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991.
Rafael Beine Alfred Rocas Valente Rebekka Biedendieck Dieter Jahn Jürgen Seibel 《Process Biochemistry》2009,44(10):1103-1114
Many biocatalysts exhibit strict stereospecificity and regioselectivity. However, their thermodynamically controlled equilibria often limit yields in industrial production processes. Herein, we describe the synthesis of fructooligosaccharides from sucrose by various fructansucrases. We previously demonstrated that transfructosylation to diverse acceptors yields d-glucose and the fructose-containing product along with diverse by-products. To streamline this reaction, we developed a procedure that allows the enhanced transfructosylation of diverse acceptors by different fructansucrases. By diverting the released glucose from the reaction via metabolism by living cells we limited the back reaction and forced the consumption of sucrose. The basic conditions for the resulting fermentation process were optimized by a genetic algorithm and integrated into a kinetic model. This strategy allows the prediction of optimal reaction parameters for the production of desired target compounds. 相似文献
992.
Rafael C. Rodrigues Cesar A. Godoy Giandra Volpato Marco A.Z. Ayub Roberto Fernandez-Lafuente Jose M. Guisan 《Process Biochemistry》2009,44(9):963-968
This paper describes the immobilization and stabilization of the lipase from Thermomyces lanuginosus (TLL) on glyoxyl agarose. Enzymes attach to this support only by the reaction between several aldehyde groups of the support and several Lys residues on the external surface of the enzyme molecules at pH 10. However, this standard immobilization procedure is unsuitable for TLL lipase due to the low stability of TLL at pH 10 and its low content on Lys groups that makes that the immobilization process was quite slow. The chemical amination of TLL, after reversible immobilization on hydrophobic supports, has been shown to be a simple and efficient way to improve the multipoint covalent attachment of this enzyme. The modification enriches the enzyme surface in primary amino groups with low pKb, thus allowing the immobilization of the enzyme at lower pH values. The aminated enzyme was rapidly immobilized at pH 9 and 10, with activities recovery of approximately 70%. The immobilization of the chemically modified enzyme improved its stability by 5-fold when compared to the non-modified enzyme during thermal inactivation and by hundreds of times when the enzyme was inactivated in the presence of organic solvents, being both glyoxyl preparations more stable than the enzyme immobilized on bromocyanogen. 相似文献
993.
Rethinking species selection for restoration of arid shrublands 总被引:1,自引:0,他引:1
Francisco M. Padilla Rafael Ortega Joaquín Snchez Francisco I. Pugnaire 《Basic and Applied Ecology》2009,10(7):640-647
Restoration is playing an increasingly important role in ecology as natural habitats become scarcer and chances to restore ecosystems damaged by human activities are more common. However, restoration of degraded Mediterranean arid ecosystems is hampered by drought and poor soils, which cause many establishment failures. To compare how species belonging to different successional stages establish in a very stressful site, we carried out a field experiment with 14 tree and shrub species differing in functional traits. After three growing seasons, mid-successional shrubs such as the leafless Ephedra fragilis and the C4 Salsola oppositifolia, or green-stemmed legumes like Coronilla juncea, Genista umbellat, and Retama sphaerocarpa, showed survival rates up to 93%, while late-successional species like Tetraclinis articulata, Pinus halepensis, Olea europaea, and Pistacia lentiscus, frequently used and recommended in regular restoration projects, hardly reached 55%. We found that survival was highest for legumes, followed by leafless species, and C4 shrubs, traits that are believed to maximize resource uptake in cleared and infertile areas while reducing water losses. Thus, selection of mid-successional species having such traits should be considered for successful restoration. These species would increase the success of restoration programs, but also would increase soil fertility, reduce soil erosion processes, and eventually facilitate establishment of other species, therefore accelerating secondary succession. We suggest a new approach for the restoration for arid shrublands in which species are carefully selected based on traits that best suit the environmental conditions. 相似文献
994.
