86,660 research outputs found
Interaction of ARF1 and ARFA with different ARF-GEFs and localization of ARFA in <i>big5</i> mutant.
No full text(A-F) Co-immunoprecipitation (Co-IP) studies. (A) Co-IP of GNOM-Myc with ARF1-YFP, using GFP-Trap-agarose beads (IP: α-GFP) followed by immunoblot analysis (IB) with α-Myc antibody. Seedlings expressing only GNOM-Myc (GN-Myc) were used as control. (B) Co-IP of endogenous and YFP-tagged ARF1 with GNOM-Myc, using α-Myc-agarose beads (IP) followed by IB with α-GFP antibody and α-ARF1 antibody to detect YFP-tagged and endogenous ARF1. (C) Co-IP of endogenous and YFP-tagged ARF1 with GNL1-Myc, using α-Myc-agarose beads (IP) followed by IB analysis with α-GFP antibody and α-ARF1 antibody. (D) Co-IP of endogenous ARF1 with BIG3-YFP, using GFP-Trap-agarose beads (IP: α-GFP) followed by IB with α-ARF1 antibody. Col, wild-type control. (E) Co-IP of ARFA (ARFB1c)-RFP and endogenous ARF1 with BIG5-YFP, using GFP-Trap-agarose beads (IP: α-GFP) followed by IB analysis with α-RFP antibody and α-ARF1 antibody. (F) Co-IP of ARFA (ARFB1c)-RFP and endogenous ARF1 with BIG4-YFP, using GFP-Trap-agarose beads (IP: α-GFP) followed by IB analysis using α-RFP antibody and α-ARF1 antibody. IN, input; IP; immunoprecipitate; IB, immunoblot; kDa, kilodalton. (G-J) ARFA-YFP localization in wild-type (WT) (G-H) and big5 mutant (I-J). Note the highly cytosolic signal of ARFA-YFP in big5. Scale bars, 10μm. (K) In-vitro GDP-GTP exchange activity of the catalytic SEC7 domain of BIG3 (blue) and BIG5 (red) on ARFA. Negative control, ARFA alone (black). See also S5 Data.</p
PENGARUH KUALITAS PELAYANAN, PERSEPSI HARGA, DAN CITRA MEREK TERHADAP KEPUASAN PELANGGAN ARFA BARBERSHOP DI YOGYAKARTA
No full textPenelitian ini bertujuan untuk mengetahui: (1) pengaruh kualitas pelayanan terhadap kepuasan pelanggan Arfa Barbershop di Yogyakarta, (2) pengaruh persepsi harga terhadap kepuasan pelanggan Arfa Barbershop di Yogyakarta, (3) pengaruh citra merek terhadap kepuasan pelanggan Arfa Barbershop di Yogyakarta, dan (4) pengaruh kualitas pelayanan, persepsi harga, dan citra merek terhadap kepuasan pelanggan Arfa Barbershop di Yogyakarta.
Penelitian ini merupakan penelitian survei. Populasi pada penelitian ini adalah seluruh pelanggan Arfa Barbershop di Yogyakarta. Teknik pengambilan sampel menggunakan purposive sampling dengan jumlah sampel sebanyak 220 orang. Teknik pengumpulan data menggunakan kuesioner yang telah diuji validitas dan reliabilitasnya. Teknik analisis data yang digunakan adalah regresi berganda.
Hasil penelitian menunjukkan bahwa: (1) terdapat pengaruh positif kualitas pelayanan terhadap kepuasan pelanggan Arfa Barbershop di Yogyakarta, dibuktikan nilai t hitung sebesar 6,565; nilai signifikansi 0,000<0,05; dan koefisien regresi sebesar 0,300; (2) terdapat pengaruh positif persepsi harga terhadap kepuasan pelanggan Arfa Barbershop di Yogyakarta, dibuktikan dari nilai t hitung sebesar 5,391; nilai signifikansi 0,000<0,05; dan koefisien regresi sebesar 0,322; (3) terdapat pengaruh positif citra merek terhadap kepuasan pelanggan Arfa Barbershop di Yogyakarta. dibuktikan dari nilai t hitung sebesar 5,771; nilai signifikansi 0,000<0,05; dan koefisien regresi sebesar 0,134; dan (4) terdapat pengaruh kualitas pelayanan, persepsi harga, dan citra merek secara bersama-sama terhadap kepuasan pelanggan Arfa Barbershop di Yogyakarta, dibuktikan dengan nilai F hitung sebesar 61,955 dan signifikansi 0,000<0,05
ARF1 but not ARFA or ARFB is crucial for primary root growth and seed germination.
