117,965 research outputs found
Podocyte incorporation of GEnC derived EV.
No full textA) Illustration of the transwell culture of GEnC and podocytes, allowing non-contact transfer of EV. Cells are cultured in exosome free FBS. B) Representative images of podocyte EV uptake post 24-hour culture with GEnC derived EV. Podocytes remained untreated or cultured with EV from GEnC exposed to FM1-43X, with or without LPS activation. GEnC were washed prior to seeding onto transwell filters, to remove any unbound dye. Confocal images were acquired on a Leica SP5 confocal microscope. Scale bar represents 10μm. Arrows indicated FM1-43X positive GEnC EV (green). Blue = podocyte nuclei C) Median fluorescence intensity in TW (transwell) treated podocytes with EV from steady state GEnC or LPS activated GEnC. Fluorescence was measured by flow cytometry. Backdrop suppressor was added to podocytes to remove any extracellular dye/signal. n = 3 D) Illustration of GEnC transfected with a C. elegans miRNA mimic using lipofectamine. GEnC were cultured (top) on to a 0.4μm transwell filter and cultured with podocytes (bottom) for 24 hours. RNAseA was added to the podocytes to remove any free extracellular RNA, prior to podocyte RNA isolation. E) Fold change (from untreated non transfected GEnC–UT) in C. elegans miRNA in TW (transwell) treated podocytes with EV from steady state GEnC or LPS activated GEnC. n = 4 ** represents P<0.01.</p
Characterisation of GEnC derived EV.
No full textA) Illustration of the UC technique for EV isolation. Briefly, GEnC cultures were supplemented with exosome free FBS and media collected 24 hour post stimulation with glucose, LPS or PAN, or GEnC were kept at steady state (control). Successive centrifugation steps at 300g, 2,000g and 10,000g removed cells, cell debris and microvesicles. UC was performed twice at 100,000g to precipitate a pure EV fraction. B) Snapshot of steady state GEnC derived EV as visualised by Nanosight 405nm excitation. Arrows indicated vesicles. Scale bar represents 1μm. C) Representative size distribution and concentration of EV isolated from steady state and LPS treated GEnC via UC, as performed on the Nanosight LM10 instrument. D). The concentration and E) size of vesicles of from steady state, glucose, LPS and PAN GEnC derived EV n = 5. F) RNA and G) protein concentration of steady state, glucose, LPS and PAN derived GEnC EV n = 5. * represents PH) Protein western blots of exosomes derived from SS (steady state), GLU (glucose), LPS or PAN treated GEnC using antibodies for GAPDH, CD63, GRP94, and TSG101 performed under reducing conditions.</p
Measurement of OCR of podocytes stimulated with steady state, glucose, LPS or PAN derived GEnC EV.
No full textA) Podocytes remained untreated or were treated with steady state GEnC EV, glucose treated GEnC EV, LPS treated GEnC E or PAN treated GEnC EV for 24 hours. OCR (oxygen consumption rate) was measured in x96 seahorse extracellular flux analyser. Oligomycin (1μm), FCCP (2μM), and antimycin A/rotenone (0.5μM) were injected at indicated time points. n = 6. B) Baseline OCR for untreated podocytes or podocytes treated with steady state GEnC EV, glucose treated GEnC EV, LPS treated GEnC EV or PAN treated GEnC EV for 24 hours. n = 6. C) Baseline ATP production for untreated podocytes or podocytes treated with steady state GEnC EV, glucose treated GEnC EV, LPS treated GEnC EV or PAN treated GEnC EV for 24 hours. n = 6. D) ECAR profile over time with same treatments as in A) with injection of 1μM oligomycin and 50mM 2-deoxyglucose (2- DG) as indicated. n = 6. E) Glycoloysis measurements at baseline after podocoyte treatments with EV or at baseline. n = 6. F) Representative images of mitochondrial activity using 100nM MitoTracker (red) and nuclei counterstained with DAPI (blue) detected with fluorescence microscopy. Scale bar = 25 μm. G) Quantitative mitochondrial activation with 100nM MitoTracker after podocyte incubation with EV from steady state GEnC, glucose treated GEnC, LPS treated GEnC or PAN treated GEnC. Each dot represents 1 mitochondrion from n = 3 experiments. H) Flow cytometry measuring podocyte intracellular ROS levels using 5μM mitoSOX after podocyte incubation with EV from steady state GEnC, glucose treated GEnC, LPS treated GEnC or PAN treated GEnC. n = 3. *, ***, **** represents P<0.05, P<0.001 or P<0.0001 respectively, groups were compared using a one-way Anova.</p
Extracellular vesicles derived from activated GEnC mediate podocyte dysfunction—A role for miRNA-200c-3p.
