Study of the isotropic contribution to the analysis of photoelectron diffraction experiments at the ALOISA beamline

The angular distribution of the intensity in photoemission experiments is affected by electron diffraction patterns and by a smoothly varying ISO contribution originated by both intrumental details and physical properties of the samples. The origin of the various contributions to the ISO component has been identified since many years. Nonetheless in this work we present original developement of the ED analysis, which arises from the evolution of instrumental performance, in terms of analyzers positioning and angular resolution, as well as collimation and size of X-ray beams in third generation synchrotron sources. The analytical treatement of the instrumental factors is presented in detail for the end station of the ALOISA beamline (Trieste Synchrotron), where a wide variety of scattering geometries is available for ED experiments. We present here the basic formulae and their application to experimental data taken on the Fe/Cu3Au(001) system in order to highlight the role of the various parameters included in the distribution function. A specific model for the surface illumination has been developed as well as the overlayer thickness and surface roughness have been considered.Comment: RevTex, nine pages with five eps figures; to be published in J. Electron Spectrosc. Relat. Pheno

New active site oriented glyoxyl-agarose derivatives of Escherichia coli penicillin G acylase

<p>Abstract</p> <p>Background</p> <p>Immobilized Penicillin G Acylase (PGA) derivatives are biocatalysts that are industrially used for the hydrolysis of Penicillin G by fermentation and for the kinetically controlled synthesis of semi-synthetic β-lactam antibiotics. One of the most used supports for immobilization is glyoxyl-activated agarose, which binds the protein by reacting through its superficial Lys residues. Since in <it>E. coli </it>PGA Lys are also present near the active site, an immobilization that occurs through these residues may negatively affect the performance of the biocatalyst due to the difficult diffusion of the substrate into the active site. A preferential orientation of the enzyme with the active site far from the support surface would be desirable to avoid this problem.</p> <p>Results</p> <p>Here we report how it is possible to induce a preferential orientation of the protein during the binding process on aldehyde activated supports. A superficial region of PGA, which is located on the opposite side of the active site, is enriched in its Lys content. The binding of the enzyme onto the support is consequently forced through the Lys rich region, thus leaving the active site fully accessible to the substrate. Different mutants with an increasing number of Lys have been designed and, when active, immobilized onto glyoxyl agarose. The synthetic performances of these new catalysts were compared with those of the immobilized wild-type (wt) PGA. Our results show that, while the synthetic performance of the wt PGA sensitively decreases after immobilization, the Lys enriched mutants have similar performances to the free enzyme even after immobilization.</p> <p>We also report the observations made with other mutants which were unable to undergo a successful maturation process for the production of active enzymes or which resulted toxic for the host cell.</p> <p>Conclusion</p> <p>The desired orientation of immobilized PGA with the active site freely accessible can be obtained by increasing the density of Lys residues on a predetermined region of the enzyme. The newly designed biocatalysts display improved synthetic performances and are able to maintain a similar activity to the free enzymes. Finally, we found that the activity of the immobilized enzyme proportionally improves with the number of introduced Lys.</p

Sexual Dimorphism in Cellular and Molecular Features in Human ACTH-Secreting Pituitary Adenomas.

Background. Cushing\u2019s disease presents gender disparities in prevalence and clinical course. Little is known, however, about sexual dimorphism at the level of the corticotrope adenoma itself. The aim of the present study was to evaluate molecular features of ACTH-secreting pituitary adenomas collected from female and male patients with Cushing\u2019s disease. (2) Methods. We analyzed 153 ACTH-secreting adenomas collected from 31 men and 122 women. Adenomas were established in culture and ACTH synthesis and secretion assessed in basal conditions as well as during incubation with CRH or dexamethasone. Concurrently, microarray analysis was performed on formalin-fixed specimens and differences in the expression profiles between specimens from male and female patients identified. (3) Results. ACTH medium concentrations in adenomas obtained from male patients were significantly lower than those observed in adenomas from female patients. This could be observed for baseline as well as modulated secretion. Analysis of corticotrope transcriptomes revealed considerable similarities with few, selected differences in functional annotations. Differentially expressed genes comprised genes with known sexual dimorphism, genes involved in tumour development and genes relevant to pituitary pathophysiology. (4) Conclusions. Our study shows for the first time that human corticotrope adenomas present sexual dimorphism and underlines the need for a gender-dependent analysis of these tumours. Differentially expressed genes may represent the basis for gender-tailored target therapy

