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Mira Fine Edna Sakal Dorit Vashdi Nava Chapnik-Cohen Violet Daniel Avigdor Levanon Orli Lipshitz Arieh Gertler 《Fish physiology and biochemistry》1993,11(1-6):353-361
Carp growth hormone (cGH) cDNA, in which Cys-123 was mutated to Ala, was prepared, transferred to the expression vector, expressed
in Escherichia coli and the mutant was purified to homogeneity. The mutation only slightly improved yield of the monomeric fraction, indicating
that Cys-123 is not involved in improper refolding. As compared to cGH, the mutant (cGH-C123A) exhibited lower binding affinity
toward homologous liver receptors and lower bioactivity in a 3T3-F442A preadipocyte bioassay despite the fact that both hormones
exhibited almost identical cross-reactivity with anti-cGH antibodies. These results, along with those of a structural comparison
to hGH, suggest that Cys-123 is located in the hydrophobic core of the hormone, and is most likely affecting the conformation
of the binding site. Dimeric forms of the hormone and its mutant were less active than their respective monomers. Homologous
binding experiments using a carp liver microsomal fraction revealed a single receptor population with Kd = 0.77 nM and Bmax = 241 fmol/mg microsomal protein.
Résumé Un ADN complémentaire (cDNA) de l'hormone de croissance de carpe (cGH), dans lequel l'acide aminé Cys-123 a été muté en Ala, a été préparé, inséré dans un vecteur d'expression, et exprimé dans Escherichia coli. Le mutant a ensuite été purifié jusqu'à homogénéité. La mutation améliore seulement faiblement la production de la fraction monomérique, indiquant que le Cys-123 n'est pas impliqué dans un repliement erroné. Comparé à la cGH, sa forme mutée (cGH-C123A) montre une plus faible affinité de liaison vis à vis de récepteurs hépatiques homologues, et une plus faible activité biologique dans un test réalisé sur des préadipocytes 3T3-F442A; cela en dépit du fait que les deux hormones présentent des réactions croisées presque identiques avec un anticorps anti-cGH polyclonale. Ces résultats, associés à une comparaison à la structure de l'hGH, suggèrent que le Cys-123 est localisé dans la partie hydrophobique de l'hormone, et affecte, le plus vraisemblablement, la conformation du site de liaison. Les formes dimériques de l'hormone et de sa forme mutée sont moins actives que leurs monomères respectifs. Les études de liaison homologue, réalisées avec des fractions microsomales de foie, révèlent une population unique de récepteurs de Kd = 0,77 nM et de Bmax = 241 fmol/mg de proteine microsomale.相似文献
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Urban habitats, particularly wastelands and brownfields, maintain rich biodiversity and offer habitat for many species, even rare and endangered taxa. However, such habitats are also under socio-economic pressures due to redevelopment for housing and industrial uses. In order to maintain urban biodiversity, it is currently unknown how much open area must be preserved and whether conservation is possible without complete exclusion from economic development. In this study, we applied a simulation model based on species distribution models for plants, grasshoppers, and leafhoppers to investigate planning options for urban conservation with special focus on business areas. Altogether, we modelled the occurrence of 81 species of the urban species pool and analysed settings of different proportions of open sites, different habitat turnover times, and different lot sizes. Our simulations demonstrated that dynamic land use supports urban biodiversity in terms of species richness and rarity. Setting aside brownfields before redevelopment for a period of on average 15 years supported the highest conservation value. Consequently, we recommend integrating the concept of ‘temporary conservation’ into urban planning for industrial and business areas. This concept requires habitat to be destroyed by redeveloping brownfield sites to built-up sites, but simultaneously creating new open spaces due to abandonment of urban land uses at other locations. This maintains a spatio-temporal mosaic of different successional stages ranging from pioneer to pre-forest communities. 相似文献
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Jun M Shao Y Ho CT Koetter U Lech S 《Journal of agricultural and food chemistry》2003,51(21):6340-6346
The degradation profile of glucosamine bulk form stressed at 100 degrees C for 2 h in an aqueous solution was studied. Column chromatography of acetylated product mixture led to isolation of two pure compounds (1b and 2b) and a mixture of at least three isomers (3b). 1a and 2a were identified as 5-(hydroxymethyl)-2-furaldehyde (5-HMF) and 2-(tetrahydroxybutyl)-5-(3',4'-dihydroxy-1'-trans-butenyl)pyrazine, respectively, by utilizing a variety of analytical techniques, such as GC-MS, LC-MS, on-line UV spectrum, (1)H and (13)C NMR, and DEPT, as well as (1)H-(1)H COSY. 3a was identified as 2-(tetrahydroxybutyl)-5-(2',3',4'-trihydroxybutyl)pyrazine, commonly known as deoxyfructosazine. In addition, glucosamine solid dosage form was exposed to 40 degrees C/75% relative humility for 10 weeks. Methanol extract of glucosamine solid dosage form was analyzed after acetylation by LC-MS, resulting in degradants 3b and 4b. 3a and 4a were, therefore, determined as deoxyfructosazine and 2,5-bis(tetrahydroxybutyl)pyrazine (fructosazine), respectively. Furthermore, the mechanisms of formation of identified degradation products are proposed and briefly discussed. 相似文献
