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Biomedical subjects

J L Alonso

Publications and source records attributed to J L Alonso.

At least 19 recordsLinked to original sources

The marine sphingolipid-derived compound ES 285 triggers an atypical cell death pathway.

The isolation of new molecules from marine sources opens the door to their possible therapeutic use against tumors and other pathological conditions. Indeed, we recently defined the cytotoxicity of ES 285, obtained from the clam Mactromeris polynima, and its affects on the cells microfilament but not the microtubule network. Considering the analogy between ES 285 and sphingosine-related lipids, we wondered whether ES 285 might affect the activity of PKC at the intracellular level. While we anticipated that ES 285 might inhibit PKC, it turns out that in contrast it serves to activate PKC at the cellular level. Indeed, like other sphingosine-related lipids, ES 285 induces the phosphorylation of MARCKS. Additionally, we further examined the cytotoxicity of ES 285 to elucidate the molecular mechanisms through which this compound triggers apoptosis. When the influence of ES 285 on "cell death markers" was assessed, it became clear that ES285 activates caspase 3 and 12, and that it modified the phosphorylation of p53. In contrast, ES 285 does not affect other pathways widely implicated in regulating cell survival/apoptosis, such as JNK, Erks or Akt. Thus, these data suggest that ES 285-triggers an atypical cell death program when compared to other sphingosine-dependent apoptosis pathways.

Alkanes↗

Quantum mechanical calculation of the effects of stiff and rigid constraints in the conformational equilibrium of the alanine dipeptide.

If constraints are imposed on a macromolecule, two inequivalent classical models may be used: the stiff and the rigid one. This work studies the effects of such constraints on the conformational equilibrium distribution (CED) of the model dipeptide HCO-L-Ala-NH(2)without any simplifying assumption. We use ab initio quantum mechanics calculations including electron correlation at the MP2 level to describe the system, and we measure the conformational dependence of all the correcting terms to the naive CED based in the potential energy surface that appear when the constraints are considered. These terms are related to mass-metric tensors determinants and also occur in the Fixman's compensating potential. We show that some of the corrections are non-negligible if one is interested in the whole Ramachandran space. On the other hand, if only the energetically lower region, containing the principal secondary structure elements, is assumed to be relevant, then, all correcting terms may be neglected up to peptides of considerable length. This is the first time, as far as we know, that the analysis of the conformational dependence of these correcting terms is performed in a relevant biomolecule with a realistic potential energy function.

Alanine↗

Definition of Systematic, Approximately Separable, and Modular Internal Coordinates (SASMIC) for macromolecular simulation.

A set of rules is defined to systematically number the groups and the atoms of polypeptides in a modular manner. Supported by this numeration, a set of internal coordinates is defined. These coordinates (termed Systematic, Approximately Separable, and Modular Internal Coordinates--SASMIC) are straightforwardly written in Z-matrix form and may be directly implemented in typical Quantum Chemistry packages. A number of Perl scripts that automatically generate the Z-matrix files are provided as supplementary material. The main difference with most Z-matrix-like coordinates normally used in the literature is that normal dihedral angles ("principal dihedrals" in this work) are only used to fix the orientation of whole groups and a different type of dihedrals, termed "phase dihedrals," are used to describe the covalent structure inside the groups. This physical approach allows to approximately separate soft and hard movements of the molecule using only topological information and to directly implement constraints. As an application, we use the coordinates defined and ab initio quantum mechanical calculations to assess the commonly assumed approximation of the free energy, obtained from "integrating out" the side chain degree of freedom chi, by the Potential Energy Surface (PES) in the protected dipeptide HCO-L-Ala-NH2. We also present a subbox of the Hessian matrix in two different sets of coordinates to illustrate the approximate separation of soft and hard movements when the coordinates defined in this work are used. (PACS: 87.14.Ee, 87.15.-v, 87.15.Aa, 87.15.Cc)

Alanine↗

Refining of autohydrolysis liquors for manufacturing xylooligosaccharides: evaluation of operational strategies.

