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11.
Capilla E Díaz M Albalat A Navarro I Pessin JE Keller K Planas JV 《American journal of physiology. Endocrinology and metabolism》2004,287(2):E348-E357
Glucose transport across the plasma membrane is mediated by a family of glucose transporter proteins (GLUTs), several of which have been identified in mammalian, avian, and, more recently, in fish species. Here, we report on the cloning of a salmon GLUT from adipose tissue with a high sequence homology to mammalian GLUT4 that has been named okGLUT4. Kinetic analysis of glucose transport following expression in Xenopus laevis oocytes demonstrated a 7.6 +/- 1.4 mM K(m) for 2-deoxyglucose (2-DG) transport measured under zero-trans conditions and 14.4 +/- 1.5 mM by equilibrium exchange of 3-O-methylglucose. Transport of 2-DG by okGLUT4-injected oocytes was stereospecific and was competed by D-glucose, D-mannose, and, to a lesser extent, D-galactose and D-fructose. In addition, 2-DG uptake was inhibited by cytochalasin B and ethylidene glucose. Moreover, insulin stimulated glucose uptake in Xenopus oocytes expressing okGLUT4 and in isolated trout adipocytes, which contain the native form of okGLUT4. Despite differences in protein motifs important for insulin-stimulated translocation of mammalian GLUT4, okGLUT4 was able to translocate to the plasma membrane from intracellular localization sites in response to insulin when expressed in 3T3-L1 adipocytes. These data demonstrate that okGLUT4 is a structural and functional fish homolog of mammalian GLUT4 but with a lower affinity for glucose, which could in part explain the lower ability of fish to clear a glucose load. 相似文献
12.
Active and inactive ecto-5'-nucleotidase variants in liver of control and dystrophic Lama2dy mice 总被引:1,自引:0,他引:1
Morote-García JC García-Ayllón MS Campoy FJ Vidal CJ Muñoz-Delgado E 《The international journal of biochemistry & cell biology》2004,36(3):422-433
The ecto-5'-nucleotidase (eNT) activity and the eNT protein content in liver of normal and merosin-deficient dystrophic Lama2dy mice were studied. After the solubilization procedure, the eNT activity in the final extract was 9.2+/-2.5U/mg (nmol of phosphate released from AMP per min and per mg protein) in normal liver, and it rose to 16.1+/-3.9U/mg (P=0.005) in dystrophic liver. The increase of activity was less pronounced in Lama2dy liver (1.7-fold) than the one reported in muscle (four-fold), which probably reflects the lower content of merosin in liver. Similarly to muscle, liver contained active and inactive eNT, as demonstrated by the higher level of immunoreactive protein in normal than in dystrophic liver in Western blots performed with samples containing the same units of eNT activity. PNGase F digestion decreased the size of liver and muscle eNT from 72 and 69kDa, to 63 and 60kDa. Oligoglycan cleavage did not alter eNT activity or the sedimentation coefficient, revealing that oligosaccharides are not required for catalysis or for maintaining the dimeric structure. The eNT protein content in samples of normal liver decreased by 55 or 80% after the trypsinolysis of native or deglycosylated enzyme, but the activity did not change. Such a high proportion of inactive eNT is unlikely to come from aged enzyme, which suggests the involvement of inactive enzyme in non-catalytic actions. 相似文献
13.
Capilla E Díaz M Gutiérrez J Planas JV 《American journal of physiology. Endocrinology and metabolism》2002,283(1):E44-E49
We have recently cloned a glucose transporter from brown trout muscle (btGLUT) with high sequence homology to mammalian GLUT4 that is predominantly expressed in red and white skeletal muscle, the two major sites of glucose uptake in trout. To study the physiological regulation of this putative fish GLUT4, we have investigated the expression of btGLUT in red and white skeletal muscle of trout in which blood insulin levels have been altered experimentally. The expression of btGLUT in red muscle increased significantly when insulin plasma levels were elevated by either insulin or arginine treatment and decreased significantly when insulin plasma levels were reduced either by fasting or by feeding a low-protein, high-carbohydrate diet. In contrast, the expression of btGLUT in white muscle was not affected by changes in the plasma levels of insulin. These results strongly suggest that insulin could be regulating the expression of btGLUT in trout red muscle in vivo and set the ground to test the hypothesis that btGLUT may be considered a GLUT4 homolog in fish. 相似文献
14.
