Botulinum toxin A and B raise blood flow and increase survival of critically ischemic skin flaps

BACKGROUND Botulinum toxin (BTX) A and B are commonly used for aesthetic indications and in neuromuscular disorders. New concepts seek to prove efficacy of BTX for critical tissue perfusion. Our aim was to evaluate BTX A and B in a mouse model of critical flap ischemia for preoperative and intraoperative application. METHODS BTX A and B were applied on the vascular pedicle of an axial pattern flap in mice preoperatively or intraoperatively. Blood flow, tissue oxygenation, tissue metabolism, flap necrosis rate, apoptosis assay, and RhoA and eNOS expression were endpoints. RESULTS Blood-flow measurements 1 d after the flap operation revealed a significant reduction to 53% in the control group, while flow was maintained or increased in all BTX groups (103%-129%). Over 5 d all BTX groups showed significant increase in blood flow to 166-187% (P < 0.01). Microdialysis revealed an increase of glucose and reduced lactate/pyruvate ratio and glycerol levels in the flap tissue of all BTX groups. This resulted in significantly improved tissue survival in all BTX groups compared with the control group (62% ± 10%; all P < 0.01): BTX A preconditioning (84% ± 5%), BTX A application intraoperatively (88% ± 4%), BTX B preconditioning (91% ± 4%), and intraoperative BTX B treatment (92% ± 5%). This was confirmed by TUNEL assay. Immunofluorescence demonstrated RhoA and eNOS expression in BTX groups. All BTX applications were similarly effective, despite pharmacologic dissimilarities and different timing. CONCLUSIONS In conclusion, we were able to show on a vascular, tissue, cell, and molecular level that BTX injection to the feeding arteries supports flap survival through ameliorated blood flow and oxygen delivery.

Venepuncture during head-up tilt testing in patients with suspected vasovagal syncope - implications for the test protocol.

BACKGROUND AND PURPOSE Head-up tilt (HUT) testing is a widely used diagnostic tool in patients with suspected vasovagal syncope (VVS). However, no gold standard exists for this examination and the various protocols used have a limited sensitivity and specificity. Our aim was to determine the sensitivity of a sequential HUT testing protocol including venepuncture (VP) and sublingual nitroglycerin application. METHODS This was a retrospective analysis of the diagnostic gain of a sequential HUT testing protocol including VP applied 10 min after the start of HUT testing and sublingual application of nitroglycerin 20 min after the start of the test protocol in 106 patients with a final diagnosis of VVS. The sensitivity of the test protocol was compared between patients with positive and negative history for VP induced VVS. RESULTS Overall, pre-syncope or syncope occurred in 68 patients (64.2%). Only 17% of all patients fainted spontaneously within 10 min of passive HUT. Another 39.6% fainted within 20 min. Application of nitroglycerin after 20 min of HUT evoked syncope in another 7.5% until the end of 45 min of HUT. The sensitivity of the test protocol for evoking (pre-)syncope was 94.4% in patients with a positive history for VP associated VVS and 58% in patients with a negative history (P < 0.01**); 85.7% of patients with a positive history and 42.9% of patients with a negative history fainted within 20 min of HUT testing (P < 0.01**). CONCLUSIONS Implementation of VP in sequential HUT testing protocols allows the sensitivity of HUT testing to be increased, especially in patients with a positive history for VP associated VVS.

Materno-fetal nutrient transfer across primary human trophoblast layer.