When xylan-containing lignocellulosic materials are subjected to the action of heat in an aqueous medium (autohydrolysis reaction) under suitable operational conditions, the heterocyclic ether bonds of the polymeric chain are split to give xylooligosaccharides (XO) and a variety of other products derived from hemicelluloses, lignin and extractives. For product applications requiring high-purity XO, subsequent purification stages have to be implemented. Various strategies have been evaluated for refining Eucalyptus wood autohydrolysis liquors, including the two-stage reaction, precipitation and solvent extraction of freeze-dried, dewaxed liquors. Data on the degree of xylooligosaccharide recovery as well as on the composition of the isolates (measured in terms of hemicellulose-derived products and non-saccharide products) are provided for the various operational procedures assayed.

Cellulose↗

Recovery of lactic acid from simultaneous saccharification and fermentation media using anion exchange resins.

The physicochemical properties (capacity, kinetics and selectivity) of the ion exchange resins Amberlite IRA900, IRA400, IRA96 and IRA67 were determined to evaluate their comparative suitability for lactic acid recovery. Both the kinetics of lactic acid sorption from aqueous solutions and the equilibrium were assessed using mathematical models, which provided a close interpretation of the experimental results. The best resins (Amberlite IRA96 and IRA67) were employed in further fixed-bed operation using aqueous lactic acid solutions as feed. In this set of experiments, parameters such as capacity, regenerant consumption, percentage of lactic acid recovery and product concentration were measured. Amberlite IRA67, a weak base resin, was selected for lactic acid recovery from SSF (simultaneous saccharification and fermentation) broths. Owing to the presence of nutrients and ions other than lactate, a slightly decreased capacity was determined when using SSF media instead aqueous lactic acid solutions, but quantitative lactic acid recoveries at constant capacities were obtained in four sequential load/regeneration cycles.

Journal Article↗

Use of fluorescent in situ hybridization to evidence the presence of Helicobacter pylori in water.

We have evaluated the use of a fluorescent in situ hybridization (FISH) technique for the detection of Helicobacter pylori in water (river and wastewater) samples. The assay was compared with PCR detection and isolation of cells on selective media. 16S rRNA and UreA+B sequence data were used as oligonucleotide probe and specific primers for FISH and PCR, respectively. Using FISH technique, H. pylori was detected in two river water and one wastewater samples, while PCR yielded only one positive result. H. pylori culture was not possible from any sample. According to these results, FISH technique has the potential to be used as a quick and sensitive method for detection of H. pylori in environmental samples.

DNA, Bacterial↗

Presence of bacterial infection in bleeding cirrhotic patients is independently associated with early mortality and failure to control bleeding.

Bacterial infection is strongly associated with gastrointestinal bleeding in cirrhotic patients and seems to be related with the failure to control bleeding. The aims of this study were to assess the influence of infections on the failure to control bleeding and death in cirrhotic patients without antibiotic prophylaxis. Ninety-one consecutive bleeding cirrhotic patients were analyzed. Bleeding was managed using somatostatin with sclerotherapy for active bleeding. Screening for bacterial infection (analysis and culture of blood, urine, ascitic and other fluids, together with chest radiography) was made at time 0 and when clinical signs suggested infection. The cause of bleeding was variceal in 72 (79%) patients. Failure to control bleeding occurred in 24 (26%) patients, and 10 (11%) of the patients died. Compared with the group without infection, failure to control bleeding (65% vs 15%; P < 0.001) and mortality (40% vs 3%; P < 0.001), were observed more frequently in patients with infection. Multivariate analysis showed that bacterial infection (OR = 9.7; P < 0.001) and the presence of shock (OR = 3.5; P < 0.05) were independently associated with failure to control bleeding. Bacterial infection (OR = 12.6; P < 0.01), encephalopathy (OR = 6.9; P < 0.05), and shock (OR = 5.8; P < 0.05) were identified as predictive of death. In conclusion, in bleeding cirrhotic patients bacterial infection is associated with failure to control bleeding as well as mortality.

Female↗

Outcomes of a health education intervention in a sample of patients infected by HIV, most of them injection drug users: possibilities and limitations.

We studied the receptivity of a population of HIV-infected patients to the development of a group educational intervention in order to enhance the adherence to therapy. We designed a group educational activity conducted by family physicians and directed to patients with HIV infection treated with antiretrovirals for at least six months. We conducted a randomized trial that compared two interventions: an educational intervention plus individual counsel or the last option alone. We studied their clinical situation, adherence to therapy and familial support. We offered 115 patients (66.9% injection drug users (IDUs), 69.6% males) the opportunity to be included in our trial, but 73.9% of them refused. No group with four or more participants was formed. Fifty-nine per cent refused to be included claiming personal reasons and 32.9% cited trouble in their jobs. There was a non-significant trend to accept in the case of women (p = 0.19), patients with AIDS (p = 0.11) and non-adherent patients (p = 0.08). This trend was significant in the group of women with AIDS (p = 0.01) and non-adherent women (p = 0.05). We concluded that HIV-infected patients, mainly IDUs, are not receptive to group educational programmes. The design of future interventions must take into account the patients' characteristics.