No full text(A-P) Activation-impaired (T31N; TN) or hydrolysis-impaired (Q71L; QL) variants of ARF1, ARFA and ARFB were expressed from the estradiol-inducible system. (A) Primary root growth was severely affected by ARF1-TN-YFP, ARF1-QL-YFP and ARFA-QL-YFP but not by ARFA-TN-YFP, ARFB-TN-YFP or ARFB-QL-YFP. Number of seedlings analyzed is indicated above each column. (+), 20μM estradiol; (-), without estradiol. See also S1 Data. (B-N) Seed germination on estradiol-containing medium is inhibited by ARF1-TN-YFP (C, D), ARF1-QL-YFP (I, J) and ARFA-QL-YFP (K, L) but not by ARFA-TN-YFP (E, F), ARFB-TN-YFP (G, H) or ARFB-QL-YFP (M, N). Scale bar, 5 mm (B-N). (O, P) Western blot of estradiol-induced expression of activation-impaired (TN) or hydrolysis-impaired (QL) variants of ARF1, ARFA and ARFB in seedlings (O). Asterisk, expected size of approx. 47kDa. (P) γCOP as loading control; two asterisks, expected size of approx. 100kDa. COL, wild-type control. (Q-S) Quantitative secretion assay. (Q) Co-expression of the secretory reporter α-amylase with wild type (WT), activation-impaired (TN) or hydrolysis-impaired (QL) ARFA variants in tobacco protoplasts. Secretion index, ratio of extracellular to intracellular reporter activity; control, expression of α-amylase alone. (R) Co-expression of α-amylase and ARFA-QL with rising concentrations of ARFA-WT, ARFA-TN or ARF1-WT. Numbers below indicate the amount of respective plasmid used for protoplast transformation relative to ARFA-QL; constant amounts of α-amylase and ARFA-QL were transformed. (S) Co-expression of α-amylase and ARFA-QL with rising concentrations of ARFA-TN,QL. Numbers below indicate the amount of respective plasmid used for protoplast transformation relative to ARFA-QL; constant amounts of α-amylase and ARFA-QL were transformed. Panels below (Q-S): ARF protein expression levels evidenced indirectly by detection of GFP. Both ARF and GFP coding sequences are under control of the bidirectional mas promoter (consisting of a mas1’ and a mas2’ part) on the same plasmid, with mas1’ directing GFP expression and mas2’ directing ARF expression in a ratio of 1 to 10 [65]. Note full recovery of secretion by ARFA-TN (R) and ARFA-TN,QL (S) overexpression. See also S2, S3 and S4 Data.</p
The effect of functional ions (Y3 +, F-, Ti4 +) on the structure, sintering and crystallization of diopside-calcium pyrophosphate bioglasses
No full textThe effects of three functional ions (yttrium Y3 +, fluorine F-, titanium Ti4 +) on the glass forming ability, sintering, crystallization, and thermo-physical properties of glasses and glass-ceramics were studied in a diopside-calcium pyrophosphate (90% CaMgSi2O6-10% Ca2P2O7) system. Three different percentages (1, 3 and 5 wt%) for each additive were tested. The structural features of glasses were assessed through FT-IR (Fourier Transform infra-red spectroscopy) and 29Si and 31P NMR (nuclear magnetic resonance), showing that the silicate network in all the investigated glasses is predominantly coordinated in Q2 (Si) units, while phosphorus tends to remain in the orthophosphate (Q0) environment. All glasses exhibited fast rates of biomineralization, making them promising candidates for biomedical applications. The sintering and crystallization behaviours of the glass powders were studied by differential thermal analysis (DTA), while the coefficient of thermal expansion (CTE) was determined by dilatometry. Glass transition temperature (Tg) values of all doped glasses were lower than the parent glass, while CTE values decreased with initial addition (1 wt%), of dopants before exhibiting an increase with further addition. In Y-doped glasses, a gradual increase was seen in the values of maximum crystallization peak temperature, Tp, up to 3 wt%, while an opposite trend was observed in Ti-doped glasses, showing an enhancement of the stability of the Y-doped glasses against devitrification. F-doped glasses exhibited a similar trend. Crystalline phase evolution was analysed by X-ray diffraction (XRD), and amorphous glass were obtained by sintering powder compacts from all the glasses at 800 °C for 1 h
Hydrolysis-impaired forms of ARF1 and ARFA block early secretion, endosomal recycling as well as post-Golgi and vacuolar trafficking.
