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81.
Marko Kreft Mateja Prebil Helena H. Chowdhury Sonja Grilc Jørgen Jensen Robert Zorec 《Protoplasma》2010,246(1-4):73-80
A line profile of fluorescent intensities in confocal images is frequently examined. We have developed the computer software tool to analyse the profiles of intensities of fluorescent probes in confocal images. The software averages neighbouring pixels, adjacent to the central line, without reducing the spatial resolution of the image. As an experimental model, we have used the skeletal muscle fibre isolated from the rat skeletal muscle extensor digitorum brevis. As a marker of myofibrils' structure, we have used phalloidin–rhodamine staining and the anti-TIM antibody to label mitochondria. We also tested the distribution of the protein kinase B/Akt. Since signalling is ordered in modules and large protein complexes appear to direct signalling to organelles and regulate specific physiological functions, a software tool to analyse such complexes in fluorescent confocal images is required. The software displays the image, and the user defines the line for analysis. The image is rotated by the angle of the line. The line profile is calculated by averaging one dimension of the cropped rotated image matrix. The spatial resolution in averaged line profile is not decreased compared with single-pixel line profile, which was confirmed by the discrete Fourier transform computed with a fast Fourier transform algorithm. We conclude that the custom software tool presented here is a useful tool to analyse line profiles of fluorescence intensities in confocal images. 相似文献
82.
Mateja Erdani Kreft Daniele Di Giandomenico Galina V. Beznoussenko Nataša Resnik Alexander A. Mironov Kristijan Jezernik 《Biology of the cell / under the auspices of the European Cell Biology Organization》2010,102(11):593-607
Background information. The GA (Golgi apparatus) has an essential role in membrane trafficking, determining the assembly and delivery of UPs (uroplakins) to the APM (apical plasma membrane) of superficial UCs (uroepithelial cells) of urinary bladder. UPs are synchronously and uniformly delivered from the GA to the APM by DFVs (discoidal‐ or fusiform‐shaped vesicles); however, the mechanism of UP delivery is not known. We have used the culture model of UCs with the capacity to undergo terminal differentiation to study the process of uniform delivery of DFVs to the APM and to elucidate the mechanisms involved. Results. By three‐dimensional localization using confocal microscopy of immunofluorescence‐labelled GA‐related markers [GM130 (cis‐Golgi matrix protein of 130 kDa), GS15 (Golgi Snare 15 kDa), GS28 and giantin], uroepithelial differentiation‐related markers (UPs), MTs (microtubules; α‐tubulin) and intermediate filaments [CK7 (cytokeratin 7) and CK20], we found that in non‐differentiated, UP‐negative UCs the GA is mostly organized as a single ribbon‐like structure close to the nucleus, whereas in differentiated, UP‐positive UCs the GA is fragmented and spread almost through the entire cell. The FRAP (fluorescence recovery after photobleaching) experiments on the UCs transfected with GalT (trans‐Golgi/TGN enzyme β1,4‐galactosyltransferase) fused to fluorescent protein showed that Golgi‐resident enzyme cycles freely within ribbon‐like GA but not within fragmented GA. By CLEM (correlative light—electron microscopy), we examined the GA fragments in cells expressing UPs. We found that GA fragments are fully functional and similar to the GA fragments that are formed after nocodazole treatment. Furthermore, we demonstrated that the reorganization of GA into a fragmented form is associated with the impairment of the MT organization in the basal, central and subapical cytoplasm and the accumulation of intermediate filaments in the apical cytoplasm that could affect the kinetics of MT star leading to the peripheral fragmentation of the GA in the differentiated UCs. Conclusions. The fragmentation of the GA and the subsequent spreading of GA to the cell periphery represent one of the key events that promote the uniform delivery of UPs over the entire APM of differentiating UCs and thus are of major importance in the final proper formation and maintenance of the blood—urine barrier. 相似文献
83.