Adult↗

Resin selection and single-step production and recovery of lactic acid from pretreated wood.

Four ion-exchange resins (Amberlite IRA 900, IRA 400, IRA 96, and IRA 67) were employed for lactic acid recovery from simultaneous saccharification and fermentation (SSF) media. The best resins (Amberlite IRA 900 and IRA 400) were assayed for capacity, regenerant consumption, percentage of lactic acid recovery, and product concentration. Almost quantitative lactic acid recoveries at constant capacities were achieved in four sequential loading/regeneration cycles. A strong-base resin (Amberlite IRA 400) was selected for intermittent lactic acid separation in a typical SSF process, in which pretreated wood was saccharified by cellulases in the presence of Lactobacillus delbrueckii. The dynamics of lactic acid generation and lactic acid recovery were established.

Biotechnology↗

Quantitative determination of total coliforms and Escherichia coli in marine waters with chromogenic and fluorogenic media.

This study compared the performance of LMX(R) broth (LMX), Chromocult Coliform(R) agar (CC) and Chromocult Coliform agar plus cefsulodin (10 microg ml-1) (CC-CFS), with standard methods multiple tube fermentation (MTF), for the enumeration of total coliforms and Escherichia coli from marine recreational waters. LMX and CC are two media designed to concurrently detect total coliform (TC) bacteria and E. coli by the specific action of beta-galactosidase (total coliforms) and beta-glucuronidase (E. coli). Overall results for the TC test showed that LMX, CC and MTF recovered 2.63, 1.95 and 1.90 times as many TCs as CC-CFS, respectively. Data from the multiple range test showed significant differences (P < 0.05) between TC counts on CC-CFS and LMX. The traditional MTF was less sensitive for E. coli enumeration. However, there was no statistically significant differences between LMX, CC, CC-CFS and the MTF method for E. coli enumeration. Background interference was reduced on CC-CFS and the counts obtained reflected more accurately the number of TCs. Therefore, the contribution of beta-galactosidase positive, non coliform bacteria (Aeromonas spp. and Vibrio spp.) to TC counts should not be neglected.

Bacteriological Techniques↗

[Neuropathies of the spinal accessory nerve secondary to cervical surgery: clinical and electrophysiological study of 7 cases].

After innervating the sternocleidomastoid muscle, the spinal accessory nerve emerges and down crosses the posterior cervical triangle to innervate the trapezius. At the posterior triangle, the nerve is closely related to local lymph nodes and may be injured by their enlargement or surgical removal. Injury to this nerve is uncommom. Most cases are iatrogenically due to surgical procedures in the posterior cervical triangle, often following lymph nodes biopsies or benign turmors approaches. We present seven cases of post-surgical related spinal accessory nerve injury and discuss its clinical and electrophysiologycal profiles. The major motor signs were shoulder drop and paresis to raise arm. Pain and sensory complaints were almost universal and most probably due to concomitant injury of cervical plexus cutaneous branches, which are in direct relation to the spinal accessory nerve in the posterior cervical triangle. Lesion type is usually axonotmesis, but varied degrees of compression by cicatricial spurs are observed in some patients. Surgical approach and nerve repair must be considered for pacients with delayed recovery.

Accessory Nerve Diseases↗

[Information program applied to the temporal study of pathological processes in animal populations].

WinEpi Tasas 2.0 is a new, globally accessible computer program, which has been developed as a research tool for the epidemiological study of the pathological processes which affect animal populations. This new version of the program, based on the experimental version, WinEpi Tasas 1.0, enables users to study the distribution and evolution of diseases in affected populations, over time. This information, combined with data on spatial distribution, is of fundamental importance for the establishment of appropriate control strategies and health policies. Such disease control measures are of particular significance in the current context of emerging and re-emerging diseases. The program can be run in either English or Spanish.

Animal Diseases↗

Fragments from alpha-actinin insert into reconstituted lipid bilayers.