MATERNO-FETAL NUTRIENT TRANSFER ACROSS PRIMARY HUMAN TROPHOBLAST MONOLAYER Objectives: Polarized trophoblasts represent the transport and metabolic barrier between the maternal and fetal circulation. Currently human placental nutrient transfer in vitro is mainly investigated unidirectionallyon cultured primary trophoblasts, or bidirectionally on the Transwell® system using BeWo cells treated with forskolin. As forskolin can induce various gene alterations (e.g. cAMP response element genes), we aimed to establish a physiological primary trophoblast model for materno-fetal nutrient exchange studies without forskolin application. Methods: Human term cytotrophoblasts were isolated by enzymatic digestion and Percoll® gradient separation. The purity of the primary cells was assessed by flow cytometry using the trophoblast-specific marker cytokeratin-7. After screening different coating matrices, we optimized the growth conditions for the primary cytotrophoblasts on Transwell/ inserts. The morphology of 5 days cultured trophoblasts was determined by scanning electron microscopy (SEM) and transmission electron microscopy (TEM). Membrane makers were visualized using confocal microscopy. Additionally transport studies were performed on the polarized trophoblasts in the Transwell® system. Results: During 5 days culture, the trophoblasts (>90% purity) developed a modest trans-epithelial electrical resistance (TEER) and a sizedependent apparent permeability coefficient (Papp) to fluorescently labeled compounds (MW ~400-70’000D). SEM analyses confirmed a confluent trophoblast layer with numerous microvilli at day six, and TEM revealed a monolayer with tight junctions. Immunocytochemistry on the confluent trophoblasts showed positivity for the cell-cell adhesion molecule E-cadherin, the tight junction protein ZO-1, and the membrane proteins ABCA1 and Na+/K+-ATPase. Vectorial glucose and cholesterol transport studies confirmed functionality of the cultured trophoblast barrier. Conclusion: Evidence from cell morphology, biophysical parameters and cell marker expressions indicate the successful and reproducible establishment of a primary trophoblast monolayer model suitable for transport studies. Application of this model to pathological trophoblasts will help to better understand the mechanism underlying gestational diseases, and to define the consequences of placental pathology on materno-fetal nutrient transport.

[Pilot Project: Improved Lung Deposition via a New Nasal Inhalation Procedure].

INTRODUCTION Inhaled drugs can only be effective if they reach the middle and small airways. This study introduces a system that combines a trans-nasal application of aerosols with noninvasive pressure support ventilation. METHODS In a pilot study, 7 COPD patients with GOLD stages II and III inhaled a radiolabeled marker dissolved in water via a trans-nasal route. The mean aerosol particle size was 5.5 µm. Each patient took part in two inhalation sessions that included two application methods and were at least 70 hours apart. During the first session ("passive method"), the patient inhaled the aerosol through an open tube system. The second session ("active method") included pressure support ventilation during the inhalation process. A gamma camera and planar scintigraphy was used to determine the distribution of aerosol particles in the patient's body and lung. RESULTS The pressure supported inhalation ("active method") results in an increased aerosol lung deposition compared to the passive method. Above all, we could demonstrate deposition in the lung periphery with relatively large aerosol particles (5.5 µm). DISCUSSION The results prove that the combination of trans-nasal inhalation with noninvasive pressure support ventilation leads to significantly increased particle deposition in the lung.

