Activation of fatty acids by non-glyoxysomal peroxisomes

Ethylene treatment (approx. 20 l ·1^-1 in air for 2 d) of tobacco (Nicotiana tabacum L. cv. Havana 425) plants markedly increases the endo--1,3-glucanase (EC 3.2.1.39) content of leaves. The antigenic form of the enzyme induced is the same one whose production is blocked by treating cultured cells with combinations of auxin (1.1 · 10^-5 M -naphthaleneacetic acid) and cytokinin (1.4 · 10^-6 M kinetin). Evidence is presented that cultured tobacco cells require ethylene for -1,3-glucanase accumulation: i) ethylene treatment increased the accumulation of \-1,3-glucanase in callus tissues >10 d after subculturing and in cell-suspension cultures; ii) callus tissues can produce ethylene; iii) conditions known to inhibit ethylene production (1 mM CoCl₂; 33° C treatment) or ethylene action (approx. 1.6 mmol · 1^-1 norbornadiene in air) inhibited -1,3-glucanase accumulation by callus tissues treated for 4 d following subculturing; and, these inhibitory effects were prevented by exogenous ethylene. Combinations of auxin and cytokinin blocked ethylene-induced accumulation of -1,3-glucanase by cell-suspension cultures. The results favor a model in which ethylene induces results favor a model in which ethylene induces 1,3-glucanase accumulation, and auxin and cytokinin inhibit this induction process.Abbreviations NAA -naphthaleneacetic acid - NDE norbornadiene     

A homologous radioimmunoassay for guinea pig corticosteroid-binding globulin

A rapid, specific, and sensitive (requiring only 20 fmole of antigen equivalent to 0.007 microliter of serum) radioimmunoassay (RIA) was developed for the measurement of guinea pig corticosteroid-binding globulin (CBG). CBG was purified to homogeneity from guinea pig serum by affinity chromatography and used for immunization, as the standard and as the radiolabeled trace in the RIA. The antiserum to CBG was raised in rabbits. It was judged specific by immunoelectrophoresis and by comparison of RIA values with steroid-binding assay profiles obtained on serum separated on the basis of size and ion-exchange properties. The results of the radioimmunoassays agree with those of a steroid-binding assay run on identical samples. The sensitivity of the assay allows detection of CBG in serial serum samples, other biologic fluids such as milk, and cell culture supernatants.     

Ice Encasement Injury to Microsomal Membranes from Winter Wheat Crowns : I. Comparison of Membrane Properties after Lethal Ice Encasement and during a Post-Thaw Period

The functional and physical properties of cellular membranes isolated from Triticum aestivum, cvs Norstar and Fredrick, were altered coincident with changes in composition after a lethal ice-encasement stress and further during a 6 hour post-thaw period. Crowns encased in ice for a duration which inhibited regrowth, exhibited enhanced rates of electrolyte leakage. Furthermore, the recovery of total microsomal protein and phospholipid declined, suggesting that some membrane degradation had been induced during the anoxic stress. The microviscosity of microsomes and liposomes prepared from such membranes increased during stress, and this was correlated with a 2- to 4-fold increase in the free fatty acid levels in the microsomal fraction. There was, however, only a relatively minor change in fatty acid unsaturation during the ice-encasement stress. The process continued during a 6 hour aerobic post-thaw treatment, but the pattern was somewhat different. During this phase, the leakage of electrolytes was further increased and the recovery of microsomal protein and phospholipid continued to decline, indicating general degradation; but, in contrast to the anoxic phase, the degree of fatty acid unsaturation declined markedly, indicating lipid peroxidation.     

