Gene expression responses of Lactobacillus rhamnosus GG to pH changes of the growth environment

Monitoroimalla organismin geeniekspressiota voidaan saada hyödyllistä informaatiota fysiologiasta bioprosessikasvatuksen eri vaiheissa. Tässä työssä on tutkittu kirjallisuudessa aiemmin mainittujen pH-stressiin liittyvien geenien atpA, aldB, cfa, groEL, hrcA ja pstS ekspressoitumista suhteellisen geeniekspression menetelmällä käyttäen vertailugeeninä ldhD:tä. Ekspressiomittaukset tehtiin käyttäen qPCR- ja TRAC-menetelmää. Tutkittuna organismina oli probioottina tunnettu Lactobacillus rhamnosus GG (LGG). Teollisen kasvatuksen pH:n säädössä esiintyviä emäsgradientteja sekä lämpötilagradientteja simuloitiin scale-down-menetelmällä. Lisäksi tutkittiin laboratorioravintoalustan ja teollisen ravintoalustan välisiä eroja panoskasvatuksilla. Scale-down-kokeissa havaittiin geenien groEL, hrcA ja atpA antavan vasteita lämpötilan ja pH:n muutoksiin. pstS-geenin ekspressio kasvoi lähes lineaarisesti kemostaattikokeissa tulppavirtausreaktoriin aiheutetun emäsgradientin kasvaessa. Työssä tutkittujen geenien ekspressioiden yksittäiset korrelaatiot kemostaattikokeiden näytteille tehtyjen happostressi- ja pakastussäilyvyyksien välillä olivat vähäisiä. Panoskasvatuksista tehtyjen geeniekspressiomittausten perusteella erilaisilla kasvualustoilla näyttäisi olevan vaikutusta tutkittujen geenien ekspressioon. Teollisella alustalla ekspressioiden suhteelliset muutokset eri kasvuvaiheiden välillä olivat pienempiä, mutta vertailugeeniin verratut ekspressiot olivat pääsääntöisesti suurempia kuin laboratorioalustalla.Monitoring organism's gene expression can provide useful information on it's physiology in different phases of cultivation. In this work expression of pH stress related genes found from the literature (atpA, aldB, cfa, groEL, hrcA and pstS) were studied as a relative gene expression study using ldhD as a reference gene. Expression measurements were carried out using qPCR and TRAC methods. Study organism was Lactobacillus rhamnosus GG, a renowned probiotic. Base gradients caused by pH control in an industrial process and temperature gradients were simulated with a scale-down method. A pH gradient was created in a plug flow reactor. Furthermore, differences between laboratory cultivation medium and industrial cultivation medium were studied with batch cultivations. In these cultivations the set point of pH control was oscillated. In the scale-down experiments the responses of groEL, hrcA and atpA genes to temperature and pH changes were studied. The expression of pstS gene was induced almost linearly in the chemostat cultivation experiments when base gradient in plug flow reactor was increased. There was rather low correlations between the expressions of studied genes and surviving from acid stress test or freeze stability test, which were carried out from the samples of the chemostat cultivation experiments. Based on gene expression measurements from the batch cultivations it seems that the type of the cultivation medium affects expressions of the studied genes. In case of the industrial cultivation medium relative changes in gene expressions were less between different growth phases but expressions related to the reference gene were mainly higher compared to the laboratory medium

The Politics of Constitutional Change

In this paper, an optimized control drum system for the European Lead Cooled Training Reactor (ELECTRA) is proposed. By changing the number of rotating drums from 6 to 12, we succeed in reducing the maximum rotational worth of a single drum from 4 $to 1.64$. As a consequence, the safety margin during reactivity insertions is significantly improved.QC 20121108. Correction in: Nuclear Engineering And Design, vol 255, Pages 376-376,Doi: 10.1016/j.nucengdes.2012.11.001, WOS:000315839700039ScopusID: 2-s2.0-84884902351</p