Estudio de la redistribución tensional en la interfase frp-hormigón

El uso de materiales compuestos para el refuerzo, reparación y rehabilitación de estructuras de hormigón se ha convertido en una técnica muy utilizada en la última década. Con independencia de la técnica del refuerzo, uno de los principales condicionantes del diseño es el fallo de la adherencia entre el hormigón y el material compuesto, atribuida generalmente a las tensiones en la interfaz de estos materiales. Las propiedades mecánicas del hormigón y de los materiales compuestos son muy distintas. Los materiales compuestos comúnmente utilizados en ingeniería civil poseen alta resistencia a tracción y tienen un comportamiento elástico y lineal hasta la rotura, lo cual, en contraste con el ampliamente conocido comportamiento del hormigón, genera una clara incompatibilidad para soportar esfuerzos de forma conjunta. Esta incompatibilidad conduce a fallos relacionados con el despegue del material compuesto del sustrato de hormigón. En vigas de hormigón reforzadas a flexión o a cortante, el despegue del material compuesto es un fenómeno que frecuentemente condiciona la capacidad portante del elemento. Existen dos zonas potenciales de iniciación del despegue: los extremos y la zona entre fisuras de flexión o de flexión-cortante. En el primer caso, la experiencia a través de los últimos años ha demostrado que se puede evitar prolongando el refuerzo hasta los apoyos o mediante el empleo de algún sistema de anclaje. Sin embargo, las recomendaciones para evitar el segundo caso de despegue aún se encuentran lejos de poder prever el fallo de forma eficiente. La necesidad de medir la adherencia experimentalmente de materiales FRP adheridos al hormigón ha dado lugar a desarrollar diversos métodos por la comunidad de investigadores. De estas campañas experimentales surgieron modelos para el pronóstico de la resistencia de adherencia, longitud efectiva y relación tensión-deslizamiento. En la presente tesis se propone un ensayo de beam-test, similar al utilizado para medir la adherencia de barras de acero, para determinar las características de adherencia del FRP al variar la resistencia del hormigón y el espesor del adhesivo. A la vista de los resultados, se considera que este ensayo puede ser utilizado para investigar diferentes tipos de adhesivos y otros métodos de aplicación, dado que representa con mayor realidad el comportamiento en vigas reforzadas. Los resultados experimentales se trasladan a la comprobación del fallo por despegue en la región de fisuras de flexión o flexión cortante en vigas de hormigón presentando buena concordancia. Los resultados condujeron a la propuesta de que la limitación de la deformación constituye una alternativa simple y eficiente para prever el citado modo de fallo. Con base en las vigas analizadas, se propone una nueva expresión para el cálculo de la limitación de la deformación del laminado y se lleva a cabo una comparación entre los modelos existentes mediante un análisis estadístico para evaluar su precisión. Abstract The use of composite materials for strengthening, repairing or rehabilitating concrete structures has become more and more popular in the last ten years. Irrespective of the type of strengthening used, design is conditioned, among others, by concrete-composite bond failure, normally attributed to stresses at the interface between these two materials. The mechanical properties of concrete and composite materials are very different. Composite materials commonly used in civil engineering possess high tensile strength (both static and long term) and they are linear elastic to failure, which, in contrast to the widely known behavior of concrete, there is a clear incompatibility which leads to bond-related failures. Bond failure in the composite material in bending- or shear-strengthened beams often controls bearing capacity of the strengthened member. Debonding failure of RC beams strengthened in bending by externally-bonded composite laminates takes place either, at the end (plate end debonding) or at flexure or flexure-shear cracks (intermediate crack debonding). In the first case, the experience over the past years has shown that this can be avoided by extending laminates up to the supports or by using an anchoring system. However, recommendations for the second case are still considered far from predicting failure efficiently. The need to experimentally measure FRP bonding to concrete has induced the scientific community to develop test methods for that purpose. Experimental campaigns, in turn, have given rise to models for predicting bond strength, effective length and the stress-slip relationship. The beam-type test proposed and used in this thesis to determine the bonding characteristics of FRP at varying concrete strengths and adhesive thicknesses was similar to the test used for measuring steel reinforcement to concrete bonding conditions. In light of the findings, this test was deemed to be usable to study different types of adhesives and application methods, since it reflects the behavior of FRP in strengthened beams more accurately than the procedures presently in place. Experimental results are transferred to the verification of peeling-off at flexure or flexure-shear cracks, presenting a good general agreement. Findings led to the conclusion that the strain limitation of laminate produces accurate predictions of intermediate crack debonding. A new model for strain limitation is proposed. Finally, a comprehensive evaluation based on a statistical analysis among existing models is carried out in order to assess their accuracy.

Using realist evaluation to assess primary healthcare teams’ responses to intimate partner violence in Spain

Objective: Few evaluations have assessed the factors triggering an adequate health care response to intimate partner violence. This article aimed to: 1) describe a realist evaluation carried out in Spain to ascertain why, how and under what circumstances primary health care teams respond to intimate partner violence, and 2) discuss the strengths and challenges of its application. Methods: We carried out a series of case studies in four steps. First, we developed an initial programme theory (PT1), based on interviews with managers. Second, we refined PT1 into PT2 by testing it in a primary healthcare team that was actively responding to violence. Third, we tested the refined PT2 by incorporating three other cases located in the same region. Qualitative and quantitative data were collected and thick descriptions were produced and analysed using a retroduction approach. Fourth, we analysed a total of 15 cases, and identified combinations of contextual factors and mechanisms that triggered an adequate response to violence by using qualitative comparative analysis. Results: There were several key mechanisms —the teams’ self-efficacy, perceived preparation, women-centred care—, and contextual factors —an enabling team environment and managerial style, the presence of motivated professionals, the use of the protocol and accumulated experience in primary health care—that should be considered to develop adequate primary health-care responses to violence. Conclusion: The full application of this realist evaluation was demanding, but also well suited to explore a complex intervention reflecting the situation in natural settings.

