Antigenic Protein In Microgravity-Grown Human Mixed Mullerian Tumor (LN1) Cells Preserved In RNA Stabilizing Agent

Cells treated with RNAlater(TradeMark) have previously been shown to contain antigenic proteins that can be visualized using Western blot analysis. These proteins seem to be stable for several months when stored in RNA stabilizer at 4 C. Antigenic protein can be recovered from cells that have been processed using an Ambion RNAqueous(Registered TradeMark) kit to remove RNA. In this set of experiments, human mixed Mullerian tumor (LN1) cells grown on the International Space Station during Expedition 3 were examined for antigenic stability after removal of RNA. The cells were stored for three months in RNAlater(TradeMark) and RNA was extracted. The RNA filtrate Containing the protein was precipitated, washed, and suspended in buffer containing sodium dodecyl sulfate (SDS). Samples containing equal concentrations of protein were loaded onto SDS-polyacrylamide gels. Proteins were separated by electrophoresis and transferred by Western blot to polyvinylidene fluoride (PVDF) membrane. The Western blots were stained with an enhanced chemiluminescent ECL(Registered TradeMark)Plus detection kit (Amersham) and scanned using a Storm 840 gel image analyzer (Amersham, Molecular Dynamics). ImageQuant(Registered TradeMark)a software was used to quantify the densities of the protein bands. The ground control and flight LN1 cell samples showed a similar staining pattern over time with antibodies to vimentin, glyceraldehyde-3-phosphate dehydrogenase, and epithelial membrane antigens

Energetics and performance of a microscopic heat engine based on exact calculations of work and heat distributions

We investigate a microscopic motor based on an externally controlled two-level system. One cycle of the motor operation consists of two strokes. Within each stroke, the two-level system is in contact with a given thermal bath and its energy levels are driven with a constant rate. The time evolution of the occupation probabilities of the two states are controlled by one rate equation and represent the system's response with respect to the external driving. We give the exact solution of the rate equation for the limit cycle and discuss the emerging thermodynamics: the work done on the environment, the heat exchanged with the baths, the entropy production, the motor's efficiency, and the power output. Furthermore we introduce an augmented stochastic process which reflects, at a given time, both the occupation probabilities for the two states and the time spent in the individual states during the previous evolution. The exact calculation of the evolution operator for the augmented process allows us to discuss in detail the probability density for the performed work during the limit cycle. In the strongly irreversible regime, the density exhibits important qualitative differences with respect to the more common Gaussian shape in the regime of weak irreversibility.Comment: 21 pages, 7 figure

Proteomic Retrieval from Nucleic Acid Depleted Space-Flown Human Cells

Compared to experiments utilizing humans in microgravity, cell-based approaches to questions about subsystems of the human system afford multiple advantages, such as crew safety and the ability to achieve statistical significance. To maximize the science return from flight samples, an optimized method was developed to recover protein from samples depleted of nucleic acid. This technique allows multiple analyses on a single cellular sample and when applied to future cellular investigations could accelerate solutions to significant biomedical barriers to human space exploration. Cell cultures grown in American Fluoroseal bags were treated with an RNA stabilizing agent (RNAlater - Ambion), which enabled both RNA and immunoreactive protein analyses. RNA was purified using an RNAqueous(registered TradeMark) kit (Ambion) and the remaining RNA free supernatant was precipitated with 5% trichloroacetic acid. The precipitate was dissolved in SDS running buffer and tested for protein content using a bicinchoninic acid assay (1) (Sigma). Equal loads of protein were placed on SDS-PAGE gels and either stained with CyproOrange (Amersham) or transferred using Western Blotting techniques (2,3,4). Protein recovered from RNAlater-treated cells and stained with protein stain, was measured using Imagequant volume measurements for rectangles of equal size. BSA treated in this way gave quantitative data over the protein range used (Fig 1). Human renal cortical epithelial (HRCE) cells (5,6,7) grown onboard the International Space Station (ISS) during Increment 3 and in ground control cultures exhibited similar immunoreactivity profiles for antibodies to the Vitamin D receptor (VDR) (Fig 2), the beta isoform of protein kinase C (PKC ) (Fig 3), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (Fig 4). Parallel immunohistochemical studies on formalin-fixed flight and ground control cultures also showed positive immunostaining for VDR and other biomarkers (Fig 5). These results are consistent with data from additional antigenic recovery experiments performed on human Mullerian tumor cells cultured in microgravity (8)

Analysis of landrace cultivation in Europe: A means to support in situ conservation of crop diversity

