Isolation and characterization of an exopolymer produced by Bacillus licheniformis: In vitro antiviral activity against enveloped viruses.

The exopolymer (EPSp) produced by the strain B. licheniformis IDN-EC was isolated and characterized using different techniques (MALDI-TOF, NMR, ATR-FTIR, TGA, DSC, SEM). The results showed that the low molecular weight EPSp contained a long polyglutamic acid and an extracellular teichoic acid polysaccharide. The latter was composed of poly(glycerol phosphate) and was substituted at the 2-position of the glycerol residues with a αGal and αGlcNH2. The αGal O-6 position was also found to be substituted by a phosphate group. The antiviral capability of this EPSp was also tested on both enveloped (herpesviruses HSV, PRV and vesicular stomatitis VSV) and non-enveloped (MVM) viruses. The EPSp was efficient at inhibiting viral entry for the herpesviruses and VSV but was not effective against non-enveloped viruses. The in vivo assay of the EPSp in mice showed no signs of toxicity which could allow for its application in the healthcare sector.

[Exposure to service user violence and job satisfaction among nursing staff in public hospitals in Murcia Region]. / Exposición a violencia de los usuarios y satisfacción laboral en el personal de enfermería de los hospitales públicos de la Región de Murcia.

BACKGROUND: Manifestations of physical and psychological violence can affect the levels of job satisfaction. The objective of this study is to determine the frequency of exposure to user violence by the nursing staff in public hospitals in the region of Murcia and its relationship with their level of job satisfaction. METHODS: Cross-sectional descriptive study through a self-administered and anonymous questionnaire sent to the nursing staff of the public hospitals in the region of Murcia (1,489 professionals). The instruments used were the Hospital Aggressive Behaviour Scale-Users (HABS-U) to rate violence and the Overall Job Satisfaction (OJS) to assess job satisfaction. RESULTS: Most workers (71%) reported suffering at least one of the manifestations of non-physical violence and 19.9% one of the physical manifestations with at least an annual frequency. Mean extrinsic satisfaction was 30.1 (SD=6.9), being coworkers the factor with higher frequency of satisfied workers (68.2%) and how the company was managed the factor with more dissatisfied workers (28.8%). Intrinsic satisfaction was lower (25.5, SD=7.2), being variety of tasks the factor with more satisfied workers (51.7%) and promotion opportunity the one with more dissatisfied workers (22.4%). Both extrinsic and intrinsic satisfaction correlated inversely with the exposure to non-physical and physical violence (higher exposure to violence lowered job satisfaction). CONCLUSIONS: In general, the scores for nurses' job satisfaction were low. One of the aspects affecting this dissatisfaction might be exposure to workplace violence; thus reducing exposure to violence could increase job satisfaction that, in turn, would improve patient care.

Catching elusive glycosyl cations in a condensed phase with HF/SbF5 superacid.

Glycosyl cations are universally accepted key ionic intermediates in the mechanism of glycosylation, the reaction that covalently links carbohydrates to other molecules. These ions have remained hypothetical species so far because of their extremely short life in organic media as a consequence of their very high reactivity. Here, we report the use of liquid hydrofluoric acid-antimony pentafluoride (HF/SbF5) superacid to generate and stabilize the glycosyl cations derived from peracetylated 2-deoxy and 2-bromoglucopyranose in a condensed phase. Their persistence in this superacid medium allows their three-dimensional structure to be studied by NMR, aided by complementary computations. Their deuteration further confirms the impact of the structure of the glycosyl cation on the stereochemical outcome of its trapping.

Review: Use of residual dipolar couplings to determine the structure of carbohydrates.

Solution nuclear magnetic resonance spectroscopy is especially useful in the carbohydrate field. The measurement of residual dipolar couplings provides long-range structural information, a valuable complement for the structural study of carbohydrates either in its free form or in the bound state to proteins. They permit to deduce the geometry and the flexibility of the glycosidic linkages, which have a major influence on the conformation of carbohydrates and their overall shape. This article reviews the current application of the residual dipolar couplings methodology to carbohydrates.

Lignin composition and structure in young versus adult Eucalyptus globulus plants.