Aplicação de métodos e técnicas sustentáveis na gestão de resíduos sólidos

O artigo aborda a temática da produção e consumo de bens pela sociedade atual que ocasionam sérios problemas ambientais e sociais devido ao grande descarte de materiais de diversas naturezas. O consumo exagerado é incentivado pelas estratégias de markting que incentivam novas compras e descartes muitas vezes prematuros de produtos e embalagens. A sustentabilidade é um fator importante que deve ser considerado nesse debate pois está diretamente vinculada ao chamado desenvolvimento sustentável, um paradigma ainda em construção. À partir dos princípios gerais do desenvolvimento sustentável podem ser estabelecidos outros que abordem exclusivamente a sustentabilidade com relação ao Gestão e Gerenciamento dos Resíduos Sólidos Urbanos abrangendo as dimensões ambiental/ecológica, econômica e social. As estratégias sustentáveis para esse gerenciamento são investigadas há anos por instituições de pesquisa/ensino, empresas e administração pública no intuito de se alcançar uma solução para os problemas advindos a produção e descarte de resíduos sólidos. O texto apresenta exemplos de ações no Brasil e Espanha no que tange às ações de gestão dos resíduos descartados nesses países, com destaque para a Comunidade de Barcelona.

Evaluation of disinfecting procedures for aseptic transfer in hospital pharmacy departments

Current practice in National Health Service (NHS) hospitals employs 70% Industrial Methylated Spirit spray for surface disinfection of components required in Grade A pharmaceutical environments. This study seeks to investigate other agents and procedures that may provide more effective sanitisation. Several methods are available to test the efficacy of disinfectants against vegetative organisms. However, no methods currently available test the efficacy of disinfectants against spores on the hard surfaces encountered in the pharmacy aseptic processing environment. Therefore, a method has been developed to test the efficacy of disinfectants against spores, modified from British Standard 13697 and Association of Analytical Chemists standards. The testing procedure was used to evaluate alternative biocides and disinfection methods for transferring components into hospital pharmacy cleanrooms, and to determine which combinations of biocide and application method have the greatest efficacy against spores of Bacillus subtilis subspecies subtilis 168, Bacillus subtilis American Type Culture Collection (ATCC) 6633, and Bacillus pumilis ATCC 27142. Stainless steel carrier test plates were used to represent the hard surfaces in hospital pharmacy cleanrooms. Plates were inoculated with 10(7)-10(8) colony-forming units per milliliter (CFU/mL) and treated with the various biocide formulations, using different disinfection methods. Sporicidal activity was calculated as log reduction in CFU. Of the biocides tested, 6% hydrogen peroxide and a quaternary ammonium compound/chlorine dioxide combination were most effective compared to a Quat/biguanide, amphoteric surfactant, 70% v/v ethanol in deionised water and isopropyl alcohol in water for injection. Of the different application methods tested, spraying followed by wiping was the most effective, followed closely by wiping alone. Spraying alone was least effective.

Applied evolution: An integrated approach to studying life history traits in response to drug selection

