Molecular allergen profiling in horses by microarray reveals Fag e 2 from buckwheat as a frequent sensitizer.

BACKGROUND: Companion animals are also affected by IgE-mediated allergies, but the eliciting molecules are largely unknown. We aimed at refining an allergen microarray to explore sensitization in horses and compare it to the human IgE reactivity profiles. METHODS: Custom-designed allergen microarray was produced on the basis of the ImmunoCAP ISAC technology containing 131 allergens. Sera from 51 horses derived from Europe or Japan were tested for specific IgE reactivity. The included horse patients were diagnosed for eczema due to insect bite hypersensitivity, chronic coughing, recurrent airway obstruction and urticaria or were clinically asymptomatic. RESULTS: Horses showed individual IgE-binding patterns irrespective of their health status, indicating sensitization. In contrast to European and Japanese human sensitization patterns, frequently recognized allergens were Aln g 1 from alder and Cyn d 1 from Bermuda grass, likely due to specific respiratory exposure around paddocks and near the ground. The most prevalent allergen for 72.5% of the tested horses (37/51) was the 2S-albumin Fag e 2 from buckwheat, which recently gained importance not only in human but also in horse diet. CONCLUSION: In line with the One Health concept, covering human health, animal health and environmental health, allergen microarrays provide novel information on the allergen sensitization patterns of the companion animals around us, which may form a basis for allergen-specific preventive and therapeutic concepts.

Pyrite surface environment drives molecular adsorption: cystine on pyrite(100) investigated by X-ray photoemission spectroscopy and low energy electron diffraction.

We have demonstrated that the annealing process for cleaning pyrite surfaces is a critical parameter in promoting ordering on the surface and driving surface reactivity. Furthermore, we describe a spectroscopic surface characterization of the presence or absence of the surface ordering, as indicated by the Low Energy Electron Diffraction (LEED) pattern, as a function of the surface annealing process. Complementary X-ray photoemission spectroscopy (XPS) results provide evidence that longer annealing processes of over 3 hours repair the sulfur vacancies in the pyrite, making FeS species partially disappear in favor of FeS2 species. These features play an important role in molecular adsorption. We show that in the case of the cystine amino acid on the (100) pyrite surface, the substrate structure is responsible for the chemical adsorption form. The presence of an ordered structure on the surface, as indicated by the LEED pattern, favors the cystine NH3+ chemical form, whereas the absence of the surface ordering promotes cystine NH2 adsorption due to the sulfur-deficient surface. The cystine molecule could then act by changing its chemical functionalities to compensate for the iron surface coordination. The chemical molecular adsorption form can be selected by the surface annealing conditions, implying that environmental conditions could drive molecular adsorption on mineral surfaces. These findings are relevant in several surface processes, and they could play a possible role in prebiotic chemistry surface reactions and iron-sulfur scenarios.

Estudio transversal sobre la prevalencia de la Enfermedad Metabólica Ósea(EMO) y Nutrición Parenteral Domiciliaria (NPD) en España: datos del Grupo NADYA / Transversal study on the Prevalence of Metabolic Bone Disease (mbd) and Home Parenteral Nutrition (hpn) in Spain: Data from NADYA Group

Los pacientes con insuficiencia intestinal que reciben NPD presentan un elevado riesgo de presentar EMO. El origen de esta afectación ósea es multifactorial y depende en gran parte de la enfermedad de base que origina la necesidaddel soporte. En nuestro medio no disponemos de datos acerca de la prevalencia de esta enfermedad, por lo que elgrupo NADYA-SENPE ha patrocinado este estudio transversal para intentar conocer la prevalencia de la EMO. Material y métodos: Se han recogido datos retrospectivos de 51 pacientes pertenecientes a 13 hospitales. La encuesta realizada incluía datos demográficos y los datos clínicos más relevantes que pudieran influir en la apariciónde EMO. También se han registrados los datos analíticosmás significativos para este proceso (calciuria, PTH,25 OH vitamina D) y los resultados de la primera y la última densitometría realizadas. Resultados: Solamente 21 pacientes tenían realizada una densitometría en el momento de iniciar la NPD. La calidad del hueso está alterada al inicio de la NPD en un porcentaje significativo de casos (52%) Tras un seguimiento medio de 6 años ese porcentaje se eleva hasta el 81%. Dado el carácter retrospectivo del estudio y el escaso número de sujetos no esposible determinar el papel que juega la NPD en la etiología de la EMO. Sólo un 35% de los pacientes presentan niveles de vitamina D por encima de los niveles recomendados y la mayoría de ellos no recibe suplementación específica. Conclusiones: La NPD se asocia a un riesgo muy elevado de presentar EMO, por tanto es necesario disponer de protocolos de actuación que permitan detectar precozmente este problema y orientar el seguimiento y tratamiento de estos pacientes (AU)

