Homeostatic proliferation and survival of naïve and memory T cells.

The immune system relies on homeostatic mechanisms in order to adapt to the changing requirements encountered during steady-state existence and activation by antigen. For T cells, this involves maintenance of a diverse repertoire of naïve cells, rapid elimination of effector cells after pathogen clearance, and long-term survival of memory cells. The reduction of T-cell counts by either cytotoxic drugs, irradiation, or certain viruses is known to lead to lymphopenia-induced proliferation and restoration of normal T-cell levels. Such expansion is governed by the interaction of TCR with self-peptide/MHC (p/MHC) molecules plus contact with cytokines, especially IL-7. These same ligands, i.e. p/MHC molecules and IL-7, maintain naïve T lymphocytes as resting cells under steady-state T-cell-sufficient conditions. Unlike naïve cells, typical "central" memory T cells rely on a combination of IL-7 and IL-15 for their survival in interphase and for occasional cell division without requiring signals from p/MHC molecules. Other memory T-cell subsets are less quiescent and include naturally occurring activated memory-phenotype cells, memory cells generated during chronic viral infections, and effector memory cells. These subsets of activated memory cells differ from central memory T cells in their requirements for homeostatic proliferation and survival. Thus, the factors controlling T-cell homeostasis can be seen to vary considerably from one subset to another as described in detail in this review.

PRPF mutations are associated with generalized defects in spliceosome formation and pre-mRNA splicing in patients with retinitis pigmentosa.

Proteins PRPF31, PRPF3 and PRPF8 (RP-PRPFs) are ubiquitously expressed components of the spliceosome, a macromolecular complex that processes nearly all pre-mRNAs. Although these spliceosomal proteins are conserved in eukaryotes and are essential for survival, heterozygous mutations in human RP-PRPF genes lead to retinitis pigmentosa, a hereditary disease restricted to the eye. Using cells from patients with 10 different mutations, we show that all clinically relevant RP-PRPF defects affect the stoichiometry of spliceosomal small nuclear RNAs (snRNAs), the protein composition of tri-small nuclear ribonucleoproteins and the kinetics of spliceosome assembly. These mutations cause inefficient splicing in vitro and affect constitutive splicing ex-vivo by impairing the removal of at least 9% of endogenously expressed introns. Alternative splicing choices are also affected when RP-PRPF defects are present. Furthermore, we show that the steady-state levels of snRNAs and processed pre-mRNAs are highest in the retina, indicating a particularly elevated splicing activity. Our results suggest a role for PRPFs defects in the etiology of PRPF-linked retinitis pigmentosa, which appears to be a truly systemic splicing disease. Although these mutations cause widespread and important splicing defects, they are likely tolerated by the majority of human tissues but are critical for retinal cell survival.

Development of fluid conduits in the auriferous shear zones of the Hutti Gold Mine, India: evidence for spatially and temporally heterogeneous fluid flow

The gold mineralization of the Hutti Mine is hosted by nine parallel, N - S trending, steeply dipping, 2 - 10 m wide shear zones, that transect Archaean amphibolites. The shear zones were formed after peak metamorphism during retrograde ductile D, shearing in the lower amphibolite facies. They were reactivated in the lower to mid greenschist facies by brittle-ductile D-3 shearing and intense quartz veining. The development of a S-2-S-3 crenulation cleavage facilitates the discrimination between the two deformation events and contemporaneous alteration and gold mineralization. Ductile D, shearing is associated with a pervasively developed distal chlorite - sed cite alteration assemblage in the outer parts of the shear zones and the proximal biotite-plagioclase alteration in the center of the shear zones. D3 is characterized by development of the inner chlorite-K-feldspar alteration, which forms a centimeter-scale alteration halo surrounding the laminated quartz veins and replaces earlier biotite along S-3. The average size of the laminated vein systems is 30-50 m along strike as well as down-dip and 2-6 m in width. Mass balance calculations suggest strong metasomatic changes for the proximal biotite-plagioclase alteration yielding mass and volume increase of ca. 16% and 12%, respectively. The calculated mass and volume changes of the distal chlorite-sericite alteration (ca. 11%, ca. 8%) are lower. The decrease in 6180 values of the whole rock from around 7.5 parts per thousand for the host rocks to 6-7 parts per thousand for the distal chlorite-sericite and the proximal biotite-plagioclase alteration and around 5 parts per thousand for the inner chlorite-K-feldspar alteration suggests hydrothermal alteration during two-stage deformation and fluid flow. The ductile D-2 deformation in the lower amphibolite facies has provided grain scale porosities by microfracturing. The pervasive, steady-state fluid flow resulted in a disseminated style of gold-sulfide mineralization and a penetrative alteration of the host rocks. Alternating ductile and brittle D3 deformation during lower to mid greenschist facies conditions followed the fault-valve process. Ductile creep in the shear zones resulted in a low permeability environment leading to fluid pressure build-up. Strongly episodic fluid advection and mass transfer was controlled by repeated seismic fracturing during the formation of laminated quartz(-gold) veins. The limitation of quartz veins to the extent of earlier shear zones indicate the importance of preexisting anisotropies for fault-valve action and economic gold mineralization. (C) 2003 Elsevier B.V. All rights reserved.

