Thermal inhibition of repair of methylmethane sulfonate-damaged DNA in chick embryo fibroblasts

Chick embryo fibroblasts were treated with the monofunctional alkylating agent methylmethane sulfonate at various concentrations for 1 h at 42°C, rinsed and then incubated post-treatment at various temperatures at which the kinetics of alkali-labile bond disappearance was followed. Growth experiments showed that these cells grew similarly at temperatures of either 37°C or 42°C. Repair as assessed by removal of alkali-labile bond was also similar for postincubation in the temperature range 37–42°C for damage due to methylmethane sulfonate treatment at concentrations less than 1.5 mM. When the postincubation temperature was raised higher than 42.5–43°C, this type of repair was stopped. The normal internal body temperature of adult chickens is about 41.6°C. Hence the present finding indicates that chick cells are much more severely restricted in DNA repair at temperatures above normal than are mammalian cells, which can function in this respect for several deg. C above 37°C.     

Cell-wall lysing enzymes and products of cell-wall digestion elicit ethylene in citrus

Ethylene production was induced in Valencia oranges [ Citrus sinensis (L.) Osbcck] by injection of the fungal enzyme mixture Pectolyase ( Aspergillus japonicus ) which contains pectolytic enzymes into the peel. The mixture also stimulated production of 1-aminocyclopropane-1-carboxylic acid (ACC). Cycloheximide partially inhibited the Pectolyase-induced ethylene response. Pectin fragments, resulting from partial acid hydrolysis or Pectolyase digestion, caused an increase in ethylene production when injected into the peel of intact orange fruits. Pectic fragments produced by fungal enzymes are known to be elicitors of phytoalexins and in this study are shown to elicit ethylene in citurs.     

Repair of benzo[a]pyrene-initiated DNA damage in human cells requires activation of DNA polymerase alpha

Normal human fibroblasts treated with r-7,t-8-dihydroxy-t-9,10-epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene (BPDE) yielded DNA polymerase alpha with elevated levels of activity, incorporated [3H]thymidine as a function of unscheduled DNA synthesis, and exhibited restoration of normal DNA-strand length as a function of unscheduled DNA synthesis. Lipoprotein-deficient fibroblasts treated with BPDE did not show elevated levels of DNA polymerase alpha activity, exhibited minimal [3H]thymidine incorporation, and had fragmented DNA after 24 h of repair in the absence of lipoprotein or phosphatidylinositol supplementation. When DNA polymerase beta activity was inhibited, cells with normal lipoprotein uptake exhibited [3H]thymidine incorporation into BPDE-damaged DNA but did not show an increase in DNA-strand length. DNA polymerase alpha activity and [3H]thymidine incorporation in lipoprotein-deficient fibroblasts increased to normal levels when the cells were permeabilized and low-density lipoproteins or phosphatidylinositol were introduced into the cells. DNA polymerase alpha isolated from normal human fibroblasts, but not from lipoprotein-deficient fibroblasts, showed increased specific activity after the cells were treated with BPDE. When BPDE-treated lipoprotein-deficient fibroblasts were permeabilized and 32P-ATP was introduced into the cells along with lipoproteins, 32P-labeled DNA polymerase alpha with significantly increased specific activity was isolated from the cells. These data suggest that treatment of human fibroblasts with BPDE initiates unscheduled DNA synthesis, as a function of DNA excision repair, which is correlated with increased activity of DNA polymerase alpha, and that increased DNA polymerase alpha activity may be correlated with phosphorylation of the enzyme in a reaction that is stimulated by low-density lipoprotein or by the lipoprotein component, phosphatidylinositol.     

The distribution of zinc and copper in plasma, erythrocytes and erythrocyte membranes of rainbow trout (Salmo gairdneri)

1. The zinc and copper concentration of plasma was determined in rainbow trout, lake trout, walleye and whitefish. 2. These fish had mean plasma zinc concentrations ranging from 9.3 to 15.1 ppm and copper concentrations from 0.6 to 1.3 ppm. 3. In rainbow trout, the concentration of zinc and copper is greater in the erythrocyte membrane than in the total erythrocyte. 4. Ultrafilterable plasma zinc and copper concentration in rainbow trout was determined to be 0.03 and 0.019 ppm, respectively. 5. Dialysis of rainbow trout plasma against 20 mM EDTA results in removal of 99% of the zinc and 88% of the copper from plasma proteins.     

The effects of ultraviolet-B radiation on loblolly pine. I. Growth, photosynthesis and pigment production in greenhouse-grown seedlings

One-year old loblolly pine ( Pinus taeda L.) seedlings were grown in an unshaded greenhouse for 7 months under 4 levels of ultraviolet-B (UV-B) radiation simulating stratospheric ozone reductions of 16, 25 and 40% and included a control with no UV-B radiation. Periodic measurements were made of growth and gas exchange characteristics and needle chlorophyll and UV-B-absorbing-compound concentrations. The effectiveness of UV-B radiation on seedling growth and physiology varied with the UV-B irradiance level. Seedlings receiving the lowest supplemental UV-B irradiance showed reductions in growth and photosynthetic capacity after only 1 month of irradiation. These reductions persisted and resulted in lower biomass production, while no increases in UV-B-absorbing compounds in needles were observed. Seedlings receiving UV-B radiation which simulated a 25% stratospheric ozone reduction showed an increase in UV-B-absorbing-compound concentrations after 6 months, which paralleled a recovery in photosynthesis and growth after an initial decrease in these characteristics. The seedlings grown at the highest UV-B irradiance (40% stratospheric ozone reduction) showed a more rapid increase in the concentration of UV-B-absorbing compounds and no effects of UV-B radiation on growth or photosynthetic capacity until after 4 months at this irradiance. Changes in photosynthetic capacity were probably the result of direct effects on light-dependent processes, since no effects were observed on either needle chlorophyll concentrations or stomatal conductance. Further studies are necessary to determine whether these responses persist and accumulate over subsequent years.