Lignin changes during plant growth were investigated in a selected Eucalyptus globulus clone. The lignin composition and structure were studied in situ by a new procedure enabling the acquisition of two-dimensional nuclear magnetic resonance (2D-NMR) spectra on wood gels formed in the NMR tube as well as by analytical pyrolysis-gas chromatography-mass spectrometry. In addition, milled-wood lignins were isolated and analyzed by 2D-NMR, pyrolysis-gas chromatography-mass spectrometry, and thioacidolysis. The data indicated that p-hydroxyphenyl and guaiacyl units are deposited at the earlier stages, whereas the woods are enriched in syringyl (S) lignin during late lignification. Wood 2D-NMR showed that ß-O-4' and resinol linkages were predominant in the eucalypt lignin, whereas other substructures were present in much lower amounts. Interestingly, open ß-1' structures could be detected in the isolated lignins. Phenylcoumarans and cinnamyl end groups were depleted with age, spirodienone abundance increased, and the main substructures (ß-O-4' and resinols) were scarcely modified. Thioacidolysis revealed a higher predominance of S units in the ether-linked lignin than in the total lignin and, in agreement with NMR, also indicated that resinols are the most important nonether linkages. Dimer analysis showed that most of the resinol-type structures comprised two S units (syringaresinol), the crossed guaiacyl-S resinol appearing as a minor substructure and pinoresinol being totally absent. Changes in hemicelluloses were also shown by the 2D-NMR spectra of the wood gels without polysaccharide isolation. These include decreases of methyl galacturonosyl, arabinosyl, and galactosyl (anomeric) signals, assigned to pectin and related neutral polysaccharides, and increases of xylosyl (which are approximately 50% acetylated) and 4-O-methylglucuronosyl signals.

El abuso de drogas en el marco de los trastornos del comportamiento perturbador / Drug abuse in the context of disruptive behavior disorders

El presente estudio es una revisión que trata de analizar las causas y la evolución de los trastornos del comportamiento perturbador en la infancia y la adolescencia, así como su repercusión en el desarrollo de un posible trastorno por abuso de sustancias. La metodología utilizada fue la búsqueda bibliográfica, utilizando los descriptores MESH, "conduct disorders and substance abuse disorders", en las principales bases de datos (Biblioteca Cochrane, Tripdatabase, Medline, LILACS, CINAHL y CUIDEN). Con esta metodología, se seleccionaron un total de 35 estudios, por su especial relevancia y relación con el tema objeto de esta revisión. El análisis y lectura crítica de los estudios revisados nos llevan a concluir que: a) existe una relación de causalidad entre los trastornos del comportamiento perturbador y el desarrollo de un trastorno por abuso de sustancias; b) la gran influencia que en esta evolución tiene el entorno familiar y comunitario requiere que la prevención y el tratamiento deban centrarse en la familia (agente socializador más importante); también son necesarias actuaciones en el ámbito escolar, social (grupo de iguales) y comunitario (AU)

This study is a review that seeks to analyze the causes and development of disruptive behavior disorders in childhood and adolescence and its impact on the development of a possible substance abuse disorder. The methodology used was literature search using the MESH descriptors, "conduct disorders and substance abuse disorders", in the main databases (Cochrane Library, Tripdatabase, Medline, LILACS, CINAHL and CUIDEN). Using this methodology, we selected a total of 35 studies, for its special relevance and relationship to the subject under review. The analysis and critical reading of the reviewed studies lead us to conclude that: a) there is a causal link between the disruptive behavior disorders and the development of a substance abuse disorder; b) the great influence in this evolution is the family and community environment, requires the prevention and treatment should focus on the family (most important socializing agent) and the necessary actions in schools, social (peer group) and community (AU)

Abordaje familiar del trastorno de comportamiento perturbador en la infancia / Family approach to disruptive behavior disorder in childhood

El presente trabajo trata de reflejar la eficacia observada en estudios recientes sobre los tratamientos psicoterapéuticos en niños y adolescentes que presentan problemas de comportamiento y agresividad, limitándonos exclusivamente a la intervención familiar. La metodología utilizada fue la búsqueda bibliográfica de estudios en metabuscadores (Trip database, Biblioteca Cochrane)y bases de datos (MEDLINE, CINAHL, EMBASE), centrándonos en metaanálisis, revisiones sistemáticas y ensayos clínicos aleatorizados publicados desde el 01/01/1999. El análisis de dichos estudios indica que la intervención familiares eficaz y necesaria para la modificación del comportamiento perturbador en los menores, y supone una optimización de la economía sanitaria pública, al reducir de forma notable las demandas a la red sanitaria de salud mental. También exponemos un ejemplo de este tipo de intervención en España, la Escuela de Padres, válida especialmente para los casos en que se observa un problema relacionado con la crianza del niño (Z.62, CIE-10). Sin embargo, tras la búsqueda bibliográfica, no encontramos estudios de relevancia sobre este tema en nuestro país (AU)

