Metabolism of the Cyanogenic Glucosides in Developing Flax: Metabolic Analysis, and Expression Pattern of Genes

Cyanogenic glucosides (CG), the monoglycosides linamarin and lotaustralin, as well as the diglucosides linustatin and neolinustatin, have been identified in flax. The roles of CG and hydrogen cyanide (HCN), specifically the product of their breakdown, differ and are understood only to a certain extent. HCN is toxic to aerobic organisms as a respiratory inhibitor and to enzymes containing heavy metals. On the other hand, CG and HCN are important factors in the plant defense system against herbivores, insects and pathogens. In this study, fluctuations in CG levels during flax growth and development (using UPLC) and the expression of genes encoding key enzymes for their metabolism (valine N-monooxygenase, linamarase, cyanoalanine nitrilase and cyanoalanine synthase) using RT-PCR were analyzed. Linola cultivar and transgenic plants characterized by increased levels of sulfur amino acids were analyzed. This enabled the demonstration of a significant relationship between the cyanide detoxification process and general metabolism. Cyanogenic glucosides are used as nitrogen-containing precursors for the synthesis of amino acids, proteins and amines. Therefore, they not only perform protective functions against herbivores but are general plant growth regulators, especially since changes in their level have been shown to be strongly correlated with significant stages of plant development.

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Strain-specific differences in age-related changes in rat susceptibility to experimental autoimmune encephalomyelitis and dendritic cell cytokine gene expression

Genetika ◽

10.2298/gensr1401287b ◽

2014 ◽

Vol 46 (1) ◽

pp. 287-301 ◽

Cited By ~ 6

Author(s):

Biljana Bufan ◽

Jasmina Djikic ◽

Mirjana Nacka-Aleksic ◽

Zorica Stojic-Vukanic ◽

Mirjana Dimitrijevic ◽

...

Keyword(s):

Dendritic Cell ◽

Rt Pcr ◽

Autoimmune Encephalomyelitis ◽

Expression Of Genes ◽

Age Related ◽

Genes Encoding ◽

Immunosuppressive Cytokines ◽

Age Related Changes ◽

Experimental Autoimmune

Experimental autoimmune encephalomyelitis (EAE) is an animal model of multiple sclerosis, a prototype of Th1/Th17-mediated organ-specific autoimmune disease. In the rat, susceptibility to development of these diseases is shown to be strain-and age-dependent. In adult rats of distinct strains, it correlates with splenic dendritic cell (DC) subset composition, which also exhibit age-related changes. The aim of this study was to examine influence of aging on: i) Albino Oxford (relatively resistant to EAE) and Dark Agouti (susceptible to EAE) rat development of EAE and ii) their splenic conventional (OX62+) DC population in respect to its subset composition and expression of mRNAs for proinflammatory and immunosuppressive cytokines. We used 3month-old (young) and 26-month-old (aged) rats of AO and DA strain. The rats were immunized for EAE with rat spinal cord homogenate in complete Freund?s adjuvant and clinical course of the disease was followed. Fresh OX62+DCs were examined for the expression of CD4 (using flow cytometry) and genes encoding cytokines influencing DC activation/maturation (TNF-? and IL-6) using RT-PCR. Additionally, in vitro lipopolysaccharide (LPS) activated/matured DCs were examined for the expression of genes encoding cytokines controlling Th1/Th17 cell polarization using RT-PCR. With aging, AO rats became more susceptible, whereas DA rats largely lose their susceptibility to the induction of EAE. In AO rats aging shifted CD4+:CD4DC ratio towards CD4-cells, producing large amount of proinflammatory cytokines, whereas in DA rats CD4+:CD4-DC ratio remained stable with aging. In fresh DCs from rats of both the strains the expression of TNF-? mRNA increased with aging, whereas that of IL-6 mRNA decreased and increased in DCs from AO and DA rats, respectively. Following in vitro LPS stimulation OX62+ DCs from aged AO rats up-regulated the expression of mRNA for IL-23p19 (specific subunit of IL-23; crucial for sustained IL-17 production) and IL-1? (positive IL-17 regulator), whereas down-regulated the expression of IL-10 (negative IL-17 regulator) when compared with young strain-matched rats. In DA rats aging incresed IL-23p19 mRNA expression in LPS-stimulated DCs, whereas exerted the opposing effects on the expression of mRNAs for IL-10 and IL-1? compared to AO rats. Irrespective of the rat strain, aging did not influence mRNA expression for IL-12p35 (driving Th1 polarization) in DCs. Overall, results suggest role of changes in the expression of genes encoding proinflammatory and immunosuppressive cytokines in development of age-related alterations in rat susceptibility to EAE induction.

