Genetic diagnostics of hereditary breast and ovarian cancer (HBOC) has been performed in Slovakia in many different forms before the year 2000. Complex HBOC genetic analysis consists of many steps, including the initial genetic consultation, laboratory testing of genes associated with HBOC, interpretation and report of DNA analysis results, secondary explanatory genetic consultation and recommendation of clinical management for pathological mutation carriers. Many clinicians are participating on this workflow, such as clinical geneticists, laboratory diagnosticians as well as gynaecologists, oncologists or radio-diagnosticians. Currently, genetic testing is still technically and financially demanding and aimed only at selected families or patients who fulfil the defined clinical indication criteria.
Positive result of DNA analysis, that is, detection of pathological mutation in genes associated with HBOC syndrome means that the risk of breast/ovarian cancer onset in mutation carriers is amplified. This predisposition markedly affects the clinical management and treatment of patient and other members of the family, thus creating the demand to establish widely accepted specific recommendations for genetic diagnostics of HBOC. In the past, the analysis of HBOC in Slovakia followed various technical approaches and indication criteria depending on the workflow of specific laboratory. The guidelines reported below adhere to the current trends in DNA analysis and clinical healthcare, define the criteria for diagnostic laboratories, conditions for genetic testing and determine indications for selection of HBOC families and further clinical management of mutation carriers.
In order to study the soil aggregate distributions and soil organic matter (SOM), we sampled top- and subsoils in four intensively farmed croplands (two organic (Org-OB and Org-LA), and two conventional (Con-OB and Con-LA)) on Haplic Chernozems located in Marchfeld in the east of Vienna (Austria). Soil structure and SOM quantity, quality and distribution between free and occluded particulate organic matter and aggregate size fractions (<20 µm, 20-250 µm, 250-5000 µm) were studied by following a density fractionation procedure with low-energy ultrasound treatment. The relation of the soil physicochemical (e.g., particle size distribution, pH, organic carbon, total nitrogen) and biological properties (e.g., fungal biomass, active fungi) with stable soil aggregate size fractions and SOM was studied. The mean weight diameter (MWD) showed no significant difference between all studied sites and was between 3.8 mm and 10.0 mm in topsoils and between 6.7 mm and 11.9 mm in subsoils. In topsoils, the contents of calcium-acetate-lactate (CAL)-extractable P, active fungal biomass, dithionite-extractable Fe and sand were significantly positively correlated with the amount of the macroaggregates and with the MWD. We observed that most soil organic carbon, depending on soil texture, was stored in the microaggregate size classes <20 µm and 20-250 µm.
To investigate the effect of macromolecular transport and the incorporation of protein aggregate impurities in growing crystals, experiments were performed on the International Space Station (ISS) and compared with control experiments performed in a 1G laboratory environment. Crystal growth experiments for hen egg-white lysozyme (HEWL) and Plasmodium falciparum glutathione S-transferase (PfGST) were monitored using the ISS Light Microscopy Module (LMM). Experiments were performed applying the liquid–liquid counter diffusion crystallization method using rectangular, optically transparent capillaries. To analyze the quantity of impurity incorporated into growing crystals, stable fluorescently labeled protein aggregates were prepared and subsequently added at different percent concentrations to nonlabeled monomeric protein suspensions. For HEWL, a covalent cross-linked HEWL dimer was fluorescently labeled, and for PfGST, a stable tetramer was prepared. Crystallization solutions containing different protein aggregate ratios were prepared. The frozen samples were launched on 19.02.2017 via SpaceX-10 mission and immediately transferred to a -80°C freezer on the ISS. Two series of crystallization experiments were performed on ISS, one during 26.02.2017 to 10.03.2017 and a second during 16.06.2017 to 23.06.2017. A comparison of crystal growth rate and size showed different calculated average growth rates as well as different dimensions for crystals growing in different positions along the capillary. The effect of macromolecular mass transport on crystal growth in microgravity was experimentally calculated. In parallel, the percentage of incorporated fluorescent aggregate into the crystals was monitored utilizing the fluorescent LMM and ground-based fluorescent microscopes.