This paper tries to reflect the performance observed in recent studies on the psychotherapeutic treatment in children and adolescents who have behavioral and aggressiveness problems, focusing exclusively on the family intervention. The methodology used in the literature bibliographical metasearchers (Trip Database, Cochrane Library), and Database Studies (MEDLINE, CINAHL, EMBASE), focusing on metaanalysis, systematic reviews and randomized clinical trials published since 01/01/1999.The analysis of said studies indicate that family intervention is effective and necessary for the modification of unruly behavior in children, and mean an optimization of the economy public health, to significantly reduce the demands on the health network of mental health. They also present an example of this type of intervention in Spain, the Parent's School, especially valid for those cases where there is a problem with the upbringing of children (Z.62,ICD-10). However, after the bibliographical search, we found no studies on this important subject in our country (AU)

Alternativa psicofamiliar al tratamiento precoz de los problemas de conducta: la escuela de padres / No disponible

Los trastornos del comportamiento perturbador(TCP) constituyen un tema de gran interés, debido alaumento en la prevalencia que se ha producido en lasúltimas décadas, la problemática que representa para eladolescente y su entorno, la complejidad del abordaje ytratamiento, preferentemente multidisciplinar, así comolas consecuencias a corto y largo plazo tanto para elpropio joven como para la sociedad en general.El presente trabajo trata de reflejar la eficacia observadaen estudios recientes sobre los tratamientos psicoterapéuticoscentrándonos en la escuela de padres (EP),especialmente para aquellos casos en los que se observaun problema relacionado con la crianza del niño (Z.62, CIE-10)(AU)

The behavioral disorders (BD) of the teenager are atopic of great interest, due to the increase in prevalencethat has occurred in recent decades, the problems it createsfor adolescents and their environment, the complexityof the management and treatment, preferablymultidisciplinary, as well as the short and long term forboth the young and himself to society in general.This study tries to reflect the effectiveness observedin recent studies, the psychotherapeutic treatments,focusing on the school parent (SP), especially for thosecases in which there is a problem with the upbringing ofchildren (Z.62, ICD-10)(AU)

Protection from Staphylococcus aureus mastitis associated with poly-N-acetyl beta-1,6 glucosamine specific antibody production using biofilm-embedded bacteria.

Staphylococcus aureus vaccines based on bacterins surrounded by slime, surface polysaccharides coupled to protein carriers and polysaccharides embedded in liposomes administered together with non-biofilm bacterins confer protection against mastitis. However, it remains unknown whether protective antibodies are directed to slime-associated known exopolysaccharides and could be produced in the absence of bacterin immunizations. Here, a sheep mastitis vaccination study was carried out using bacterins, crude bacterial extracts or a purified exopolysaccharide from biofilm bacteria delivered in different vehicles. This polysaccharide reacted specifically with antibodies to poly-N-acetyl-beta-1,6-glucosamine (PNAG) and not with antibodies to other capsular antigens or bacterial components. Following intra-mammary challenge with biofilm-producing bacteria, antibody production against the polysaccharide, milk bacterial counts and mastitis lesions were determined. Bacterins from strong biofilm-producing bacteria triggered the highest production of antibodies to PNAG and conferred the highest protection against infection and mastitis, compared with weak biofilm-producing bacteria and non-cellular inocula. Thus, bacterins from strong biofilm bacteria, rather than purified polysaccharide, are proposed as a cost-efficient vaccination against S. aureus ruminant mastitis.

The interaction of microtubules with stabilizers characterized at biochemical and structural levels.

Since the discovery of paclitaxel and its peculiar mechanism of cytotoxicity, which has made it and its analogues widely used antitumour drugs, great effort has been made to understand the way they produce their effect in microtubules and to find other products that share this effect without the undesired side effects of low solubility and development of multidrug resistance by tumour cells. This chapter reviews the actual knowledge about the biochemical and structural mechanisms of microtubule stabilization by microtubule stabilizing agents, and illustrates the way paclitaxel and its biomimetics induce microtubule assembly, the thermodynamics of their binding, the way they reach their binding site and the conformation they have when bound.

