Fluorescence-activated cell sorting and cloning of bona fide CD8+ CTL with reversible MHC-peptide and antibody Fab' conjugates.

The isolation of subsets of Ag-specific T cells for in vitro and in vivo studies by FACS is compromised by the fact that the soluble MHC-peptide complexes and Abs used for staining, especially when combined, induce unwanted T cell activation and eventually apoptosis. This is especially a problem for CD8+ CTL, which are susceptible to activation-dependent cell death. In this study, we show that reversible MHC-peptide complexes (tetramers) can be prepared by conjugating MHC-peptide monomers with desthiobiotin (DTB; also called dethiobiotin) and multimerization by reaction with fluorescent streptavidin. While in the cold these reagents are stable and allow good staining, they rapidly dissociate in monomers at elevated temperatures, especially in the presence of free biotin. FACS cloning of Melan-A (MART-1)-specific CTL from a melanoma-infiltrated lymph node with reversible HLA-A2 Melan-A26-35 multimers yielded over two times more clones than when using the conventional biotin-containing multimers. CTL clones obtained by means of reversible multimers killed Melan-A-positive tumor cells more efficiently as compared with clones obtained with the stable multimers. Among the CTL obtained with the reversible multimers, but much less among those obtained with the stable multimers, a high proportion of clones exhibited high functional and physical avidity and died upon incubation with soluble MHC-peptide complexes. Finally, we show that Fab' of an anti-CD8 Ab can be converted in reversible DTB streptavidin conjugates the same way. These DTB reagents efficiently and reversibly stained murine and human CTL without affecting their viability.

Cab45 is required for Ca(2+)-dependent secretory cargo sorting at the trans-Golgi network

Ca(2+) import into the lumen of the trans-Golgi network (TGN) by the secretory pathway calcium ATPase1 (SPCA1) is required for the sorting of secretory cargo. How is Ca(2+) retained in the lumen of the Golgi, and what is its role in cargo sorting? We show here that a soluble, lumenal Golgi resident protein, Cab45, is required for SPCA1-dependent Ca(2+) import into the TGN; it binds secretory cargo in a Ca(2+)-dependent reaction and is required for its sorting at the TGN.

Repeatability in automatic sorting of curved Norway spruce saw logs

Abstract

Technical Progress, Sorting, and Early Retirement

[cat] El progrés tecnològic afecta a la jubilació anticipada a través de dues forces oposades. Per una banda, augmenta el salari real i, per tant, crea incentius per posposar la jubilació. Per altra banda, causa l’erosió de les habilitats dels treballadors, el qual augmenta la probabilitat de jubilació anticipada. En aquest article reexaminem l’efecte del progrés tecnològic en la jubilació anticipada tenint en compte que, al principi de la vida laboral, els individus es distribueixen en diferents sectors segons la seva habilitat. Obtenim dos resultats principals: 1) per petits (grans) canvis tecnològics, l’efecte salari (erosió) domina, i 2) els individus més capaços resisteixen millor l’efecte erosió.

Ras-association domain of sorting nexin 27 is critical for regulating expression of GIRK potassium channels

G protein-gated inwardly rectifying potassium (GIRK) channels play an important role in regulating neuronal excitability. Sorting nexin 27b (SNX27b), which reduces surface expression of GIRK channels through a PDZ domain interaction, contains a putative Ras-association (RA) domain with unknown function. Deleting the RA domain in SNX27b (SNX27b-DRA) prevents the down-regulation of GIRK2c/GIRK3 channels. Similarly, a point mutation (K305A) in the RA domain disrupts regulation of GIRK2c/GIRK3 channels and reduces H-Ras binding in vitro. Finally, the dominant-negative H-Ras (S17N) occludes the SNX27b-dependent decrease in surface expression of GIRK2c/GIRK3 channels. Thus, the presence of a functional RA domain and the interaction with Ras-like G proteins comprise a novel mechanism for modulating SNX27b control of GIRK channel surface expression and cellular excitability.

