A PiggyBac-mediated approach for muscle gene transfer or cell therapy.

An emerging therapeutic approach for Duchenne muscular dystrophy is the transplantation of autologous myogenic progenitor cells genetically modified to express dystrophin. The use of this approach is challenged by the difficulty in maintaining these cells ex vivo while keeping their myogenic potential, and ensuring sufficient transgene expression following their transplantation and myogenic differentiation in vivo. We investigated the use of the piggyBac transposon system to achieve stable gene expression when transferred to cultured mesoangioblasts and into murine muscles. Without selection, up to 8% of the mesoangioblasts expressed the transgene from 1 to 2 genomic copies of the piggyBac vector. Integration occurred mostly in intergenic genomic DNA and transgene expression was stable in vitro. Intramuscular transplantation of mouse Tibialis anterior muscles with mesoangioblasts containing the transposon led to sustained myofiber GFP expression in vivo. In contrast, the direct electroporation of the transposon-donor plasmids in the mouse Tibialis muscles in vivo did not lead to sustained transgene expression despite molecular evidence of piggyBac transposition in vivo. Together these findings provide a proof-of-principle that piggyBac transposon may be considered for mesoangioblast cell-based therapies of muscular dystrophies.

Shallow-transfer rule-based machine translation for Czech to Polish

This article describes the developmentof an Open Source shallow-transfer machine translation system from Czech to Polish in theApertium platform. It gives details ofthe methods and resources used in contructingthe system. Although the resulting system has quite a high error rate, it is still competitive with other systems.

Regulation of the vitellogenin gene B1 promoter after transfer into hepatocytes in primary cultures.

The estrogen-dependent and tissue-specific regulation of the Xenopus laevis vitellogenin gene B1 promoter has been studied by lipid-mediated DNA transfer into Xenopus hepatocytes in primary culture. Hepatocytes achieve an efficient hormonal control of this promoter through a functional interaction between the estrogen responsive elements and a promoter proximal region upstream of the TATA box, which is characterized by a high density of binding sites for the transcription factors CTF/NF-1, C/EBP and HNF3. DNA accessibility to restriction enzymes within the chromosomal copy of the vitellogenin gene B1 promoter shows that the estrogen responsive unit and the promoter proximal region are sensitive to digestion in uninduced and estrogen-induced hepatocytes but not in erythrocyte nuclei. Together, these findings support the notion that chromatin configuration as well as the interplay of promoter elements mediate proper hormone-dependent and tissue-specific expression of the B1 vitellogenin gene.

Effect of antifoams on oxygen mass transfer in pulp and paper mill effluents

Vaahdonestoaineiden haitallinen vaikutus hapen liukenemisnopeuteen biologisen puhdistamon jätevesissä on yleisesti tunnettua. Aineiden eri vaikutusmekanismien takia on silti vaikea etukäteen arvioida, miten ja kuinka paljon aineensiirto muuttuu. Työn tavoitteena oli saada tietoa vaahdonestoaineiden ja muiden pinta-aktiivisten aineiden vaikutuksesta kuplakokoon, kaasun tilavuusosuuteen ja kaasu-neste aineensiirtoon. Työn teoriaosassa on kuvailtu vaahdon muodostumiseen vaikuttavia tekijöitä sekä eri vaahdonestoaineiden vaikutusmekanismeja sellu- ja paperitehtaan jätevedessä. Edelleen on esitetty useita hapen siirtoa estäviä ja parantavia aineita. Työn kokeellisessa osassa tutkittiin kahdenkymmenenviiden eri pinta-aktiivisen aineen vaikutusta hapen liukenemisnopeuteen yksivaiheisessa kuplakolonnissa. Kokeet tehtiin kahdella pitoisuudella, kahdella eri kaasunjakolaitteella ja kolmella eri kaasun tyhjäputkinopeudella. Aineensiirtokokeiden rinnalla tutkittiin jätevesien laatu- ja fysikaalisiaominaisuuksia, niiden vaikutusta hapen liukenemisnopeuteen sekä testattavien koeaineiden vaikutusta fysikaalisin ominaisuuksiin. Kokeet osoittavat että pinta-aktiivisten aineiden vaikutus hapen aineensiirtoon vaihtelee riippuen kaasunjakolaitteesta ja aineen pitoisuuksista. Testatuista vaahdonestonaineista pienin negatiivinen vaikutus oli aineella AT 35 ja positiivinen vaikutustodettiin olevan vaahdonestoaineiden komponenteilla: P2, S1, F4 ja T9.

