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  • 1
    Book
    Book
    2012
    ISBN: 1869221796  ISBN: 9781869221799 
    Language: English
    Subject(s): Manpower planning ; Personnel management
    ISBN: 1869221796
    ISBN: 9781869221799
    Source: Ebook Central - Academic Complete
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  • 2
    Language: English
    In: Journal of experimental biology, 2018-07-19, Vol.221
    Description: Optical imaging of gene expression by fluorescence hybridisation (FISH) in insects is often impeded by their pigmented cuticle. Since most chemical bleaching agents are incompatible with FISH, we developed a RNA interference-based method for clearing cuticular pigmentation which enables using whole-mount body appendages for RNA FISH. Silencing or in two leaf beetles species ( ) cleared their pigmented cuticle and decreased light absorbance. Subsequently, intact appendages (palps, antennae, legs) from RNAi-cleared individuals were used to image expression and spatial distribution of antisense mRNA of two chemosensory genes (gustatory receptor, odorant-binding protein). Imaging did neither work for RNAi-controls due to retained pigmentation, nor for FISH-controls (sense mRNA). Several bleaching agents were incompatible with FISH, either due to degradation of RNA, lack of clearing efficacy or long incubation times. Overall, silencing pigmentation genes is a significant improvement over bleaching agents enabling FISH in intact appendages.
    ISSN: 0022-0949
    E-ISSN: 1477-9145
    Source: HighWire Press (Free Journals)
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  • 3
    Language: English
    In: Oncotarget, 2016-07-05, Vol.7 (27), p.41959-41973
    Description: The classification of bronchopulmonary neuroendocrine neoplasms (BP-NEN) into four tumor entities (typical carcinoids (TC), atypical carcinoids (AC), small cell lung cancers (SCLC), large cell neuroendocrine lung carcinomas (LCNEC)) is difficult to perform accurately, but important for prognostic statements and therapeutic management decisions. In this regard, we compared the expression of three proliferation markers, Ki-67, Topoisomerase II alpha (TOP2A), and RacGAP1, in a series of tumor samples from 104 BP-NEN patients (24 TC, 21 AC, 52 SCLC, 7 LCNEC) using different evaluation methods (immunohistochemistry (IHC): Average evaluation, Hotspot evaluation, digital image analysis; RT-qPCR).The results indicated that all three markers had increased protein and mRNA expression with poorer differentiation and correlated well with each other, as well as with grading, staging, and poor survival. Compared with Ki-67 and TOP2A, RacGAP1 allowed for a clearer prognostic statement. The cut-off limits obtained for Ki-67-Average (IHC) were TC-AC 1.5, AC-SCLC 19, and AC-LCNEC 23.5. The Hotspot evaluation generated equal to higher, the digital image analysis generally lower between-entity cut-off limits.All three markers enabled a clear-cut differentiation between the BP-NEN entities, and all methods evaluated were suitable for marker assessment. However, to define optimal cut-off limits, the Ki-67 evaluation methods should be standardized. RacGAP1 appeared to be a new marker with great potential.