Enhanced Lignocellulolytic Enzyme Activities on Hardwood and Softwood during Interspecific Interactions of White- and Brown-Rot Fungi

Wood decomposition is a sophisticated process where various biocatalysts act simultaneously and synergistically on biopolymers to efficiently break down plant cell walls. In nature, this process depends on the activities of the wood-inhabiting fungal communities that co-exist and interact during wood decay. Wood-decaying fungal species have traditionally been classified as white-rot and brown-rot fungi, which differ in their decay mechanism and enzyme repertoire. To mimic the species interaction during wood decomposition, we have cultivated the white-rot fungus, Bjerkandera adusta, and two brown-rot fungi, Gloeophyllum sepiarium and Antrodia sinuosa, in single and co-cultivations on softwood and hardwood. We compared their extracellular hydrolytic carbohydrate-active and oxidative lignin-degrading enzyme activities and production profiles. The interaction of white-rot and brown-rot species showed enhanced (hemi)cellulase activities on birch and spruce-supplemented cultivations. Based on the enzyme activity profiles, the combination of B. adusta and G. sepiarium facilitated birch wood degradation, whereas B. adusta and A. sinuosa is a promising combination for efficient degradation of spruce wood, showing synergy in β-glucosidase (BGL) and α-galactosidase (AGL) activity. Synergistic BGL and AGL activity was also detected on birch during the interaction of brown-rot species. Our findings indicate that fungal interaction on different woody substrates have an impact on both simultaneous and sequential biocatalytic activities

Simultaneous Measurement of Tricarboxylic Acid Cycle Intermediates in Different Biological Matrices Using Liquid Chromatography–Tandem Mass Spectrometry; Quantitation and Comparison of TCA Cycle Intermediates in Human Serum, Plasma, Kasumi-1 Cell and Murine Liver Tissue

The tricarboxylic acid (TCA) cycle is a central part of carbon and energy metabolism, also connecting to glycolysis, amino acid, and lipid metabolism. The quantitation of the TCA cycle intermediate within one method is lucrative due to the interest in central carbon metabolism profiling in cells and tissues. In addition, TCA cycle intermediates in serum have been discovered to correspond as biomarkers to various underlying pathological conditions. In this work, an Liquid Chromatography-Mass Spectrometry/Mass Spectrometry-based quantification method is developed and validated, which takes advantage of fast, specific, sensitive, and cost-efficient precipitation extraction. Chromatographic separation is achieved while using Atlantis dC18 2.1 mm × 100 mm, particle size 3-μm of Waters column with a gradient elution mobile phase while using formic acid in water (0.1% v/v) and acetonitrile. Linearity was clearly seen over a calibration range of: 6.25 to 6400 ng/mL (r2 > 0.980) for malic acid; 11.72 to 12,000 ng/mL (r2 > 0.980) for cis-aconitic acid and L-aspartic acid; 29.30 to 30,000 ng/mL (r2 > 0.980) for isocitric acid, l-serine, and l-glutamic acid; 122.07 to 125,000 ng/mL (r2 > 0.980) for citric acid, glycine, oxo-glutaric acid, l-alanine, and l-glutamine; 527.34 to 540,000 ng/mL (r2 > 0.980) for l-lactic acid; 976.56 to 1,000,000 ng/mL (r2 > 0.980) for d-glucose; 23.44 to 24,000 ng/mL (r2 > 0.980) for fumaric acid and succinic acid; and, 244.14 to 250,000 ng/mL (r2 > 0.980) for pyruvic acid. Validation was carried out, as per European Medicines Agency (EMA) “guidelines on bioanalytical method validation”, for linearity, precision, accuracy, limit of detection (LOD), limit of quantification (LLOQ), recovery, matrix effect, and stability. The recoveries from serum and tissue were 79–119% and 77–223%, respectively. Using this method, we measured TCA intermediates in serum, plasma (NIST 1950 SRM), and in mouse liver samples. The concentration found in NIST SRM 1950 (n = 6) of glycine (246.4 µmol/L), l-alanine (302.4 µmol/L), and serine (92.9 µmol/L)