NMR structural studies of oligosaccharides related to cancer processes.

A summary of spectroscopic methods for structural and conformational elucidation of bioactive carbohydrates based on nuclear magnetic resonance (NMR) is described. The formation of carbohydrate-protein complexes is often the initial step of biological responses. Therefore, knowledge about the structural factors that stabilize the complex may be relevant and contribute to predict the structural/conformational requirements of new drugs acting as agonists. Two examples of medical significance in the cancer research field are discussed (1) conformational studies of glycoconjugates related to antitumour vaccines (2) conformational analysis of glycosaminoglycans and the interaction heparin-fibroblast growth factor (FGF).

Chemical structure of a polysaccharide isolated from the cell wall of Arachniotus verruculosus and A. ruber.

The structure of a cell wall alkali-extractable and water-soluble polysaccharide isolated from two species of Arachniotus has been established by reductive cleavage and NMR spectroscopy. The linear polysaccharide consists of a regular disaccharide-repeating unit with the structure: [-->6)-beta-D-Galf-(1-->5)-beta-D-Galf-(1-->](n)-->mannan core.

Structure of a cell wall polysaccharide isolated from Hypocrea gelatinosa.

The structure of a polysaccharide isolated from the cell wall of Hypocrea gelatinosa has been investigated by means of chemical analyses and 1D and 2D NMR spectroscopy. The polysacharide has an irregular structure, idealized as follows: [carbohydrate structure in text].

Synthesis and solution conformational analysis of 2,3-anhydro-3-C-[(1R)-2,6-anhydro-1-deoxy-1-fluoro-D-glycero-D-gulo-heptitol-1-C-yl]-beta-D-gulo-furanose: first example of a monofluoromethylene-linked C-disaccharide.

Condensation of 2,3,4,6-tetra-O-benzyl-beta-D-glucopyranosylcarbaldehyde with isolevoglucosenone induced by Et(2)AlI, followed by epoxidation, gave an aldol that was fluorinated into a monofluoromethylene C-glucopyranoside that was converted into the title C-disaccharide 1. Its conformational behavior in water has been studied by using a combination of NMR spectroscopy (J and NOE data) and molecular mechanics calculations.

Carrier protein-modulated presentation and recognition of an N-glycan: observations on the interactions of Man(8) glycoform of ribonuclease B with conglutinin.

Conglutinin is a serum lectin of the innate immune system, which binds high mannose N-glycans when these are appropriately presented on proteins. Here we use the conglutinin-ribonuclease B (RNaseB)-recognition system as a model to investigate the structural basis of selective recognition of protein-bound oligosaccharides by this carbohydrate-binding receptor. Conglutinin shows little binding to the isolated RNaseB-Man(8 )glycoform, and no binding to Man(5-6) glycoforms. In contrast, when the protein moiety is reduced and denatured we observe that conglutinin binds strongly to the isolated RNaseB-Man(8) glycoform and weakly to the Man(5-6) glycoforms. These results are in accord with observations on the binding to the N-glycans in the absence of carrier protein. NMR analyses of native RNaseB-Man(8) and -Man(5-6) glycoforms reveal that the three-dimensional structure of the protein moiety is essentially identical to that of non-glycosylated RNase (RNaseA). Thus there are no perceptible differences between the RNase protein forms that could account for differential availability of the N-glycan for conglutinin-binding. After reduction and denaturation, the NMR spectrum became typical of a non-structured polypeptide, although the conformational preferences of the N-glycosidic linkage were unchanged, and most importantly, the Man(8 )oligosaccharide retained the average conformational behavior of the free oligosaccharide irrespective of the carrier protein fold. This conformational freedom is clearly not translated into full availability of the oligosaccharide for the carbohydrate-recognition protein. We propose, therefore, that the differing bioactivity of the N-glycan is a reflection of the existence of different geometries of presentation of the carbohydrate determinant in relation to the protein surface within the glycan:carrier protein ensemble.

Towards defining the role of glycans as hardware in information storage and transfer: basic principles, experimental approaches and recent progress.