No full textExpression of YFP-tagged hydrolysis-impaired Q71L (QL; green) variants of ARF1 and ARFA was induced by 20μM estradiol for 5-6h and trafficking markers (magenta) were analyzed in immunostaining (A-N) or live-cell imaging (O-I1). (A-G) The COPI subunit γCOP (A, B, E) was recruited to the Golgi in wild-type control (A; Col). Expression of ARF1-QL-YFP (C) and ARFA-QL-YFP (F) induced aggregation of γCOP (B, E) co-localizing with ARF1 and ARFA (D, G). (H-N) Clathrin coat (H, I, L) was recruited to the TGN in wild-type control (H). In contrast, clathrin formed aggregates and co-localized with ARF1-QL-YFP (I-K) and ARFA-QL-YFP (L-N). (O-U) BFA treatment (50μM for 1h) was used to visualize endocytosed FM4-64 (O, P, S) in BFA-compartments. Endocytosis of FM4-64 in ARF1-QL-YFP (P-R) and ARFA-QL-YFP (S-U) was comparable to wild-type control (O). (V-B1) Seedlings were treated with 20μM estradiol and 50μM BFA for 5h. Recycling of H4::RFP-PEN1 was analyzed after BFA wash-out for 2h. H4::RFP-PEN1 (V, W, Z) returned to the plasma membrane in wild-type control (V). ARF1-QL-YFP (W-Y) and ARF A-QL-YFP (Z-B1) interfered with recycling. (C1-I1) Trafficking of the vacuolar cargo, AFVY-RFP (C1, D1, G1), expressed from the estradiol-inducible system, was inhibited in ARF1-QL-YFP (D1-F1) and in ARFA-QL-YFP (G1-I1) expressing lines but not in wild-type (C1). Scale bar, 10μm. The same wild-type controls were used as in Figs 3 and S3 and S5.</p
Ultrastructural defects of endomembrane compartments caused by interference with ARF1 or ARFA function.
No full textSeedling root cells were ultrastructurally analyzed after high-pressure freezing, freeze-substitution and embedding in epoxy resin. (A-E) ARF1-TN-YFP. (A) Tubular ER (er) connected to large compartments (erc) and circular structures (*). (B, C) Circular structures (*) connected to ER contain several membranes including ER membranes. (D) Circular structures are heavily labeled with gold-conjugated anti-ARF1 antibodies (arrows). (E) Disintegrated Golgi stack (arrowhead). (F, G) ARF1-QL-YFP. (F) Large vesicle aggregates with Golgi(-derived) cisternae (arrowheads). (G) Golgi cisternae (arrowheads) producing unusually large (secretory) vesicles. (H, I) ARFA-TN-YFP. (H) Golgi stacks (arrowheads) and TGN structures (t). (I) Golgi stack (arrowhead) with slightly enlarged TGN compartment (t) at higher magnification. (K-M) ARFA-QL-YFP. (K) Aberrant Golgi stacks (arrowheads) and TGN structures (t) in two adjacent cells. (L) Cell-wall stubs (cw) and aberrant Golgi stack (arrowhead) and TGN structures (t). (M) Aberrant rounded Golgi stacks (arrowheads) with budding vesicles and large TGN structures (t) at higher magnification. Abbreviations: cw, cell wall; er, endoplasmic reticulum (ER); erc, ER-connected compartment; m, mitochondrion; n, nucleus; t, trans-Golgi network (TGN); v, vacuole. Scale bars, 500 nm.</p
Variations on the Author
No full text“Variations on the Author” discusses two of Eduardo Coutinho’s recent films (Um Dia na Vida, from 2010, and Últimas Conversas, posthumously released in 2015) and their contribution to the general question of documentary authorship. The director’s filmography is characterized by a consistent yet self-effacing form of authorial self-inscription: Coutinho often features as an interviewer that rather than express opinions propels discourses; an interviewer that is good at listening. This mode of self-inscription characterizes him as an author who is not expressive but who is nonetheless markedly present on the screen. In Um Dia na Vida, however, Coutinho is completely absent form the image, while Últimas Conversas, on the contrary, includes a confessional prologue that moves the director from the margins to the center of his films. This article examines the ways in which these works stand out in the filmography of a director who offers new insights into the notion of cinematic authorship
