Late expression of FosB transcription factor in 4-aminopyridine-induced seizures in the rat cerebral cortex

In order to deliver services to citizens, public sector agencies of today offer various types of channels for interaction. In this article we aim to develop an understanding of the reciprocal relation between multi‑channel services and work prac tice. By showing how actors at different organizational levels in an agency differ in their interpretation of multi‑channel services we have generated knowledge that is essential in multi‑channel strategy formulation. The study illustrates an example of a n agency that works actively and in a mature way with e‑government and e‑services. This qualitative case study illustrates that the top level management, middle management and the case officers need to confront and discuss their understanding of e‑governm ent and their work practice in order to reach a situation where strategies actually are influencing daily work more explicitly. An important aspect is the decision to adopt an internal or external perspective on multi‑channel service management. The resul t of this decision might be to view multi‑channel service management as a way of reaching either internal agency efficiency or external citizen benefits

Gabonafélék petesejtjeinek és zigótáinak krioprezerválása = Cryopreservation of gametes in cereals

A pályázat célja a növényi petesejtek ill. zigóták krioprezelválási módszerének kidolgozása gabonaféléken. Meghatároztuk azokat a módszertani feltételeket (adaptációs közeg összetétele, ideje, krioprotektáns anyagok minősége, a fagyasztás módja és a rehidratáció körülményei), melyek segítségével 2 módszert is sikeresen alkalmaztunk növényi petesejtek és zigóták mélyhűtésére. Megállapítottuk, hogy a krioprezervációs módszer sikeressége nagymértékben függ a búza fajtájától. A hidegérézékeny Siete cerros petesejtek csak lassú hűtési eljárással, a hidegtűrő Mironovszkaja az un. vitrifikációs módszerrel is fagyaszthatók. A petesejtekre kidolgozott krioprezervációs módszert sikeresen alkalmaztuk zigóták mélyhűtésére is. Zigóták esetében 10-15%-al nagyobb hatékonyság érhető el. Kimutattuk továbbá, hogy - az aszkorbinsav kedvezően befolyásolja a petesejtek ill. zigóták túlélési képességét úgy a dehidratációs, mind a rehidratációs fázisban. - a krioprotektáns anyagok közül a DMSO, ellentétben számos szomatikus növényi szövettel és sejttel, toxikus hatású a petesejtekre. Helyette a PG sikeresen alkalmzaható. - az előkezelések alacsony hőmérsékleten (T=0°C) végzése mérsékli a krioprotektáns anyagok kedvezőtlen hatását. Strukturális vizsgálatokkal kimutattuk, hogy a krioprezerválás során a petesejtek ill. zigóták belső citoplazmaszerkezete jelentős mértékben árendeződik: az organellumok a plazma sejtmag körüli részén koncentrálódtak, míg a perifériális részen megjelent egy világosabban festődő, organellumokat alig tartalmazó citoplazmarégió. | The aim of the project was to elaborate cryopreservation methods for plant egg-cells and zygotes. The methodological conditions (composition of the adaptation medium, duration of dehydration, quality of cryoprotectants, freezing method, rehydration conditions) were elaborated for deep freezing of plant egg-cells and zygotes. The type of cryopreservation method required and the success with which it could be applied were found to depend greatly on the variety of wheat used in the experiments. While the egg-cells of the cold-sensitive variety Siete Cerros could only be frozen at a slow rate, the vitrification technique was more efficient for the cold-tolerant variety Mironovskaya 808. The cryopreservation method elaborated for egg-cells was also successfully applied to zygotes, where the success rate was 10-15% higher. Moreover, it was demonstrated that -ascorbic acid has a favourable influence on the survival rate of egg-cells and zygotes in both the dehydration and rehydration stages, -among the cryoprotectants tested, DMSO, which can be successfully applied to somatic plant tissues and cells, was found to be toxic to egg-cells, and had to be replaced by PG, -carrying out treatment at low temperature (0°C) reduced the negative effects of the cryoprotectants. Structural analyses revealed a rearrangement of the internal cytoplasm structure in egg-cells and zygotes during cryopreservation: the cell organelles became concentrated in the plasma region around the nucleus, while a more lightly stained cytoplasm region containing hardly any organelles appeared in the peripheral regions

A versatile modular vector set for optimizing protein expression among bacterial, yeast, insect and mammalian hosts

We have developed a unified, versatile vector set for expression of recombinant proteins, fit for use in any bacterial, yeast, insect or mammalian cell host. The advantage of this system is its versatility at the vector level, achieved by the introduction of a novel expression cassette. This cassette contains a unified multi-cloning site, affinity tags, protease cleavable linkers, an optional secretion signal, and common restriction endonuclease sites at key positions. This way, genes of interest and all elements of the cassette can be switched freely among the vectors, using restriction digestion and ligation without the need of polymerase chain reaction (PCR). This vector set allows rapid protein expression screening of various hosts and affinity tags. The reason behind this approach was that it is difficult to predict which expression host and which affinity tag will lead to functional expression. The new system is based on four optimized and frequently used expression systems (Escherichia coli pET, the yeast Pichia pastoris, pVL and pIEx for Spodoptera frugiperda insect cells and pLEXm based mammalian systems), which were modified as described above. The resulting vector set was named pONE series. We have successfully applied the pONE vector set for expression of the following human proteins: the tumour suppressor RASSF1A and the protein kinases Aurora A and LIMK1. Finally, we used it to express the large multidomain protein, Rho-associated protein kinase 2 (ROCK2, 164 kDa) and demonstrated that the yeast Pichia pastoris reproducibly expresses the large ROCK2 kinase with identical activity to the insect cell produced counterpart. To our knowledge this is among the largest proteins ever expressed in yeast. This demonstrates that the cost-effective yeast system can match and replace the industry-standard insect cell expression system even for large and complex mammalian proteins. These experiments demonstrate the applicability of our pONE vector set

Az UV sugárzás hatása a La és Ro antigén-mRNS-ek expressziójára humán keratinocitákban: kapcsolata autoimmun betegségekkel = Effect of the UV irradiation on the expression of La and Ro antigens in human keratinocytes relationship with autoimmune diseases

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Influence of Fe concentration in the medium on multicellular pollen grains and haploid plants induced by mannitol pretreatment in barley (Hordeum vulgare L.)

This study aims to clarify the short- and long-term effects of the iron concentration in the medium on androgenesis induced in barley by isolated microspore culture. The ultrastructural features and pectin composition of the intine wall were studied in the initial stages of androgenesis. The evolution of electron-dense iron deposits on the intine was analysed in multicellular pollen grains obtained by isolated microspore culture performed for 3, 6, and 9 days using various concentrations of FeNa2 EDTA. Finally, the number of embryo-like structures and green plants obtained by microspore culture using different Fe concentrations was evaluated in order to estimate the optimum concentration for isolated microspore culture.This study was supported by a Spain-Hungary Co-operation Project, nr. REF98HU0014. A. Pulido received a fellowship from the Junta de Andalucía, Spain.Peer Reviewe