Encarnación Mellado M. Teresa García Esther Roldán Joaquín J. Nieto A. Ventosa 《Extremophiles : life under extreme conditions》1998,2(4):435-438
The genomes of 11 moderately halophilic bacteria belonging to the genera Halomonas and Chromohalobacter have been analyzed by restriction endonuclease digestion and pulsed-field gel electrophoresis (PFGE). By using the infrequently
cutting restriction endonucleases SpeI and SwaI, highly characteristic fingerprintings were obtained for each of the isolates studied. On the basis of the lengths of the
SpeI and SwaI fragments, separated by PFGE, the genome size of the 11 strains studied was estimated. The genome size for 8 Halomonas strains ranged from 1450 to 2830 kb, whereas for the 3 Chromohalobacter strains studied it ranged from 1770 to 2295 kb. Finally, we show that macrorestriction fingerprints could be a useful tool
to elucidate the taxonomic position of bacteria belonging to the Halomonas–Deleya complex.
Received: October 13, 1997 / Accepted: May 12, 1998 相似文献
15.
Patricia Garcia de Olalla Ema Molas María Jesús Barberà Silvia Martín Encarnació Arellano Mercè Gosch Pilar Saladie Teresa Carbonell Hernando Knobel Elia Diez Joan A Caylà 《PloS one》2015,10(4)
Background
An estimated 30% of HIV cases in the European Union are not aware of their serological status. This study aimed to assess the effectiveness of a pilot HIV partner notification program.Methods
HIV cases diagnosed between January 2012 and June 2013 at two healthcare settings in Barcelona were invited to participate in a prospective survey. We identified process and outcome measures to evaluate this partner notification program, including the number of partners identified per interviewed index case, the proportion of partners tested for HIV as a result of the partner notification, and the proportion of new HIV diagnoses among their sex or needle-sharing partners.Results
Of the 125 index cases contacted, 108 (86.4%) agreed to provide information about partners. A total of 199 sexual partners were identified (1.8 partners per interviewed index case). HIV outcome was already known for 58 partners (70.7% were known to be HIV-positive), 141 partners were tested as result of partner notification, and 26 were newly diagnosed with HIV. The case-finding effectiveness of the program was 18.4%.Conclusion
This pilot program provides evidence of the effectiveness of a partner notification program implemented in healthcare settings. This active partner notification program was feasible, acceptable to the user, and identified a high proportion of HIV-infected patients previously unaware of their status. 相似文献16.
Melchor Juan J. Jesús Casas M. Antonia Elorrieta Santiago Bonachela Irene Gallego Francisca Fuentes-Rodríguez Encarnación Fenoy 《Hydrobiologia》2014,732(1):183-196
Irrigation ponds may act as a source of phytopathogenic species that might infest crops through the irrigation systems. Many studies have shown that submerged macrophytes can improve water clarity by out-competing phytoplankton by means of various mechanisms: favoured phytoplankton-grazing zooplankton, reduced nutrient and light availability, increased sinking losses and the release of allelopathically active substances. However, less information is available on the effects of submerged macrophytes on heterotrophic aquatic organisms such as pathogenic bacteria, fungi or oomycete species. This paper studies the effects of three submerged macrophytes—Chara fragilis, Potamogeton pectinatus and Najas marina—on the viability in water of propagules of two phytopathogenic isolates of the oomycetes Pythium aphanidermatum and Pythium ultimum. Moreover, we tested general antimicrobial properties (against bacteria and fungi in water) for these macrophytes. The results showed clear inhibitory effects of all three macrophytes on bacterial density in water and of C. fragilis on the viability of Pythium. Thus, preserving aquatic vegetation in irrigation ponds (i.e. charophytes), besides its purely environmental interest, may have important agronomic benefits owing to their role as biological control against some phytopathogenic agents. 相似文献
17.
Gary Tresadern Jose María Cid Gregor J. Macdonald Juan Antonio Vega Ana Isabel de Lucas Aránzazu García Encarnación Matesanz María Lourdes Linares Daniel Oehlrich Hilde Lavreysen Ilse Biesmans Andrés A. Trabanco 《Bioorganic & medicinal chemistry letters》2010,20(1):175-179
Imidazo[1,2-a]pyridines were identified via their shape and electrostatic similarity as novel positive allosteric modulators of the metabotropic glutamate 2 receptor. The subsequent synthesis and SAR are described. Potent, selective and metabolically stable compounds were found representing a promising avenue for current further studies. 相似文献
18.
