Flow Cytometric Quantification of All Phases of the Cell Cycle and Apoptosis in a Two-Color Fluorescence Plot

An optimal technology for cell cycle analysis would allow the concomitant measurement of apoptosis, G0, G1, S, G2 and M phases in combination with cell surface phenotyping. We have developed an easy method in flow cytometry allowing this discrimination in an only two-color fluorescent plot. It is based on the concomitant use of 7-amino-actinomycin D and the antibodies anti-Ki67 and anti-phospho(Ser10)-histone H3, both conjugated to Alexa Fluor®488 to discriminate G0 and M phases, respectively. The method is particularly valuable in a clinical setting as verified in our laboratory by analyzing human leukemic cells from marrow samples or after exposure to cell cycle modifiers.     

Plant virus infection development as affected by heavy metal stress

The work was focused on the investigation of possible dependencies between the development of viral infection in plants and the presence of high heavy metal concentrations in soil. Field experiments have been conducted in order to study the development of systemic tobacco mosaic virus (TMV) infection in Lycopersicon esculentum L. cv. Miliana plants under effect of separate salts of heavy metals Cu, Zn and Pb deposited in soil. As it is shown, simultaneous effect of viral infection and heavy metals in tenfold maximum permissible concentration leads to decrease of total chlorophyll content in experiment plants mainly due to the degradation of chlorophyll a. The reduction of chlorophyll concentration under the combined influence of both stress factors was more serious comparing to the separate effect of every single factor. Plants' treatment with toxic concentrations of lead and zinc leaded to slight delay in the development of systemic TMV infection together with more than twofold increase of virus content in plants that may be an evidence of synergism between these heavy metal's and virus' effects. Contrary, copper although decreased total chlorophyll content but showed protective properties and significantly reduced amount of virus in plants.     

The Genographic Project public participation mitochondrial DNA database

The Genographic Project is studying the genetic signatures of ancient human migrations and creating an open-source research database. It allows members of the public to participate in a real-time anthropological genetics study by submitting personal samples for analysis and donating the genetic results to the database. We report our experience from the first 18 months of public participation in the Genographic Project, during which we have created the largest standardized human mitochondrial DNA (mtDNA) database ever collected, comprising 78,590 genotypes. Here, we detail our genotyping and quality assurance protocols including direct sequencing of the mtDNA HVS-I, genotyping of 22 coding-region SNPs, and a series of computational quality checks based on phylogenetic principles. This database is very informative with respect to mtDNA phylogeny and mutational dynamics, and its size allows us to develop a nearest neighbor-based methodology for mtDNA haplogroup prediction based on HVS-I motifs that is superior to classic rule-based approaches. We make available to the scientific community and general public two new resources: a periodically updated database comprising all data donated by participants, and the nearest neighbor haplogroup prediction tool.     

Biodiversity in eutrophicated shallow lakes: determination of tipping points and tools for monitoring

Nutrient-rich freshwater ecosystems are generally considered as having low ecological quality and low associated biodiversity. In such systems we analysed the effects of water quality on biodiversity of several species groups, to determine tipping points and tools for monitoring. We investigated the water quality of 99 eutrophic and hypertrophic shallow lakes with extensive fish culture during a 3-year study, through the measures of physico-chemical parameters, phytoplankton biomass and structure. In a second step, we related the water quality with richness of aquatic plants, macroinvertebrates and dragonflies. With concentrations of chlorophyll-a above 30 or 70 μg l^?1, shallow lakes are normally classified, respectively, in a poor or bad ecological state. However, our results show that chlorophyll-a concentrations up to 78 μg l^?1 could be found together with relatively high species or family richness of aquatic plants, invertebrates and dragonflies. We identified most tipping points with 50–60 μg l^?1 of chlorophyll-a, values above which a significant decrease of species diversity was found. For monitoring of these shallow lakes we propose to use chlorophyll-a concentrations in combination with water transparency during spring. These parameters are easily applicable and cheap and they yield a good forecast of the biodiversity for the species groups studied.