Erčulj F Blas M Bračič M 《Journal of strength and conditioning research / National Strength & Conditioning Association》2010,24(11):2970-2978
The aim of the study was to determine and analyze the level of certain motor abilities (acceleration and agility, the explosive strength of arms, and take-off power) of young elite European female basketball players. We also wanted to establish whether there were any differences between 3 groups of female basketball players who differed in terms of their playing performance. The sample of subjects consists of 65 female basketball players aged 14.49 (± 0.61) years who were divided into 3 groups (divisions A, B, and C of the European Championships). We compare the groups by using 8 motor tests. p Values <0.05 were considered statistically significant. The results show that the division C players achieved below-average results in all tests and thus differ from the players from divisions A and B whose test results were relatively homogeneous. The division C players differ from those from divisions A and B mainly in the 6 × 5-m sprint dribble (discriminant ratio coefficients [DRC] = 0.435), medicine ball throw (DRC = 0.375), and 20-m sprint (DRC = 0.203). Discriminatory power in the 6 × 5-m sprint dribble and 20-m sprint tests is preserved even after eliminating the effect of body height. We assume that, besides the deficit in body height and training status, this is also 1 of the key reasons for these players' lower playing efficiency compared to those from divisions A and B. We hope the findings of this study will enable the generation of model values, which can assist basketball coaches for this age category in basketball clubs, high schools, national teams, and basketball camps. 相似文献
84.
Amit Kapoor Natasha Mehta Frank Esper Mateja Poljsak-Prijatelj Phenix-Lan Quan Natasha Qaisar Eric Delwart W. Ian Lipkin 《PloS one》2010,5(7)
A novel parvovirus, provisionally named Gorilla Bocavirus species 1 (GBoV1), was identified in four stool samples from Western gorillas (Gorilla gorilla) with acute enteritis. The complete genomic sequence of the new parvovirus revealed three open reading frames (ORFs) with an organization similar to that of known bocaviruses. Phylogenetic analysis using complete capsid and non structural (NS) gene sequence suggested that the new parvovirus is most closely related to human bocaviruses (HBoV). However, the NS ORF is more similar in length to the NS ORF found in canine minute virus and bovine parvovirus than in HBoV. Comparative genetic analysis using GBoV and HBoV genomes enabled characterization of unique splice donor and acceptor sites that appear to be highly conserved among all four HBoV species, and provided evidence for expression of two different NS proteins in all primate bocaviruses. GBoV is the first non-human primate bocavirus identified and provides new insights into the genetic diversity and evolution of this highly prevalent and recently discovered group of parvoviruses. 相似文献
85.
The influence of ambient and filtered solar ultraviolet-B (UV)-B radiation and of selenium treatment was determined on photochemical efficiency, respiratory potential measured by electron transport system (ETS) activity, and yield in pumpkins, Cucurbita pepo L. Yield and ETS activity were higher when solar UV-B was filtered out. The results suggested that the decreased yield was related to the UV-B impaired flow of electrons in the respiratory chain. Selenium increased yield under ambient radiation conditions. However, no significant effect of excluding UV-B radiation or of treatment with selenium was observed on the photochemical efficiency of photosystem II (PSII). 相似文献
86.
Distribution of junction- and differentiation-related proteins in urothelial cells at the leading edge of primary explant outgrowths 总被引:2,自引:2,他引:0
Leading edge cells, which are located at the forefront of a wound margin, play a significant role in coordinating the wound healing process. In this study, leading edge cells of the urothelial explant outgrowth, resembling leading edge cells during urothelial full-thickness wound healing in vivo, were analyzed for expression and distribution of junction and differentiation-related proteins. Ultrastructural and immunofluorescence studies revealed that urothelial cells at the leading edge expressed ZO-1, claudin-4, occludin, E-cadherin, cytokeratin 7 and cytokeratin 20, while no expression of claudin-8 was noted. ZO-1, claudin-4, occludin and E-cadherin were localized along the cell membranes where neighbouring leading edge cells were in contact. Cytokeratin 7 was detected as filaments and cytokeratin 20 as small dots and sparse filaments. In conclusion, we detected early expression of ZO-1, claudin-4 and occludin at the urothelial leading edge, predicating the later formation of tight junctions as a necessary stage for the differentiation process that subsequently begins. The expression of occludin and cytokeratin 20 in urothelial cells at the leading edge suggests that leading edge cells may develop into fully differentiated superficial cells. 相似文献
87.