Muñoz-Delgado E Montenegro MF Campoy FJ Moral-Naranjo MT Cabezas-Herrera J Kovacs G Vidal CJ 《The FEBS journal》2010,277(21):4519-4529
Despite the aberrant expression of cholinesterases in tumours, the question of their possible contribution to tumorigenesis remains unsolved. The identification in kidney of a cholinergic system has paved the way to functional studies, but details on renal cholinesterases are still lacking. To fill the gap and to determine whether cholinesterases are abnormally expressed in renal tumours, paired pieces of normal kidney and renal cell carcinomas (RCCs) were compared for cholinesterase activity and mRNA levels. In studies with papillary RCC (pRCC), conventional RCC, chromophobe RCC, and renal oncocytoma, acetylcholinesterase activity increased in pRCC (3.92 ± 3.01 mU·mg(-1), P = 0.031) and conventional RCC (2.64 ± 1.49 mU·mg(-1), P = 0.047) with respect to their controls (1.52 ± 0.92 and 1.57 ± 0.44 mU·mg(-1)). Butyrylcholinesterase activity increased in pRCC (5.12 ± 2.61 versus 2.73 ± 1.15 mU·mg(-1), P = 0.031). Glycosylphosphatidylinositol-linked acetylcholinesterase dimers and hydrophilic butyrylcholinesterase tetramers predominated in control and cancerous kidney. Acetylcholinesterase mRNAs with exons E1c and E1e, 3'-alternative T, H and R acetylcholinesterase mRNAs and butyrylcholinesterase mRNA were identified in kidney. The levels of acetylcholinesterase and butyrylcholinesterase mRNAs were nearly 1000-fold lower in human kidney than in colon. Whereas kidney and renal tumours showed comparable levels of acetylcholinesterase mRNA, the content of butyrylcholinesterase mRNA was increased 10-fold in pRCC. The presence of acetylcholinesterase and butyrylcholinesterase mRNAs in kidney supports their synthesis in the organ itself, and the prevalence of glycosylphosphatidylinositol-anchored acetylcholinesterase explains the splicing to acetylcholinesterase-H mRNA. The consequences of butyrylcholinesterase upregulation for pRCC growth are discussed. 相似文献
19.
Robledo M Jiménez-Zurdo JI Soto MJ Velázquez E Dazzo F Martínez-Molina E Mateos PF 《Molecular plant-microbe interactions : MPMI》2011,24(7):798-807
The establishment of rhizobia as nitrogen-fixing endosymbionts within legume root nodules requires the disruption of the plant cell wall to breach the host barrier at strategic infection sites in the root hair tip and at points of bacterial release from infection threads (IT) within the root cortex. We previously found that Rhizobium leguminosarum bv. trifolii uses its chromosomally encoded CelC2 cellulase to erode the noncrystalline wall at the apex of root hairs, thereby creating the primary portal of its entry into white clover roots. Here, we show that a recombinant derivative of R. leguminosarum bv. trifolii ANU843 that constitutively overproduces the CelC2 enzyme has increased competitiveness in occupying aberrant nodule-like root structures on clover that are inefficient in nitrogen fixation. This aberrant symbiotic phenotype involves an extensive uncontrolled degradation of the host cell walls restricted to the expected infection sites at tips of deformed root hairs and significantly enlarged infection droplets at termini of wider IT within the nodule infection zone. Furthermore, signs of elevated plant host defense as indicated by reactive oxygen species production in root tissues were more evident during infection by the recombinant strain than its wild-type parent. Our data further support the role of the rhizobial CelC2 cell wall-degrading enzyme in primary infection, and show evidence of its importance in secondary symbiotic infection and tight regulation of its production to establish an effective nitrogen-fixing root nodule symbiosis. 相似文献
20.
Sergio?EncarnaciónEmail author Magdalena?Hernández Gabriel?Martínez-Batallar Sandra?Contreras María?del?Carmen?Vargas Jaime?Mora 《Biological procedures online》2005,7(1):117-135
We propose two-dimensional gel electrophoresis (2-DE) and mass spectrometry to define the protein components of regulons and
stimulons in bacteria, including those organisms where genome sequencing is still in progress. The basic 2-DE protocol allows
high resolution and reproducibility and enables the direct comparison of hundreds or even thousands of proteins simultaneously.
To identify proteins that comprise stimulons and regulons, peptide mass fingerprint (PMF) with matrix-assisted laser desorption
ionization/time-of-flight mass spectrometry (MALDI-TOF-MS) analysis is the first option and, if results from this tool are
insufficient, complementary data obtained with electrospray ionization tandem-MS (ESI-MS/MS) may permit successful protein
identification. ESI-MS/MS and MALDI-TOF-MS provide complementary data sets, and so a more comprehensive coverage of a proteome
can be obtained using both techniques with the same sample, especially when few sequenced proteins of a particular organism
exist or genome sequencing is still in progress. 相似文献