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Schneider, Thomas,Schellenberg, Maja,Meyer, Stefan,Keller, Felix,Gehrig, Peter,Riedel, Kathrin,Lee, Youngsook,Eberl, Leo,Martinoia, Enrico WILEY-VCH Verlag 2009 Proteomics Vol.9 No.10
<P>Although the vacuole is the most important final store for toxic heavy metals like cadmium (Cd<SUP>2+</SUP>), our knowledge on how they are transported into the vacuole is still insufficient. It has been suggested that Cd<SUP>2+</SUP> can be transported as phytochelatin-Cd<SUP>2+</SUP> by an unknown ABC transporter or in exchange with protons by cation/proton exchanger (CAX) transporters. To unravel the contribution of vacuolar transporters to Cd<SUP>2+</SUP> detoxification, a quantitative proteomics approach was performed. Highly purified vacuoles were isolated from barley plants grown under minus, low (20 μM), and high (200 μM) Cd<SUP>2+ </SUP>conditions and protein levels of the obtained tonoplast samples were analyzed using isobaric tag for relative and absolute quantitation (iTRAQ™). Although 56 vacuolar transporter proteins were identified, only a few were differentially expressed. Under low-Cd<SUP>2+</SUP> conditions, an inorganic pyrophosphatase and a γ-tonoplast intrinsic protein (γ-TIP) were up-regulated, indicating changes in energization and water fluxes. In addition, the protein ratio of a CAX1a and a natural resistance-associated macrophage protein (NRAMP), responsible for vacuolar Fe<SUP>2+</SUP> export was increased. CAX1a might play a role in vacuolar Cd<SUP>2+</SUP> transport. An increase in NRAMP activity leads to a higher cytosolic Fe<SUP>2+</SUP> concentration, which may prevent the exchange of Fe<SUP>2+</SUP> by toxic Cd<SUP>2+</SUP>. Additionally, an ABC transporter homolog to AtMRP3 showed up-regulation. Under high Cd<SUP>2+</SUP> conditions, the plant response was more specific. Only a protein homologous to AtMRP3 that showed already a response under low Cd<SUP>2+</SUP> conditions, was up-regulated. Interestingly, AtMRP3 is able to partially rescue a Cd<SUP>2+</SUP>-sensitive yeast mutant. The identified transporters are good candidates for further investigation of their roles in Cd<SUP>2+</SUP> detoxification.</P>
Lapauw, Thomas,Tytko, Darius,Vanmeensel, Kim,Huang, Shuigen,Choi, Pyuck-Pa,Raabe, Dierk,Caspi, El’ad N.,Ozeri, Offir,to Baben, Moritz,Schneider, Jochen M.,Lambrinou, Konstantina,Vleugels, Jozef American Chemical Society 2016 Inorganic Chemistry Vol.55 No.11
<P>The solubility of zirconium (Zr) in the Nb<SUB>4</SUB>AlC<SUB>3</SUB> host lattice was investigated by combining the experimental synthesis of (Nb<SUB><I>x</I></SUB>, Zr<SUB>1–<I>x</I></SUB>)<SUB>4</SUB>AlC<SUB>3</SUB> solid solutions with density functional theory calculations. High-purity solid solutions were prepared by reactive hot pressing of NbH<SUB>0.89</SUB>, ZrH<SUB>2</SUB>, Al, and C starting powder mixtures. The crystal structure of the produced solid solutions was determined using X-ray and neutron diffraction. The limited Zr solubility (maximum of 18.5% of the Nb content in the host lattice) in Nb<SUB>4</SUB>AlC<SUB>3</SUB> observed experimentally is consistent with the calculated minimum in the energy of mixing. The lattice parameters and microstructure were evaluated over the entire solubility range, while the chemical composition of (Nb<SUB>0.85</SUB>, Zr<SUB>0.15</SUB>)<SUB>4</SUB>AlC<SUB>3</SUB> was mapped using atom probe tomography. The hardness, Young’s modulus, and fracture toughness at room temperature as well as the high-temperature flexural strength and E-modulus of (Nb<SUB>0.85</SUB>, Zr<SUB>0.15</SUB>)<SUB>4</SUB>AlC<SUB>3</SUB> were investigated and compared to those of pure Nb<SUB>4</SUB>AlC<SUB>3</SUB>. Quite remarkably, an appreciable increase in fracture toughness was observed from 6.6 ± 0.1 MPa/m<SUP>1/2</SUP> for pure Nb<SUB>4</SUB>AlC<SUB>3</SUB> to 10.1 ± 0.3 MPa/m<SUP>1/2</SUP> for the (Nb<SUB>0.85</SUB>, Zr<SUB>0.15</SUB>)<SUB>4</SUB>AlC<SUB>3</SUB> solid solution.