Inhibition by interferon-gamma of human mononuclear cell-mediated low density lipoprotein oxidation. Participation of tryptophan metabolism along the kynurenine pathway

In this study we examined the potential inhibition by interferon-gamma (IFN gamma) of the early stages of low density lipoprotein (LDL) oxidation mediated by human peripheral blood mononuclear cells (PBMC) and monocyte-derived macrophages (MDM) in Ham's F-10 medium supplemented with physiological amounts of L-tryptophan (Trp). We assessed LDL oxidation by measuring the consumption of LDL's major antioxidant (i.e., alpha-tocopherol) and targets for oxidation (cholesteryllinoleate and cholesterylarachidonate), together with the accumulation of cholesterylester hydroperoxides and the increase in relative electrophoretic mobility of the lipoprotein particle. Exposure of PBMC or MDM to IFN gamma induced the degradation of extracellular Trp with concomitant accumulation of kynurenine, anthranilic and 3-hydroxyanthranilic acid (3HAA) in the culture medium. Formation of 3HAA, but neither Trp degradation nor formation of kynurenine and anthranilic acid, was inhibited by low amounts of diphenylene iodonium (DPI) in a concentration-dependent manner. In contrast to oxidative Trp metabolism, exposure of human PBMC or MDM to IFN gamma failed to induce degradation of arginine, and nitrite was not detected in the cell supernatant, indicating that nitric oxide synthase was not induced under these conditions. Incubation of LDL in Trp-supplemented F-10 medium resulted in a time-dependent oxidation of the lipoprotein that was accelerated in the presence of PBMC or MDM but inhibited strongly in the presence of both cells and IFN gamma, i.e., when Trp degradation and formation of 3HAA were induced. In contrast, when IFN gamma was added to PBMC or MDM in F-10 medium that was virtually devoid of Trp, inhibition of cell-accelerated LDL oxidation was not observed. Exogenous 3HAA added to PBMC or purified monocytes in the absence of IFN gamma also strongly and in a concentration-dependent manner inhibited LDL oxidation. Selective inhibition of IFN gamma-induced formation of 3HAA by DPI caused reversion of the inhibitory action of this cytokine on both PBMC- and MDM-mediated LDL oxidation. These results show that IFN gamma treatment of human PBMC or MDM in vitro attenuates the extent of LDL oxidation caused by these cells, and indicate that Trp degradation with formation of 3HAA is a major contributing factor to this inhibitory activity.

Porous silicon technology for integrated microsystems

With the development of micro systems, there is an increasing demand for integrable porous materials. In addition to those conventional applications, such as filtration, wicking, and insulating, many new micro devices, including micro reactors, sensors, actuators, and optical components, can benefit from porous materials. Conventional porous materials, such as ceramics and polymers, however, cannot meet the challenges posed by micro systems, due to their incompatibility with standard micro-fabrication processes. In an effort to produce porous materials that can be used in micro systems, porous silicon (PS) generated by anodization of single crystalline silicon has been investigated. In this work, the PS formation process has been extensively studied and characterized as a function of substrate type, crystal orientation, doping concentration, current density and surfactant concentration and type. Anodization conditions have been optimized for producing very thick porous silicon layers with uniform pore size, and for obtaining ideal pore morphologies. Three different types of porous silicon materials: meso porous silicon, macro porous silicon with straight pores, and macro porous silicon with tortuous pores, have been successfully produced. Regular pore arrays with controllable pore size in the range of 2µm to 6µm have been demonstrated as well. Localized PS formation has been achieved by using oxide/nitride/polysilicon stack as masking materials, which can withstand anodization in hydrofluoric acid up to twenty hours. A special etching cell with electrolytic liquid backside contact along with two process flows has been developed to enable the fabrication of thick macro porous silicon membranes with though wafer pores. For device assembly, Si-Au and In-Au bonding technologies have been developed. Very low bonding temperature (~200 degrees C) and thick/soft bonding layers (~6µm) have been achieved by In-Au bondi ng technology, which is able to compensate the potentially rough surface on the porous silicon sample without introducing significant thermal stress. The application of the porous silicon material in micro systems has been demonstrated in a micro gas chromatograph system by two indispensable components: an integrated vapor source and an inlet filter, wherein porous silicon performs the basic functions of porous media: wicking and filtration. By utilizing a macro porous silicon wick, the calibration vapor source was able to produce a uniform and repeatable vapor generation for n-decane with less than a 0.1% variation in 9 hours, and less than a 0.5% variation in rate over 7 days. With engineered porous silicon membranes the inlet filter was able to show a depth filtration with nearly 100% collection efficiency for particles larger than 0.3µm in diameter, a low pressure-drop of 523Pa at 20sccm flow rate, and a filter capacity of 500µg/cm2.

