Age-correlated DNA damage in human muscle tissue.

This investigation represents the largest study so far published on human DNA damage and aging. The subject of this investigation is damage, determined as DNA alterations which give rise to complete molecular breaks in the course of treatment of purified DNA solutions with single-strand-specific nucleases. The DNA is derived from milligram samples of human muscle of individuals mostly undergoing surgical treatment. Care has been taken to bring the muscle samples, once shut off from blood circulation to liquid nitrogen temperatures within few seconds. The DNA is prepared by a procedure keeping breaks by handling and by DNAase attack as low as possible, however pushing DNA purity, especially with respect to protein as high as possible. Highly purified DNA treated in this way has some sites which are susceptible to single-strand (ss) specific DNAase splitting (ss-events). Three different deoxyribonucleases have been used: Nuclease S1, Nuclease BAL31 and Pea Endo-Nuclease. They give very similar results, i.e. splitting of the DNA so as to yield DNA pieces of given distribution. The lengths of these double-strand (ds) pieces have been determined from their electron microscopical pictures, either by following the image contours with a magnetostrictive stylo of the projected photo on a pad, by following the contours with a mileage ruler, or by integrating the silver grains on the photo. The molecular weight averages of the ds DNA threads between two ss-events for each individual have been determined from 20 to 200 molecules. The 470 individuals contributing their data were from age groups from 1 to 91 years. The molecular weights show a considerable scatter with an average molecular weight of the DNA ds pieces between two ss-events of 43.93 MDa and a standard deviation of 17.99 MDa. Among the single-strand breaks (ssb) that split the DNA into such pieces is a fraction, the number of which increases in a highly significant fashion with the age of the donor. From this derives the fact that the average molecular weight of the DNA strand pieces between two ssb decreases with age. It is remarkable that the standard deviation of the molecular weights of such pieces increases with age significantly, too. On the basis of additional information mainly supplied by the DNA donor himself or by his parents the 470 members of the main group M where grouped according to their life-style, into: (1) abstinent people, essentially non-smokers and refraining from use of licit or illicit drugs, sub-group N.(ABSTRACT TRUNCATED AT 400 WORDS)

The effect of benzo[a]pyrene on sponges as model organisms in marine pollution.

The majority of the investigations were performed with the marine sponge Tethya lyncurium at concentrations of 2 X 10(-8) to 1 X 10(-11) g/ml of benzo[a]pyrene (BaP). Sea-pollution was characterized as BaP equivalent activity in the Ames test. Increased activity of ornithine decarboxylase (ODC) was observed when sponges were artificially exposed at polluted marine areas for 3 weeks. In contrast to the situation in higher animals no ODC induction of the fast type was observed. Mixed function oxygenases (MFO) were not detected in sponges nor could they be induced as in vertebrates. BaP was absorbed by Tethya and concentrated 30--60-fold. In live, but not dead, artificially perfused sponges [3H]- and [14C]BaP-radiolabeled became firmly associated with DNA, RNA and protein of the sponges. The association persisted in isolated fractions, in nucleotides, in nucleosides and in protein hydrolysates. The BaP binding ratio to DNA was found to be strongly correlated to the concentration of BaP. Light modifies BaP and thus enables binding. In the dark only very low association, if any, is observed. The possible consequences of these findings are discussed.

Electron microscopical characterization of sponge aggregation factors.

Aggregation factors, purified from 14 sponges which belong to the classes Tetraxonida and Cornacuspongia, Were visualized electron microscopically. Two types of basic structural forms were detected; first, circular structures from the species Ancorina cerebrum, Mycale massa, Hemimycale columella, Crella rosea, Clathria coralloides, Axinella cannabina, Pellina semitubulosa, Ircinia muscarum, Hippospongia communis, Verongia aerophoba and second, rod-like structures from Tethya lyncurium, Tedania anhelans, Hymeniacidon sanguinea, Dysidea tupha. In most of the cases the structures carry side chains.

Species-specific aggregation factor in sponges. XIII. Entire and core structure of the large circular proteid particle from Geodia cydonium.

High-molecular weight particles have been isolated from the sponge Geodica cydonium. In the "native" from these particles consist of a spherical center and have 25-30 filaments attached to it. The core structure of the particles is assembled of a central circle and 25 radially-arranged filaments. The core structure is obtained from the entire structure by incubation in a medium, containing a non-ionic detergent and EDTA. The molecular weight of the enitre structure was in the range of 1.4 X 10(9) daltons or more and of the core structure 6.1 x 10(8) daltons. Two functional proteins are released from the "native" particles: the aggregation factor and the sialytransferase.