β1- and β3- voltage-gated sodium channel subunits modulate cell surface expression and glycosylation of Nav1.7 in HEK293 cells.

Voltage-gated sodium channels (Navs) are glycoproteins composed of a pore-forming α-subunit and associated β-subunits that regulate Nav α-subunit plasma membrane density and biophysical properties. Glycosylation of the Nav α-subunit also directly affects Navs gating. β-subunits and glycosylation thus comodulate Nav α-subunit gating. We hypothesized that β-subunits could directly influence α-subunit glycosylation. Whole-cell patch clamp of HEK293 cells revealed that both β1- and β3-subunits coexpression shifted V ½ of steady-state activation and inactivation and increased Nav1.7-mediated I Na density. Biotinylation of cell surface proteins, combined with the use of deglycosydases, confirmed that Nav1.7 α-subunits exist in multiple glycosylated states. The α-subunit intracellular fraction was found in a core-glycosylated state, migrating at ~250 kDa. At the plasma membrane, in addition to the core-glycosylated form, a fully glycosylated form of Nav1.7 (~280 kDa) was observed. This higher band shifted to an intermediate band (~260 kDa) when β1-subunits were coexpressed, suggesting that the β1-subunit promotes an alternative glycosylated form of Nav1.7. Furthermore, the β1-subunit increased the expression of this alternative glycosylated form and the β3-subunit increased the expression of the core-glycosylated form of Nav1.7. This study describes a novel role for β1- and β3-subunits in the modulation of Nav1.7 α-subunit glycosylation and cell surface expression.

Dynamic Model of a Small Once-Through Boiler

In this study, a model for the unsteady dynamic behaviour of a once-through counter flow boiler that uses an organic working fluid is presented. The boiler is a compact waste-heat boiler without a furnace and it has a preheater, a vaporiser and a superheater. The relative lengths of the boiler parts vary with the operating conditions since they are all parts of a single tube. The present research is a part of a study on the unsteady dynamics of an organic Rankine cycle power plant and it will be a part of a dynamic process model. The boiler model is presented using a selected example case that uses toluene as the process fluid and flue gas from natural gas combustion as the heat source. The dynamic behaviour of the boiler means transition from the steady initial state towards another steady state that corresponds to the changed process conditions. The solution method chosen was to find such a pressure of the process fluid that the mass of the process fluid in the boiler equals the mass calculated using the mass flows into and out of the boiler during a time step, using the finite difference method. A special method of fast calculation of the thermal properties has been used, because most of the calculation time is spent in calculating the fluid properties. The boiler was divided into elements. The values of the thermodynamic properties and mass flows were calculated in the nodes that connect the elements. Dynamic behaviour was limited to the process fluid and tube wall, and the heat source was regarded as to be steady. The elements that connect the preheater to thevaporiser and the vaporiser to the superheater were treated in a special way that takes into account a flexible change from one part to the other. The model consists of the calculation of the steady state initial distribution of the variables in the nodes, and the calculation of these nodal values in a dynamic state. The initial state of the boiler was received from a steady process model that isnot a part of the boiler model. The known boundary values that may vary during the dynamic calculation were the inlet temperature and mass flow rates of both the heat source and the process fluid. A brief examination of the oscillation around a steady state, the so-called Ledinegg instability, was done. This examination showed that the pressure drop in the boiler is a third degree polynomial of the mass flow rate, and the stability criterion is a second degree polynomial of the enthalpy change in the preheater. The numerical examination showed that oscillations did not exist in the example case. The dynamic boiler model was analysed for linear and step changes of the entering fluid temperatures and flow rates.The problem for verifying the correctness of the achieved results was that there was no possibility o compare them with measurements. This is why the only way was to determine whether the obtained results were intuitively reasonable and the results changed logically when the boundary conditions were changed. The numerical stability was checked in a test run in which there was no change in input values. The differences compared with the initial values were so small that the effects of numerical oscillations were negligible. The heat source side tests showed that the model gives results that are logical in the directions of the changes, and the order of magnitude of the timescale of changes is also as expected. The results of the tests on the process fluid side showed that the model gives reasonable results both on the temperature changes that cause small alterations in the process state and on mass flow rate changes causing very great alterations. The test runs showed that the dynamic model has no problems in calculating cases in which temperature of the entering heat source suddenly goes below that of the tube wall or the process fluid.