Respiratory potential and Se compounds in pea (Pisum sativum L.) plants grown from Se-enriched seeds
Selenium (Se) has been proved to be an essential element for humans and animals. However, less is known about its effects on plants. Pea plants were treated foliarly once (OT) and twice (TT) with Se solution during their flowering period. Seeds obtained from these plants contained 383 and 743 ng Se g(-1), respectively, and, together with control seeds from untreated plants (UT) containing 21 ng Se g(-1), were sown in soil in a greenhouse. Se content and its chemical form in young plants were studied, and its impact on plant respiratory potential, measured as terminal electron transport system (ETS) activity, determined. ETS activity was highest in young pea leaves with the highest Se content. Higher ETS activity possibly reflected increased glutathione peroxidase (GSH-Px) activity in mitochondria. The Se content of leaves and stems of plants grown from control seeds was similar to that in the seed, being around 40 ng Se g(-1). Se concentration in leaves of young plants grown from OT and TT seeds was 605%, and 1340% higher, respectively, than the control, and in their stems 355%, and 680% higher, respectively. The ratio of Se concentrations in OT and TT seeds was the same as in the leaves and stems in the young plants grown from them. SeMet was the major Se compound in Se-rich pea seeds and leaves, comprising 49% and 67% of the total Se content in OT and TT seeds, respectively, and 85% and 79% in the corresponding leaves. 相似文献
88.
Antonenko YN Roginsky VA Pashkovskaya AA Rokitskaya TI Kotova EA Zaspa AA Chernyak BV Skulachev VP 《The Journal of membrane biology》2008,222(3):141-149
The antioxidant activity of mitochondria-targeted small molecules, SkQ1 and MitoQ (conjugates of a lipophilic decyltriphenylphosphonium cation with an antioxidant moiety of a plastoquinone and ubiquinone, respectively), was studied in aqueous solution and in a lipid environment, i.e., micelles, liposomes and planar bilayer lipid membranes. Reactive oxygen species (ROS) were generated by azo initiators or ferrous ions with or without tert-butyl-hydroperoxide (t-BOOH). Chemiluminescence, fluorescence, oxygen consumption and inactivation of gramicidin peptide channels were measured to detect antioxidant activity. In all of the systems studied, SkQ1 was shown to effectively scavenge ROS. The scavenging was inherent to the reduced form of the quinone (SkQ1H(2)). In the majority of the above model systems, SkQ1 exhibited higher antioxidant activity than MitoQ. It is concluded that SkQ1H(2) operates as a ROS scavenger in both aqueous and lipid environments, being effective at preventing ROS-induced damage to membrane lipids as well as membrane-embedded peptides. 相似文献
89.
The degree of enzyme deactivation for lipases from Candida rugosa and Pseudomonas sp., hydroxynitrile lyase and mandelate racemase upon exposure to organic solvents can be correlated to their respective partition coefficients (log P values). However, three unexpected results were obtained: (1) the deactivation exerted by protic solvents, e.g., methanol, is severely underestimated; (2) little deactivation by an organic solvent cannot neccessarily be correlated to catalytic activity in this medium, and (3) in contrast to other enzymes, hydroxynitrile lyase is exceptionally stable towards deactivation by DMF. 相似文献
90.
David J. Zorko Karen Choong Jonathan Gilleland Barbara Agar Shawn Baker Cindy Brennan Eleanor Pullenayegum 《PloS one》2013,8(6)