Patients with intestinal failure who receive HPN are at high risk of developing MBD. The origin of this bonealteration is multifactorial and depends greatly on theunderlying disease for which the nutritional support is required. Data on the prevalence of this disease in our environment is lacking, so NADYA-SEMPE group has sponsored this transversal study with the aim of knowing the actual MBD prevalence.Material and methods: Retrospective data from 51 patients from 13 hospitals were collected. The questionnaire included demographic data as well as the most clinically relevant for MBD data. Laboratory data (calciuria,PTH, 25 -OH -vitamin D) and the results from the first and last bone densitometry were also registered. Results: Bone mineral density had only been assessed by densitometry in 21 patients at the moment HPN was started. Bone quality is already altered before HPN in a significant percentage of cases (52%). After a mean follow up of 6 years, this percentage increases up to 81%. Due toretrospective nature of the study and the low number of subjects included it has not been possible to determine the role that HPN plays in MBD etiology. Only 35% of patients have vitamin D levels above the recommended limits and the majority of them is not on specific supplementation. Conclusions: HPN is associated with very high risk ofMBD, therefore, management protocols that can lead toearly detection of the problem as well as guiding for followup and treatment of these patients are needed (AU)

[Transversal study on the prevalence of Metabolic Bone Disease (MBD) and Home Parenteral Nutrition (HPN) in Spain: data from NADYA group]. / Estudio transversal sobre la prevalencia de la Enfermedad Metabólica Ósea (EMO) y Nutrición Parenteral Domiciliaria (NPD) en España: datos del Grupo NADYA.

UNLABELLED: Patients with intestinal failure who receive HPN are at high risk of developing MBD. The origin of this bone alteration is multifactorial and depends greatly on the underlying disease for which the nutritional support is required. Data on the prevalence of this disease in our environment is lacking, so NADYA-SEMPE group has sponsored this transversal study with the aim of knowing the actual MBD prevalence. MATERIAL AND METHODS: Retrospective data from 51 patients from 13 hospitals were collected. The questionnaire included demographic data as well as the most clinically relevant for MBD data. Laboratory data (calciuria, PTH, 25 -OH -vitamin D) and the results from the first and last bone densitometry were also registered. RESULTS: Bone mineral density had only been assessed by densitometry in 21 patients at the moment HPN was started. Bone quality is already altered before HPN in a significant percentage of cases (52%). After a mean follow up of 6 years, this percentage increases up to 81%. Due to retrospective nature of the study and the low number of subjects included it has not been possible to determine the role that HPN plays in MBD etiology. Only 35% of patients have vitamin D levels above the recommended limits and the majority of them is not on specific supplementation. CONCLUSIONS: HPN is associated with very high risk of MBD, therefore, management protocols that can lead to early detection of the problem as well as guiding for follow up and treatment of these patients are needed.

Self-assembled monolayers of peptide nucleic acids on gold surfaces: a spectroscopic study.

We have characterized self-assembled monolayers (SAMs) of thiol-derivatized peptide nucleic acid (PNA) chains adsorbed on gold surfaces by using reflection absorption infrared spectroscopy (RAIRS) and X-ray photoemission spectroscopy (XPS) techniques. We have found that the molecular orientation of PNAs strongly depends on surface coverage. At low coverage, PNA chains lie flat on the surface, while at high coverage, PNA molecules realign their molecular axes with the surface normal and form SAMs without the need of co-immobilization of spacers or other adjuvant molecules. The change in the molecular orientation has been studied by infrared spectroscopy and it has been confirmed by atomic force microscopy (AFM). PNA immobilization has been followed by analyzing the N(1s) XPS core-level peak. We show that the fine line shape of the N(1s) core-level peak at optimal concentration for biosensing is due to a chemical shift. A combination of the above-mentioned techniques allow us to affirm that the structure of the SAMs is stabilized by molecule-molecule interactions through noncomplementary adjacent nucleic bases.

[Training of inspiratory muscles in chronic obstructive lung disease. Its impact on functional changes and exercise tolerance]. / Entrenamiento de los músculos inspiratorios en la enfermedad pulmonar obstructiva crónica. Su impacto sobre las alteraciones funcionales y sobre la tolerancia al ejercicio.