Horizontal Governance and Governance Discourses in Career Guidance and Counselling in Finland

In the 2000s, horizontal governance has become a growing mode of public sector governance. In this study, we examined horizontal governance in the context of career guidance and counselling in Finland. Our empirical research data consisted of ten (10) thematic interviews with public sector officials and other experts working in the governance of guidance at the national level. Our research aim was to scrutinize how the three ideal-types in public sector governance – bureaucratic (Weberian), new public management, and new public governance – were manifested in the informants’ discourses. Our results showed that ideologically, there was a shared understanding of the need to increase horizontal governance in guidance. However, in terms of legislation, finance or evaluation of guidance, existing rules and beliefs built in previous ideal-types tend to challenge the implementation of horizontal governance in practice.Peer reviewe

Interface interactions in UN-X-UO2 systems (X = V, Nb, Ta, Cr, Mo, W) by pressure-assisted diffusion experiments at 1773 K

QC 20220524</p

Oxidation of UN-U2N3/UO2 composites: an evaluation of UO2 as an oxidation barrier for the nitride phases

Composite fuels such as UN-UO2 are being considered to address the lower oxidation resistance of the UN fuel from a safety perspective for use in light water reactors, whilst improving the in-reactor behaviour of the more ubiquitous UO2 fuel. An innovative UN-UO2 accident tolerant fuel has recently been fabricated and studied: UN microspheres embedded in UO2 matrix. In the present study, detailed oxidative thermogravimetric investigations (TGA/DSC) of high-density UN/U2N3-UO2 composite fuels (91-97 %TD), as well as post oxidised microstructures obtained by SEM, are reported and analysed. Triplicate TGA measurements of each specimen were carried out at 5 K/min up to 973 K in a synthetic air atmosphere to assess their oxidation kinetics. The mass variation due to the oxidation reactions (%), the oxidation onset temperatures (OOTs), and the maximum reaction temperatures (MRTs) are also presented and discussed. The results show that all composites have similar post oxidised microstructures with mostly intergranular cracking and spalling. The oxidation resistance of the pellet with initially 10 wt% of UN microspheres is surprisingly better than the UO2 reference. Moreover, there is no significant difference in the OOT (~557 K) and MRT (~615 K) when 30 wt% or 50 wt% of embedded UN microspheres are used. Therefore, the findings in this article demonstrate that the UO2 matrix acts as a barrier to improve the oxidation resistance of the nitride phases at the beginning of life conditions

UN microspheres embedded in UO2 matrix: an innovative accident tolerant fuel

Uranium nitride (UN)-uranium dioxide (UO2) composite fuels are being considered as an accident tolerant fuel (ATF) option for light water reactors. However, the complexity related to the chemical interactions between UN and UO(2 )during sintering is still an open problem. Moreover, there is a lack of knowledge regarding the influence of the sintering parameters on the amount and morphology of the alpha-U2N3 phase formed. In this study, a detailed investigation of the interaction between UN and UO2 is provided and a formation mechanism for the resulting alpha-U2N3 phase is proposed. Coupled with these analyses, an innovative ATF concept was investigated: UN microspheres and UO2,13 powder were mixed and subsequently sintered by spark plasma sintering. Different temperatures, pressures, times and cooling rates were evaluated. The pellets were characterised by complementary techniques, including XRD, DSC, and SEM-EDS/WDS/EBSD. The UN and UO2 interaction is driven by O diffusion into the UN phase and N diffusion in the opposite direction, forming a long-range solid solution in the UO2 matrix, that can be described as UO2-xNx. The cooling process decreases the N solubility in UO2-xNx, causing then N redistribution and precipitation as alpha-U2N3 phase along and inside the UO2 grains. This precipitation mechanism occurs at temperatures between 1273 K and 973 K on cooling, following specific crystallographic grain orientation patterns. (C) 2020 The Authors. Published by Elsevier B.V