</P><P>The solubility of Zr in nanolaminated (Nb<SUB>1−<I>x</I></SUB>, Zr<SUB><I>x</I></SUB>)<SUB>4</SUB>AlC<SUB>3</SUB> is investigated. Experimentally, a maximum at <I>x</I> = 18.5% is found, which is consistent with a minimum in the calculated energy of mixing at low Zr contents. The mechanical behavior of this solid solution shows a significant improvement in fracture toughness and temperature stability compared to those of Nb<SUB>4</SUB>AlC<SUB>3</SUB>.</P><P><B>Graphic Abstract</B> <IMG SRC='http://pubs.acs.org/appl/literatum/publisher/achs/journals/content/inocaj/2016/inocaj.2016.55.issue-11/acs.inorgchem.6b00484/production/images/medium/ic-2016-00484u_0007.gif'></P>
Rohana P. Dassanayake,Thomas C. Truscott,Dongyue Zhuang,David A. Schneider,Sally A. Madsen-Bouterse,Alan J. Young,James B. Stanton,William C. Davis,Katherine I. O’Rourke 대한수의학회 2015 Journal of Veterinary Science Vol.16 No.2
Scrapie is diagnosed antemortem in sheep by detecting misfolded isoforms of prion protein (PrPSc) in lymphoid follicles of the rectal mucosa and nictitating membranes. Assay sensitivity is limited if (a) the biopsy is collected early during disease development, (b) an insufficient number of follicles is collected, or (c) peripheral accumulation of PrPSc is reduced or delayed. A blood test would be convenient for mass live animal scrapie testing. Currently approved techniques, however, have their own detection limits. Novel detection methods may soon offer a non-animal-based, rapid platform with detection sensitivities that rival the prion bioassay. In anticipation, we sought to determine if diseased animals could be routinely identified with a bioassay using B lymphocytes isolated from blood sample volumes commonly collected for diagnostic purposes in small ruminants. Scrapie transmission was detected in five of six recipient lambs intravenously transfused with B lymphocytes isolated from 5∼10 mL of blood from a naturally scrapie-infected sheep. Additionally, scrapie transmission was observed in 18 ovinized transgenic Tg338 mice intracerebrally inoculated with B lymphocytes isolated from 5∼10 mL of blood from two naturally scrapie-infected sheep. Based on our findings, we anticipate that these blood sample volumes should be of diagnostic value.
Jeong, Jae‐,Ho,Kim, Hyun‐,Ju,Kim, Kun‐,Hee,Shin, Minsang,Hong, Yeongjin,Rhee, Joon Haeng,Schneider, Thomas D.,Choy, Hyon E. Blackwell Publishing Ltd 2012 Molecular microbiology Vol.83 No.3
<P><B>Summary</B></P><P>Transcription start points in bacteria are influenced by the nature of the RNA polymerase·promoter interaction. For <I>Escherichia coli</I> RNA polymerase holoenzyme containing σ70, it is presumed that specific sequence in one or more of the −10, extended −10 and −35 elements of the promoter guides the RNAP to select the cognate start point. Here, we investigated the promoter driving expression of the <I>LEE1</I> operon in enteropathogenic <I>E. coli</I> and found two promoters separated by 10 bp, <I>LEE1</I> P1A (+1) and <I>LEE1</I> P1B (+10) using various <I>in vitro</I> biochemical tools. A unique feature of P1B was the presence of multiple transcription starts from five neighbouring As at the initial transcribed region. The multiple products did not arise from stuttering synthesis. Analytical software based on information theory was employed to determine promoter elements. The concentration of the NTP pool altered the preferred transcription start points, albeit the underlying mechanism is elusive. Under <I>in vivo</I> conditions, dominant P1B, but not P1A, was subject to regulation by IHF.</P>