No full textA) Podocytes were treated for 24 hrs with EV from steady state GEnC, glucose treated GEnC, LPS treated GEnC or PAN treated GEnC and VEGF secretion measured by ELISA. Presented as change from untreated podocytes (steady state). n = 6. B) Representative images of actin microfilaments detected using phalloidin staining with fluorescence microscopy. Nuclei were stained with DAPI. Podocytes were treated for 24 hrs with EV from steady state GEnC, glucose treated GEnC, LPS treated GEnC or PAN treated GEnC. Scale bar represents 25μm. Arrows indicate stress fibres during steady state. C) Increased expression of miRNA-200c-3p in podocytes treated with EV derived from GEnC treated with glucose or PAN versus podocytes treated with steady state GEnC derived EV (from miRNA sequencing) D) Effect of miR-200c-3p transfection on podocyte VEGF sprotein expression as measured by ELISA. Negative control mimic was used as a control. n = 3. *, ** and *** represents P<0.05, P<0.01,P<0.001 respectively, groups were compared using an unpaired t-test.</p
GLU, LPS and PAN GEnC EV alter mRNA and miRNA expression in podocytes.
No full textA) Volcano plots comparing podocyte mRNA fold change in UT (untreated GEnC derived EV) vs SS (steady state podocytes), SS vs GLU (glucose treated GEnC derived EV) and SS vs PAN (PAN treated GEnC derived EV). Change in miRNA expression represented as blue and red spots for significantly upregulated and downregulated miRNA respectively. Significance calculated as fold change >1.5 and PB) Venn diagram of overlapping or treatment specific significant (fold change >1.5 and PC) GO analysis of pathways enriched from mRNA sequencing analysis (see Methods). Bar plot illustrating GO associations upregulated from SS vs GLU and SS vs PAN treatments. D) Volcano plots comparing podocyte miRNA fold change in UT (untreated GEnC derived EV) vs SS (steady state podocytes), SS vs GLU (glucose treated GEnC derived EV), SS vs PAN (PAN treated GEnC derived EV) and SS vs LPS (LPS treated GEnC derived EV). Change in miRNA expression represented as blue and red spots for significantly upregulated and downregulated miRNA respectively. Significance calculated as fold change >1.5 and PE) Venn diagram of overlapping or treatment specific significant (fold change >1.5 and PF) GO analysis of pathways enriched from mRNA sequencing analysis (see Methods). Bar plot illustrating GO associations upregulated from SS vs GLU and SS vs PAN treatments.</p
Screening for zinc efficiency in barley (Hordeum vulgare L.) / Yusuf Genc.
