Previous illumination of a water soluble chlorine photosensitizer increases its cytotoxicity

Photodithazine (PDZ) is an N-methyl-D-glucosamine derivative of chlorine e6 that is water soluble and has an intense absorption in the range of 650-680 nm. PDZ photobleaching and photoproduct formation were induced by illumination with laser at two wavelengths: 514 nm (ion argon laser) as well as in 630 nm (dye laser). The time constants of PDZ photobleaching were: 18 min for 630 nm irradiation and 50 min for 514 nm irradiation, suggesting that degradation after irradiation with red light is faster than with green light. Photoproducts formation was evidenced by the appearance of a new absorption band at 668 nm with slight broaden of the Soret band, suggesting that there was no break of the macrocycle. The cytotoxicity of the photodegradated PDZ was investigated and showed to be lower in the dark and higher than non irradiated PDZ. These results may have important clinical implications for PDT such as the possibility to use the previously irradiated PDZ just before clinical application in order to get increased efficiency.

Safety assessment of oral photodynamic therapy in rats

Photodynamic therapy (PDT) is based on the synergism of a photosensitive drug (a photosensitizer) and visible light to destroy target cells (e.g., malignant, premalignant, or bacterial cells). The aim of this study was to investigate the response of normal rat tongue mucosa to PDT following the topical application of hematoporphyrin derivative (Photogem®), Photodithazine®, methylene blue (MB), and poly(lactic-co-glycolic acid) (PLGA) nanoparticles loaded with MB. One hundred and thirty three rats were randomly divided in various groups: the PDT groups were treated with the photosensitizers for 10 min followed by exposure to red light. Those in control groups received neither photosensitizer nor light, and they were subjected to light exposure alone or to photosensitizer alone. Fluorescent signals were obtained from tongue tissue immediately after the topical application of photosensitizers and 24 h following PDT. Histological changes were evaluated at baseline and at 1, 3, 7, and 15 days post-PDT treatment. Fluorescence was detected immediately after the application of the photosensitizers, but not 24 h following PDT. Histology revealed intact mucosa in all experimental groups at all evaluation time points. The results suggest that there is a therapeutic window where PDT with Photogem®, Photodithazine®, MB, and MB-loaded PLGA nanoparticles could safely target oral pathogenic bacteria without damaging normal oral tissue.

Influencia de la longitud de onda de la luz en el nivel de actividad de Holothuria sanctori, Holothuria dakarensis y Holothuria arguinensis

Se estudió el papel de la longitud de onda de la luz (color) en el grado de actividad motriz de tres especies de pepino de mar (Holothuria sanctori, H. dakarensis y H. arguinensis). Con independencia del tamaño de los individuos, H. sanctori prefiere los ambientes oscuros, permaneciendo inactiva en zonas iluminadas con independencia del color de la luz. H. dakarensis muestra actividad bajo luz roja, y H. arguinensis prefiere ambientes oscuros pero su desplazamiento es mayor en presencia de luz azul. ABSTRACT: The effect of the light wave length on the degree of activity of three species of sea cucumbers (Holothuria sanctori, H. dakarensis and H. arguinensis) was studied. Regardless of individuals sizes, H. sanctori prefered dark environments; this species remained inactive in illuminated areas regardless of the light colour. H. dakarensis showed activity under red light and H. arguinensis preferred dark environments, though its displacement was larger under blue light.

Microclima de la canopia de la vid : influencia del manejo del suelo y coberturas vegetales.

Para determinar la influencia del manejo del suelo sobre el microclima de la canopia se estableció un experimento en un viñedo de Cabernet Sauvignon conducido en doble cordón de pitones en espladero alto. Se aplicaron tres tratamientos: Testigo (TR)-suelo sin maleza; (CVP)-cobertura de suelo espontánea y control del desarrollo vegetativo por desbrozado y (CA)-cobertura de flora espontánea y control del desarrollo vegetativo con aplicación de herbicida de contacto desecante. Se midió la radiación fotosintéticamente activa (PAR), la composición espectral de la radiación directa y reflejada, y la temperatura a nivel de racimos en el interior de la canopia. El TR reflejó más radiación en la zona del UV y del visible. El CVP reflejó entre un 8 y 9%, el CA entre el 16 y 18% y el TR entre 18 y 19%. En la radiación reflejada, la relación rojo/rojo lejano fue mayor en CA y TR, mientras que la relación azul/rojo fue mayor en CVP. La temperatura en el interior de la canopia fue mayor en TR (30,8°C) que en CVP (26,7°C). La utilización de coberturas vegetales de raíces superficiales y permanentemente desbrozadas sería de utilidad para reducir la radiación reflejada y la temperatura de la canopia sin competir con el viñedo por agua y nutrimentos.