The term 'code' in biological information transfer appears to be tightly and hitherto exclusively connected with the genetic code based on nucleotides and translated into functional activities via proteins. However, the recent appreciation of the enormous coding capacity of oligosaccharide chains of natural glycoconjugates has spurred to give heed to a new concept: versatile glycan assembly by the genetically encoded glycosyltransferases endows cells with a probably not yet fully catalogued array of meaningful messages. Enciphered by sugar receptors such as endogenous lectins the information of code words established by a series of covalently linked monosaccharides as letters for example guides correct intra- and intercellular routing of glycoproteins, modulates cell proliferation or migration and mediates cell adhesion. Evidently, the elucidation of the structural frameworks and the recognition strategies within the operation of the sugar code poses a fascinating conundrum. The far-reaching impact of this recognition mode on the level of cells, tissues and organs has fueled vigorous investigations to probe the subtleties of protein-carbohydrate interactions. This review presents information on the necessarily concerted approach using X-ray crystallography, molecular modeling, nuclear magnetic resonance spectroscopy, thermodynamic analysis and engineered ligands and receptors. This part of the treatise is flanked by exemplarily chosen insights made possible by these techniques.

NMR investigations of protein-carbohydrate interactions: insights into the topology of the bound conformation of a lactose isomer and beta-galactosyl xyloses to mistletoe lectin and galectin-1.

A hallmark of oligosaccharides is their often limited spatial flexibility, allowing them to access a distinct set of conformers in solution. Viewing each individual or even the complete ensemble of conformations as potential binding partner(s) for lectins in protein-carbohydrate interactions, it is pertinent to address the question on the characteristics of bound state conformation(s) in solution. Also, it is possible that entering the lectin's binding site distorts the low-energy topology of a glycosidic linkage. As a step to delineate the strategy of ligand selection for galactosides, a common physiological docking point, we have performed a NMR study on two non-homologous lectins showing identical monosaccharide specificity. Thus, the conformation of lactose analogues bound to bovine heart galectin-1 and to mistletoe lectin in solution has been determined by transferred nuclear Overhauser effect measurements. It is demonstrated that the lectins select the syn conformation of lactose and various structural analogues (Galbeta(1-->4)Xyl, Galbeta(1-->3)Xyl, Galbeta(1-->2)Xyl, and Galbeta(1-->3)Glc) from the ensemble of presented conformations. No evidence for conformational distortion was obtained. Docking of the analogues to the modeled binding sites furnishes explanations, in structural terms, for exclusive recognition of the syn conformer despite the non-homologous design of the binding sites.

Conformational studies of the Man8 oligosaccharide on native ribonuclease B and on the reduced and denatured protein.

Site-specific presentation of oligosaccharides in the context of carrier proteins can influence markedly their recognition by carbohydrate-binding proteins. On RNaseB, the Man5-9 N-glycans at Asn-34 are bound by the serum lectin conglutinin when the glycoprotein is reduced and denatured, but there is no binding to the N-glycans on the native form of RNaseB. The RNaseB Man8, which is a glycoform preferentially bound by conglutinin, is the subject of the present study. The conformational behavior of the protein-linked oligosaccharide Man8 is investigated on the native and on the reduced and denatured RNaseB, using a combination of NMR and theoretical calculations. Quantitative data on the NOESY crosspeaks have been obtained, thereby allowing the comparison of mobilities of homologous linkages within the glycan chain. Oligosaccharide conformations compatible with the NMR data have been explored by molecular modeling of the free oligosaccharide, using two different force fields (AMBER and SYBYL). There are some differences between the results produced by the two force fields, the AMBER simulations providing a better agreement with the experimental data. The results indicate that both on the native and on the reduced heat-denatured glycoprotein, the RNase Man8 oligosaccharide exhibits a conformational behavior very similar to that of the free oligosaccharide. However, this conformational freedom of the N-glcyan does not amount to full availability for carbohydrate-recognition proteins and enzymes.

Studies on the solution conformation and dynamics of a polysaccharide from Sinorhizobium fredii HH103 and its monosaccharide repeating unit.

The conformational behavior of the homopolysaccharide isolated from Sinorhizobium fredii HH103 and its monosaccharide repeating unit (5-acetamido-3,5,7,9-tetradeoxy-7-(3-hydroxybutyramido)-L-glycero- L- manno-nonulosonic acid) was analyzed by nuclear magnetic resonance (NMR) spectroscopy and extensive molecular dynamics simulations (MD). The results indicate that the glycosidic linkages and lateral chains may adopt a variety of conformations. MD simulations using the generalized Born solvent-accessible surface area (GB/SA) continuum solvent model for water and the MM3* force field provide a population distribution of conformers that satisfactorily agrees with the experimental NMR data for the torsional degrees of freedom of the molecule.