The aim of this study was to evaluate the impact of inspiratory muscle training on lung function and exercise tolerance in patients with chronic obstructive pulmonary disease (COPD). Thirty-five patients with stable COPD were enrolled. We measured lung function variables and peak inspiratory and expiratory pressures (PImax and PEmax). Tests of progressive maximal exercise tolerance and stable submaximal exercise tolerance were administered. Two study groups were formed. Group A patients (n = 20) were enrolled in a respiratory muscle training program lasting four months. Group B (n = 15) was the control group. At the end of the study period the patients underwent testing similar to the first battery of tests. All showed moderate to severe obstruction with no significant differences between groups (FEV1: group A 37.6 +/- 13%, group B 36.6 +/- 12%; FVC: group A 80.4 +/- 15%, group B 80 +/- 12%). Nor were there any significant differences between the two groups in initial results of either maximal respiratory pressures or exercise tolerance. No lung function changes were observed in either group. PImax in group A increased significantly at the end of the study (from 54 +/- 9 to 78 +/- 16 cmH2O; p < 0.001); there were no changes in group B. No changes were seen in VO2max or ventilatory response and/or gasometry during exercise in any of the groups. The trained group, on the other hand, experienced a significant decrease in dyspnea evaluated on the Borg scale exercise in maximal (5.7 +/- 1.1 versus 4.7 +/- 1.2, p < 0.005) and submaximal (5.9 +/- 0.9 versus 4.9 +/- 1.3, p < 0.005) and an increase in time of submaximal exercise tolerance (5.5 +/- 2 versus 7 +/- 3 min, p < 0.05), changes that were not observed in the control group. Based on these results, and although specific training of inspiratory muscles does not appear to improve lung function in patients with COPD, it is accompanied by a decreased sense of dyspnea during exercise and greater tolerance.

Chicken antibodies to a recombinant fragment of the equine immunoglobulin epsilon heavy-chain recognising native horse IgE.

An equine immunoglobulin E (IgE) heavy-chain cDNA fragment (CH3-CH4, nucleotides 1132 to 1592) was cloned, expressed in Escherichia coli as a fusion protein with a [His]6-tag and purified over a Ni-NTA column. The recombinant protein was used to immunise hens. Testing of the raised egg yolk immunoglobulin G (IgG) in Western-blot and ELISA revealed high titres against the recombinant equine IgE fragment (reqIgEf). The reqIgEf-specific IgG was successfully affinity-purified on an unconventional affinity matrix: the [His]6-tagged recombinant IgE fragment was bound to Ni-NTA agarose and used to adsorb specific immunoglobulins. In Western-blot of ammonium sulphate precipitated horse serum and bronchoalveolar lavage fluid, separated by SDS-PAGE under denaturing-reducing conditions, the raised antibodies reacted with a protein of approximately 80 kDa. A reaction of the reqIgEf-specific IgG was seen with a 190-200 kDa band when the same horse serum or bronchoalveolar fluid (BALF) was separated under non-reducing conditions. These reactions could be inhibited by preincubation of the immune IgG with reqIgEf, while preincubation with horse IgG did not inhibit the reaction. Antibody-affinity chromatography of horse serum with the reqIgEf-specific chicken IgG resulted in an enrichment of the 80 kDa protein in denaturing Western-blot. Determination of the amino acid composition of this protein and comparison with the equine IgE heavy- chain sequence strongly indicates that the 80 kDa band corresponds to the heavy chain of the horse IgE. The reqIgEf-specific chicken IgG was further characterised in an ELISA for the detection of allergen-specific horse IgE. It was demonstrated that heating IgE positive horse sera at 54 degrees C for 10 min drastically diminished the recognition by the reqIgEf-specific chicken IgG. The reaction is inhibitable by preincubation with reqIgEf in a concentration dependent manner. In addition, preincubation with horse IgG, a nonrelevant [His]6-tagged protein or 2% equine colostrum had no influence on the reqIgEf-specific chicken IgG binding characteristic. This antibody recognising horse IgE will be useful for further studies on the pathogenesis of equine allergic diseases.

Distribution of neuropeptide Y-like immunoreactivity in the brain of the lizard Gallotia galloti.

The distribution of neuropeptide Y (NPY)-like immunoreactivity was studied in the brain of the lizard Gallotia galloti, in order to gain insight into the comparative topography of this peptide. Antisera against both NPY and its C-terminal flanking peptide (C-PON) were used, demonstrating a general coexistence of both peptides, as described in other vertebrates. Most NPY-like immunoreactive (NPY-LI) cell bodies were observed in the telencephalon, specifically in various olfactory structures, all cortices, septum, basal ganglia (except for the globus pallidus), the nucleus of the diagonal band of Broca, the amygdaloid complex, and the bed nucleus of the anterior commissure. NPY-LI cells were also seen in the preoptic and hypothalamic regions and the dorsal thalamus (mainly in the perirotundal belt), as well as in the mesencephalic tegmentum (in the ventral tegmental area, the substantia nigra, and the retrorubral area). NPY-LI fibers and terminals were widely distributed in the brain. All visual and auditory neuropiles were densely innervated. Specially dense plexuses were seen in the nucleus accumbens, the ventral pallidum, the suprachiasmatic and ventromedial hypothalamic nuclei, the nucleus medialis thalami, the left habenula, and the central nucleus of the torus semicircularis. Our analysis shows that the distribution of NPY-like immunoreactivity in the forebrain of Gallotia largely resembles that of other vertebrates, whereas differences are mainly observed in the brainstem. The widespread distribution of NPY in the lizard brain suggests several modulatory functional roles, either in local-circuit systems of the forebrain, or in various limbic, neuroendocrine, and sensory pathways.