    Subject(s): Biomarkers, Tumor - genetics ; Biomarkers, Tumor - metabolism ; Carcinoid Tumor - diagnosis ; Carcinoid Tumor - genetics ; Carcinoid Tumor - metabolism ; Carcinoma, Large Cell - diagnosis ; Carcinoma, Large Cell - genetics ; Carcinoma, Large Cell - metabolism ; DNA Topoisomerases, Type II - genetics ; DNA Topoisomerases, Type II - metabolism ; Gene Expression Regulation, Neoplastic - drug effects ; Gene Expression Regulation, Neoplastic - radiation effects ; GTPase-Activating Proteins - genetics ; GTPase-Activating Proteins - metabolism ; Humans ; Kaplan-Meier Estimate ; Ki-67 Antigen - genetics ; Ki-67 Antigen - metabolism ; Lung - drug effects ; Lung - pathology ; Lung - radiation effects ; Lung Neoplasms - diagnosis ; Lung Neoplasms - genetics ; Lung Neoplasms - metabolism ; Neoplasm Grading ; Neoplasm Staging ; Neuroendocrine Tumors - diagnosis ; Neuroendocrine Tumors - genetics ; Neuroendocrine Tumors - metabolism ; Poly-ADP-Ribose Binding Proteins - genetics ; Poly-ADP-Ribose Binding Proteins - metabolism ; Prognosis ; Small Cell Lung Carcinoma - diagnosis ; Small Cell Lung Carcinoma - genetics ; Small Cell Lung Carcinoma - metabolism
    E-ISSN: 1949-2553
    Source: PubMed Central
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  • 4
    Language: English
    In: BMC plant biology, 2015-10-28, Vol.15 (1), p.262-262
    Description: Labdane-related diterpenoids form the largest group among the diterpenes. They fulfill important functions in primary metabolism as essential plant growth hormones and are known to function in secondary metabolism as, for example, phytoalexins. The biosynthesis of labdane-related diterpenes is mediated by the action of class II and class I diterpene synthases. Although terpene synthases have been well investigated in poplar, little is known about diterpene formation in this woody perennial plant species. The recently sequenced genome of Populus trichocarpa possesses two putative copalyl diphosphate synthase genes (CPS, class II) and two putative kaurene synthase genes (KS, class I), which most likely arose through a genome duplication and a recent tandem gene duplication, respectively. We showed that the CPS-like gene PtTPS17 encodes an ent-copalyl diphosphate synthase (ent-CPS), while the protein encoded by the putative CPS gene PtTPS18 showed no enzymatic activity. The putative kaurene synthases PtTPS19 and PtTPS20 both accepted ent-copalyl diphosphate (ent-CPP) as substrate. However, despite their high sequence similarity, they produced different diterpene products. While PtTPS19 formed exclusively ent-kaurene, PtTPS20 generated mainly the diterpene alcohol, 16α-hydroxy-ent-kaurane. Using homology-based structure modeling and site-directed mutagenesis, we demonstrated that one amino acid residue determines the different product specificity of PtTPS19 and PtTPS20. A reciprocal exchange of methionine 607 and threonine 607 in the active sites of PtTPS19 and PtTPS20, respectively, led to a complete interconversion of the enzyme product profiles. Gene expression analysis revealed that the diterpene synthase genes characterized showed organ-specific expression with the highest abundance of PtTPS17 and PtTPS20 transcripts in poplar roots. The poplar diterpene synthases PtTPS17, PtTPS19, and PtTPS20 contribute to the production of ent-kaurene and 16α-hydroxy-ent-kaurane in poplar. While ent-kaurene most likely serves as the universal precursor for gibberellins, the function of 16α-hydroxy-ent-kaurane in poplar is not known yet. However, the high expression levels of PtTPS20 and PtTPS17 in poplar roots may indicate an important function of 16α-hydroxy-ent-kaurane in secondary metabolism in this plant organ.