The use of chemical control measures to reduce the impact of parasite and pest species has frequently resulted in the development of resistance. Thus, resistance management has become a key concern in human and veterinary medicine, and in agricultural production. Although it is known that factors such as gene flow between susceptible and resistant populations, drug type, application methods, and costs of resistance can affect the rate of resistance evolution, less is known about the impacts of density-dependent eco-evolutionary processes that could be altered by drug-induced mortality. The overall aim of this thesis was to take an experimental evolution approach to assess how life history traits respond to drug selection, using a free-living dioecious worm (Caenorhabditis remanei) as a model. In Chapter 2, I defined the relationship between C. remanei survival and Ivermectin dose over a range of concentrations, in order to control the intensity of selection used in the selection experiment described in Chapter 4. The dose-response data were also used to appraise curve-fitting methods, using Akaike Information Criterion (AIC) model selection to compare a series of nonlinear models. The type of model fitted to the dose response data had a significant effect on the estimates of LD50 and LD99, suggesting that failure to fit an appropriate model could give misleading estimates of resistance status. In addition, simulated data were used to establish that a potential cost of resistance could be predicted by comparing survival at the upper asymptote of dose-response curves for resistant and susceptible populations, even when differences were as low as 4%. This approach to dose-response modeling ensures that the maximum amount of useful information relating to resistance is gathered in one study. In Chapter 3, I asked how simulations could be used to inform important design choices used in selection experiments. Specifically, I focused on the effects of both within- and between-line variation on estimated power, when detecting small, medium and large effect sizes. Using mixed-effect models on simulated data, I demonstrated that commonly used designs with realistic levels of variation could be underpowered for substantial effect sizes. Thus, use of simulation-based power analysis provides an effective way to avoid under or overpowering a study designs incorporating variation due to random effects. In Chapter 4, I 3 investigated how Ivermectin dosage and changes in population density affect the rate of resistance evolution. I exposed replicate lines of C. remanei to two doses of Ivermectin (high and low) to assess relative survival of lines selected in drug-treated environments compared to untreated controls over 10 generations. Additionally, I maintained lines where mortality was imposed randomly to control for differences in density between drug treatments and to distinguish between the evolutionary consequences of drug treatment versus ecological processes affected by changes in density-dependent feedback. Intriguingly, both drug-selected and random-mortality lines showed an increase in survivorship when challenged with Ivermectin; the magnitude of this increase varied with the intensity of selection and life-history stage. The results suggest that interactions between density-dependent processes and life history may mediate evolved changes in susceptibility to control measures, which could result in misleading conclusions about the evolution of heritable resistance following drug treatment. In Chapter 5, I investigated whether the apparent changes in drug susceptibility found in Chapter 4 were related to evolved changes in life-history of C. remanei populations after selection in drug-treated and random-mortality environments. Rapid passage of lines in the drug-free environment had no effect on the measured life-history traits. In the drug-free environment, adult size and fecundity of drug-selected lines increased compared to the controls but drug selection did not affect lifespan. In the treated environment, drug-selected lines showed increased lifespan and fecundity relative to controls. Adult size of randomly culled lines responded in a similar way to drug-selected lines in the drug-free environment, but no change in fecundity or lifespan was observed in either environment. The results suggest that life histories of nematodes can respond to selection as a result of the application of control measures. Failure to take these responses into account when applying control measures could result in adverse outcomes, such as larger and more fecund parasites, as well as over-estimation of the development of genetically controlled resistance. In conclusion, my thesis shows that there may be a complex relationship between drug selection, density-dependent regulatory processes and life history of populations challenged with control measures. This relationship could have implications for how resistance is monitored and managed if life histories of parasitic species show such eco-evolutionary responses to drug application.

RICORDACI: Research on the Conservation, Restoration and Diagnosis of Cinematographic Films

This thesis is the result of the RICORDACI project, a three-year European-funded initiative involving the collaboration between the University of Bologna and the restoration laboratory of the Cineteca di Bologna, L'immagine Ritrovata, which aimed to develop innovative solutions and technologies for the preservation of cinematographic film heritage. In particular, this thesis presents new analytical methodologies to exploit two types of portable miniaturized Near Infrared spectrometers working in Diffuse Reflectance over the Short Wave Infrared (SWIR) range, to study the near infrared (NIR) spectral behavior of film base materials for an accurate, non-invasive and fast characterization of the polymer type; and for films with cellulose acetate supports, they can be employed as a diagnostic tool for monitoring the Degree of substitution (DS) affected by the loss of acetyl groups. The proposed methods offer non-invasive, fast, inexpensive and simple alternatives for the characterization and diagnosis of film bases to help the strategic planning and decision-making regarding storage, digitalization and intervention of film collections. Secondly, the thesis includes the evaluation of new green cleaning systems and solvents for the effective, fast and innocuous removal of undesired substances from degraded cinematographic films bases; these tests compared the efficiency of traditional systems and solvents against the new proposals. Firstly, the use of Deep Eutectic Solvent formulations for removing softened gelatin residues from cellulose nitrate bases; and secondly, the employment of green volatile solvents with different application methods, including the use of new electrospun nylon mats, for avoiding the dangerous use of friction for the removal of Triphenyl Phosphate blooms from the surface of cellulose acetate bases. The results obtained will help improving the efficiency of the interventions needed before the digitalization of historical cinematographic films and will pave the way for further investigation on the use of green solvents for cleaning polymeric heritage objects.