Development and characterization of fluorescent pH sensors based on porous silica and hydrogel support matrices

The hydrogen ion activity (pH) is a very important parameter in environment monitoring, biomedical research and other applications. Optical pH sensors have several advantages over traditional potentiometric pH measurement, such as high sensitivity, no need of constant calibration, easy for miniaturization and possibility for remote sensing. Several pH indicators has been successfully immobilized in three different solid porous materials to use as pH sensing probes. The fluorescent pH indicator fluorescein-5-isothiocyanate (FITC) was covalently bound onto the internal surface of porous silica (pore size ~10 nm) and retained its pH sensitivity. The excited state pK* a of FITC in porous silica (5.58) was slightly smaller than in solution (5.68) due to the free silanol groups (Si-OH) on the silica surface. The pH sensitive range for this probe is pH 4.5 - 7.0 with an error less than 0.1 pH units. The probe response was reproducible and stable for at least four month, stored in DI water, but exhibit a long equilibrium of up to 100 minutes. Sol-gel based pH sensors were developed with immobilization of two fluorescent pH indicators fluorescein-5-(and-6)-sulfonic acid, trisodium salt (FS) and 8-hydroxypyrene- 1,3,6-trisulfonic acid (HPTS) through physical entrapment. Prior to immobilization, the indicators were ion-paired with a common surfactant hexadecyltrimethylammonium bromide (CTAB) in order to prevent leaching. The sol-gel films were synthesized through the hydrolysis of two different precursors, ethyltriethoxysilane (ETEOS) and 3- glycidoxypropyltrimethoxysilane (GPTMS) and deposited on a quartz slide through spin coating. The pK a of the indicators immobilized in sol-gel films was much smaller than in solutions due to silanol groups on the inner surface of the sol-gel films and ammonium groups from the surrounding surfactants. Unlike in solution, the apparent pK a of the indicators in sol-gel films increased with increasing ionic strength. The equilibrium time for these sensors was within 5 minutes (with film thickness of ~470 nm). Polyethylene glycol (PEG) hydrogel was of interest for optical pH sensor development because it is highly proton permeable, transparent and easy to synthesize. pH indicators can be immobilized in hydrogel through physical entrapment and copolymerization. FS and HPTS ion-pairs were physically entrapped in hydrogel matrix synthesized via free radical initiation. For covalent immobilization, three indicators, 6,8-dihydroxypyrene-1,3- disulfonic acid (DHPDS), 2,7-dihydroxynaphthalene-3,6-disulfonic acid (DHNDS) and cresol red were first reacted with methacrylic anhydride (MA) to form methacryloylanalogs for copolymerization. These hydrogels were synthesized in aqueous solution with a redox initiation system. The thickness of the hydrogel film is controlled as ~ 0.5 cm and the porosity can be adjusted with the percentage of polyethylene glycol in the precursor solutions. The pK a of the indicators immobilized in the hydrogel both physically and covalently were higher than in solution due to the medium effect. The sensors are stable and reproducible with a short equilibrium time (less than 4 minutes). In addition, the color change of cresol red immobilized hydrogel is vivid from yellow (acidic condition) to purple (basic condition). Due to covalently binding, cresol red was not leaching out from the hydrogel, making it a good candidate of reusable "pH paper".

Novel cell-free strategy for therapeutic angiogenesis: in vitro generated conditioned medium can replace progenitor cell transplantation