Manejo de suelo mediante coberturas vegetales establecidas

El objetivo de esta investigación fue determinar los efectos de las coberturas vegetales en el microclima de la planta de vid. Se compararon cinco coberturas de diferente ciclo vegetativo con respecto al manejo de suelo sin labranza mediante aplicación de herbicidas. El estudio se desarrolló en un viñedo cv. Malbec conducido en espaldera alta, situado en Agrelo, Luján de Cuyo, Mendoza, Argentina. Se determinaron parámetros microclimáticos, temperatura, humedad relativa y radiación a nivel de racimos, temperatura del suelo, cantidad y calidad de la radiación reflejada por la cobertura. También se midió la expresión vegetativa y de uvas y el potencial enológico. Se verificó una significativa disminución de la radiación fotosintéticamente activa (RFA) reflejada por las coberturas con una menor relación “Rojo/Rojo lejano" comparada con el suelo descubierto. Sin embargo, el efecto no se percibió dentro de la canopia debido a que las coberturas permanentes de trébol rojo (Trifolium pratensis) y agropiro alargado (Agropyron elongatum) restringieron el vigor de las cepas, disminuyendo el crecimiento de brotes y el tamaño de hojas, lo cual se tradujo en una mayor recepción directa de la RFA a nivel de racimos. No hubo una significativa variación en cuanto a temperatura máxima, mínima y amplitud térmica a nivel de racimos. No obstante ello, los tratamientos con mayor cobertura de suelo tendieron a reducir levemente la temperatura mínima a nivel de racimos. La humedad relativa en la canopia no fue significativamente afectada. El trébol rojo, el agropiro alargado, la mezcla centeno-cebadilla (Secale cereale-Bromus catharticus) y el sorgo del Sudán (Sorghum sudanensis) redujeron notablemente la amplitud térmica del suelo. El efecto fue determinado principalmente por la disminución de la temperatura máxima. Las coberturas vegetales con alguna dificultad para desarrollarse durante su ciclo vegetativo tuvieron un comportamiento intermedio o uno muy similar al de un suelo descubierto. La introducción de una cobertura permanente con buena invasión del sitio interfilar permitió modificar indirectamente las características microclimáticas de la canopia, a través del control del crecimiento vegetativo y de los rendimientos de la planta de vid, modificando el equilibrio vigor / producción del viñedo, y por lo tanto la composición de las uvas y del vino elaborado.

Impact studies from different factors on root development from Stevia rebaudiana bertoni

The demand for an alternative and a high potency sweetener to substitute sugar increases year in year out, more so as a high percentage of the world population becomes increasingly diabetic. The alternative natural sweetener at hand has been Stevia rebaudiana Bertoni, a plant species, native to Paraguay and a member of the family compositae. Stevia is usually propagated by stem cuttings due to low percentage (10 %) seed germination, thus limiting large scales cultivation. To cultivate this crop en mass therefore, there is need to evolve efficient rooting techniques. Influences of irradiation from light, and hormones on rooting have been reported. The rooting efficacy in stem cuttings of this crop under varying light wavelengths, dark and hormone factors was investigated. Evaluated parameters include- (i) day of root emergent, (ii) percentage of rooted cuttings, (iii) average number, (iv) length and (v) width, of roots. Analysis of variance at p<.05 revealed that the number, length and width, of roots differed significantly in each case at p<0.000. Light irradiation was highly effective and a necessary factor for rooting in stems cuttings of this crop. The red light-IBA combined factors served best in stem micro-cutting practice and facilitation of effective mass cultivation in stevia crop.