Stochasticity and bistability in horizontal transfer control of a genomic island in Pseudomonas.

Genomic islands (GEI) comprise a recently recognized large family of potentially mobile DNA elements and play an important role in the rapid differentiation and adaptation of bacteria. Most importantly, GEIs have been implicated in the acquisition of virulence factors, antibiotic resistances or toxic compound metabolism. Despite detailed information on coding capacities of GEIs, little is known about the regulatory decisions in individual cells controlling GEI transfer. Here, we show how self-transfer of ICEclc, a GEI in Pseudomonas knackmussii B13 is controlled by a series of stochastic processes, the result of which is that only a few percent of cells in a population will excise ICEclc and launch transfer. Stochastic processes have been implicated before in producing bistable phenotypic transitions, such as sporulation and competence development, but never before in horizontal gene transfer (HGT). Bistability is instigated during stationary phase at the level of expression of an activator protein InrR that lays encoded on ICEclc, and then faithfully propagated to a bistable expression of the IntB13 integrase, the enzyme responsible for excision and integration of the ICEclc. Our results demonstrate how GEI of a very widespread family are likely to control their transfer rates. Furthermore, they help to explain why HGT is typically confined to few members within a population of cells. The finding that, despite apparent stochasticity, HGT rates can be modulated by external environmental conditions provides an explanation as to why selective conditions can promote DNA exchange.

Simulation of Boundary Layers and Heat Transfer in the Entrance Region of Pipes

The study of fluid flow in pipes is one of the main topic of interest for engineers in industries. In this thesis, an effort is made to study the boundary layers formed near the wall of the pipe and how it behaves as a resistance to heat transfer. Before few decades, the scientists used to derive the analytical and empirical results by hand as there were limited means available to solve the complex fluid flow phenomena. Due to the increase in technology, now it has been practically possible to understand and analyze the actual fluid flow in any type of geometry. Several methodologies have been used in the past to analyze the boundary layer equations and to derive the expression for heat transfer. An integral relation approach is used for the analytical solution of the boundary layer equations and is compared with the FLUENT simulations for the laminar case. Law of the wall approach is used to derive the empirical correlation between dimensionless numbers and is then compared with the results from FLUENT for the turbulent case. In this thesis, different approaches like analytical, empirical and numerical are compared for the same set of fluid flow equations.

Flow phenomena, heat and mass transfer in microchannel reactors

The studies of flow phenomena, heat and mass transfer in microchannel reactors are beneficial to estimate and evaluate the ability of microchannel reactors to be operated for a given process reaction such as Fischer-Tropsch synthesis. The flow phenomena, for example, the flow regimes and flow patterns in microchannel reactors for both single phase and multiphase flow are affected by the configuration of the flow channel. The reviews of the previous works about the analysis of related parameters that affect the flow phenomena are shown in this report. In order to predict the phenomena of Fischer-Tropsch synthesis in microchannel reactors, the 3-dimensional computational fluid dynamic simulation with commercial software package FLUENT was done to study the flow phenomena and heat transfer for gas phase Fischer-Tropsch products flow in rectangular microchannel with hydraulic diameter 500 ¿m and length 15 cm. Numerical solution with slip boundary condition was used in the simulation and the flowphenomena and heat transfer were determined.

An operon of three transcriptional regulators controls horizontal gene transfer of the integrative and conjugative element ICEclc in Pseudomonas knackmussii B13.

The integrative and conjugative element ICEclc is a mobile genetic element in Pseudomonas knackmussii B13, and an experimental model for a widely distributed group of elements in Proteobacteria. ICEclc is transferred from specialized transfer competent cells, which arise at a frequency of 3-5% in a population at stationary phase. Very little is known about the different factors that control the transfer frequency of this ICE family. Here we report the discovery of a three-gene operon encoded by ICEclc, which exerts global control on transfer initiation. The operon consists of three consecutive regulatory genes, encoding a TetR-type repressor MfsR, a MarR-type regulator and a LysR-type activator TciR. We show that MfsR autoregulates expression of the operon, whereas TciR is a global activator of ICEclc gene expression, but no clear role was yet found for MarR. Deletion of mfsR increases expression of tciR and marR, causing the proportion of transfer competent cells to reach almost 100% and transfer frequencies to approach 1 per donor. mfsR deletion also caused a two orders of magnitude loss in population viability, individual cell growth arrest and loss of ICEclc. This indicates that autoregulation is an important feature maintaining ICE transfer but avoiding fitness loss. Bioinformatic analysis showed that the mfsR-marR-tciR operon is unique for ICEclc and a few highly related ICE, whereas tciR orthologues occur more widely in a large variety of suspected ICE among Proteobacteria.