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Aviram M Volkova N Coleman R Dreher M Reddy MK Ferreira D Rosenblat M 《Journal of agricultural and food chemistry》2008,56(3):1148-1157
We have analyzed in vivo and in vitro the antiatherogenic properties and mechanisms of action of all pomegranate fruit parts: peels (POMxl, POMxp), arils (POMa), seeds (POMo), and flowers (POMf), in comparison to whole fruit juice (PJ). Atherosclerotic E 0 mice consumed POM extracts [200 microg of gallic acid equivalents (GAE)/mouse/day] for 3 months. Blood samples, peritoneal macrophages (MPM), and aortas were then collected. All POM extracts possess antioxidative properties in vitro. After consumption of PJ, POMxl, POMxp, POMa, or POMf by E (0) mice, the atherosclerotic lesion area was significantly decreased by 44, 38, 39, 6, or 70%, respectively, as compared to placebo-treated group, while POMo had no effect. POMf consumption reduced serum lipids, and glucose levels by 18-25%. PJ, POMxl, POMxp, POMf, or POMa consumption resulted in a significant decrement, by 53, 42, 35, 27, or 13%, respectively, in MPM total peroxides content, and increased cellular paraoxonase 2 (PON2) activity, as compared to placebo-treated mice. The uptake rates of oxidized-LDL by E (0)-MPM were significantly reduced by approximately 15% after consumption of PJ, POMxl, or POMxp. Similar results were obtained on using J774A.1 macrophage cell line. Finally, pomegranate phenolics (punicalagin, punicalin, gallic acid, and ellagic acid), as well as pomegranate unique complexed sugars, could mimic the antiatherogenic effects of pomegranate extracts. We conclude that attenuation of atherosclerosis development by some of the POM extracts and, in particular, POMf, could be related to the combined beneficial effects on serum lipids levels and on macrophage atherogenic properties. 相似文献
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José M. Herrera Isa de Sá Teixeira Javier Rodríguez-Pérez António Mira 《Landscape Ecology》2016,31(4):731-743
Context
Seed dispersal is recognized as having profound effects on the distribution, dynamics and structure of plant populations and communities. However, knowledge of how landscape structure shapes carnivore-mediated seed dispersal patterns is still scarce, thereby limiting our understanding of large-scale plant population processes.Objectives
We aim to determine how the amount and spatial configuration of forest cover impacted the relative abundance of carnivorous mammals, and how these effects cascaded through the seed dispersal kernels they generated.Methods
Camera traps activated by animal movement were used for carnivore sampling. Colour-coded seed mimics embedded in common figs were used to know the exact origin of the dispersed seed mimics later found in carnivore scats. We applied this procedure in two sites differing in landscape structure.Results
We did not find between-site differences in the relative abundance of the principal carnivore species contributing to seed dispersal patterns, Martes foina. Mean dispersal distance and the probability of long dispersal events were higher in the site with spatially continuous and abundant forest cover, compared to the site with spatially aggregated and scarcer forest cover. Seed deposition closely matched the spatial patterning of forest cover in both study sites, suggesting behaviour-based mechanisms underpinning seed dispersal patterns generated by individual frugivore species.Conclusions
Our results provide the first empirical evidence of the impact of landscape structure on carnivore-mediated seed dispersal kernels. They also indicate that seed dispersal kernels generated strongly depend on the effect that landscape structure exerts on carnivore populations, particularly on habitat-use preferences.10.
NR Salvetti C Baravalle GA Mira EJ Gimeno BE Dallard F Rey HH Ortega 《Reproduction in domestic animals》2009,44(5):805-814
The purpose of this study was to estimate the expression and relative amounts of estrogen (ER) and progesterone receptors (PR) and their isoforms as well as heat shock protein 70 (HSP70) in ovaries of rats with induced cystic ovarian disease (COD). Primary, secondary, tertiary, atretic and cystic follicles were evaluated by immunohistochemistry and total ovarian proteins were analyzed by Western blot. In the granulosa layer, growing and cystic follicles in the treated group have a higher expression of ERα than growing follicles of control individuals. In the theca interna layer, tertiary follicles presented a significantly higher expression of ERα in the treated group. An increase in total ERα protein was detected in the treated group. Granulosa cells of all growing, atretic and cystic follicles show a lower expression of ERβ in animals with COD, and the total protein expression of ERβ was lower in this group. The expression of PR was lower in the granulosa cell layer of tertiary and cystic follicles in treated animals, and theca interna layer had less intense immunostaining in this group. Although there were no differences in the expression of PR-B by Western blotting, the expression of PR-A was higher and the expression of PR-C was smaller in the treated group. An intense HSP70 immunostaining was observed in the cells of cystic follicles. By Western blotting, higher protein expression of HSP70 was detected in the ovarian samples of the control group than those of the treated ones. Ovaries of animals with COD exhibited an altered steroid receptor expression and subtype balance as compared with control animals, and an increase in HSP70 immunoexpression. 相似文献