Technical Progress, Sorting, and Early Retirement

[cat] El progrés tecnològic afecta a la jubilació anticipada a través de dues forces oposades. Per una banda, augmenta el salari real i, per tant, crea incentius per posposar la jubilació. Per altra banda, causa l’erosió de les habilitats dels treballadors, el qual augmenta la probabilitat de jubilació anticipada. En aquest article reexaminem l’efecte del progrés tecnològic en la jubilació anticipada tenint en compte que, al principi de la vida laboral, els individus es distribueixen en diferents sectors segons la seva habilitat. Obtenim dos resultats principals: 1) per petits (grans) canvis tecnològics, l’efecte salari (erosió) domina, i 2) els individus més capaços resisteixen millor l’efecte erosió.

Use of scenario analysis in studying emerging technology – Case Grid computing

Tutkimuksen selvitettiin miten skenaarioanalyysia voidaan käyttää uuden teknologian tutkimisessa. Työssä havaittiin, että skenaarioanalyysin soveltuvuuteen vaikuttaa eniten teknologisen muutoksen taso ja saatavilla olevan tiedon luonne. Skenaariomenetelmä soveltuu hyvin uusien teknologioiden tutkimukseen erityisesti radikaalien innovaatioiden kohdalla. Syynä tähän on niihin liittyvä suuri epävarmuus, kompleksisuus ja vallitsevan paradigman muuttuminen, joiden takia useat muut tulevaisuuden tutkimuksen menetelmät eivät ole tilanteessa käyttökelpoisia. Työn empiirisessä osiossa tutkittiin hilaverkkoteknologian tulevaisuutta skenaarioanalyysin avulla. Hilaverkot nähtiin mahdollisena disruptiivisena teknologiana, joka radikaalina innovaationa saattaa muuttaa tietokonelaskennan nykyisestä tuotepohjaisesta laskentakapasiteetin ostamisesta palvelupohjaiseksi. Tällä olisi suuri vaikutus koko nykyiseen ICT-toimialaan erityisesti tarvelaskennan hyödyntämisen ansiosta. Tutkimus tarkasteli kehitystä vuoteen 2010 asti. Teorian ja olemassa olevan tiedon perusteella muodostettiin vahvaan asiantuntijatietouteen nojautuen neljä mahdollista ympäristöskenaariota hilaverkoille. Skenaarioista huomattiin, että teknologian kaupallinen menestys on vielä monen haasteen takana. Erityisesti luottamus ja lisäarvon synnyttäminen nousivat tärkeimmiksi hilaverkkojen tulevaisuutta ohjaaviksi tekijöiksi.

A conceptual model for grid learning services automatic composition

Peer-reviewed

Modelling of North-West Russian market behavior considering technical restrictions of transmission grid

Nowadays the Finnish-Russian electric energy interaction is carried out through the back-to-back DC Vyborg substation and several power plants working synchronously with Finnish power system. Constant amount of energy flows in one direction — from Russia to Finland. But the process of electricity market development in Russian energy system makes the new possibilities of electrical cooperation available. The goal of master's thesis is to analyze the current state and possible evolution trends of North-West Russian system in relation with future possible change in power flow between Russia and Finland. The research is done by modelling the market of North-West Russia and examination of technical grid restrictions. The operational market models of North-West region of Russia for the years 2008 and 2015 were created during the research process. The description of prepared market models together with modelling results and their analysis are shown in the work. The description of power flow study process and results are also presented.

Development of power flows around Baltic countries with new grid links

In the Thesis main focus is done on power flow development paths around the Baltic States as well as on market-based requirements for creation of the common Baltic electricity market. Current market regulations between the countries are presented; barriers for creating competitive common Baltic power market and for electricity trading with third countries are clarified; solutions are offered and corresponding road map is developed. Future power development paths around the Baltic States are analysed. For this purpose the 330 kV transmission grid of Estonia, Latvia and Lithuania is modelled in a power flow tool. Power flow calculations are carried out for winter and summer peak and off-peak load periods in 2020 with different combinations of interconnections. While carrying out power balance experiments several power flow patterns in the Baltic States are revealed. Conclusions are made about security of supply, grid congestion and transmission capacity availability for different scenarios.

Improving the oscillating grid method to characterize aquaculture biosolids using a laser beam and image analysis

Quickremovalofbiosolidsinaquaculturefacilities,andspeciallyinrecirculatingaquaculturesystems(RAS),isoneofthemostimportantstepinwastemanagement.Sedimentationdynamicsofbiosolidsinanaquaculturetankwilldeterminetheiraccumulationatthebottomofthetank.