Heat Transfer Analysis of the EPR Core Catcher Test Facility Volley

Uusi EPR-reaktorikonsepti on suunniteltu selviytymään tapauksista, joissa reaktorinsydän sulaa ja sula puhkaisee paineastian. Suojarakennuksen sisälle on suunniteltu alue, jolle sula passiivisesti kerätään, pidätetään ja jäähdytetään. Alueelle laaditaan valurautaelementeistä ns.sydänsieppari, joka tulvitetaan vedellä. Sydänsulan tuottama jälkilämpö siirtyyveteen, mistä se poistetaan suojarakennuksen jälkilämmönpoistojärjestelmän kautta. Suuri osa lämmöstä poistuu sydänsulasta sen yläpuolella olevaan veteen, mutta lämmönsiirron tehostamiseksi myös sydänsiepparin alapuolelle on sijoitettu vedellä täytettävät jäähdytyskanavat. Jotta sydänsiepparin toiminta voitaisiin todentaa, on Lappeenrannan Teknillisellä Yliopistolla rakennettu Volley-koelaitteisto tätä tarkoitusta varten. Koelaitteisto koostuu kahdesta täysimittaisesta valuraudasta tehdystä jäähdytyskanavasta. Sydänsulan tuottamaa jälkilämpöä simuloidaan koelaitteistossa sähkövastuksilla. Tässä työssä kuvataan simulaatioiden suorittaminen ja vertaillaan saatuja arvoja mittaustuloksiin. Työ keskittyy sydänsiepparista jäähdytyskanaviin tapahtuvan lämmönsiirron teoriaan jamekanismeihin. Työssä esitetään kolme erilaista korrelaatiota lämmönsiirtokertoimille allaskiehumisen tapauksessa. Nämä korrelaatiot soveltuvat erityisesti tapauksiin, joissa vain muutamia mittausparametreja on tiedossa. Työn toinen osa onVolley 04 -kokeiden simulointi. Ensin käytettyä simulointitapaa on kelpoistettuvertaamalla tuloksia Volley 04 ja 05 -kokeisiin, joissa koetta voitiin jatkaa tasapainotilaan ja joissa jäähdytteen käyttäytyminen jäähdytyskanavassa on tallennettu myös videokameralla. Näiden simulaatioiden tulokset ovat hyvin samanlaisiakuin mittaustulokset. Korkeammilla lämmitystehoilla kokeissa esiintyi vesi-iskuja, jotka rikkoivat videoinnin mahdollistavia ikkunoita. Tämän johdosta osassa Volley 04 -kokeita ikkunat peitettiin metallilevyillä. Joitakin kokeita jouduttiin keskeyttämään laitteiston suurten lämpöjännitysten johdosta. Tällaisten testien simulaatiot eivät ole yksinkertaisia suorittaa. Veden pinnan korkeudesta ei ole visuaalista havaintoa. Myöskään jäähdytteen tasapainotilanlämpötiloista ei ole tarkkaa tietoa, mutta joitakin oletuksia voidaan tehdä samoilla parametreilla tehtyjen Volley 05 -kokeiden perusteella. Mittaustulokset Volley 04 ja 05 -kokeista, jotka on videoitu ja voitu ajaa tasapainotilaan saakka, antoivat simulaatioiden kanssa hyvin samankaltaisia lämpötilojen arvoja. Keskeytettyjen kokeiden ekstrapolointi tasapainotilaan ei onnistunut kovin hyvin. Kokeet jouduttiin keskeyttämään niin paljon ennen termohydraulista tasapainoa, ettei tasapainotilan reunaehtoja voitu ennustaa. Videonauhoituksen puuttuessa ei veden pinnan korkeudesta saatu lisätietoa. Tuloksista voidaan lähinnä esittää arvioita siitä, mitä suuruusluokkaa mittapisteiden lämpötilat tulevat olemaan. Nämä lämpötilat ovat kuitenkin selvästi alle sydänsiepparissa käytettävän valuraudan sulamislämpötilan. Joten simulaatioiden perusteella voidaan sanoa, etteivät jäähdytyskanavien rakenteet sula, mikäli niissä on pienikin jäähdytevirtaus, eikä useampia kuin muutama vierekkäinen kanava ole täysin kuivana.