Sliding Mode Control for Switched Mode Power Supplies

Control applications of switched mode power supplies have been widely investigated. The main objective ofresearch and development (R&D) in this field is always to find the most suitable control method to be implemented in various DC/DC converter topologies. Inother words, the goal is to select a control method capable of improving the efficiency of the converter, reducing the effect of disturbances (line and load variation), lessening the effect of EMI (electro magnetic interference), and beingless effected by component variation. The main objective of this research work is to study different control methods implemented in switched mode power supplies namely (PID control, hysteresis control, adaptive control, current programmed control, variable structure control (VSC), and sliding mode control (SMC). The advantages and drawbacks of each control method are given. Two control methods, the PID and the SMC are selected and their effects on DC/DC (Buck, Boost, and Buck-Boost) converters are examined. Matlab/SimulinkTM is used to implement PID control method in DC/DC Buck converter and SMC in DC/DC (Buck, and Buck Boost) converters. For the prototype, operational amplifiers (op-amps) are used to implement PID control in DC/DC Buck converter. For SMC op-amps are implemented in DC/DC Buck converter and dSPACETM is used to control the DC/DC Buck-Boost converter. The SMC can be applied to the DC/DC (Buck, Boost, and Buck-Boost) converters. A comparison of the effects of the PID control and the SMC on the DC/DC Buck converter response in steady state, under line variations, load variations, and different component variations is performed. Also the Conducted RF-Emissions between the PID and SMC DC/DC Buck Converter are compared. The thesis shows that, in comparison with the PID control, the SMC provides better steady-state response, better dynamic response, less EMI, inherent order reduction, robustness against system uncertainty disturbances, and an implicit stability proof. Giving a better steady-state and dynamic response, the SMC is implemented in a DC/DC resonant converter. The half-wave zero current switching (HWZCS) DC/DC Buck converter is selected as a converter topology. A general guideline to select the tank component values, needed for the designing of a HWZCS DC/DC Buck, is obtained. The implementation of the SMC to a HWZCS DC/DC Buck converter is analysed. The converter response is investigated in the steady-state region and in the dynamic region.

Liver-specific loss of lipin-1-mediated phosphatidic acid phosphatase activity does not mitigate intrahepatic TG accumulation in mice.

Lipin proteins (lipin 1, 2, and 3) regulate glycerolipid homeostasis by acting as phosphatidic acid phosphohydrolase (PAP) enzymes in the TG synthesis pathway and by regulating DNA-bound transcription factors to control gene transcription. Hepatic PAP activity could contribute to hepatic fat accumulation in response to physiological and pathophysiological stimuli. To examine the role of lipin 1 in regulating hepatic lipid metabolism, we generated mice that are deficient in lipin-1-encoded PAP activity in a liver-specific manner (Alb-Lpin1(-/-) mice). This allele of lipin 1 was still able to transcriptionally regulate the expression of its target genes encoding fatty acid oxidation enzymes, and the expression of these genes was not affected in Alb-Lpin1(-/-) mouse liver. Hepatic PAP activity was significantly reduced in mice with liver-specific lipin 1 deficiency. However, hepatocytes from Alb-Lpin1(-/-) mice had normal rates of TG synthesis, and steady-state hepatic TG levels were unaffected under fed and fasted conditions. Furthermore, Alb-Lpin1(-/-) mice were not protected from intrahepatic accumulation of diacylglyerol and TG after chronic feeding of a diet rich in fat and fructose. Collectively, these data demonstrate that marked deficits in hepatic PAP activity do not impair TG synthesis and accumulation under acute or chronic conditions of lipid overload.