BACKGROUND: Current evidence suggests that endothelial progenitor cells (EPC) contribute to ischemic tissue repair by both secretion of paracrine factors and incorporation into developing vessels. We tested the hypothesis that cell-free administration of paracrine factors secreted by cultured EPC may achieve an angiogenic effect equivalent to cell therapy. METHODOLOGY/PRINCIPAL FINDINGS: EPC-derived conditioned medium (EPC-CM) was obtained from culture expanded EPC subjected to 72 hours of hypoxia. In vitro, EPC-CM significantly inhibited apoptosis of mature endothelial cells and promoted angiogenesis in a rat aortic ring assay. The therapeutic potential of EPC-CM as compared to EPC transplantation was evaluated in a rat model of chronic hindlimb ischemia. Serial intramuscular injections of EPC-CM and EPC both significantly increased hindlimb blood flow assessed by laser Doppler (81.2+/-2.9% and 83.7+/-3.0% vs. 53.5+/-2.4% of normal, P<0.01) and improved muscle performance. A significantly increased capillary density (1.62+/-0.03 and 1.68+/-0.05/muscle fiber, P<0.05), enhanced vascular maturation (8.6+/-0.3 and 8.1+/-0.4/HPF, P<0.05) and muscle viability corroborated the findings of improved hindlimb perfusion and muscle function. Furthermore, EPC-CM transplantation stimulated the mobilization of bone marrow (BM)-derived EPC compared to control (678.7+/-44.1 vs. 340.0+/-29.1 CD34(+)/CD45(-) cells/1x10(5) mononuclear cells, P<0.05) and their recruitment to the ischemic muscles (5.9+/-0.7 vs. 2.6+/-0.4 CD34(+) cells/HPF, P<0.001) 3 days after the last injection. CONCLUSIONS/SIGNIFICANCE: Intramuscular injection of EPC-CM is as effective as cell transplantation for promoting tissue revascularization and functional recovery. Owing to the technical and practical limitations of cell therapy, cell free conditioned media may represent a potent alternative for therapeutic angiogenesis in ischemic cardiovascular diseases.

Compressive strength of interbody cages in the lumbar spine: the effect of cage shape, posterior instrumentation and bone density

One goal of interbody fusion is to increase the height of the degenerated disc space. Interbody cages in particular have been promoted with the claim that they can maintain the disc space better than other methods. There are many factors that can affect the disc height maintenance, including graft or cage design, the quality of the surrounding bone and the presence of supplementary posterior fixation. The present study is an in vitro biomechanical investigation of the compressive behaviour of three different interbody cage designs in a human cadaveric model. The effect of bone density and posterior instrumentation were assessed. Thirty-six lumbar functional spinal units were instrumented with one of three interbody cages: (1) a porous titanium implant with endplate fit (Stratec), (2) a porous, rectangular carbon-fibre implant (Brantigan) and (3) a porous, cylindrical threaded implant (Ray). Posterior instrumentation (USS) was applied to half of the specimens. All specimens were subjected to axial compression displacement until failure. Correlations between both the failure load and the load at 3 mm displacement with the bone density measurements were observed. Neither the cage design nor the presence of posterior instrumentation had a significant effect on the failure load. The loads at 3 mm were slightly less for the Stratec cage, implying lower axial stiffness, but were not different with posterior instrumentation. The large range of observed failure loads overlaps the potential in vivo compressive loads, implying that failure of the bone-implant interface may occur clinically. Preoperative measurements of bone density may be an effective tool to predict settling around interbody cages.

Synthesis of bone-like micro-porous calcium phosphate/iota-carrageenan composites by gel diffusion

Brushite and octacalcium phosphate (OCP) crystals are well-known precursors of hydroxylapatite (HAp), the main mineral found in bone. In this report, we present a new method for biomimicking brushite and OCP using single and double diffusion techniques. Brushite and OCP crystals were grown in an iota-carrageenan gel. The aggregates were analyzed by scanning electron microscopy (SEM), X-ray diffraction (XRD), infrared spectroscopy (IR) and thermal gravimetric analysis (TGA). SEM revealed different morphologies of brushite crystals from highly porous aggregates to plate-shaped forms. OCP crystals grown in iota-carrageenan showed a porous spherical shape different from brushite growth forms. The XRD method demonstrated that the single-diffusion method favors the formation of monoclinic brushite. In contrast, the double diffusion method was found to promote the formation of the triclinic octacalcium phosphate OCP phase. By combining the different parameters for crystal growth in carrageenan, such as ion concentration, gel pH and gel density, it is possible to modify the morphology of composite crystals, change the phase of calcium phosphate and modulate the amount of carrageenan inclusion in crystals. This study suggests that iota-carrageenan is a high-molecular-weight polysaccharide that is potentially applicable for controlling calcium phosphate crystallization.