Transfer of penicillin resistance between Neisseriae in microcosm

Horizontal gene transfer between commensal and pathogenic Neisseriae is the mechanism proposed to explain how pathogenic species acquire altered portions of the penA gene, which encodes penicillin binding protein 2. These changes resulted in a moderately penicillin-resistant phenotype in the meningococci, whose frequency of isolation in Spain increased at the end of the 1980s. Little has been published about the possibility of this gene transfer in nature or about its simulation in the laboratory. We designed a simple microcosm, formed by solid and liquid media, that partially mimics the upper human respiratory tract. In this microcosm, penicillin-resistant commensal strains and the fully susceptible meningococcus were co-cultivated. The efficiency of gene transfer between the strains depended on the phase of bacterial growth and the conditions of culture. Resistance of penicillin was acquired in different steps irrespective of the source of the DNA. The presence of DNase in the medium had no effect on gene transfer, but it was near zero when nicked DNA was used. Cell-to-cell contact or membrane blebs could explain these results. The analysis of sequences of the transpeptidase domain of PBP2 from transformants, and from donor and recipient strains demonstrated that the emergence of moderately resistant transformants was due to genetic exchange between the co-cultivated strains. Finally, mechanisms other than penA modification could be invoked to explain decreased susceptibility

Interfirm Knowledge Transfer in Coopetitive Relationships

Tämä tutkimuskeskittyy yritysten väliseen samanaikaiseen kilpailuun ja yhteistyöhön. Suurin mielenkiinto tässä tutkimuksessa kohdistuu yritysten väliseen tietämyksen siirtoon. Tietämyksen siirto samanaikaisissa kilpailu- ja yhteistyösuhteissa voi olla erityisen hyödyllistä samankaltaisten toimintalogiikoiden ja ymmärryksen takia. Toisaalta, mahdollinen kriittisen tiedon vuotaminen kilpailevalle yritykselle onriski, joka täytyy ottaa huomioon, kun yhteistyötä kilpailijan kanssa suunnitellaan. Tämän tutkimuksen tavoitteena on selvittää, kuinka tietämystä siirretään samanaikaisissa kilpailu- ja yhteistyösuhteissa välttäen kuitenkin tähän liittyvät riskit. Tutkimuksen empiirinen osa sisältää kolme erilaista case-esimerkkiä samanaikaisista kilpailu- ja yhteistyösuhteista. Empiirisessä osassa tehdyt havainnot tukevat pääasiassa teoriaosuudessa käsiteltyjä asioita. Tietämyksen siirron motiivit, suhdekohtaiset tietämyksen omaksumiskyvyt, tietojohtamiskäytännöt ja luottamus ovat tekijöitä, jotka vaikuttavat tietämyksen siirron tehokkuuteen. Pääasialliset keinot tietämyksen siirron riskin välttämiseksi samanaikaisissa kilpailu- ja yhteistyösuhteissa ovat hierarkkisen hallintomuodon käyttäminen tai horisontaalisesti ja vertikaalisesti rajattu yhteistyön laajuus.

Modeling of non-isothermalvapor membrane separation with thermodynamic models and generalized mass transfer equations

A rigorous unit operation model is developed for vapor membrane separation. The new model is able to describe temperature, pressure, and concentration dependent permeation as wellreal fluid effects in vapor and gas separation with hydrocarbon selective rubbery polymeric membranes. The permeation through the membrane is described by a separate treatment of sorption and diffusion within the membrane. The chemical engineering thermodynamics is used to describe the equilibrium sorption of vapors and gases in rubbery membranes with equation of state models for polymeric systems. Also a new modification of the UNIFAC model is proposed for this purpose. Various thermodynamic models are extensively compared in order to verify the models' ability to predict and correlate experimental vapor-liquid equilibrium data. The penetrant transport through the selective layer of the membrane is described with the generalized Maxwell-Stefan equations, which are able to account for thebulk flux contribution as well as the diffusive coupling effect. A method is described to compute and correlate binary penetrant¿membrane diffusion coefficients from the experimental permeability coefficients at different temperatures and pressures. A fluid flow model for spiral-wound modules is derived from the conservation equation of mass, momentum, and energy. The conservation equations are presented in a discretized form by using the control volume approach. A combination of the permeation model and the fluid flow model yields the desired rigorous model for vapor membrane separation. The model is implemented into an inhouse process simulator and so vapor membrane separation may be evaluated as an integralpart of a process flowsheet.

