An easy method for quantification of anaerobic and microaerobic gene expression with fluorescent reporter proteins

Fluorescent proteins, such as green fluorescent proteins, are invaluable tools for detecting and quantifying gene expression in high-throughput reporter gene assays. However, they introduce significant inaccuracies in studies involving microaerobiosis or anaerobiosis, as oxygen is required for the maturation of these proteins' chromophores. In this study, the authors highlight the errors incurred by using fluorescent proteins under limited oxygenation by comparing standard fluorescence-based reporter gene assays to quantitative real-time PCR data in the study of a complex oxygen-regulated gene network. Furthermore, a solution to perform quantification of anaerobic and microaerobic gene expression with fluorescent reporter proteins using a microplate reader with an oxygen control system and applying pulses of full oxygenation before fluorescence measurements is provided

Line-Strength Indices in Bright Spheroidals: Evidence for a Stellar Population Dichotomy between Spheroidal and Elliptical Galaxies

We present new measurements of central line-strength indices (namely Mg2, , and Hbeta and gradients for a sample of 6 bright spheroidal galaxies (Sph's) in the Virgo cluster. Comparison with similar measurements for elliptical galaxies (E's), galactic globular clusters (GGC's), and stellar population models yield the following results: (1) In contrast with bright E's, bright Sph's are consistent with solar abundance [Mg/Fe] ratios; (2) Bright Sph's exhibit metallicities ranging from values typical for metal-rich GGC's to those for E's; (3) Although absolute mean ages are quite model dependent, we find evidence that the stellar populations of some (if not all) Sph's look significantly younger than GGC's; and (4) Mg2 gradients of bright Sph's are significantly shallower than those of E galaxies. We conclude that the dichotomy found in the structural properties of Sph and E galaxies is also observed in their stellar populations. A tentative interpretation in terms of differences in star formation histories is suggested.Comment: 14 pages, LaTeX file + 2 PostScript figures, aasms4.sty require

CAFE: Calar Alto Fiber-fed Echelle spectrograph

We present here CAFE, the Calar Alto Fiber-fed Echelle spectrograph, a new instrument built at the Centro Astronomico Hispano Alem\'an (CAHA). CAFE is a single fiber, high-resolution ($R\sim$70000) spectrograph, covering the wavelength range between 3650-9800\AA. It was built on the basis of the common design for Echelle spectrographs. Its main aim is to measure radial velocities of stellar objects up to $V\sim$13-14 mag with a precision as good as a few tens of $m s^{-1}$. To achieve this goal the design was simplified at maximum, removing all possible movable components, the central wavelength is fixed, so the wavelentgth coverage; no filter wheel, one slit and so on, with a particular care taken in the thermal and mechanical stability. The instrument is fully operational and publically accessible at the 2.2m telescope of the Calar Alto Observatory. In this article we describe (i) the design, summarizing its manufacturing phase; (ii) characterize the main properties of the instrument; (iii) describe the reduction pipeline; and (iv) show the results from the first light and commissioning runs. The preliminar results indicate that the instrument fulfill the specifications and it can achieve the foreseen goals. In particular, they show that the instrument is more efficient than anticipated, reaching a $S/N\sim$20 for a stellar object as faint as $V\sim$14.5 mag in $\sim$2700s integration time. The instrument is a wonderful machine for exoplanetary research (by studying large samples of possible systems cotaining massive planets), galactic dynamics (high precise radial velocities in moving groups or stellar associations) or astrochemistry.Comment: 12 pages, 23 figures; Acepted for publishing in A&A, 201

Internal Kinematics of Luminous Compact Blue Galaxies

We describe the dynamical properties which may be inferred from HST/STIS spectroscopic observations of luminous compact blue galaxies (LCBGs) between 0.1<z<0.7. While the sample is homogeneous in blue rest-frame color, small size and line-width, and high surface-brightness, their detailed morphology is eclectic. Here we determine the amplitude of rotation versus random, or disturbed motions of the ionized gas. This information affirms the accuracy of dynamical mass and M/L estimates from Keck integrated line-widths, and hence also the predictions of the photometric fading of these unusual galaxies. The resolved kinematics indicates this small subset of LCBGs are dynamically hot, and unlikely to be embedded in disk systems.Comment: To appear in "Starbursts: from 30 Doradus to Lyman Break Galaxies" 2005, eds. R. de Grijs and R. M. Gonzalez Delgado (Kluwer

Microesferas de plga: un sistema para la liberación controlada de moléculas con actividad inmunogénica

En el campo de la tecnología farmacéutica se presenta actualmente considerable interés en el desarrollo de micropartículas biodegradables como un sistema para la liberación controlada de moléculas con actividad biológica. Son una buena alternativa, teniendo en cuenta que una de las principales desventajas de los productos disponibles es la necesidad de realizar varias administraciones para cumplir totalmente con el tratamiento. Esta contribución examina el papel actual de la microencapsulación de moléculas con actividad antigénica en microesferas elaboradas con polímeros biodegradables derivados de los ácidos láctico y glicólico (PLGA). Aquí se describen las propiedades fisicoquímicas de los biopolímeros empleados, los métodos de obtención de las micropartículas y de algunos de las variables que influyen en las propiedades del producto final. Además, se realiza una breve descripción del modo de acción propuesto para este sistema microparticular.At the present time, there is a great interest in pharmaceutical technology focused on the development of biodegradable microparticles as a controlled release system of molecules with biological activity. This strategy arises as a good alternative; mainly if it is keep in mind that one of the principal disadvantages of the available products currently is that those products require several administrations to fulfilled the treatment. Here in is examined the role of microencapsulated antigens into microespheres performed with biodegradable copolymers derived from lactic and glycolic acid (PLGA). Physicochemical properties of biopolymers used, microspheres formulation as well as the parameters that affected the properties of the final product are also described. Additionally, there is a brief review of the action mechanism raised for this microparticular system