Recent experiments have indicated that alpha-actinin interacts with phospholipid membranes. Using computer analysis methods we determined two possible lipid binding sites capable of membrane attachment/insertion, residues 281-300 and 720-739 of the primary amino acid sequence on smooth muscle alpha-actinin. Having expressed these regions as fusion proteins with schistosomal GST (glutathione S-transferase), we used differential scanning calorimetry (DSC) to investigate their interaction with mixtures of zwitterionic (dimyristoyl-l-alpha-phosphatidylcholine, DMPC) and anionic (dimyristoyl-l-alpha-phosphatidylglycerol, DMPG) phospholipids in reconstituted lipid bilayers. Calorimetric measurements showed that as fusion protein concentration increased, the main chain transition enthalpy decreased and chain melting temperatures shifted, which is indicative of partial protein insertion into the hydrophobic region of the lipid membranes. Centrifugation assay and subsequent SDS/Page chromatography confirmed this finding.

Actinin↗

The shedding of membrane-anchored heparin-binding epidermal-like growth factor is regulated by the Raf/mitogen-activated protein kinase cascade and by cell adhesion and spreading.

Heparin-binding epidermal-like growth factor (HB-EGF) is synthesized as a transmembrane precursor (HB-EGF(TM)). The addition of phorbol ester (PMA, phorbol 12-myristate 13-acetate) to cells expressing HB-EGF(TM) results in the metalloproteinase-dependent release (shedding) of soluble HB-EGF. To analyze mechanisms that regulate HB-EGF shedding, a stable cell line was established expressing HB-EGF(TM) in which the ectodomain and the cytoplasmic tail were tagged with hemagglutinin (HA) and Myc epitopes, respectively (HB-EGF(TM)HA/Myc). HB-EGF(TM)HA/Myc cleavage was followed by the appearance of soluble HB-EGFHA in conditioned medium, the loss of biotinylated cell-surface HB-EGF(TM)HA/Myc, and the appearance of a Myc-tagged cytoplasmic tail fragment in cell lysates. By using this approach, several novel metalloproteinase-dependent regulators of HB-EGF(TM) shedding were identified as follows. (i) HB-EGF(TM)HA/Myc shedding induced by PMA was blocked by the mitogen-activated protein (MAP) kinase kinase inhibitor, PD98059. PMA activated MAP kinase within 5 min, but HB-EGF(TM)HA/Myc shedding did not occur until 20 min, suggesting that MAP kinase activation was a necessary step in the pathway of PMA-induced HB-EGF(TM) cleavage. (ii) Activation of an inducible Raf-1 kinase, DeltaRaf-1:estrogen receptor, resulted in a rapid MAP kinase activation within 10 min and shedding of HB-EGF(TM)HA/Myc within 20-40 min. (iii) Serum induced MAP kinase activation and HB-EGF(TM)HA/Myc shedding that were inhibited by PD98059. (iv) Whereas PMA induced HB-EGF(TM)HA/Myc shedding in attached cells, no shedding occurred when the cells were placed in suspension. Shedding was fully restored shortly after cells were allowed to spread on fibronectin, and the extent of PMA-induced shedding increased with the extent of cell spreading. PMA induced the same level of MAP kinase activation whether the cells were attached or in suspension suggesting that although MAP kinase activation might be necessary for shedding, it was not sufficient. Taken together, these results suggest that there are two components of cell regulation that contribute to the shedding process, not previously recognized, the Raf-1/MAP kinase signal transduction pathway and cell adhesion and spreading.

Animals↗

Neuropilin-1 mediates collapsin-1/semaphorin III inhibition of endothelial cell motility: functional competition of collapsin-1 and vascular endothelial growth factor-165.