Self-navigated isotropic three-dimensional cardiac T2 mapping.

PURPOSE: To implement and characterize an isotropic three-dimensional cardiac T2 mapping technique. METHODS: A self-navigated three-dimensional radial segmented balanced steady-state free precession pulse sequence with an isotropic 1.7-mm spatial resolution was implemented at 3T with a variable T2 preparation module. Bloch equation and Monte Carlo simulations were performed to determine the influence of the heart rate, B1 inhomogeneity and noise on the T2 fitting accuracy. In a phantom study, the accuracy of the pulse sequence was studied through comparison with a gold-standard spin-echo T2 mapping method. The robustness and homogeneity of the technique were ascertained in a study of 10 healthy adult human volunteers, while first results obtained in patients are reported. RESULTS: The numerical simulations demonstrated that the heart rate and B1 inhomogeneity cause only minor deviations in the T2 fitting, whereas the phantom study showed good agreement of the technique with the gold standard. The volunteer study demonstrated an average myocardial T2 of 40.5 ± 3.3 ms and a <15% T2 gradient in the base-apex and anterior-inferior direction. In three patients, elevated T2 values were measured in regions with expected edema. CONCLUSION: This respiratory self-navigated isotropic three-dimensional technique allows for accurate and robust in vitro and in vivo T2 quantification. Magn Reson Med 73:1549-1554, 2015. © 2014 Wiley Periodicals, Inc.

Imaging of prolonged BOLD response in the somatosensory cortex of the rat.

Blood oxygenation level-dependent (BOLD) functional MRI is a widely employed methodology in experimental and clinical neuroscience, although its nature is not fully understood. To gain insights into BOLD mechanisms and take advantage of the new functional methods, it is of interest to investigate prolonged paradigms of activation suitable for long experimental protocols and to observe any long-term modifications induced by these functional challenges. While different types of sustained stimulation paradigm have been explored in human studies, the BOLD response is typically limited to a few minutes in animal models, due to fatigue, anesthesia effects and physiological instability. In the present study, the rat forepaw was electrically stimulated for 2 h, which resulted in a prolonged and localized cortical BOLD response over that period. The stimulation paradigm, including an inter-stimulus interval (ISI) of 10 s, that is 25% of the total time, was applied at constant or variable frequency over 2 h. The steady-state level of the BOLD response was reached after 15-20 min of stimulation and was maintained until the end of the stimulation. On average, no substantial loss in activated volume was observed at the end of the stimulation, but less variability in the fraction of remaining activated volume and higher steady-state BOLD amplitude were observed when stimulation frequency was varied between 2 and 3 Hz every 5 min. We conclude that the combination of ISI and variable stimulus frequency reproducibly results in robust, prolonged and localized BOLD activation. Copyright © 2015 John Wiley & Sons, Ltd.

Optimization of KOH etching process to obtain textured substrates suitable for heterojunction solar cells fabricated by HWCVD

In this work, we have studied the texturization process of (100) c-Si wafers using a low concentration potassium hydroxide solution in order to obtain good quality textured wafers. The optimization of the etching conditions have led to random but uniform pyramidal structures with good optical properties. Then, symmetric heterojunctions were deposited by Hot-Wire CVD onto these substrates and the Quasi-Steady-State PhotoConductance technique was used to measure passivation quality. Little degradation in the effective lifetime and implicit open circuit voltage of these devices (< 20 mV) was observed in all cases. It is especially remarkable that for big uniform pyramids, the open-circuit voltage is comparable to the values obtained on flat substrates.

