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.     

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 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.     

Interrelationship of kernel water activity,soil temperature,maturity, and phytoalexin production in preharvest aflatoxin contamination of drought-stressed peanuts

Samples of Florunner peanuts were collected throughout a period of late-season drought stress with mean geocarposphere temperatures of 29 and 25 °C, and determinations of maturity, kernel water activity (a_w), percent moisture, capacity for phytoalexin production, and aflatoxin contamination were made. Results showed an association between the loss of the capacity of kernels to produce phytoalexins and the appearance of aflatoxin contamination. Kernel a_w appeared to be the most important factor controlling the capacity of kernels to produce phytoalexins. Mature peanuts possessed additional resistance to contamination that could not be attributed solely to phytoalexin production. Kernel moisture loss was accelerated in the 29 °C treatment compared to the 25 °C treatment, and data indicated that the higher soil temperature also favored growth and aflatoxin production by Aspergillus flavus in peanuts susceptible to contamination.Mention of a trademark or proprietary product does not constitute a guarantee or warranty of the product by the U.S. Department of Agriculture and does not imply its approval to the exclusion of other products that may also be suitable.     

Higher order structure is present in the yeast nucleus: autoantibody probes demonstrate that the nucleolus lies opposite the spindle pole body

A panel of sera from 892 autoimmune patients was screened by indirect immunofluorescence on mammalian cells. Seventy-three sera were identified that recognize the nucleolus. Three of these sera appear to stain the nucleolus in yeast, suggesting that they recognize highly conserved antigens. These three sera also immunoprecipitate mammalian U3 snRNA-containing particles, which reside in the nucleolus and have been implicated in rRNA processing. Double immunofluorescence experiments with anti-nucleolus and anti-tubulin antibodies revealed a novel form of non-random nuclear organization in yeast. The spindle pole body and the nucleolus — both of which are associated with the nuclear envelope — preferentially localize at opposite ends of the nucleus. Organization of these and other components into specific regions of the nucleus may be important for optimizing their proper function.     

Urinary elastase 1 in chronic pancreatic disease

Serum and urine elastase 1, its renal output and clearance and urinary gamma-glutamyltransferase and ribonuclease excretions were measured in 16 patients with pancreatic cancer, 23 with chronic pancreatitis and in 22 healthy controls in order to evaluate elastase 1 plasma-urine transfer in chronic pancreatic disease and to investigate any factors that might influence the clearance of this enzyme. In an additional group of 17 patients with different pancreatic diseases the serum molecular size distribution of elastase 1 after chromatography was ascertained. An increased urinary elastase 1 output was found in 4/16 patients with pancreatic cancer and in 6/23 with chronic pancreatitis. No correlation was found between circulating elastase 1 and its urinary output; a negative correlation was detected between the serum levels of this enzyme and its clearance. The excretion of ribonuclease and gamma-glutamyltransferase was correlated with elastase 1 output and clearance. While the majority of elastase 1 in serum was accounted for by high molecular forms, probably the expression of complexes with serum inhibitors, free circulating enzyme was present in all patients with high serum elastase 1. Our findings suggest that elastase 1 urinary excretion increases in some patients with chronic pancreatic disease regardless of the neoplastic or inflammatory nature of the illness. Although the availability of different amounts of ultrafiltrable enzyme may play a role in influencing elastase 1 plasma-urine transfer, renal tubular damage appears to be the most important factor influencing the increase in the urinary output of elastase 1.