Image enhancement with MatLab algorithms

Nowadays, a lot of applications use digital images. For example in face recognition to detect and tag persons in photograph, for security control, and a lot of applications that can be found in smart cities, as speed control in roads or highways and cameras in traffic lights to detect drivers ignoring red light. Also in medicine digital images are used, such as x-ray, scanners, etc. These applications depend on the quality of the image obtained. A good camera is expensive, and the image obtained depends also on external factor as light. To make these applications work properly, image enhancement is as important as, for example, a good face detection algorithm. Image enhancement also can be used in normal photograph, for pictures done in bad light conditions, or just to improve the contrast of an image. There are some applications for smartphones that allow users apply filters or change the bright, colour or contrast on the pictures. This project compares four different techniques to use in image enhancement. After applying one of these techniques to an image, it will use better the whole available dynamic range. Some of the algorithms are designed for grey scale images and others for colour images. It is used Matlab software to develop and present the final results. These algorithms are Successive Means Quantization Transform (SMQT), Histogram Equalization, using Matlab function and own implemented function, and V transform. Finally, as conclusions, we can prove that Histogram equalization algorithm is the simplest of all, it has a wide variability of grey levels and it is not suitable for colour images. V transform algorithm is a good option for colour images. The algorithm is linear and requires low computational power. SMQT algorithm is non-linear, insensitive to gain and bias and it can extract structure of the data. RESUMEN. Hoy en día incontable número de aplicaciones usan imágenes digitales. Por ejemplo, para el control de la seguridad se usa el reconocimiento de rostros para detectar y etiquetar personas en fotografías o vídeos, para distintos usos de las ciudades inteligentes, como control de velocidad en carreteras o autopistas, cámaras en los semáforos para detectar a conductores haciendo caso omiso de un semáforo en rojo, etc. También en la medicina se utilizan imágenes digitales, como por ejemplo, rayos X, escáneres, etc. Todas estas aplicaciones dependen de la calidad de la imagen obtenida. Una buena cámara es cara, y la imagen obtenida depende también de factores externos como la luz. Para hacer que estas aplicaciones funciones correctamente, el tratamiento de imagen es tan importante como, por ejemplo, un buen algoritmo de detección de rostros. La mejora de la imagen también se puede utilizar en la fotografía no profesional o de consumo, para las fotos realizadas en malas condiciones de luz, o simplemente para mejorar el contraste de una imagen. Existen aplicaciones para teléfonos móviles que permiten a los usuarios aplicar filtros y cambiar el brillo, el color o el contraste en las imágenes. Este proyecto compara cuatro técnicas diferentes para utilizar el tratamiento de imagen. Se utiliza la herramienta de software matemático Matlab para desarrollar y presentar los resultados finales. Estos algoritmos son Successive Means Quantization Transform (SMQT), Ecualización del histograma, usando la propia función de Matlab y una nueva función que se desarrolla en este proyecto y, por último, una función de transformada V. Finalmente, como conclusión, podemos comprobar que el algoritmo de Ecualización del histograma es el más simple de todos, tiene una amplia variabilidad de niveles de gris y no es adecuado para imágenes en color. El algoritmo de transformada V es una buena opción para imágenes en color, es lineal y requiere baja potencia de cálculo. El algoritmo SMQT no es lineal, insensible a la ganancia y polarización y, gracias a él, se puede extraer la estructura de los datos.

A Model for Signal Transduction during Gamete Release in the Fucoid Alga Pelvetia compressa1

Fucoid algae release gametes into seawater following an inductive light period (potentiation), and gamete expulsion from potentiated receptacles of Pelvetia compressa began about 2 min after a light-to-dark transition. Agitation of the medium reversed potentiation, with an exponential time course completed in about 3 h. Light regulated two signaling pathways during potentiation and gamete expulsion: a photosynthetic pathway and a photosynthesis-independent pathway in which red light was active but blue light was not. Uptake of K+ appears to have an important role in potentiation, because a 50% inhibition of potentiation occurred in the presence of the tetraethylammonium ion, a K+-channel blocker. A central role of anion channels in the maintenance of potentiation is suggested by the premature release of gametes in the light when receptacles were incubated with inhibitors of slow-type anion channels. An inhibitor of tyrosine kinases, tyrphostin A63, also inhibited potentiation. A model for gamete release from P. compressa is presented that proposes that illumination results in the accumulation of ions (e.g. K+) throughout the cells of the receptacle during potentiation, which then move into the extracellular matrix during gamete expulsion to generate osmomechanical force, resulting in gamete release.