Optimum Electron Temperature and Density for Short-Wavelength Plasma-Lasing from Nickel-like ions

Soft X-ray lasing across a Ni-like plasma gain-medium requires optimum electron temperature and density for attaining to the Ni-like ion stage and for population inversion in the View the MathML source3d94d1(J=0)→3d94p1(J=1) laser transition. Various scaling laws, function of operating parameters, were compared with respect to their predictions for optimum temperatures and densities. It is shown that the widely adopted local thermodynamic equilibrium (LTE) model underestimates the optimum plasma-lasing conditions. On the other hand, non-LTE models, especially when complemented with dielectronic recombination, provided accurate prediction of the optimum plasma-lasing conditions. It is further shown that, for targets with Z equal or greater than the rare-earth elements (e.g. Sm), the optimum electron density for plasma-lasing is not accessible for pump-pulses at View the MathML sourceλ=1ω=1μm. This observation explains a fundamental difficulty in saturating the wavelength of plasma-based X-ray lasers below 6.8 nm, unless using 2ω2ω pumping.

Permittivity measurements of porous matter in support of investigations of the surface and interior of 67P/Churyumov-Gerasimenko

Aims. Permittivity measurements on porous samples of volcanic origin have been performed in the 0.05-190 GHz range under laboratory conditions in support of the Rosetta mission to comet 67P/Churyumov-Gerasimenko, specifically with the MIRO radiometric experiment and CONSERT radar experiment. Methods. The samples were split into several subsamples with different size ranges covering a few mu m to 500 mu m. Bulk densities of the subsamples were estimated to be in the 800 to 1500 kg/m(3) range. The porosities were in the range of 48% to 65%. From 50 MHz to 6 GHz and at 190 GHz, permittivity has been determined with a coaxial cell and with a quasi-optical bench, respectively. Results. Without taking into account the volume-scattering effect at 190 GHz, the real part of the permittivity, normalized by the bulk density, is in the range of 2.1 to 2.6. The results suggest that the real part of the permittivity of an ice-free dust mantle covering the nucleus is in the 1.5-2.2 range at 190 GHz. From these values, a lower limit for the absorption length for the millimeter receiver of MIRO has been estimated to be between 0.6 and 2 cm, in agreement with results obtained from MIRO in September 2014. At frequencies of interest for CONSERT experiment, the real part of the permittivity of a suspected ice-free dust mantle should be below 2.2. It may be in the range of 1.2 to 1.7 for the nucleus, in agreement with first CONSERT results, taking into account a mean temperature of 110 K and different values for the dust-to-ice volumetric ratio. Estimations of contributions of the different parameters to the permittivity variation may indicate that the porosity is the main parameter.

SAMPLING METHODOLOGY FOR AIRBORNE SEMIVOLATILE ORGANIC POLLUTANTS USING POLYURETHANE FOAMS (COLLECTION, PESTICIDES, VOLATILE)

Indoor and ambient air organic pollutants have been gaining attention because they have been measured at levels with possible health effects. Studies have shown that most airborne polychlorinated biphenyls (PCBs), pesticides and many polycyclic aromatic hydrocarbons (PAHs) are present in the free vapor state. The purpose of this research was to extend recent investigative work with polyurethane foam (PUF) as a collection medium for semivolatile compounds. Open-porous flexible PUFs with different chemical makeup and physical properties were evaluated as to their collection affinities/efficiencies for various classes of compounds and the degree of sample recovery. Filtered air samples were pulled through plugs of PUF spiked with various semivolatiles under different simulated environmental conditions (temperature and humidity), and sampling parameters (flow rate and sample volume) in order to measure their effects on sample breakthrough volume (V(,B)). PUF was also evaluated in the passive mode using organo-phosphorus pesticides. Another major goal was to improve the overall analytical methodology; PUF is inexpensive, easy to handle in the field and has excellent airflow characteristics (low pressure drop). It was confirmed that the PUF collection apparatus behaves as if it were a gas-solid chromatographic system, in that, (V(,B)) was related to temperature and sample volume. Breakthrough volumes were essentially the same using both polyether and polyester type PUF. Also, little change was observed in the V(,B)s after coating PUF with common chromatographic liquid phases. Open cell (reticulated) foams gave better recoveries than closed cell foams. There was a slight increase in (V(,B)) with an increase in the number of cells/pores per inch. The high-density polyester PUF was found to be an excellent passive and active collection adsorbent. Good recoveries could be obtained using just solvent elution. A gas chromatograph equipped with a photoionization detector gave excellent sensitivities and selectivities for the various classes of compounds investigated. ^

