Microsatellite DNA in peach (Prunus persica L. Batsch) and its use in fingerprinting and testing the genetic origin of cultivars.

We isolated and sequenced 26 microsatellites from two genomic libraries of peach cultivar 'Redhaven', enriched for AC/GT and AG/CT repeats, respectively. For 17 of these microsatellites, it was possible to demonstrate Mendelian inheritance. Microsatellite polymorphism was assayed in 50 peach and nectarine cultivars. Of the 1300 PCRs carried out, all but two produced amplified products of the expected size. All microsatellites were polymorphic, showing 2-8 alleles per locus. Heterozygosity ranged from 0.04-0.74 (mean 0.47); the discrimination power (PD) ranged from 0.04-0.84 (mean 0.60). Cultivar heterozygosity varied greatly, with one cultivar ('Independence') being homozygous at all loci. The set of microsatellites discriminated all cultivars investigated, except several sport mutations, i.e., 'Dixitime' vs. 'Springcrest', 'Compact Redhaven' vs. 'Redhaven', and two pairs of cultivars, 'Venus' vs. 'Orion' and 'Elegant Lady' vs. 'Rome Star', whose pedigrees are controversial. We were able to analyze the paternity of several cultivars. In most cases, the parenthood was confirmed. The comparison of three long-living 'Redhaven' accessions supplied by different repositories did not provide any evidence of somatic instability of microsatellites. Hence, microsatellites, ranked according to their information content, are recommended as markers of choice for peach fingerprinting and suggestions are provided for interpreting band profiles and the correct sizing of alleles.     

The biological effects in animals related to the accident at the Chernobyl Atomic Electric Power Station. 1. The experimental model. The radiation loads on animals kept continuously under external and internal radiation exposure in the area of the Chernobyl Atomic Electric Power Station]

Irradiation conditions in which laboratory animals were kept in experimental laboratories of Chernobyl and Kiev after the accident at the Chernobyl A.P.S. are described. The data are presented on the spectral structural and activity of radionuclides in the diet as well as in the organs and tissues of the animals. The radiation loads have been estimated with regard to an external gamma component and the internal one contributed by the incorporated radionuclides. It has been shown that radiation doses received by the animals during their lifetime due to these contributions do not exceed units of cGy.     

The biological action of low-level radiation

On the basis of their own data and the literature the authors have analyzed the mechanism of biological action of low-level radiation on cells. In studying the harmful effect of gamma-radiation (10-40 cGy) on HeLa cells it was shown that damages occurred in 12 postirradiation generations. Analysis of cell distribution within the colonies has demonstrated that the share of colonies with low cell content and the number of giant cell increase. These data indicate that low-level radiation causes reproductive cell death that can occur not immediately but in later generations.     

Topochemistry of blood cells of the Gallus domesticus (Aves, Galliforme).

Blood smears of both male and female chicken Gallus domesticus were analysed by using the following topochemical methods: a) Periodic acid-Schiff (PAS) for glycogen. b) Mercury-bromophenol blue for protein. c) O-Toluidine for myeloperoxidase. d) Sudan black B for lipid. The PAS reaction revealed glycogen in the cytoplasm of all thrombocytes and in a few heterophils. The presence of proteins was evidenced in all types of cells. However variation in the intensity of staining of protein granules was observed in the fusiform structures of the heterophils. A negative reaction for myeloperoxidase was found in all cells. Although some evidence of myeloperoxidase activity was show in the polymorphonuclears it was not enough to ascertain a positive reaction. Lipids were detected in the cytoplasm of few heterophils, eosinophils and monocytes.     

The influence of alkyl pyridinium sponge toxins on membrane properties, cytotoxicity, transfection and protein expression in mammalian cells

The ability of two alkyl pyridinium sponge toxin preparations (poly-APS and halitoxin) to form transient pores/lesions in cell membranes and allow transfection of plasmid cDNA have been investigated using HEK 293 cells. Poly-APS and halitoxin preparations caused a collapse in membrane potential, reductions in input resistance and increased Ca²⁺ permeability. At least partial recovery was observed after poly-APS application but recovery was more rarely seen with halitoxin. The transfection with plasmid cDNAs for an enhanced green fluorescent protein (EGFP) and human tumour necrosis factor receptor 2 (TNFR2) was assessed for both toxin preparations and compared with lipofectamine. Stable transfection was achieved with poly-APS although it was less efficient than lipofectamine. These results show that viable cells transfected with alien cDNA can be obtained using novel transient pore-forming alkyl pyridinium sponge toxins and a simple pre-incubation protocol. This provides the first proof of principle that pore-forming alkyl pyridinium compounds can be used to deliver cDNA to the intracellular environment without permanently compromising the plasma membrane.     

In vivo protein synthesis from 14C-substrates in porcine tissues during the transition to postnatal development

[2-14C] leucine, [1-14C] alanine, [1-14C] glucose, [1-14C] lactate and [1-14C] pyruvate utilization in the protein synthesis has been studied in vivo at early stages of postnatal development of piglets. It has been established, that during the first 24 hours after birth the protein synthesis intensity, judging by [2-14C] leucine incorporation, in liver, skeletal muscle, duodenal wall and subcutaneous tissue of piglets increases 5, 7, 6.5 and 2.1 times respectively. At the age of 1-2 h the radioactive carbon incorporation from [1-14C] glucose into the brain proteins is more pronounced than into the proteins of liver and skeletal muscle. During the first days of life the intensity of the label incorporation from [1-14C] glucose into liver and skeletal muscle proteins of piglets is enhanced, whereas in brain it remains at the same level. The degree of 14C carbon incorporation from [1-14C]-alanine, [1-14C] pyruvate and [1-14C] lactate into the liver and skeletal muscle proteins of 5-days-old piglets is approximately the same, 14C substrates of protein synthesis in brain and subcutaneous adipose tissue having some peculiarities.     

A non-invasive micronucleus assay in the rat liver

The potent rat-liver mitogen 4-acetylaminofluorene (4AAF) is shown here to provide an effective replacement for the surgical procedure of 2/3 partial hepatectomy (2/3PH) in the in vivo rat-liver micronucleus assay described by Tates and his colleagues. This protocol modification enables the assay to be conducted on a routine basis. Control observations for both 2/3PH and 4AAF-treated rats are presented, together with evidence indicating 4AAF itself to be without activity in the assay, irrespective of the mitogenic stimulus. The activities of the rat carcinogens DMN, 2AAF, DMH and 6BT, and of the non-carcinogens 4AAF and 4N are demonstrated. Recommendations for the conduct of the modified assay are made.