No full textBibliography: leaves 229-250.xxi, 250 leaves : ill. (chiefly col.) ; 30 cm.The aims of the study were to develop a reliable method for screening for Zn efficiency as an alternative to the current field-based methods, and to determine the extent of genotypic variation in tolerance to Zn deficiency in barley.Thesis (Ph.D.)--University of Adelaide, Dept. of Plant Science, 199
Going Beyond Counting First Authors in Author Co-citation Analysis
No full textThe present study examines one of the fundamental aspects of author co-citation analysis (ACA) - the way co-citation
counts are defined. Co-citation counting provides the data on which all subsequent statistical analyses and mappings
are based, and we compare ACA results based on two different types of co-citation counting - the traditional type that
only counts the first one among a cited work's authors on the one hand and a non-traditional type that takes into
account the first 5 authors of a cited work on the other hand. Results indicate that the picture produced through this non-traditional author co-citation counting contains more coherent author groups and is therefore considerably clearer. However, this picture represents fewer specialties in the research field being studied than that produced through the traditional first-author co-citation counting when the same number of top-ranked authors is selected and analyzed. Reasons for these effects are discussed
Screening of Organophosphate Resistance in the Acetycholinesterase Gene of Field Collected Olive Fruit Fly, Bactrocera Oleae Rossi (Diptera: Tephritidae)
No full textOlives infested with the olive fruit fly were collected by monitoring olive orchards in Canakkale area during 2006 and 2007, and in 2013 to study the level of organophosphate insecticide resistance in the acetyl cholinesterase gene. Single fly was used for genomic DNA isolation. All genomic DNA's were monitored by PCR-RFLP in the presence of resistance alleles. The assay was used to test the Glycine488Serine resistance mutation occurred in exon VI. The technique was established previously by mutagenesis due to introduced reverse primer that just reaches codon 488, creating Mbi I restriction site. The specimens screened in 2006 carried a resistance mutation (31.70%), and heterozygote mutation (65.21%), while only 3.10% of the specimens were organophosphate susceptible. The specimens screened in 2007carried a resistance mutation (54.14%), and heterozygote mutation (44.75%), while 1.10% of the specimens were susceptible. The specimens screened in 2013 carried a resistance mutation (81.77%) and heterozygote mutation (18.23%). There were no homozygous susceptible individuals found. This study presented the status and frequency of acetyl cholinesterase point mutation in the field collected olive flies against used organophosphate insecticides.Scientific and Technical Research Council of Turkey, TUBITAK [105 O 706, Greece 877]This research was financially supported by The Scientific and Technical Research Council of Turkey, TUBITAK, (Projects Grant No: 105 O 706 and Greece 877). The author thanks Dr. George Skavdis and Dr. John Vontas for establishing the PCR-RFLP assay in our laboratory and Dr. Kemal Melik Taskin for providing support. The author thanks Prof. Dr. James L. Nation for useful suggestions on an earlier version of the manuscript and Prof. Dr. Levent Genc for help in maping collection sites, as well as the owners of the respective olive orchards in Canakkale province. The part of this study was published as an abstract in the 3rd Plant Protection Congress, July 15-18, 2009 in Van, Turkey
Square Dancing with the Stars to Enhance Dynamic Hirschman Linkages?
No full textIn this Presidential Address, the author takes the reader on a reconnaissance of his life and time as a regional scientist. He points out scenery he found scintillating along the way, hoping that some may pick up the banner and chew on a few of the ideas for a while. He suggests a revisit to Albert O. Hirschman’s notion of key sectors and more empirical analysis related to Marcus Berliant’s and Masahisa Fujita’s notion of knowledge creation and transfer.Presidential Address, San Antonio, Texas, March 29, 2014 (53rd Meetings of the Southern Regional Science Association
Appropriate Similarity Measures for Author Cocitation Analysis
No full textWe provide a number of new insights into the methodological discussion about author cocitation analysis. We first argue that the use of the Pearson correlation for measuring the similarity between authors’ cocitation profiles is not very satisfactory. We then discuss what kind of similarity measures may be used as an alternative to the Pearson correlation. We consider three similarity measures in particular. One is the well-known cosine. The other two similarity measures have not been used before in the bibliometric literature. Finally, we show by means of an example that our findings have a high practical relevance.information science;Pearson correlation;cosine;similarity measure;author cocitation analysis
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