Sara Rodríguez‐Enríquez Alvaro Marín‐Hernández Juan Carlos Gallardo‐Pérez Rafael Moreno‐Sánchez 《Journal of cellular physiology》2009,221(3):552-559
Metabolic control analysis of tumor glycolysis has indicated that hexokinase (HK) and glucose transporter (GLUT) exert the main flux control (71%). To understand why they are the main controlling steps, the GLUT and HK kinetics and the contents of GLUT1, GLUT2, GLUT3, GLUT4, HKI, and HKII were analyzed in rat hepatocarcinoma AS‐30D and HeLa human cervix cancer. An improved protocol to determine the kinetic parameters of GLUT was developed with D ‐[2‐3H‐glucose] as physiological substrate. Kinetic analysis revealed two components at low‐ and high‐glucose concentrations in both tumor cells. At low glucose and 37°C, the Vmax was 55 ± 20 and 17.2 ± 6 nmol (min × mg protein)?1, whereas the Km was 0.52 ± 0.7 and 9.3 ± 3 mM for hepatoma and HeLa cells, respectively. GLUT activity was partially inhibited by cytochalasin B (IC50 = 0.44 ± 0.1; Ki = 0.3 ± 0.1 µM) and phloretin (IC50 = 8.7 µM) in AS‐30D hepatocarcinoma. At physiological glucose, GLUT1 and GLUT3 were the predominant active isoforms in HeLa cells and AS‐30D cells, respectively. HK activity in HeLa cells was much lower (60 mU/mg protein) than that in AS‐30D cells (700 mU/mg protein), but both HKs were strongly inhibited by G6P. HKII was the predominant isoform in AS‐30D carcinoma and HeLa cells. The much lower GLUT Vmax and catalytic efficiency (Vmax/Km) values in comparison to those of G6P‐sensitive HK suggested the transporter exerts higher control on the glycolytic flux than HK in cancer cells. Thus, GLUT seems a more adequate therapeutic target. J. Cell. Physiol. 221: 552–559, 2009. © 2009 Wiley‐Liss, Inc. 相似文献
995.
Rafael Gonzalo-Gobernado Diana Reimers Antonio S. Herranz Juan Jos�� D��az-Gil Cristina Osuna Mar��a Jos�� Asensio Silvia Baena Macarena Rodr��guez-Serrano Eulalia Baz��n 《The journal of histochemistry and cytochemistry》2009,57(5):491-502
Neural stem cells with self-renewal and multilineage potential persist in the subventricular zone of the adult mammalian forebrain. These cells remain relatively quiescent but, under certain conditions, can be stimulated, giving rise to new neurons. Liver growth factor (LGF) is a mitogen for liver cells that shows biological activity in extrahepatic sites and is useful for neuroregenerative therapies. The aim of this study was to investigate the potential neurogenic activity of LGF in the 6-hydroxydopamine rat model of Parkinson''s disease. Proliferation was significantly increased in the subventricular zone and denervated striatum of rats receiving ICV LGF infusions, and 25% of the proliferating cells were doublecortin-positive neurons. Doublecortin-positive cells with the morphology of migrating neuroblasts were also observed in the dorsal and ventral regions of the striatum of LGF-infused animals. Moreover, some newly generated cells were neuronal nuclei-positive mature neurons. LGF also stimulated microglia and induced astrogliosis, both phenomena associated with generation and migration of new neurons in the adult brain. In summary, our study shows that LGF stimulates neurogenesis when applied intraventricularly in 6-hydroxydopamine–lesioned rats. Considering that this factor also promotes neuronal migration into damaged tissue, we propose LGF as a novel factor useful for neuronal replacement in neurodegenerative diseases. (J Histochem Cytochem 57:491–502, 2009) 相似文献
996.
997.