    Subject(s): 16α-hydroxy-ent-kaurane ; Alkyl and Aryl Transferases - chemistry ; Alkyl and Aryl Transferases - genetics ; Alkyl and Aryl Transferases - metabolism ; Amino Acid Sequence ; Amino Acids - metabolism ; Diterpene synthases ; Diterpenes, Kaurane - metabolism ; Ent-kaurene ; Gas Chromatography-Mass Spectrometry ; Gene duplication ; Gene Expression Regulation, Plant - drug effects ; Genes, Plant ; Gibberellin biosynthesis ; Magnesium - pharmacology ; Molecular Sequence Data ; Phylogeny ; Populus - drug effects ; Populus - enzymology ; Populus - genetics ; Populus trichocarpa ; RNA, Messenger - genetics ; RNA, Messenger - metabolism ; Sequence Homology, Amino Acid ; Substrate Specificity - drug effects
    ISSN: 1471-2229
    E-ISSN: 1471-2229
    Source: BioMedCentral Open Access
    Source: Academic Search Ultimate
    Source: PubMed Central
    Source: DOAJ Directory of Open Access Journals - Not for CDI Discovery
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  • 5
    Language: English
    In: BMC plant biology, 2015-10-28, Vol.15 (1)
    Description: Labdane-related diterpenoids form the largest group among the diterpenes. They fulfill important functions in primary metabolism as essential plant growth hormones and are known to function in secondary metabolism as, for example, phytoalexins. The biosynthesis of labdane-related diterpenes is mediated by the action of class II and class I diterpene synthases. Although terpene synthases have been well investigated in poplar, little is known about diterpene formation in this woody perennial plant species. The recently sequenced genome of Populus trichocarpa possesses two putative copalyl diphosphate synthase genes (CPS, class II) and two putative kaurene synthase genes (KS, class I), which most likely arose through a genome duplication and a recent tandem gene duplication, respectively. We showed that the CPS-like gene PtTPS17 encodes an ent-copalyl diphosphate synthase (ent-CPS), while the protein encoded by the putative CPS gene PtTPS18 showed no enzymatic activity. The putative kaurene synthases PtTPS19 and PtTPS20 both accepted ent-copalyl diphosphate (ent-CPP) as substrate. However, despite their high sequence similarity, they produced different diterpene products. While PtTPS19 formed exclusively ent-kaurene, PtTPS20 generated mainly the diterpene alcohol, 16[alpha]-hydroxy-ent-kaurane. Using homology-based structure modeling and site-directed mutagenesis, we demonstrated that one amino acid residue determines the different product specificity of PtTPS19 and PtTPS20. A reciprocal exchange of methionine 607 and threonine 607 in the active sites of PtTPS19 and PtTPS20, respectively, led to a complete interconversion of the enzyme product profiles. Gene expression analysis revealed that the diterpene synthase genes characterized showed organ-specific expression with the highest abundance of PtTPS17 and PtTPS20 transcripts in poplar roots. The poplar diterpene synthases PtTPS17, PtTPS19, and PtTPS20 contribute to the production of ent-kaurene and 16[alpha]-hydroxy-ent-kaurane in poplar. While ent-kaurene most likely serves as the universal precursor for gibberellins, the function of 16[alpha]-hydroxy-ent-kaurane in poplar is not known yet. However, the high expression levels of PtTPS20 and PtTPS17 in poplar roots may indicate an important function of 16[alpha]-hydroxy-ent-kaurane in secondary metabolism in this plant organ.
    Subject(s): Genes ; Gibberellins ; Phytoalexin
    ISSN: 1471-2229
    E-ISSN: 1471-2229
    Source: DOAJ Directory of Open Access Journals - Not for CDI Discovery
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  • 6
    Dissertation
    Dissertation
    2017
    ISBN: 9798708788191 
    Language: English
    Description: Microorganisms, such as bacteria and fungi produce odors, also known as volatile organic compounds (VOCs). VOCs are small, carbon-containing molecules that exhibit high vapor pressure under ambient conditions and have low boiling points. Especially in soil and in the rhizosphere, these unique properties provide the advantage to travel over long distances, mediating interaction and communication amongst physically separated microorganisms. In this thesis, we zoom for the first time into the mechanisms of volatile mediated fungalbacterial interactions. For this, in vitro bioassays were combined with “omic” approaches, such as genomics, transcriptomics, proteomics and metabolomics. Whilst so far most attention has been paid to bacterial VOCs, the first step of the thesis was to investigate the diversity and ecological role of fungal VOCs. Comparing the “volatilome” of a range