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Frequency and expression of mutacin biosynthesis genes in isolates of Streptococcus mutans with different mutacin-producing phenotypes

Journal of Medical Microbiology ◽

10.1099/jmm.0.47749-0 ◽

2008 ◽

Vol 57 (5) ◽

pp. 626-635 ◽

Cited By ~ 16

Author(s):

Regianne Umeko Kamiya ◽

José Francisco Höfling ◽

Reginaldo Bruno Gonçalves

Keyword(s):

Streptococcus Mutans ◽

Staphylococcus Epidermidis ◽

High Frequency ◽

Phenotypic Diversity ◽

Genetic Determinants ◽

Rt Pcr ◽

Expression Of Genes ◽

Genes Encoding ◽

Spearman Correlation Test ◽

Expression Frequency

The aim of this study was to analyse the frequency and expression of biosynthesis genes in 47 Streptococcus mutans isolates with different mutacin-producing phenotypes. Detection of the frequency and expression of genes encoding mutacin types I, II, III and IV were carried out by PCR and semi-quantitative RT-PCR, respectively, using primers specific for each type of biosynthesis gene. In addition, a further eight genes encoding putative bacteriocins, designated bsm 283, bsm 299, bsm 423, bsm 1889c, bsm 1892c, bsm 1896, bsm 1906c and bsm 1914, were also screened. There was a high phenotypic diversity; some Streptococcus mutans isolates presented broad antimicrobial spectra against other Streptococcus mutans clinical isolates, including bacteria resistant to common antibiotics, as well as Staphylococcus aureus, Staphylococcus epidermidis, Enterococcus faecalis and Streptococcus pyogenes. The expression frequency of the bsm gene was higher than that of the previously characterized mutacins (I–IV). There was no positive correlation between the number of indicator strains inhibited (antimicrobial spectra) and the number of biosynthesis genes expressed (Spearman correlation test, r=−0.03, P>0.05). In conclusion, the high diversity of mutacin-producing phenotypes, associated with high frequency of expression of the biosynthesis genes screened, reveals a broad repertoire of genetic determinants encoding antimicrobial peptides that can act in different combinations.

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Primary structure, expression and localization of two intermediate subunit lectins ofEntamoeba disparthat contain multiple CXXC motifs

Parasitology ◽

10.1017/s0031182007003459 ◽

2007 ◽

Vol 134 (14) ◽

pp. 1989-1999 ◽

Cited By ~ 7

Author(s):

H. TACHIBANA ◽

X.-J. CHENG ◽

S. KOBAYASHI ◽

Y. OKADA ◽

J. ITOH ◽

...

Keyword(s):

Amino Acids ◽

Amino Acid ◽

Confocal Microscopy ◽

Phenotypic Expression ◽

Surface Proteins ◽

Rt Pcr ◽

Entamoeba Dispar ◽

Genes Encoding ◽

Sequence Identities

SUMMARYWe have recently identified 2 surface proteins inEntamoeba histolyticaas intermediate subunits of galactose- andN-acetyl-D-galactosamine-inhibitable lectin (EhIgl1 and EhIgl2); these proteins both contain multiple CXXC motifs. Here, we report the molecular characterization of the corresponding proteins inEntamoeba dispar, which is neither pathogenic nor invasive. TwoIglgenes encoding 1110 and 1106 amino acids (EdIgl1 and EdIgl2) were cloned from 2 strains ofE. dispar. The amino acid sequence identities were 79% between EdIgl1 and EdIgl2, 75–76% between EdIgl1 and EhIgl1, and 73–74% between EdIgl2 and EhIgl2. However, all the CXXC motifs were conserved in the EdIgl proteins, suggesting that the fold conferred by this motif is important for function. Comparison of the expression level of theIglgenes by real-time RT-PCR showed 3–5 times higher expression ofEdIgl1compared toEdIgl2. Most EdIgl1 and EdIgl2 proteins were co-localized on the surface and in the cytoplasm of trophozoites, based on confocal microscopy. However, a different localization of EdIgl1 and EdIgl2 in intracellular vacuoles and a different level of phenotypic expression of the two Igls were also observed. These results demonstrate that Igls are important proteins even in non-pathogenic amoeba and that Igl1 and Igl2 may possess different functions.