Further refinements on the triangular flap closure of the cleft lip

Refinements in the triangular flap closure of the unilateral cleft lip are presented. Randall's mathematical interpretation of the Tennison repair has been extended by using a series of arcs to determine the crucial points which form the triangular flap. Using this easily taught method, an isosceles triangular flap can be plotted which will interdigitate into the noncleft side of the lip. Two symmetrical vertical distances on either side of the cleft are thus formed. This allows for a standardized repair which may be more readily taught than the rotation-advancement technique. In addition, flaps are created which are turned medically toward the cleft and are used to accentuate the philtral pout, close the nostril floor, and reinforce the lip repair, allowing closure of even extremely wide clefts in one stage. These refinements in using the triangular flap closure for the repair of the unilateral cleft lip are diagrammatically presented. We believe that these refinements enhance the results of this closure.     

The effects of ionizing radiation on the pulmonary vasculature of intact rats and isolated pulmonary endothelium

We studied the effects of ionizing radiation on the morphology of the pulmonary circulation using an in vivo rat model and an in vitro pulmonary artery endothelial cell model. Gamma radiation was given as either an acute (30 Gy) or fractionated (5 X 6 Gy) dose to one hemithorax of rats. An acute 30-Gy dose delivered resulted in a 70% decrease in pulmonary arterial perfusion, using technetium-99m microaggregated albumin (99mTc-MAA), in the irradiated lung by 2-3 weeks after irradiation. Pulmonary microradiographs, using a barium sulfate perfusion method, obtained 2-3 weeks after irradiation demonstrated widespread loss of capillary filling and segmentation of the vessels. Histologic examination demonstrated intact capillaries, suggesting that the alterations in pulmonary perfusion were at the precapillary level. Similar abnormalities in lung perfusion and morphology were found after delivery of fractionated doses of radiation, but the onset of the changes was delayed, occurring 4-6 weeks postirradiation. Using cultured pulmonary endothelial cell monolayers, cell sloughing and retraction from the surface substrate were observed within 24 h after in vitro delivery of 30 Gy. Similar findings occurred in monolayers given fractionated doses (5 X 6 Gy) of radiation 2-3 days after the final dose. The in vivo animal and in vitro endothelial cell models offer a useful means of examining the morphologic alterations involved in radiation lung vascular damage.     

Relative biological effectiveness (RBE) of neutrons produced by 50 MeV deuterons and by 34, 45, 65 and 75 MeV protons

RBE of p(34) + Be, p(45) + Be, p(65), + Be, p(75) + Be and d(50) + Be neutron beams produced at the cyclotron "Cyclone" of Louvain-la-Neuve were measured. The biological criterion was the regeneration of the crypts of the intestinal mucosa (50 regenerated crypts per circumference) after abdominal irradiation in mice. Taking the p(65) + Be neutrons as reference, RBE values were found equal to 1.12, 1.07, 1.00 (Ref.), 0.96 and 1.02 respectively. These results are consistent with those published for cell lethality in vitro. However, the RBE variation is smaller than this previously obtained in the laboratory for growth inhibition in Vicia faba.     

The use of slotted grids for electron microscopic radioautography

A method for electron microscopic radioautography on slotted grids is presented which allows examination of the distribution of silver grains over sections of entire structural units without interference by grid bars. Tissue sections of a size such as to fit the opening of the grid slot are placed on slides coated with a Formvar film of sufficient strength to permit transfer of the completed radioautograph onto the grid and to support it over the slot. Sections are block stained prior to radioautography to minimize the risk of loss of the radioautograph during the procedure.     

Characteristics of the ultrastructural organization of the nuclei of somatic cell hybrids

A study was made of the ultrastructure of cell nuclei of two types of hybrid clones obtained from the fusion of Chinese hamster with human skin fibroblasts, and from that of mouse hepatoma cells with mink fibroblasts. In cell nuclei of the eight hybrid clones deep invaginations of the inner membrane, not characteristic of the parent cells, were revealed. Analysis of serial sections, and application of electron microscopic radioautography and histochemistry have suggested that these structures are associated with the nuclear envelope which is necessary for regulating the superfluous chromosome localization in the hybrid nucleus.