Sediment description, CaCO3, density and porosity at DSDP Site 89-585

Siliceous sediments and sedimentary rocks occur as chert and silicified chalk, limestone, and claystone in Site 585 lower Miocene to Campanian sediments, with one older occurrence of chert near the Cenomanian/Turonian boundary. The recovered drill breccia in the Miocene to middle Eocene interval is dominated by bright red, orange, yellow, and brown chips and fragments of chert. In early Eocene and older sediments gray silicified limestone and yellowish brown chert fragments predominate. Recovery is poor in cores with chert because chert tends to fracture into smaller pieces that escape the drill and because the hard chert fragments grind away other sediments during rotary drilling. Thin-section and hand-sample studies show complex diagenetic histories of silicification (silica pore infill) and chertification (silica replacement of host rock). Multiple events of silicification can occur in the same rocks, producing chert from silicified limestone. Despite some prior silicification, silicified limestone is porous enough to provide conduits for dissolved silica-charged pore waters. Silicification and chert are more abundant in the coarser parts of the sedimentary section. These factors reflect the importance of porosity and permeability as well as chemical and lithologic controls in the process of silica diagenesis.

Theoretical study of band alignment in nano-porous ZnO interacting with substituted Phthalocyanines

The aim of this work is the theoretical study of the band alignment between the two components of a hybrid organic-inorganic solar-cell. The working organic molecules are metal tetra-sulphonated phthalocyanines (M-Pc) and the inorganic material is nano-porous ZnO growth in the 001 direction. The theoretical calculations are being made using the density functional theory (DFT) using a GGA functional with the SIESTA code, which projects electron wave functions and density onto a real space grid and uses as basis set a linear combination of numerical, finite-range localized atomic orbitals. We also used the DFT+U method included in the code that allows a semi-empirical inclusion of electronic correlations in the description of electronic spectra for systems such as zinc oxide.