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The effect of indole-3-carbinol on the expression of CYP1A1, CYP1B1 and AhR genes and proliferation of MCF-7 cells.

Acta Biochimica Polonica ◽

10.18388/abp.2007_3276 ◽

2007 ◽

Vol 54 (1) ◽

pp. 113-117 ◽

Cited By ~ 15

Author(s):

Marta Ociepa-Zawal ◽

Błazej Rubiś ◽

Mariusz Laciński ◽

Wiesław H Trzeciak

Keyword(s):

Breast Cancer ◽

Cell Cycle ◽

Breast Cancer Cell ◽

Pcr Analysis ◽

Estrogen Metabolism ◽

Rt Pcr ◽

P21 Protein ◽

Expression Of Genes ◽

Genes Encoding ◽

Mcf 7

The influence of an antiestrogen, indole-3-carbinol (I3C) on the expression of CYP1A1, CYP1B1 and AhR genes was investigated in an attempt to establish whether I3C could increase the expression of genes involved in estrone metabolism. Another purpose was to examine the proliferation of an estrogen-dependent breast cancer cell (MCF-7 line) under the influence of I3C and both I3C and DDT. In MCF-7 cells incubated with I3C or I3C and DDT combined, quantitative RT-PCR analysis revealed a significant increase in the level of CYP1A1, AhR, and CYP1B1 transcripts. The proliferation rate of MCF-7 cells was increased by treatment with DDT or estradiol (E2), whereas I3C did not affect the proliferation of MCF-7 cells but greatly reduced the stimulatory effect of DDT, and abolished the effect of E2. The level of p21 transcript, encoding p21 protein involved in the cell cycle, was increased several-fold by I3C comparing to its level in cells incubated with estradiol or DDT. The results suggest that the proliferation of MCF-7 cells is accompanied not only by expression of genes encoding cytochromes involved in estrogen metabolism, but also by changes in the expression of other genes including that encoding p21 protein involved in the cell cycle.

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Dependence of expression of genes controlling calcium signaling on gravistimulation in cells of tomato apical leaves and treatment by etephon

Doklady of the National Academy of Sciences of Belarus ◽

10.29235/1561-8323-2021-65-4-456-465 ◽

2021 ◽

Vol 65 (4) ◽

pp. 456-465

Author(s):

S. V. Sukhaveyeva ◽

A. М. Kаbachevskaya ◽

I. D. Volotovski

Keyword(s):

Signal Transduction ◽

Calcium Signaling ◽

Calcium Metabolism ◽

Early Response ◽

Calcium Signal ◽

Rt Pcr ◽

Expression Of Genes ◽

Genes Encoding ◽

Exogenous Ethylene

The sensitivity of expression at the level of transcription of genes encoding proteins involved in calcium signal transduction to gravistimulation was revealed using real-time RT-PCR. For three genes SCA2, РВР2, САМ2, the increase in the transcript formation was shown at early response stages, starting from 15–60 minute gravistimulus. The treatment of plants before the start of gravistimulation with an ethephon (source of exogenous ethylene) led to a change in the modulation of expression of the studied genes in response to gravistimulus. The role of calcium metabolism in realization of final steps of gravitropism reaction is considered.