Enhanced bioactivity of a rapidly-dried sol-gel derived quaternary bioglass
No full textNovel quaternary (67Si-24Ca-10Na-8P) glass powders were successfully synthesised by sol-gel followed by two alternative drying schedules, conventional drying (CD) and an innovative fast drying (FD) process (200 times quicker). The glasses were thermally stabilised at 550 °C, and then characterised by different complementary techniques. The samples showed very similar silica network structures, with the FD one having slightly lower degree of polymerisation than the CD sample. This less polymerised, more open, network structure exhibited an improved bioactivity in simulated body fluid (SBF), probably also due to the apparent presence of poorly crystalline HAp in the stabilised glass powder. In contrast, the CD glass exhibited an unwanted secondary crystalline silica phase. Both glasses showed excellent biomineralisation upon immersion in SBF, being more pronounced in the case of FD with clear evidence of HAp formation after 4 h, while equivalent signs in the CD samples were only noticed after longer immersion periods between 8 h and 1 week
The influence of processing parameters on morphology and granulometry of a wet-milled sol-gel glass powder
No full textA quaternary bioactive sol-gel glass of high silica content was heat treated at different temperatures, and then wet ball milled under different balls-to-powder ratios. A total of sixteen experiments were performed to study in detail the effects of both experimental variables on the structure, morphology, particle size distributions and nitrogen adsorption isotherms. The balls–to–powder ratio exerts a tremendous influence on the final particle size distribution of the powders, while its effects on the pore volume and morphology are minimal. These structural features are mostly governed by the changes in calcination temperature. Therefore, understanding the specific roles of each experimental parameter is of paramount importance towards achieving optimum powders with the desired properties. This work sheds light on the importance of using a suitable combination of these two parameters for tuning the morphology and the granulometry of the sol-gel derived bioactive glass powders
Clove and cinnamon: Novel anti–oxidant fuels for preparing magnetic iron oxide particles by the sol–gel auto–ignition method
No full textMagnetic spinel ferrite particles were successfully prepared from only iron (III) nitrate, using clove and cinnamon plant extracts for the first time as eco-fuels for auto–ignition synthesis. Both natural substances were shown to play dual roles in the synthesis of the magnetic particles; i.e. as fuels (agents for biogenic reduction), and as anti–oxidants for preserving the magnetic phases during subsequent heating. In this regard, no special protective environments such as H2/N2 or argon gases were required during the heat stabilisation process, and samples could be stabilised by heating to 550 °C in air. The magnetic crystalline phase is mostly maghemite (γ-Fe2O3) rather than magnetite (Fe3O4), as revealed by Raman, since no Raman active phonon modes were detected for magnetite, and most of the bands were related to maghemite, with traces of hematite. Both fuels produced strongly magnetic soft ferrites, with magnetisation values of 75 A m2 kg−1, around the expected ones for maghemite Moreover, TG curves showed that maghemite degradation only occurs at relatively high temperatures, namely at 550 °C for Clove, and at 744 °C for Cinnamon, confirming the sustainable nature of the as-obtained materials against oxidation
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