Dynamic analysis of payloads and structures with intermediate modal density

La necesidad de desarrollar técnicas para predecir la respuesta vibroacústica de estructuras espaciales lia ido ganando importancia en los últimos años. Las técnicas numéricas existentes en la actualidad son capaces de predecir de forma fiable el comportamiento vibroacústico de sistemas con altas o bajas densidades modales. Sin embargo, ambos rangos no siempre solapan lo que hace que sea necesario el desarrollo de métodos específicos para este rango, conocido como densidad modal media. Es en este rango, conocido también como media frecuencia, donde se centra la presente Tesis doctoral, debido a la carencia de métodos específicos para el cálculo de la respuesta vibroacústica. Para las estructuras estudiadas en este trabajo, los mencionados rangos de baja y alta densidad modal se corresponden, en general, con los rangos de baja y alta frecuencia, respectivamente. Los métodos numéricos que permiten obtener la respuesta vibroacústica para estos rangos de frecuencia están bien especificados. Para el rango de baja frecuencia se emplean técnicas deterministas, como el método de los Elementos Finitos, mientras que, para el rango de alta frecuencia las técnicas estadísticas son más utilizadas, como el Análisis Estadístico de la Energía. En el rango de medias frecuencias ninguno de estos métodos numéricos puede ser usado con suficiente precisión y, como consecuencia -a falta de propuestas más específicas- se han desarrollado métodos híbridos que combinan el uso de métodos de baja y alta frecuencia, intentando que cada uno supla las deficiencias del otro en este rango medio. Este trabajo propone dos soluciones diferentes para resolver el problema de la media frecuencia. El primero de ellos, denominado SHFL (del inglés Subsystem based High Frequency Limit procedure), propone un procedimiento multihíbrido en el cuál cada subestructura del sistema completo se modela empleando una técnica numérica diferente, dependiendo del rango de frecuencias de estudio. Con este propósito se introduce el concepto de límite de alta frecuencia de una subestructura, que marca el límite a partir del cual dicha subestructura tiene una densidad modal lo suficientemente alta como para ser modelada utilizando Análisis Estadístico de la Energía. Si la frecuencia de análisis es menor que el límite de alta frecuencia de la subestructura, ésta se modela utilizando Elementos Finitos. Mediante este método, el rango de media frecuencia se puede definir de una forma precisa, estando comprendido entre el menor y el mayor de los límites de alta frecuencia de las subestructuras que componen el sistema completo. Los resultados obtenidos mediante la aplicación de este método evidencian una mejora en la continuidad de la respuesta vibroacústica, mostrando una transición suave entre los rangos de baja y alta frecuencia. El segundo método propuesto se denomina HS-CMS (del inglés Hybrid Substructuring method based on Component Mode Synthesis). Este método se basa en la clasificación de la base modal de las subestructuras en conjuntos de modos globales (que afectan a todo o a varias partes del sistema) o locales (que afectan a una única subestructura), utilizando un método de Síntesis Modal de Componentes. De este modo es posible situar espacialmente los modos del sistema completo y estudiar el comportamiento del mismo desde el punto de vista de las subestructuras. De nuevo se emplea el concepto de límite de alta frecuencia de una subestructura para realizar la clasificación global/local de los modos en la misma. Mediante dicha clasificación se derivan las ecuaciones globales del movimiento, gobernadas por los modos globales, y en las que la influencia del conjunto de modos locales se introduce mediante modificaciones en las mismas (en su matriz dinámica de rigidez y en el vector de fuerzas). Las ecuaciones locales se resuelven empleando Análisis Estadístico de Energías. Sin embargo, este último será un modelo híbrido, en el cual se introduce la potencia adicional aportada por la presencia de los modos globales. El método ha sido probado para el cálculo de la respuesta de estructuras sometidas tanto a cargas estructurales como acústicas. Ambos métodos han sido probados inicialmente en estructuras sencillas para establecer las bases e hipótesis de aplicación. Posteriormente, se han aplicado a estructuras espaciales, como satélites y reflectores de antenas, mostrando buenos resultados, como se concluye de la comparación de las simulaciones y los datos experimentales medidos en ensayos, tanto estructurales como acústicos. Este trabajo abre un amplio campo de investigación a partir del cual es posible obtener metodologías precisas y eficientes para reproducir el comportamiento vibroacústico de sistemas en el rango de la media frecuencia. ABSTRACT Over the last years an increasing need of novel prediction techniques for vibroacoustic analysis of space structures has arisen. Current numerical techniques arc able to predict with enough accuracy the vibro-acoustic behaviour of systems with low and high modal densities. However, space structures are, in general, very complex and they present a range of frequencies in which a mixed behaviour exist. In such cases, the full system is composed of some sub-structures which has low modal density, while others present high modal density. This frequency range is known as the mid-frequency range and to develop methods for accurately describe the vibro-acoustic response in this frequency range is the scope of this dissertation. For the structures under study, the aforementioned low and high modal densities correspond with the low and high frequency ranges, respectively. For the low frequency range, deterministic techniques as the Finite Element Method (FEM) are used while, for the high frequency range statistical techniques, as the Statistical Energy Analysis (SEA), arc considered as more appropriate. In the mid-frequency range, where a mixed vibro-acoustic behaviour is expected, any of these numerical method can not be used with enough confidence level. As a consequence, it is usual to obtain an undetermined gap between low and high frequencies in the vibro-acoustic response function. This dissertation proposes two different solutions to the mid-frequency range problem. The first one, named as The Subsystem based High Frequency Limit (SHFL) procedure, proposes a multi-hybrid procedure in which each sub-structure of the full system is modelled with the appropriate modelling technique, depending on the frequency of study. With this purpose, the concept of high frequency limit of a sub-structure is introduced, marking out the limit above which a substructure has enough modal density to be modelled by SEA. For a certain analysis frequency, if it is lower than the high frequency limit of the sub-structure, the sub-structure is modelled through FEM and, if the frequency of analysis is higher than the high frequency limit, the sub-structure is modelled by SEA. The procedure leads to a number of hybrid models required to cover the medium frequency range, which is defined as the frequency range between the lowest substructure high frequency limit and the highest one. Using this procedure, the mid-frequency range can be define specifically so that, as a consequence, an improvement in the continuity of the vibro-acoustic response function is achieved, closing the undetermined gap between the low and high frequency ranges. The second proposed mid-frequency solution is the Hybrid Sub-structuring method based on Component Mode Synthesis (HS-CMS). The method adopts a partition scheme based on classifying the system modal basis into global and local sets of modes. This classification is performed by using a Component Mode Synthesis, in particular a Craig-Bampton transformation, in order to express the system modal base into the modal bases associated with each sub-structure. Then, each sub-structure modal base is classified into global and local set, fist ones associated with the long wavelength motion and second ones with the short wavelength motion. The high frequency limit of each sub-structure is used as frequency frontier between both sets of modes. From this classification, the equations of motion associated with global modes are derived, which include the interaction of local modes by means of corrections in the dynamic stiffness matrix and the force vector of the global problem. The local equations of motion are solved through SEA, where again interactions with global modes arc included through the inclusion of an additional input power into the SEA model. The method has been tested for the calculation of the response function of structures subjected to structural and acoustic loads. Both methods have been firstly tested in simple structures to establish their basis and main characteristics. Methods are also verified in space structures, as satellites and antenna reflectors, providing good results as it is concluded from the comparison with experimental results obtained in both, acoustic and structural load tests. This dissertation opens a wide field of research through which further studies could be performed to obtain efficient and accurate methodologies to appropriately reproduce the vibro-acoustic behaviour of complex systems in the mid-frequency range.