Electroretinogram analysis of relative spectral sensitivity in genetically identified dichromatic macaques

The retinas of macaque monkeys usually contain three types of photopigment, providing them with trichromatic color vision homologous to that of humans. However, we recently used molecular genetic analysis to identify several macaques with a dichromatic genotype. The affected X chromosome of these animals contains a hybrid gene of long-wavelength-sensitive (L) and middle-wavelength-sensitive (M) photopigments instead of separate genes encoding L and M photopigments. The product of the hybrid gene exhibits a spectral sensitivity close to that of M photopigment; consequently, male monkeys carrying the hybrid gene are genetic protanopes, effectively lacking L photopigment. In the present study, we assessed retinal expression of L photopigment in monkeys carrying the hybrid gene. The relative sensitivities to middle-wavelength (green) and long-wavelength (red) light were measured by electroretinogram flicker photometry. We found the sensitivity to red light to be extremely low in protanopic male monkeys compared with monkeys with the normal genotype. In female heterozygotes, sensitivity to red light was intermediate between the genetic protanopes and normal monkeys. Decreased sensitivity to long wavelengths was thus consistent with genetic loss of L photopigment.

Mutational analysis of phytochrome B identifies a small COOH-terminal-domain region critical for regulatory activity.

Overexpression of phytochrome B (phyB) in transgenic Arabidopsis results in enhanced deetiolation in red light. To define domains of phyB functionally important for its regulatory activity, we performed chemical mutagenesis of a phyB-overexpressing line and screened for phenotypic revertants in red light. Four phyB-transgene-linked revertants that retain parental levels of full-length, dimeric, and spectrally normal overexpressed phyB were identified among 101 red-light-specific revertants. All carry single amino acid substitutions in the transgene-encoded phyB that reduce activity by 40- to 1000-fold compared to the nonmutagenized parent. The data indicate that the mutant molecules are fully active in photosignal perception but defective in the regulatory activity responsible for signal transfer to downstream components. All four mutations fall within a 62-residue region in the COOH-terminal domain of phyB, with two independent mutations occurring in a single amino acid, Gly-767. Accumulating evidence indicates that the identified region is a critical determinant in the regulatory function of both phyB and phyA.

Feedback from luminosity horizontal cells mediates depolarizing responses of chromaticity horizontal cells in the Xenopus retina.

It has been proposed that the depolarizing responses of chromaticity horizontal cells (C-HCs) to red light depend on a feedback signal from luminosity horizontal cells (L-HCs) to short-wavelength-sensitive cones in the retinas of lower vertebrates. In this regard we studied the C-HCs of the Xenopus retina. C-HCs and L-HCs were identified by physiological criteria and then injected with neurobiotin. The retina then was incubated with peanut agglutinin, which stains red-but not blue-sensitive cones. Electron microscopic examination revealed that L-HCs contact all cone classes, whereas C-HCs contact only blue-sensitive cones. Simultaneous recordings from C-HC/L-HC pairs established that when the L-HC was saturated by a steady bright red light, C-HCs alone responded to a superimposed blue stimulus. In response to red test flashes, the C-HC response was delayed by approximately 30 msec with respect to the L-HC response. Isolated HCs of both subtypes were examined by whole-cell patch clamp. Both responded to kainate with sustained inward currents and to quisqualate or alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) with desensitizing currents from a negative holding potential; i.e., both have AMPA-type glutamate receptors. gamma-Aminobutyric acid or glycine opened a chloride channel in the L-HC, whereas the C-HC was unresponsive to either inhibitory amino acid. Since glycine has been shown to abolish selectively the depolarizing response of the C-HC, this finding and other pharmacological data strongly implicate the L-HC in the underlying circuit. Moreover, because the C-HC does not respond to gamma-aminobutyric acid, the neurotransmitter of the L-HC, by elimination, a feedback synapse from L-HC to blue cone is the most plausible mechanism for the creation of depolarizing responses in C-HCs.