Assembling surfactants-mucoadhesive polymer nanomicelles (ASMP-nano) for ocular delivery of cyclosporine-A

The physiological protective mechanisms of the eye reduce the bioavailability of topically administered drugs above all for those with high molecular weight and /or lipophilic characteristics, such as Cyclosporine A (CyA). The combined strategy based on the association of nanomicelles and mucoadhesive polymer seems promising since a limited number of commercial products containing CyA have been recently approved. The scope of this investigation was the design of Assembling Surfactants-Mucoadhesive Polymer Nanomicelles (ASMP-Nano), based on a binary system of two surfactants in combination with hyaluronic acid, and their biopharmaceutical evaluation. The optimisation of the ASMP-Nano in term of the amount of surfactants, CyA-loading and size determined the selection of the clear and stable Nano1HAB-CyA formulation containing 0.105% w/w CyA loaded-nanomicelles with a size of 14.41 nm. The nanostructured system had a protective effect towards epithelial corneal cells with a cell viability of more than 80%. It interacted with cellular barriers favouring the uptake and the accumulation of CyA into the cells as evidenced by fluorescent probe distribution, by hindering CyA permeation through reconstituted corneal epithelial tissue. In pharmacokinetics study on rabbits, the nanomicellar carrier prolonged the CyA retention time in the precorneal area mainly in presence of hyaluronic acid (HA), a mucoadhesive polymer

Surfactant-like Effect and Dissolution of Ultrathin Fe Films on Ag(001)

The phase immiscibility and the excellent matching between Ag(001) and Fe(001) unit cells (mismatch 0.8 %) make Fe/Ag growth attractive in the field of low dimensionality magnetic systems. Intermixing could be drastically limited at deposition temperatures as low as 140-150 K. The film structural evolution induced by post-growth annealing presents many interesting aspects involving activated atomic exchange processes and affecting magnetic properties. Previous experiments, of He and low energy ion scattering on films deposited at 150 K, indicated the formation of a segregated Ag layer upon annealing at 550 K. Higher temperatures led to the embedding of Fe into the Ag matrix. In those experiments, information on sub-surface layers was attained by techniques mainly sensitive to the topmost layer. Here, systematic PED measurements, providing chemical selectivity and structural information for a depth of several layers, have been accompanied with a few XRD rod scans, yielding a better sensitivity to the buried interface and to the film long range order. The results of this paper allow a comparison with recent models enlightening the dissolution paths of an ultra thin metal film into a different metal, when both subsurface migration of the deposit and phase separation between substrate and deposit are favoured. The occurrence of a surfactant-like stage, in which a single layer of Ag covers the Fe film is demonstrated for films of 4-6 ML heated at 500-550 K. Evidence of a stage characterized by the formation of two Ag capping layers is also reported. As the annealing temperature was increased beyond 700 K, the surface layers closely resembled the structure of bare Ag(001) with the residual presence of subsurface Fe aggregates.Comment: 4 pages, 3 figure

The spectrum of small-bowel lesions of AL-type amyloidosis at capsule endoscopy

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Refertazione dell'emoglobina glicata in presenza di varianti emoglobiniche

Measurement of glycated hemoglobin (HbA1c) is a key-role laboratory test in the management of diabetic patients. Clinicians need reliable and accurate measurements, with negligible pre-analytical and post-analytical errors. Among the pre-analytical variables, the presence of hemoglobin variants is a challenge to the laboratorians: the purpose of this document is to give some practical advices on how to report HbA1c values in presence of hemoglobin variants. Reporting is quite simple if measurement are carried on by HPLC and other similar analytical methods, when the Hb variant is well separated and does not interfere with HbA or HbA1c, such as in case of heterozygosis for HbS, HbC, HbD and HbE. In case of homozygosis or double heterozygosis for such variants the HbA1c values should not to be reported. Unusual HbA1c values, typically 15% (142 mmol/mol), could indicate the presence of hemoglobin variants. In these cases other approaches to the retrospective evaluation of glycemic control, based on the measurement of the glycation of different proteins, such as albumin, should be followed. Copyright original