During the last century, the progressive substitution of landraces with modern, high yielding varieties, led to a dramatic reduction of in situ conserved crop diversity in Europe. Nowadays there is limited and scattered information on where landraces are cultivated. To fill this gap and lay the groundwork for a regional landrace in situ conservation strategy, information on more than 19,335 geo-referenced landrace cultivation sites were collated from 14 European countries. According to collected data, landraces of 141 herbaceous and 48 tree species are cultivated across Europe: Italy (107 species), Greece (93), Portugal (45) and Spain (44) hold the highest numbers. Common bean, onion, tomato, potato and apple are the species of main interest in the covered countries. As from collected data, about 19.8% of landrace cultivation sites are in protected areas of the Natura 2000 network. We also got evidence that 16.7% and 19.3% of conservation varieties of agricultural species and vegetables are currently cultivated, respectively. Results of the GIS analysis allowed the identification of 1261 cells (25 km × 25 km) including all the cultivation sites, distributed across all European biogeographical regions. Data of this study constitute the largest ever produced database of in situ-maintained landraces and the first attempt to create an inventory for the entire Europe. The availability of such resource will serve for better planning of actions and development of policies to protect landraces and foster their use

Welwitindolinone C synthetic studies. Construction of the welwitindolinone carbon skeleton via a transannular nitrone cycloaddition

Described is the construction of the N-methylwelwitindolinone C core via an efficient strategy that employs a sequential rhodium carbenoid-mediated O–H insertion, Claisen rearrangement and transannular [3+2] nitrone cycloaddition

CREATION OF A WEB MAP AND MOBILE APPLICATION BASED ON A PRINTED BOOK

The project describes a process of conversion of printed books into a web map and mobile application. The goal of the project is to make spatial data in the book accessible to wide public using GIS especially on web in order to spread the information about this topic. Moreover, as a result of the analysis and of the new perspectives gained from the data context, historians will be able to find new connections. The books that serve as sources of the project (two books with the scope of about 1400 pages featuring hundreds of locations where each location is associated with more events of different types) refer to places with many addresses in Prague and some villages in the Czech Republic which are related to events that took place during the World War II. The paper describes the steps of conversion, the design of the data model in Esri geodatabase and examples of outputs. The historical data are connected to actual addresses and thanks to such a combination of historical and actual locations, the project will help to discover a part of the history of the Czech Republic and it will show new context in data via GIS capabilities. This project is a continuation of a project which recorded a march of death on a map. This is a unique project created in cooperation with Academia Publishing. The outputs of the project will serve as a core resource for a multimedia history portal. The author of the book is currently writing sequels from the post-war period and at least two other books are envisioned, so the future of the project is ensured

MORPHOLOGICAL AND MOLECULAR EVALUATION OF THE FAR EAST FRUIT GENETIC RESOURCES OF <i> LONICERA CAERULEA </i> L.

Field studies of honeysuckle (Lonicera caerulea L. s.l.) and material for investigation (91 herbarium accessions; 20 scion samples; and fruits for detailed morphological description, organoleptic and molecular analyses) have been collected in the north-eastern part of the area of honeysuckle distribution, in Kamchatka and Sakhalin regions. All coenopopulations have been characterized ecologically, geobotanically, and phytosociologically. Factors threatening natural coenopopulations have been estimated, and these data preconditioned recommendations for honeysuckle in situ conservation. Fruits of 20 Russian honeysuckle cultivars have been included in comparative molecular assessment together with fruits from natural ecotypes. As a result of AFLP analysis, all investigated material was divided into three groups: two groups include samples L. caerulea, collected in Kamchatka, and the majority of the investigated honeysuckle cultivated forms, while the third group unites only genotypes of the sakhalin plants

MONITORING FRUIT LANDRACES IN THE CZECH REPUBLIC, TRACING THEIR ORIGIN AND POTENTIAL FOR THEIR CONSERVATION

Landraces of cultivated plants originated from the beginning of agriculture through selection from wild ecotypes and cultivated by farmers in their domestic region were investigated with the goal to conserve fragments of still existing fruit landraces, including restoration of variable countryside with scattered and roadside trees, wind-breaking and snow-catching functions including fruit consumption. Individual trees were identified, attributed to cultivars, localized by GPS and mapped in the Geobase Map of the Czech Republic. The inventory listed 388 fruits out of total number of 2866 accessions. It is a basis for searching lost and extinct materials. The database was analyzed for cultivars, regionality and their longevity by the length of registration. The best indigenous materials were proposed for in situ conservation mainly in the territory of National Parks and Protected Landscape Areas