Progress in a-Si:H/c-Si heterojunction emitters obtained by Hot-Wire CVD at 200°C

In this work, we investigate heterojunction emitters deposited by Hot-Wire CVD on p-type crystalline silicon. The emitter structure consists of an n-doped film (20 nm) combined with a thin intrinsic hydrogenated amorphous silicon buffer layer (5 nm). The microstructure of these films has been studied by spectroscopic ellipsometry in the UV-visible range. These measurements reveal that the microstructure of the n-doped film is strongly influenced by the amorphous silicon buffer. The Quasy-Steady-State Photoconductance (QSS-PC) technique allows us to estimate implicit open-circuit voltages near 700 mV for heterojunction emitters on p-type (0.8 Ω·cm) FZ silicon wafers. Finally, 1 cm 2 heterojunction solar cells with 15.4% conversion efficiencies (total area) have been fabricated on flat p-type (14 Ω·cm) CZ silicon wafers with aluminum back-surface-field contact.

Rehufosfaattitehtaan raaka-ainesyöttöjen säätöpiirien kehitys

Diplomityö tehtiin Kemira Chemicals Oy:n Kokkolan tehtaiden rehufosfaattitehtaalla. Työn tavoitteena oli uusien reseptiohjauksien suunnittelu ja käyttöönotto sekä tasaussäiliön pinnansäädön ja rakeistusrummun suhdesäädön suunnittelu. Käyttöhenkilöstön koulutus uuteen prosessinohjaukseen kuului myös osaksi diplomityötä. Uuden reseptiohjauksen avulla pyrittiin saamaan prosessin ohjaus havainnollisemmaksi ja ennustettavammaksi käyttäjille, yhdenmukaistamaan käyttäjien ajotapaa sekä suunnitella tuotereseptit vastaamaan paremmin todellista tuotantoa. Uusi reseptiohjaus mahdollistaa myös lajikekohtaisten parametrien käytön ja hyväksi havaittujen ajoreseptien tallennuksen. Tasaussäiliön pinnankorkeuden säädön tavoitteena on poistaa tuotantokatkokset, jotka johtuivat säiliön tyhjenemisestä tai täyttymisestä. Rakeistusrummun suhdesäädön tavoitteena on reaktoriliemen ja kuiva-ainesyöttöjen oikean suhteen säilyttäminen. Uuden reseptiohjauksen käyttöönoton jälkeen ajoreseptit ovat vastanneet todellista tuotantoa ja tuotantovauhti on jäänyt pysyvästi korkeammalle tasolle. Tasaussäiliön pinnansäätö toimii suunnitellulla tavalla ja on myös otettu käyttöön. Reaktoriliemen ja kuivasyöttöjen suhdesäätö on testattu käytännössä ja toimii halutulla tavalla. Suhdesäädön paremman toiminnan kannalta epätarkkoja toimilaitteita tulisi kuitenkin uusia.

