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José Luis Alonso

Publications and source records attributed to José Luis Alonso.

10 recordsLinked to original sources
PubMed2006

The beta-tail domain (betaTD) regulates physiologic ligand binding to integrin CD11b/CD18.

Crystallographic and electron microscopy studies revealed genuflexed (bent) integrins in both unliganded (inactive) and physiologic ligandbound (active) states, suggesting that local conformational changes are sufficient for activation. Herein we have explored the role of local changes in the contact region between the membrane-proximal beta-tail domain (betaTD) and the ligand-binding betaA domain of the bent conformation in regulating interaction of integrin CD11b/CD18 (alphaMbeta2) with its physiologic ligand iC3b. We replaced the betaTD CD loop residues D658GMD of the CD18 (beta2) subunit with the equivalent D672SSG of the beta3 subunit, with AGAA or with NGTD, expressed the respective heterodimeric receptors either transiently in epithelial HEK293T cells or stably in leukocytes (K562), and measured their ability to bind iC3b and to conformation-sensitive mAbs. In the presence of the physiologic divalent cations Ca(2+) plus Mg(2+) (at 1 mM each), the modified integrins showed increased (in HEK293) or constitutive (in K562) binding to iC3b compared with wild-type receptors. K562 expressing the betaTD-modified integrins bound in Ca(2+)Mg(2+) to the betaA-directed high-affinity reporter mAb 24 but not to mAb KIM127, a reporter of the genu-straightened state. These data identify a role for the membrane proximal betaTD as an allosteric modulator of integrin activation.

Amino Acid Substitution↗
PubMed2006

Membrane-assisted processing of xylooligosaccharide-containing liquors.

Liquors from rice husk autohydrolyis, containing xylooligosaccharides, other saccharides, and nonsaccharide compounds, were subjected to two selected processing schemes to increase the proportion of substituted xylooligosaccharides in refined liquors. Nanofiltration through a ceramic membrane with a molecular mass cutoff of 1000 Da allowed simultaneous concentration and purification; this latter derived from the preferential removal of monosaccharides and nonsaccharide compounds. When liquors were nanofiltered to achieve a volume reduction factor of 5 operating at a transmembrane pressure of 14 bar, 58.6% of the nonsaccharide components and 20.9-46.9% of monosaccharides were kept in retentate, in comparison with 92% of xylooligosaccharides and glucooligosaccharides. When nanofiltered liquors were subjected to double ion-exchange processing, a final product with a nonsaccharide content near 9 kg/100 kg of nonvolatile components was obtained at a yield of 10.90 kg/100 kg oven dry rice husks. Alternatively, when nanofiltered liquors were subjected to ethyl acetate extraction and further double ion-exchange processing, a purified product with a nonsaccharide content of 5.66 kg/100 kg of nonvolatile components was obtained at a yield of 9.94 kg/100 kg oven dry rice husks. The nonsaccharide components remaining in the final concentrate were mainly made up of phenolic and nitrogen-containing compounds.

Filtration↗
PubMed2006

A physically meaningful method for the comparison of potential energy functions.

In the study of the conformational behavior of complex systems, such as proteins, several related statistical measures are commonly used to compare two different potential energy functions. Among them, the Pearson's correlation coefficient r has no units and allows only semiquantitative statements to be made. Those that do have units of energy and whose value may be compared to a physically relevant scale, such as the root-mean-square deviation (RMSD), the mean error of the energies (ER), the standard deviation of the error (SDER) or the mean absolute error (AER), overestimate the distance between potentials. Moreover, their precise statistical meaning is far from clear. In this article, a new measure of the distance between potential energy functions is defined that overcomes the aforementioned difficulties. In addition, its precise physical meaning is discussed, the important issue of its additivity is investigated, and some possible applications are proposed. Finally, two of these applications are illustrated with practical examples: the study of the van der Waals energy, as implemented in CHARMM, in the Trp-Cage protein (PDB code 1L2Y) and the comparison of different levels of the theory in the ab initio study of the Ramachandran map of the model peptide HCO-L-Ala-NH2.