Neuropilin-1 (NRP1) is a receptor for two unrelated ligands with disparate activities, vascular endothelial growth factor-165 (VEGF165), an angiogenesis factor, and semaphorin/collapsins, mediators of neuronal guidance. To determine whether semaphorin/collapsins could interact with NRP1 in nonneuronal cells, the effects of recombinant collapsin-1 on endothelial cells (EC) were examined. Collapsin-1 inhibited the motility of porcine aortic EC (PAEC) expressing NRP1 alone; coexpressing KDR and NRP1 (PAEC/KDR/NRP1), but not parental PAEC; or PAEC expressing KDR alone. The motility of PAEC expressing NRP1 was inhibited by 65-75% and this inhibition was abrogated by anti-NRP1 antibody. In contrast, VEGF165 stimulated the motility of PAEC/KDR/NRP1. When VEGF165 and collapsin-1 were added simultaneously to PAEC/KDR/NRP1, dorsal root ganglia (DRG), and COS-7/NRP1 cells, they competed with each other in EC motility, DRG collapse, and NRP1-binding assays, respectively, suggesting that the two ligands have overlapping NRP1 binding sites. Collapsin-1 rapidly disrupted the formation of lamellipodia and induced depolymerization of F-actin in an NRP1-dependent manner. In an in vitro angiogenesis assay, collapsin-1 inhibited the capillary sprouting of EC from rat aortic ring segments. These results suggest that collapsin-1 can inhibit EC motility as well as axon motility, that these inhibitory effects on motility are mediated by NRP1, and that VEGF165 and collapsin-1 compete for NRP1-binding sites.

Actins↗

Risk factors associated with the seroprevalence of leptospirosis among students at the veterinary school of Zaragoza University.

The prevalence and risk factors associated with leptospirosis were studied in veterinary students in Zaragoza. Sera were collected at the beginning and end of the academic year 1994 to 1995 and were tested by ELISA against a pool of Leptospira interrogans serovars bratislava, canicola, grippotyphosa, hardjo, icterohaemorrhagiae and pomona antigens. At the beginning of the study the prevalence was 8.14 per cent and at the end it was 11.4 per cent. The incidence of the disease during the study was 0.0394. Risk factors associated with leptospirosis included: taking the course specialising in food inspection and technology, on-farm work, contact with pets in general, and particularly carnivores, and contact with animal traders. The symptoms associated with the disease were myalgia and fever, and the treatment of the symptoms provided some protection.

Antibodies, Bacterial↗

Cogeneration of cellobiose and glucose from pretreated wood and bioconversion to lactic acid: a kinetic study.

Samples of Eucalyptus globulus wood were subjected to sequential treatments of delignification and swelling, and the solid residues were hydrolysed with a cellulase complex deficient in beta-glucosidase activity. In experiments carried out at 45 degrees C, the highest cellulose conversion (61.9%) was achieved in media containing 28 filter paper units/g using a liquor/solid ratio of 30 g/g. Models for the generation of cellobiose and glucose were developed assuming two sequential reactions (conversion of cellulose into cellobiose and hydrolysis of cellobiose to glucose) with end-product that results in competitive inhibition. Selected regression parameters were correlated with the operational conditions by means of empirical models (including linear and interaction terms) and a generalized calculation procedure that allows a close reproduction of data and shows a satisfactory ability for quantitative predictions was developed. Further experiments were carried out to study the bioconversion of hydrolysates by Lactobacillus delbrueckii into lactic acid. The kinetic modelling of the fermentation stage allowed a close interpretation of the experimental data.

Journal Article↗

Comparison and recovery of Escherichia coli and thermotolerant coliforms in water with a chromogenic medium incubated at 41 and 44.5 degrees C.

This study compared the performance of a commercial chromogenic medium, CHROMagarECC (CECC), and CECC supplemented with sodium pyruvate (CECCP) with the membrane filtration lauryl sulfate-based medium (mLSA) for enumeration of Escherichia coli and non-E. coli thermotolerant coliforms (KEC). To establish that we could recover the maximum KEC and E. coli population, we compared two incubation temperature regimens, 41 and 44.5 degrees C. Statistical analysis by the Fisher test of data did not demonstrate any statistically significant differences (P = 0.05) in the enumeration of E. coli for the different media (CECC and CECCP) and incubation temperatures. Variance analysis of data performed on KEC counts showed significant differences (P = 0.01) between KEC counts at 41 and 44.5 degrees C on both CECC and CECCP. Analysis of variance demonstrated statistically significant differences (P = 0.05) in the enumeration of total thermotolerant coliforms (TTCs) on CECC and CECCP compared with mLSA. Target colonies were confirmed to be E. coli at a rate of 91.5% and KEC of likely fecal origin at a rate of 77.4% when using CECCP incubated at 41 degrees C. The results of this study showed that CECCP agar incubated at 41 degrees C is efficient for the simultaneous enumeration of E. coli and KEC from river and marine waters.

Bacteriological Techniques↗