins from higher eukaryotes. Comparison of available structural and functional data leads us to postulate that the myosin I family of proteins result from the fusion of a conserved myosin headlike motor domain, with variable COOH-terminal domains responsible for binding to specific intracellular structures.