DNA polymerase epsilon: the latest member in the family of mammalian DNA polymerases

DNA polymerase epsilon is a mammalian polymerase that has a tightly associated 3'----5' exonuclease activity. Because of this readily detectable exonuclease activity, the enzyme has been regarded as a form of DNA polymerase delta, an enzyme which, together with DNA polymerase alpha, is in all probability required for the replication of chromosomal DNA. Recently, it was discovered that DNA polymerase epsilon is both catalytically and structurally distinct from DNA polymerase delta. The most striking difference between the two DNA polymerases is that processive DNA synthesis by DNA polymerase delta is dependent on proliferating cell nuclear antigen (PCNA), a replication factor, while DNA polymerase epsilon is inherently processive. DNA polymerase epsilon is required at least for the repair synthesis of UV-damaged DNA. DNA polymerases are highly conserved in eukaryotic cells. Mammalian DNA polymerases alpha, delta and epsilon are counterparts of yeast DNA polymerases I, III and II, respectively. Like DNA polymerases I and III, DNA polymerase II is also essential for the viability of cells, which suggests that DNA polymerase II (and epsilon) may play a role in DNA replication.     

Vitamin E Status of Healthy Swedish Cattle*

Using high pressure liquid chromatography the serum concentration of vitamin E was measured in dairy cows fed either hay or silage as their main roughage, in calves fed milk-replacer, and in young intensively fed bulls. The concentrates fed to the cows, calves and bulls were supplemented with 5–10, 25 and 5–10 mg DL-α-tocopheryl acetate per kg, respectively, and the milk-replacer for the calves was supplemented with 50 mg DL-α-tooopheryl acetate per kg powder. Cows fed silage as their main roughage had higher serum vitamin E concentrations ( $${\rm{\bar x}}$$ : 3.8–5.2 mg/l) than cows fed only hay ( $${\rm{\bar x}}$$ : 2.5–4.1 mg/1). Lactating cows had higher vitamin E concentrations than dry cows ( $${\rm{\bar x}}$$ : 4.1–5.2 and 2.5–3.8 mg/l, respectively) and calves and bulls had much lower vitamin E concentrations ( $${\rm{\bar x}}$$ : 1.4 and 1.2 mg/l, respectively) than cows. Thirty per cent of the calves and 41 % of the bulls had serum vitamin E concentrations less than 1.0 mg/l, suggesting that in these animals the conventional level of supplementation of feeds with DL-α-tocopheryl acetate in Sweden is probably inadequate for the prevention of nutritional muscular degeneration and other negative effects.     

Strain while skiing and hauling a sledge or carrying a backpack

Eight soldiers on skis transported three loads of different weights on the level, uphill and downhill. The load was placed either on a cargo sledge or in a backpack or divided between the sledge and the backpack. The sledge had a new type of haulage-shaft, which was fixed to both sides of the pelvis. A service belt spread the pull over the whole upper body. The physical stain of different transport methods and the serviceability of the sledge was studied by measuring heart rate (HR), oxygen consumption, ventilation, and perceived exertion. The results indicate that both absolute and relative strain were systematically lower when pulling the load on the sledge than when carrying it in the backpack and on the sledge. HR when pulling a load equal to the human body weight on the sledge was on average 133 beats X min-1; HR was significantly higher 144 beats X min-1 when the load was divided between backpack and sledge. At the lower load level the differences between the transport methods were not significant for HR, oxygen consumption or ventilation. Uphill travel increased oxygen consumption by about 50% over that on the level. Perceived exertion at all load levels was significantly lower with the sledge than with the backpack alone or in combination. The estimated maximal allowable working time emphasized the advantage of the sledge and the importance of high physical working capacity. The maneuverability of the sledge with the new haulage shaft was good and the braking mechanisms worked well.(ABSTRACT TRUNCATED AT 250 WORDS)     

Chloroplast DNA variation inAnemone s. lato (Ranunculaceae)

Chloroplast DNA of seven species belonging toAnemone (sectt.Omalocarpus, Anemonidium, andAnemonanthea),Hepatica, andPulsatilla have been analyzed by restriction enzymes. According to the dendrogram constructed, the sections ofAnemone and the generaHepatica andPulsatilla seem to be evolutionary approximately equidistant to each others. This supports the concept that these groups should be treated on a similar taxonomic level, either as genera or subgenera.     