of soil/rhizospheric fungi and oomycete revealed that each microbe has its own chemical signature and that the growth stage and the nutritional status (rich vs. poor media) have a strong effect on VOC emission. This makes it tempting to speculate that filamentous microbes adjust the production of VOCs for interactions and communication with other microbes depending on the nutrient status. Linking the individual “volatilomes” with their effects on the behavior of bacteria showed that in particular terpene volatiles of the fungus F. culmorum affect the motility of two bacterial isolates, Collimonas pratensis and Serratia plymuthica. Curiosity about the underlying mechanisms of VOC perception and responses lead to the next step in which we analyzed the transcriptome and proteome of S. plymuthica when exposed to VOCs emitted by the fungal plant pathogen F. culmorum. We found that the bacterium responds to fungal VOCs with changes in gene and protein expression related to motility, signal transduction, energy metabolism, cell envelope biogenesis, and secondary metabolite production. Metabolomic analysis of the bacterium exposed to the fungal VOCs and heterologous co-expression of a terpene synthase and a methyltransferase revealed the production of the unusual terpene sodorifen in response to fungal VOCs. These results lead to the proposition that terpenes are a lingua franca in long-distance communication between bacteria and fungi. However, it remains to be elucidated in future studies what the ecological roles of sodorifen are in respect of the context of microbe-microbe and microbe-plant interactions. Intrigued by the newly discovered “language”, we went on a quest for more secret terpene producers in the belowground world by comparing the genomes of several Collimonas species. In this way we identified two terpene synthase genes that encode for the production of germacrene D-4-ol by, and several sesquiterpens, including δ-cadinene. Not only bacteria are a rich source of terpenes, but fungi are promising producers too. Thus, we sequenced the fungal genome of our previous studies, Fusarium culmorum, to look for genes encoding for terpenes and other secondary metabolites that play important roles in fungal pathogenicity and in the interaction with other microorganisms. Searching through the genome we identified two terpene synthase genes, longiborneol synthase and trichodiene synthase that encode for the production of a whole array of terpenes. We propose that there may be many new, undiscovered terpenes hiding in the genomes of bacteria and fungi. However, also this quest remains to be continued in future studies. The obtained results of this thesis set the basis for further understanding the natural roles of terpene-mediated microbe-microbe as well as microbe-plant interactions. Ultimately, this knowledge can be translated into innovative strategies for developments towards ecofriendly and sustainable agricultural practices.
    Subject(s): Genomics ; Metabolites ; Plant sciences ; Soil sciences ; Sulfur ; Volatile organic compounds--VOCs
    ISBN: 9798708788191
    Source: ProQuest Dissertations & Theses Global
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  • 7
    Book chapter
    Book chapter
    2012
    ISBN: 1869222075  ISBN: 9781869222079  ISBN: 9781869221799  ISBN: 1869221796 
    Language: English
    In: Workforce Planning Toolkit, 2012-01-01
    Subject(s): Business conditions ; Economic conditions ; Human resource management ; Leadership ; Manpower planning ; Personnel management
    ISBN: 1869222075
    ISBN: 9781869222079
    ISBN: 9781869221799
    ISBN: 1869221796
    Source: Ebook Central - Academic Complete
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  • 8
    Article
    Article
    1918
    ISSN: 0036-6773 
    Language: English
    In: School review, 1918-02, Vol.26 (2), p.141-142
    ISSN: 0036-6773
    E-ISSN: 1940-1213
    Source: JSTOR Arts & Sciences IV
    Source: JSTOR Early Journal Content
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  • 9
    Article
    Article
    1918
    ISSN: 0036-6773 
    Language: English
    In: School review, 1918-02, Vol.26 (2), p.137-138
    ISSN: 0036-6773
    E-ISSN: 1940-1213
    Source: JSTOR Arts & Sciences IV
    Source: JSTOR Early Journal Content
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  • 10
    Article
    Article
    1918
    ISSN: 0036-6773 
    Language: English
    In: School review, 1918-02, Vol.26 (2), p.141-141
    ISSN: 0036-6773
    E-ISSN: 1940-1213
    Source: JSTOR Arts & Sciences IV
    Source: JSTOR Early Journal Content
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