HPLC-FLD methods to quantify chloroaluminum phthalocyanine in nanoparticles, plasma and tissue: application in pharmacokinetic and biodistribution studies

Analytical and bioanalytical methods of high-performance liquid chromatography with fluorescence detection (HPLC-FLD) were developed and validated for the determination of chloroaluminum phthalocyanine in different formulations of polymeric nanocapsules, plasma and livers of mice. Plasma and homogenized liver samples were extracted with ethyl acetate, and zinc phthalocyanine was used as internal standard. The results indicated that the methods were linear and selective for all matrices studied. Analysis of accuracy and precision showed adequate values, with variations lower than 10% in biological samples and lower than 2% in analytical samples. The recoveries were as high as 96% and 99% in the plasma and livers, respectively. The quantification limit of the analytical method was 1.12 ng/ml, and the limits of quantification of the bioanalytical method were 15 ng/ml and 75 ng/g for plasma and liver samples, respectively. The bioanalytical method developed was sensitive in the ranges of 15-100 ng/ml in plasma and 75-500 ng/g in liver samples and was applied to studies of biodistribution and pharmacokinetics of AlClPc. (C) 2011 Elsevier B.V. All rights reserved.

An investigation of the anti-pronation effect of two taping methods after application and exercise

This study investigated the effect of two anti-pronation taping techniques on vertical navicular height, an indicator of foot pronation, after its application and 20 min of exercise. The taping techniques were: the low dye (LD) and low dye with the addition of calcaneal slings and reverse sixes (LDCR). A repeated measures study was used. It found that LDCR was superior to LD and control immediately after application and exercise. LD was better than control immediately after application but not after exercise. These findings provide practical directions to clinicians regularly using anti-pronation taping techniques.

Application of Petrov-Galerkin methods to transient boundary value problems in chemical engineering: adsorption with steep gradients in bidisperse solids

Petrov-Galerkin methods are known to be versatile techniques for the solution of a wide variety of convection-dispersion transport problems, including those involving steep gradients. but have hitherto received little attention by chemical engineers. We illustrate the technique by means of the well-known problem of simultaneous diffusion and adsorption in a spherical sorbent pellet comprised of spherical, non-overlapping microparticles of uniform size and investigate the uptake dynamics. Solutions to adsorption problems exhibit steep gradients when macropore diffusion controls or micropore diffusion controls, and the application of classical numerical methods to such problems can present difficulties. In this paper, a semi-discrete Petrov-Galerkin finite element method for numerically solving adsorption problems with steep gradients in bidisperse solids is presented. The numerical solution was found to match the analytical solution when the adsorption isotherm is linear and the diffusivities are constant. Computed results for the Langmuir isotherm and non-constant diffusivity in microparticle are numerically evaluated for comparison with results of a fitted-mesh collocation method, which was proposed by Liu and Bhatia (Comput. Chem. Engng. 23 (1999) 933-943). The new method is simple, highly efficient, and well-suited to a variety of adsorption and desorption problems involving steep gradients. (C) 2001 Elsevier Science Ltd. All rights reserved.

Development of electrochemical methods for determination of tramadol - analytical application to pharmaceutical dosage forms

A square-wave voltammetric (SWV) method and a flow injection analysis system with amperometric detection were developed for the determination of tramadol hydrochloride. The SWV method enables the determination of tramadol over the concentration range of 15-75 µM with a detection limit of 2.2 µM. Tramadol could be determined in concentrations between 9 and 50 µM at a sampling rate of 90 h-1, with a detection limit of 1.7 µM using the flow injection system. The electrochemical methods developed were successfully applied to the determination of tramadol in pharmaceutical dosage forms, without any pre-treatment of the samples. Recovery trials were performed to assess the accuracy of the results; the values were between 97 and 102% for both methods.