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Contribution of OmpK36 to carbapenem susceptibility in KPC-producing Klebsiella pneumoniae

Journal of Medical Microbiology ◽

10.1099/jmm.0.012575-0 ◽

2009 ◽

Vol 58 (10) ◽

pp. 1303-1308 ◽

Cited By ~ 78

Author(s):

David Landman ◽

Simona Bratu ◽

John Quale

Keyword(s):

New York ◽

Klebsiella Pneumoniae ◽

Efflux Pump ◽

Ionization Mass ◽

Rt Pcr ◽

Clonal Group ◽

Expression Of Genes ◽

Sds Page ◽

Genes Encoding ◽

Zones Of Inhibition

Isolates of Klebsiella pneumoniae harbouring the carbapenemase KPC may have carbapenem MICs that remain in the susceptible range, and may therefore go unrecognized. To understand the mechanisms contributing to the variability in carbapenem MICs, 20 clinical isolates, all belonging to either of two clonal groups of KPC-possessing K. pneumoniae endemic to New York City, were examined. Expression of genes encoding KPC, the porins OmpK35 and OmpK36, and the efflux pump AcrAB was examined by real-time RT-PCR. Outer-membrane profiles of selected KPC-producing isolates were examined by SDS-PAGE, and proteins were identified by matrix-assisted laser desorption/ionization mass spectrometry. The identification of SHV and TEM β-lactamases and the genomic sequences of ompK35 and ompK36 were determined by PCR and DNA sequencing, respectively. For one clonal group, carbapenem MICs increased with decreasing expression of ompK36. A second clonal group also had carbapenem MICs that correlated with ompK36 expression. However, all of the isolates in this latter group continued to produce OmpK36, suggesting that porin configuration may affect entry of carbapenems. For isolates that had the greatest expression of ompK36, carbapenem MICs tended to be lower when determined by the broth microdilution technique, and scattered colonies were seen around the Etest zones of inhibition. All of the KPC-producing isolates were highly resistant to ertapenem, regardless of ompK36 expression. In conclusion, isolates of KPC-possessing K. pneumoniae that express ompK36 tend to have lower MICs to carbapenems and therefore may be more difficult to detect by clinical laboratories. Regardless of ompK36 expression, all of the KPC producers were consistently resistant to ertapenem.

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Characterization of Two Newly Identified Genes, vgaD and vatG, Conferring Resistance to Streptogramin A in Enterococcus faecium

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.00798-09 ◽

2010 ◽

Vol 54 (11) ◽

pp. 4744-4749 ◽

Cited By ~ 16

Author(s):

Young-Hee Jung ◽

Eun Shim Shin ◽

Okgene Kim ◽

Jung Sik Yoo ◽

Kyeong Min Lee ◽

...

Keyword(s):

Amino Acids ◽

Resistance Genes ◽

Enterococcus Faecium ◽

Efflux Pump ◽

Open Reading Frames ◽

Open Reading Frame ◽

Strongly Correlated ◽

Reading Frame ◽

Genes Encoding ◽

New Gene

ABSTRACT We characterized two new streptogramin A resistance genes from quinupristin-dalfopristin-resistant Enterococcus faecium JS79, which was selected from 79 E. faecium isolates lacking known genes encoding streptogramin A acetyltransferase. A 5,650-bp fragment of HindIII-digested plasmid DNA from E. faecium JS79 was cloned and sequenced. The fragment contained two open reading frames carrying resistance genes related to streptogramin A, namely, genes for an acetyltransferase and an ATP efflux pump. The first open reading frame comprised 648 bp encoding 216 amino acids with a predicted left-handed parallel β-helix domain structure; this new gene was designated vatG. The second open reading frame consisted of 1,575 bp encoding 525 amino acids with two predicted ATPase binding cassette transporters comprised of Walker A, Walker B, and LSSG motifs; this gene was designated vgaD. vgaD is located 65 bp upstream from vatG, was detected together with vatG in 12 of 179 quinupristin-dalfopristin-resistant E. faecium isolates, and was located on the same plasmid. Also, the 5.6-kb HindIII-digested fragment which was observed in JS79 was detected in nine vgaD- and vatG-containing E. faecium isolates by Southern hybridization. Therefore, it was expected that these two genes were strongly correlated with each other and that they may be composed of a transposon. Importantly, vgaD is the first identified ABC transporter conferring resistance to streptogramin A in E. faecium. Pulsed-field gel electrophoresis patterns and sequence types of vgaD- and vatG-containing E. faecium isolates differed for isolates from humans and nonhumans.

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