Disc Filter External Models in an Integrated Simulation Environment

Työssä tutkittiin kiekkosuodattimeen liittyviä ulkoisia simulointimalleja integroidussa simulointiympäristössä. Työn tarkoituksena oli parantaa olemassa olevaa mekanistista kiekkosuodatinmallia. Malli laadittiin dynaamiseen paperiteollisuuden tarpeisiin tehtyyn simulaattoriin (APMS), jossa olevaan alkuperäiseen mekanistiseen malliin tehtiin ulkoinen lisämalli, joka käyttää hyväkseen kiekkosuodatinvalmistajan mittaustuloksia. Laitetiedon saatavuutta suodattimien käyttäjille parannettiin luomalla Internetissä sijaitsevalle palvelimelle kiekkosuodattimen laitetietomäärittelyt. Suodatinvalmistaja voi palvella asiakkaitaan viemällä laitetiedot palvelimelle ja yhdistämällä laitetiedon simulointimalliin. Tämä on mahdollista Internetin ylitse käytettävän integroidun simulointiympäristön avulla, jonka on tarkoitus kokonaisvaltaisesti yhdistää simulointi ja prosessisuunnittelu. Suunnittelijalle tarjotaan työkalut, joilla dynaaminen simulointi, tasesimulointi ja kaavioiden piirtäminen onnistuu prosessilaitetiedon ollessa saatavilla. Nämä työkalut on tarkoitus toteuttaa projektissa nimeltä Galleria, jossa luodaan prosessimalli- ja laitetietopalvelin Internetiin. Gallerian käyttöliittymän avulla prosessisuunnittelija voi käyttää erilaisia simulointiohjelmistoja ja niihin luotuja valmiita malleja, sekä saada käsiinsä ajan tasalla olevaa laitetietoa. Ulkoinen kiekkosuodatinmalli laskee suodosvirtaamat ja suodosten pitoisuudet likaiselle, kirkkaalle ja superkirkkaalle suodokselle. Mallin syöttöparametrit ovat kiekkojen pyörimisnopeus, sisään tulevan syötön pitoisuus, suotautuvuus (freeness) ja säätöparametri, jolla säädetään likaisen ja kirkkaan suodoksen keskinäinen suhde. Suotautuvuus kertoo mistä massasta on kyse. Mitä suurempi suotautuvuus on, sitä paremmin massa suodattuu ja sitä puhtaampia suodokset yleensä ovat. Mallin parametrit viritettiin regressioanalyysillä ja valmistajan palautetta apuna käyttäen. Käyttäjä voi valita haluaako hän käyttää ulkoista vai alkuperäistä mallia. Alkuperäinen malli täytyy ensin alustaa antamalla sille nominaaliset toimintapisteet virtaamille ja pitoisuuksille tietyllä pyörimisnopeudella. Ulkoisen mallin yhtälöitä voi käyttää alkuperäisen mallin alustamiseen, jos alkuperäinen malli toimii ulkoista paremmin. Ulkoista mallia voi käyttää myös ilman simulointiohjelmaa Galleria-palvelimelta käsin. Käyttäjälle avautuu näin mahdollisuus tarkastella kiekkosuodattimien parametreja ja nähdä suotautumistulokset oman työasemansa ääreltä mistä tahansa, kunhan Internetyhteys on olemassa. Työn tuloksena kiekkosuodattimien laitetiedon saatavuus käyttäjille parani ja alkuperäisen simulointimallin rajoituksia ja puutteita vähennettiin.

Parallel feedback loops control the basal activity of the HOG MAPK signaling cascade.

Tight regulation of the MAP kinase Hog1 is crucial for survival under changing osmotic conditions. Interestingly, we found that Hog1 phosphorylates multiple upstream components, implying feedback regulation within the signaling cascade. Taking advantage of an unexpected link between glucose availability and Hog1 activity, we used quantitative single cell measurements and computational modeling to unravel feedback regulation operating in addition to the well-known adaptation feedback triggered by glycerol accumulation. Indeed, we found that Hog1 phosphorylates its activating kinase Ssk2 on several sites, and cells expressing a non-phosphorylatable Ssk2 mutant are partially defective for feedback regulation and proper control of basal Hog1 activity. Together, our data suggest that Hog1 activity is controlled by intertwined regulatory mechanisms operating with varying kinetics, which together tune the Hog1 response to balance basal Hog1 activity and its steady-state level after adaptation to high osmolarity.

Metabolic origin of δ15N values in nitrogenous compounds from Brassica napus L. leaves

Nitrogen isotope composition (δ15N) in plant organic matter is currently used as a natural tracer of nitrogen acquisition efficiency. However, the δ15N value of whole leaf material does not properly reflect the way in which N is assimilated because isotope fractionations along metabolic reactions may cause substantial differences among leaf compounds. In other words, any change in metabolic composition or allocation pattern may cause undesirable variability in leaf δ15N. Here, we investigated the δ15N in different leaf fractions and individual metabolites from rapeseed (Brassica napus) leaves. We show that there were substantial differences in δ15N between nitrogenous compounds (up to 30 ) and the content in (15N enriched) nitrate had a clear influence on leaf δ15N. Using a simple steady-state model of day metabolism, we suggest that the δ15N value in major amino acids was mostly explained by isotope fractionation associated with isotope effects on enzyme-catalysed reactions in primary nitrogen metabolism. δ15N values were further influenced by light versus dark conditions and the probable occurrence of alternative biosynthetic pathways. We conclude that both biochemical pathways (that fractionate between isotopes) and nitrogen sources (used for amino acid production) should be considered when interpreting the δ15N value of leaf nitrogenous compounds