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  • 1
    Book
    Book
    2012
    ISBN: 1869221796  ISBN: 9781869221799 
    Language: English
    Subject(s): Personnel management ; Manpower planning
    ISBN: 1869221796
    ISBN: 9781869221799
    Source: Ebook Central - Academic Complete
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  • 2
    Language: English
    In: Journal of experimental biology, 2018-09-25, Vol.221 (Pt 18), p.jeb185710
    Description: Optical imaging of gene expression by fluorescence hybridisation (FISH) in insects is often impeded by their pigmented cuticle. As most chemical bleaching agents are incompatible with FISH, we developed an RNA interference (RNAi)-based method for clearing cuticular pigmentation which enables the use of whole-mount body appendages for RNA FISH (termed RNA-i-FISH). Silencing or in two leaf beetles species ( and ) cleared their pigmented cuticle and decreased light absorbance. Subsequently, intact appendages (palps, antennae, legs) from RNAi-cleared individuals were used to image the expression and spatial distribution of antisense mRNA of two chemosensory genes encoding gustatory receptor and odorant-binding protein. Imaging did not work for RNAi controls because the pigmentation was retained, or for FISH controls (sense mRNA). Several bleaching agents were incompatible with FISH, because of 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 insect appendages.
    Subject(s): Index Medicus
    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): Prognosis ; Poly-ADP-Ribose Binding Proteins - metabolism ; Humans ; Lung Neoplasms - metabolism ; Ki-67 Antigen - metabolism ; Neuroendocrine Tumors - diagnosis ; GTPase-Activating Proteins - metabolism ; Carcinoid Tumor - metabolism ; Small Cell Lung Carcinoma - metabolism ; Neoplasm Grading ; Biomarkers, Tumor - metabolism ; Lung - radiation effects ; Carcinoma, Large Cell - genetics ; Gene Expression Regulation, Neoplastic - drug effects ; Lung Neoplasms - genetics ; DNA Topoisomerases, Type II - metabolism ; Lung - pathology ; Carcinoma, Large Cell - metabolism ; Carcinoma, Large Cell - diagnosis ; Neuroendocrine Tumors - metabolism ; Kaplan-Meier Estimate ; Poly-ADP-Ribose Binding Proteins - genetics ; Small Cell Lung Carcinoma - genetics ; Neuroendocrine Tumors - genetics ; Carcinoid Tumor - genetics ; Ki-67 Antigen - genetics ; Lung - drug effects ; Small Cell Lung Carcinoma - diagnosis ; DNA Topoisomerases, Type II - genetics ; Biomarkers, Tumor - genetics ; Gene Expression Regulation, Neoplastic - radiation effects ; GTPase-Activating Proteins - genetics ; Lung Neoplasms - diagnosis ; Neoplasm Staging ; Carcinoid Tumor - diagnosis ; Index Medicus
    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): Alkyl and Aryl Transferases - chemistry ; Amino Acid Sequence ; Diterpenes, Kaurane - metabolism ; Gas Chromatography-Mass Spectrometry ; Genes, Plant ; RNA, Messenger - genetics ; Alkyl and Aryl Transferases - metabolism ; Molecular Sequence Data ; Populus - genetics ; Phylogeny ; Magnesium - pharmacology ; RNA, Messenger - metabolism ; Sequence Homology, Amino Acid ; Amino Acids - metabolism ; Gene Expression Regulation, Plant - drug effects ; Populus - enzymology ; Substrate Specificity - drug effects ; Alkyl and Aryl Transferases - genetics ; Populus - drug effects ; Index Medicus
    ISSN: 1471-2229
    E-ISSN: 1471-2229
    Source: BioMedCentral Open Access
    Source: Academic Search Ultimate
    Source: PubMed Central
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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): Gibberellins ; Phytoalexin ; Genes
    ISSN: 1471-2229
    E-ISSN: 1471-2229
    Source: BioMedCentral Open Access
    Source: Academic Search Ultimate
    Source: PubMed Central
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  • 6
    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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  • 7
    Article
    Article
    1918
    ISSN: 0036-6773 
    Language: English
    In: School review, 1918-02, Vol.26 (2), p.140-140
    ISSN: 0036-6773
    E-ISSN: 1940-1213
    Source: JSTOR Arts & Sciences IV
    Source: JSTOR Early Journal Content
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  • 8
    Article
    Article
    1918
    ISSN: 0036-6773 
    Language: English
    In: School review, 1918-02, Vol.26 (2), p.140-140
    ISSN: 0036-6773
    E-ISSN: 1940-1213
    Source: JSTOR Arts & Sciences IV
    Source: JSTOR Early Journal Content
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  • 9
    Language: English
    In: School review, 1918-02, Vol.26 (2), p.138-138
    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.137-138
    ISSN: 0036-6773
    E-ISSN: 1940-1213
    Source: JSTOR Arts & Sciences IV
    Source: JSTOR Early Journal Content
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