Echolocation detections and digital video surveys provide reliable estimates of the relative density of harbour porpoises

Acknowledgements We would like to thank Erik Rexstad and Rob Williams for useful reviews of this manuscript. The collection of visual and acoustic data was funded by the UK Department of Energy & Climate Change, the Scottish Government, Collaborative Offshore Wind Research into the Environment (COWRIE) and Oil & Gas UK. Digital aerial surveys were funded by Moray Offshore Renewables Ltd and additional funding for analysis of the combined datasets was provided by Marine Scotland. Collaboration between the University of Aberdeen and Marine Scotland was supported by MarCRF. We thank colleagues at the University of Aberdeen, Moray First Marine, NERI, Hi-Def Aerial Surveying Ltd and Ravenair for essential support in the field, particularly Tim Barton, Bill Ruck, Rasmus Nielson and Dave Rutter. Thanks also to Andy Webb, David Borchers, Len Thomas, Kelly McLeod, David L. Miller, Dinara Sadykova and Thomas Cornulier for advice on survey design and statistical approache. Data Accessibility Data are available from the Dryad Digital Repository: http://dx.doi.org/10.5061/dryad.cf04g

Continuous axenic cultivation of Pneumocystis carinii

Continuous axenic culture of Pneumocystis carinii has been achieved. A culture vessel is used that allows for frequent medium exchange without disturbance of organisms that grow attached to a collagen-coated porous membrane. The growth medium is based on Minimal Essential Medium with Earle’s salt supplemented with S-adenosyl-l-methionine, putrescine, ferric pyrophosphate, N-acetyl glucosamine, putrescine, p-aminobenzoic acid, l-cysteine and l-glutamine, and horse serum. Incubation is in room air at 31°C. The pH of the medium begins at 8.8 and rises to ≈9 as the cells grow. Doubling times calculated from growth curves obtained from cultures inoculated at moderate densities ranged from 35 to 65 hours. With a low-density inoculum, the doubling time is reduced to 19 hours. The morphology of cultured organisms in stained smears and in transmission electron micrographs is that of P. carinii, and P. carinii-specific mAbs label the cultured material. Cultured organisms are infective for immunosuppressed rats and can be stored frozen and used to reinitiate culture.

Adsorbent density impact on gas storage capacities

In the literature, different approaches, terminologies, concepts and equations are used for calculating gas storage capacities. Very often, these approaches are not well defined, used and/or determined, giving rise to significant misconceptions. Even more, some of these approaches, very much associated with the type of adsorbent material used (e.g., porous carbons or new materials such as COFs and MOFs), impede a suitable comparison of their performances for gas storage applications. We review and present the set of equations used to assess the total storage capacity for which, contrarily to the absolute adsorption assessment, all its experimental variables can be determined experimentally without assumptions, ensuring the comparison of different porous storage materials for practical application. These material-based total storage capacities are calculated by taking into account the excess adsorption, the bulk density (ρbulk) and the true density (ρtrue) of the adsorbent. The impact of the material densities on the results are investigated for an exemplary hydrogen isotherm obtained at room temperature and up to 20 MPa. It turns out that the total storage capacity on a volumetric basis, which increases with both, ρbulk and ρtrue, is the most appropriate tool for comparing the performance of storage materials. However, the use of the total storage capacities on a gravimetric basis cannot be recommended, because low material bulk densities could lead to unrealistically high gravimetric values.