Rebeca Dieguez-Gonzalez Manuel Calaza Eva Perez-Pampin Alejandro Balsa Francisco J Blanco Juan D Cañete Rafael Caliz Luis Carreño Arturo R de la Serna Benjamin Fernandez-Gutierrez Ana Maria Ortiz Gabriel Herrero-Beaumont Jose L Pablos Javier Narvaez Federico Navarro Jose L Marenco Juan J Gomez-Reino Antonio Gonzalez 《Arthritis research & therapy》2009,11(2):1-8
Introduction
Genome-wide association studies of rheumatoid arthritis (RA) have identified an association of the disease with a 6q23 region devoid of genes. TNFAIP3, an RA candidate gene, flanks this region, and polymorphisms in both the TNFAIP3 gene and the intergenic region are associated with systemic lupus erythematosus. We hypothesized that there is a similar association with RA, including polymorphisms in TNFAIP3 and the intergenic region.Methods
To test this hypothesis, we selected tag-single nucleotide polymorphisms (SNPs) in both loci. They were analyzed in 1,651 patients with RA and 1,619 control individuals of Spanish ancestry.Results
Weak evidence of association was found both in the 6q23 intergenic region and in the TNFAIP3 locus. The rs582757 SNP and a common haplotype in the TNFAIP3 locus exhibited association with RA. In the intergenic region, two SNPs were associated, namely rs609438 and rs13207033. The latter was only associated in patients with anti-citrullinated peptide antibodies. Overall, statistical association was best explained by the interdependent contribution of SNPs from the two loci TNFAIP3 and the 6q23 intergenic region.Conclusions
Our data are consistent with the hypothesis that several RA genetic factors exist in the 6q23 region, including polymorphisms in the TNFAIP3 gene, like that previously described for systemic lupus erythematosus. 相似文献998.
Treatment for vitiligo is difficult and prolonged. Nevertheless, at present considerable knowledge accumulated during several decades on the pathogenic mechanisms, revealed important clues for designing new strategies to improve vitiligo depigmentation. With available medical therapies, high repigmentation percentages mostly on facial and neck lesions are achieved, although they are less effective on trunk and limbs and poor on the acral parts of the extremities. Narrow band UVB and psoralens and UVA are the two most important treatments for generalized vitiligo affecting more than 10–20% of the cutaneous surface, and topical corticosteroids, or calcineurin inhibitors are the most valuable treatments for localized vitiligo. Persistence of achieved regimentation is variable and an undefined percentage of patients may have variable recurrence. When vitiligo becomes refractory, surgical methods may improve depigmentation as effectively as with medical therapy; in segmental (unilateral) or long standing, non‐segmental (bilateral) stable vitiligo, repigmentation with surgical methods is usually permanent. 相似文献
999.
Practice on castration of piglets in Europe 总被引:1,自引:0,他引:1
Fredriksen B Font I Furnols M Lundström K Migdal W Prunier A Tuyttens FA Bonneau M 《Animal : an international journal of animal bioscience》2009,3(11):1480-1487
PIGCAS (Attitudes, practices and state of the art regarding piglet castration in Europe) is to our knowledge the first project that has focused on castration practice across European countries (European Union minus Bulgaria, Malta and Romania, plus Norway and Switzerland). About 250 million pigs are slaughtered in Europe each year. Of the 125 million male pigs, approximately 20% are left entire, less than 3% are castrated with anaesthesia and the rest is castrated without anaesthesia. The study identified large variations in castration procedures, both within and between countries. In females, castration is very rare, but is practiced without anaesthesia in special breeds/production systems in some of the southern countries. 相似文献
1000.
J?natas S. Abrah?o Maria Isabel M. Guedes Giliane S. Trindade Flávio G. Fonseca Rafael K. Campos Bruno F. Mota Zélia I. P. Lobato André T. Silva-Fernandes Gisele O. L. Rodrigues Larissa S. Lima Paulo C. P. Ferreira Cláudio A. Bonjardim Erna G. Kroon 《PloS one》2009,4(10)