Chemistry, Physical↗
PubMed2004

Processing of rice husk autohydrolysis liquors for obtaining food ingredients.

Rice husks are of a lignocellulosic nature, with a hemicellulose fraction made up of substituted arabinoxylan. Rice husks were treated with hot, compressed water (autohydrolysis reaction) under optimized conditions to cause the hydrolytic degradation of arabinoxylan. The reaction products contained volatile components and nonvolatile components (NVC), which were made up of hemicellulose-derived products (substituted oligosaccharides and monosaccharides) and other nonvolatile solutes (ONVS). To decrease the content of ONVS, concentrated autohydrolysis liquors were first subjected to ethyl acetate extraction and then subjected to various alternative treatments (solvent precipitation, freeze-drying solvent extraction, or ion exchange). The resulting liquors were assayed for composition and yield determination. Material balances are presented for the several processes considered. The best results in terms of purification were obtained with sequential stages of ethyl acetate extraction and ion exchange, which led to concentrates with hemicellulose-derived compounds (sugars and substituted sugar oligomers) accounting for 92 wt % of the NVCs.

Acetates↗
PubMed2004

Survival and injury of Arcobacter after artificial inoculation into drinking water.

The aim of this work was to assess the effect of chlorine water treatment on Arcobacter butzleri and to study the survival strategies of this organism in chlorinated and non-chlorinated drinking water. A. butzleri NCTC 12481 was inoculated into chlorinated and non-chlorinated water and samples were removed aseptically, immediately and periodically during the next 2 days (for chlorinated drinking water) or 35 days (for non-chlorinated drinking water). The membrane integrity (Live/Dead BacLight kit), 16S rRNA (FISH technique), DNA content (23S rRNA PCR-RFLPs) and culturability changes in A. butzleri cells were analyzed. Culturability of the cells was lost at 5 min in chlorinated drinking water. At that time the cells showed membrane damage, although fluorescent intensity of 16S rRNA hybridization was constant throughout the chlorine treatment. After 48 h the amplicon specific for the 23S rRNA gene was weakly detected. In non-chlorinated drinking water cells lost their culturability after 16 days but the other factors measured indicated that Arcobacter remained viable throughout the experiment.

Arcobacter↗
PubMed2003

Feeling the forces: atomic force microscopy in cell biology.

Atomic force microscopy allows three-dimensional imaging and measurements of unstained and uncoated biological samples in air or fluid. Using this technology it offers resolution on the nanometer scale and detection of temporal changes in the mechanical properties, i.e. surface stiffness or elasticity in live cells and membranes. Various biological processes including ligand-receptor interactions, reorganization, and restructuring of the cytoskeleton associated with cell motility that are governed by intermolecular forces and their mode of detection will be discussed.

Animals↗
PubMed2003

Production of D(-)-lactic acid from cellulose by simultaneous saccharification and fermentation using Lactobacillus coryniformis subsp. torquens.

D(-)-Lactic acid was produced from cellulose by simultaneous saccharification and fermentation (SSF) in media containing cellulolytic enzymes and Lactobacillus coryniformis subsp. torquens ATCC 25600 at 39 degrees C and pH 5.4, yielding 0.89 g D(-)-lactic acid g(-1) cellulose at a mean volumetric productivity of 0.5 g l(-1) h(-1). No L(+)-lactic acid was found in the medium.

Cellulose↗
PubMed2003

Specific detection of Arcobacter and Campylobacter strains in water and sewage by PCR and fluorescent in situ hybridization.

The aim of this study was to evaluate PCR and fluorescent in situ hybridization (FISH) techniques for detecting Arcobacter and Campylobacter strains in river water and wastewater samples. Both 16S and 23S rRNA sequence data were used to design specific primers and oligonucleotide probes for PCR and FISH analyses, respectively. In order to assess the suitability of the methods, the assays were performed on naturally and artificially contaminated samples and compared with the isolation of cells on selective media. The detection range of PCR and FISH assays varied between 1 cell/ml (after enrichment) to 10(3) cells/ml (without enrichment). According to our results, both rRNA-based techniques have the potential to be used as quick and sensitive methods for detection of campylobacters in environmental samples.

Arcobacter↗