Significance of Ca2+ and K+ in Micrasterias Growth and Morphogenesis

Significance of Ca²⁺ and K⁺ for the complex morphogenesis ofMicrasterias, which takes place through multipolar tip growth,was investigated. Studies with different external Ca²⁺ concentrationsand Ca²⁺ channel inhibitors LaCl₃ and verapamil indicate thatCa²⁺ and Ca²⁺ channels are essential in the development, whiletreatments with different K⁺ concentrations and K⁺ channel inhibitorTEA demonstrate that potassium or K⁺ channels are not neededin the process, albeit the existence of K⁺ channels. K⁺ is notneeded even for the regulation of turgor pressure, which wasfound to decrease clearly during cell development. The plasmamembrane ATPase inhibitors diethylstilbesterol (DES) and Na-orthovanadatestop morphogenesis and indicate the importance of ion pumpsin the developmental process. Both supraoptimal, external K⁺and Ca²⁺ cause abundant Ca²⁺ precipitate formation in chloroplasts,which shows that chloroplasts are important in regulation ofcytoplasmic Ca²⁺ metabolism and that K⁺ activates the uptakeof Ca²⁺ through Ca²⁺ channels. (Received June 13, 1995; Accepted September 13, 1996)     

The effects of continuous illumination on southern and northern Micrasterias strains

Different strains of Micrasterias (Chlorophyta, Conjugatophyceae); M. rotata (Grev.) Ralfs ex. Ralfs and M. denticulata Breb. ex. Ralfs var. angulosa (Hantzsch) W. & G. S. West from northern and southern Finland were treated with continuous illumination in order to study the cellular effects of the treatment and whether the tolerance to continuous light of the northern Finnish strains is related to the different daylenght conditions in northern and southern areas. During the growing season the Finnish strains normally live in long-day conditions or even in continuous light (between 60 and 70°N), and they also tolerated continuous illumination in the laboratory. Ultrastructural changes were found especially in the chloroplasts, where formation of calcium precipitates of different forms and sizes and also formation of plastoglobuli containing lipids appeared. However, even in 4-week treatments the ultrastructure of cells of these northern strains was not totally disrupted, contrary to what was found in southern M. torreyi , studied earlier. Southern and northern strains tolerated continuous illumination in different ways. They seem to differ from each other physiologically, and the differences are possibly located in their ionic metabolism and regulation. The injuries sustained during continuous illumination of Micrasterias may largely be caused by the accumulation of Ca²⁺ in cytoplasm and organelles, especially in the chloroplasts.     

Serum Glutathione Peroxidase Activity and Blood Selenium in Pigs

Blood serum glutathione peroxidase activity and blood selenium concentration were measured in blood samples from pigs subjected to experimentally induced selenium deficiency and dietary selenium supplementation on graded levels. A highly significant correlation between blood selenium and serum GSH-Px activity in pigs, especially in selenium deficient pigs, was demonstrated. There was also a strong relationship between blood selenium concentration and serum GSH-Px activity in pigs receiving dietary selenium at graded levels. Serum GSH-Px activity exhibited an excellent close-response relationship to dietary selenium. Linear regression analysis showed that the increased serum GSH-Px activity was a function of the dietary selenium concentration. The fitness of serum in monitoring slight changes of the selenium status of pigs with help of the estimation of GSH-Px activity was discussed. The measurement of serum GSH-Px activity seems to provide a useful and rapid means for defining selenium requirements and for identifying selenium deficiency in growing pigs.     

Combined Therapeutic Effect of Dietary Selenium and Vitamin Ε on Manifested Vesd Syndrome in Weaned Pigs

By using a therapeutic dietary supplementation in pigs, which had developed the vitamin Ε and selenium deficiency (VESD) syndrome, the same amounts of α-tocopheryl acetate and selenium were found to be effective as under prophylactic conditions. The experiment thus supported the conclusions that the addition of 5 mg DL-α-tocopheryl acetate/kg and 135 μg selenium/kg to a diet, which contained only traces of vitamin Ε and selenium, represents a level of minimal requirement. Glutathione peroxidase activity in blood serum was used to evaluate the selenium status in pigs. A modified method for determination of tocopherol in fat tissue was described. The addition of 15 mg α-tocopheryl acetate/kg diet was demonstrated to be sufficient to maintain the tocopherol stores in body fat at an unchanged level.     

Replacement of natural polyamines by cadaverine and its aminopropyl derivatives in Ehrlich ascites carcinoma cells

Ehrlich ascites carcinoma cells were cultured in the presence of difluoromethyl ornithine (DFMO) and micromolar concentrations of cadaverine for several months. This treatment resulted in a complete disappearance of putrescine and spermidine and reduced spermine content to traces of its normal content. The natural polyamines were replaced by cadaverine (about 40% of total polyamines), N-(3-aminopropyl)cadaverine (about 50%) and N,N′-bis(3-aminopropyl)cadaverine (about 5%). In comparison with untreated cells or cells grown in the presence of DFMO and putrescine, the “cadaverine cells” grew definitely slower, their protein synthesis was depressed while DNA and RNA syntheses proceeded at near normal rate. In spite of the high intracellular concentrations of cadaverine and its aminopropyl derivatives, the tumor cells grown in the presence of DFMO and cadaverine, behaved exactly like cells severly depleted of putrescine and spermidine. Though exposed to DFMO, ornithine decarboxylase activity was almost 10 times higher than that in untreated cells. S-Adenosyl-L-methionine decarboxylase activity was likewise strikingly elevated, and these cells transported methylglyoxal strikingly elevated, and these cells transported methylglyoxal bis(guanylhydrazone) (MGBG) at a rate that was more than 5 times faster than that in untreated cells. Furthermore, these cells exhibited arginase activity, which was less than one fifth of that found in untreated cells.