Numerical and ExperimentalModelling of Gas Flow and Heat Transfer in the Air Gap of An Electric Machine

This work deals with the cooling of high-speed electric machines, such as motors and generators, through an air gap. It consists of numerical and experimental modelling of gas flow and heat transfer in an annular channel. Velocity and temperature profiles are modelled in the air gap of a high-speed testmachine. Local and mean heat transfer coefficients and total friction coefficients are attained for a smooth rotor-stator combination at a large velocity range. The aim is to solve the heat transfer numerically and experimentally. The FINFLO software, developed at Helsinki University of Technology, has been used in the flow solution, and the commercial IGG and Field view programs for the grid generation and post processing. The annular channel is discretized as a sector mesh. Calculation is performed with constant mass flow rate on six rotational speeds. The effect of turbulence is calculated using three turbulence models. The friction coefficient and velocity factor are attained via total friction power. The first part of experimental section consists of finding the proper sensors and calibrating them in a straight pipe. After preliminary tests, a RdF-sensor is glued on the walls of stator and rotor surfaces. Telemetry is needed to be able to measure the heat transfer coefficients at the rotor. The mean heat transfer coefficients are measured in a test machine on four cooling air mass flow rates at a wide Couette Reynolds number range. The calculated values concerning the friction and heat transfer coefficients are compared with measured and semi-empirical data. Heat is transferred from the hotter stator and rotor surfaces to the coolerair flow in the air gap, not from the rotor to the stator via the air gap, althought the stator temperature is lower than the rotor temperature. The calculatedfriction coefficients fits well with the semi-empirical equations and precedingmeasurements. On constant mass flow rate the rotor heat transfer coefficient attains a saturation point at a higher rotational speed, while the heat transfer coefficient of the stator grows uniformly. The magnitudes of the heat transfer coefficients are almost constant with different turbulence models. The calibrationof sensors in a straight pipe is only an advisory step in the selection process. Telemetry is tested in the pipe conditions and compared to the same measurements with a plain sensor. The magnitudes of the measured data and the data from the semi-empirical equation are higher for the heat transfer coefficients than thenumerical data considered on the velocity range. Friction and heat transfer coefficients are presented in a large velocity range in the report. The goals are reached acceptably using numerical and experimental research. The next challenge is to achieve results for grooved stator-rotor combinations. The work contains also results for an air gap with a grooved stator with 36 slots. The velocity field by the numerical method does not match in every respect the estimated flow mode. The absence of secondary Taylor vortices is evident when using time averagednumerical simulation.

Applicability of Power-Line Communications to Data Transfer of On-line Condition Monitoring of Electrical Drives

The research of power-line communications has been concentrated on home automation, broadband indoor communications and broadband data transfer in a low voltage distribution network between home andtransformer station. There has not been carried out much research work that is focused on the high frequency characteristics of industrial low voltage distribution networks. The industrial low voltage distribution network may be utilised as a communication channel to data transfer required by the on-line condition monitoring of electric motors. The advantage of using power-line data transfer is that it does not require the installing of new cables. In the first part of this work, the characteristics of industrial low voltage distribution network components and the pilot distribution network are measured and modelled with respect topower-line communications frequencies up to 30 MHz. The distributed inductances, capacitances and attenuation of MCMK type low voltage power cables are measured in the frequency band 100 kHz - 30 MHz and an attenuation formula for the cables is formed based on the measurements. The input impedances of electric motors (15-250 kW) are measured using several signal couplings and measurement based input impedance model for electric motor with a slotted stator is formed. The model is designed for the frequency band 10 kHz - 30 MHz. Next, the effect of DC (direct current) voltage link inverter on power line data transfer is briefly analysed. Finally, a pilot distribution network is formed and signal attenuation in communication channels in the pilot environment is measured. The results are compared with the simulations that are carried out utilising the developed models and measured parameters for cables and motors. In the second part of this work, a narrowband power-line data transfer system is developed for the data transfer ofon-line condition monitoring of electric motors. It is developed using standardintegrated circuits. The system is tested in the pilot environment and the applicability of the system for the data transfer required by the on-line condition monitoring of electric motors is analysed.