Tunable injectable alginate-based hydrogel for cell therapy in Type 1 Diabetes Mellitus

Islet transplantation has the potential of reestablishing naturally-regulated insulin production in Type 1 diabetic patients. Nevertheless, this procedure is limited due to the low islet survival after transplantation and the lifelong immunosuppression to avoid rejection. Islet embedding within a biocompatible matrix provides mechanical protection and a physical barrier against the immune system thus, increasing islet survival. Alginate is the preferred biomaterial used for embedding insulin-producing cells because of its biocompatibility, low toxicity and ease of gelation. However, alginate gelation is poorly controlled, affecting its physicochemical properties as an injectable biomaterial. Including different concentrations of the phosphate salt Na2HPO4 in alginate hydrogels, we can modulate their gelation time, tuning their physicochemical properties like stiffness and porosity while maintaining an appropriate injectability. Moreover, these hydrogels showed good biocompatibility when embedding a rat insulinoma cell line, especially at low Na2HPO4 concentrations, indicating that these hydrogels have potential as injectable biomaterials for Type 1 Diabetes Mellitus treatment

Advances in the slow freezing cryopreservation of microencapsulated cells

Over the past few decades, the use of cell microencapsulation technology has been promoted for a wide range of applications as sustained drug delivery systems or as cells containing biosystems for regenerative medicine. However, difficulty in their preservation and storage has limited their availability to healthcare centers. Because the preservation in cryogenic temperatures poses many biological and biophysical challenges and that the technology has not been well understood, the slow cooling cryopreservation, which is the most used technique worldwide, has not given full measure of its full potential application yet. This review will discuss the different steps that should be understood and taken into account to preserve microencapsulated cells by slow freezing in a successful and simple manner. Moreover, it will review the slow freezing preservation of alginate-based microencapsulated cells and discuss some recommendations that the research community may pursue to optimize the preservation of microencapsulated cells, enabling the therapy translate from bench to the clinic

Force Spectroscopy Imaging and Constriction Assays Reveal the Effects of Graphene Oxide on the Mechanical Properties of Alginate Microcapsules

Microencapsulation of cells in hydrogel-based porous matrices is an approach that has demonstrated great success in regenerative cell therapy. These microcapsules work by concealing the exogenous cells and materials in a robust biomaterial that prevents their recognition by the immune system. A vast number of formulations and additives are continuously being tested to optimize cell viability and mechanical properties of the hydrogel. Determining the effects of new microcapsule additives is a lengthy process that usually requires extensive in vitro and in vivo testing. In this paper, we developed a workflow using nanoindentation (i.e., indentation with a nanoprobe in an atomic force microscope) and a custom-built microfluidic constriction device to characterize the effect of graphene oxide (GO) on three microcapsule formulations. With our workflow, we determined that GO modifies the microcapsule stiffness and surface properties in a formulation-dependent manner. Our results also suggest, for the first time, that GO alters the conformation of the microcapsule hydrogel and its interaction with subsequent coatings. Overall, our workflow can infer the effects of new additives on microcapsule surfaces. Thus, our workflow can contribute to diminishing the time required for the validation of new microcapsule formulations and accelerate their clinical translation

Design of a nasal spray based on cardiospermum halicacabum extract loaded in phospholipid vesicles enriched with gelatin or chondroitin sulfate

The extract of Cardiospermum halicacabum L. (C. halicacabum) obtained from flower, leaf and vine was loaded into modified phospholipid vesicles aiming at obtaining sprayable, biocompatible and effective nasal spray formulations for the treatment of nasopharyngeal diseases. Penetration enhancer-containing vesicles (PEVs) and hyalurosomes were formulated, and stabilized by adding a commercial gelatin from fish (20 mg/mL) or chondroitin sulfate from catshark cartilages (Scyliorhi-nus canicula, 20 mg/mL). Cryo-TEM images confirmed the formation of spherical vesicles, while photon correlation spectroscopy analysis disclosed the formation of small and negatively-charged vesicles. PEVs were the smaller vesicles (~100 nm) along with gelatin-hyalurosomes (~120 nm), while chondroitin-PEVs and chondroitin-hyalurosomes were larger (~160 nm). Dispersions prepared with chondroitin sulfate were more homogeneous, as the polydispersity index was ~0.15. The in vitro analysis of the droplet size distribution, average velocity module and spray cone angle suggested a good spray-ability and deposition of formulations in the nasal cavity, as the mean diameter of the droplets was in the range recommended by the Food and Drug Administration for nasal targets. The spray plume analysis confirmed the ability of PEVs, gelatin-PEVs, hyalurosomes and gelatin-hyalurosomes to be atomized in fine droplets homogenously distributed in a full cone plume, with an angle ranging from 25 to 30◦ . Moreover, vesicles were highly biocompatible and capable of protecting the epithelial cells against oxidative damage, thus preventing the inflammatory state

Harassment Origin for Kinematic Substructures in Dwarf Elliptical Galaxies?

We have run high resolution N-body models simulating the encounter of a dwarf galaxy with a bright elliptical galaxy. The dwarf absorbs orbital angular momentum and shows counter-rotating features in the external regions of the galaxy. To explain the core-envelope kinematic decoupling observed in some dwarf galaxies in high-density environments requires nearly head-on collisions and very little dark matter bound to the dwarf. These kinematic structures appear under rather restrictive conditions. As a consequence, in a cluster like Virgo ~1% of dwarf galaxies may present counter-rotation formed by harassment.Comment: 10 pages, 7 figures; Accepted for publication in Astronomy and Astrophysic