The young open cluster Berkeley 55

We present UBV photometry of the highly reddened and poorly studied open cluster Berkeley 55, revealing an important population of B-type stars and several evolved stars of high luminosity. Intermediate-resolution far-red spectra of several candidate members confirm the presence of one F-type supergiant and six late supergiants or bright giants. The brightest blue stars are mid-B giants. Spectroscopic and photometric analyses indicate an age 50 ± 10 Myr. The cluster is located at a distance d ≈ 4 kpc, consistent with other tracers of the Perseus Arm in this direction. Berkeley 55 is thus a moderately young open cluster with a sizable population of candidate red (super)giant members, which can provide valuable information about the evolution of intermediate-mass stars.

Fast X-ray transients towards the Galactic bulge with the Rossi X-ray Timing Explorer

In X-ray binaries, rapid variability in X-ray flux of greater than an order of magnitude on time-scales of a day or less appears to be a signature of wind accretion from a supergiant companion. When the variability takes the form of rare, brief, bright outbursts with only faint emission between them, the systems are called supergiant fast X-ray transients (SFXTs). We present data from twice-weekly scans of the Galactic bulge by the Rossi X-ray Timing Explorer that allow us to compare the behaviour of known SFXTs and possible SFXT candidates with the persistently bright supergiant X-ray binary 4U 1700−377. We independently confirm the orbital periods reported by other groups for SFXTs SAX J1818.6−1703 and IGR J17544−2619. The new data do not independently reproduce the orbital period reported for XTE J1739−302, but slightly improve the significance of the original result when the data are combined. The bulge source XTE J1743−363 shows a combination of fast variability and a long-term decline in activity, the latter behaviour not being characteristic of supergiant X-ray binaries. A far-red spectrum of the companion suggests that it is a symbiotic neutron star binary rather than a high-mass binary, and the reddest known of this class: the spectral type is approximately M8 III.

Factors affecting N2 fixation by the cyanobacterium Trichodesmium sp. GBRTRLI101

Various factors affecting N-2 fixation of a cultured strain of Trichodesmium sp. (GBRTRLI101) from the Great Barrier Reef Lagoon were investigated. The diurnal pattern of N2 fixation demonstrated that it was primarily light-induced although fixation continued to occur for at least 1 h in the dark in samples that had been actively fixing N-2. N-2 fixation was dependent on the light intensity and stimulated more by white light when compared with blue, green, yellow and red light whereas rates of N-2 fixation decreased most under red light. Inorganic phosphorous concentrations in the lower range of treatments up to 1.2 muM significantly stimulated N-2 fixation and further additions promoted little or no increase in N-2 fixation. Organic phosphorous (Na-glycerophosphate) also stimulated N-2 fixation rates. Added combined nitrogen (NH4+, NO3-, urea) of 10 muM did not inhibit N-2 fixation in short-term studies (first generation), however it was depressed in the long-term studies (fifth generation). (C) 2003 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved.

Synthesis and characterization of dual-functionalized core-shell fluorescent microspheres for bioconjugation and cellular delivery

The efficient transport of micron-sized beads into cells, via a non-endocytosis mediated mechanism, has only recently been described. As such there is considerable scope for optimization and exploitation of this procedure to enable imaging and sensing applications to be realized. Herein, we report the design, synthesis and characterization of fluorescent microsphere-based cellular delivery agents that can also carry biological cargoes. These core-shell polymer microspheres possess two distinct chemical environments; the core is hydrophobic and can be labeled with fluorescent dye, to permit visual tracking of the microsphere during and after cellular delivery, whilst the outer shell renders the external surfaces of the microspheres hydrophilic, thus facilitating both bioconjugation and cellular compatibility. Cross-linked core particles were prepared in a dispersion polymerization reaction employing styrene, divinylbenzene and a thiol-functionalized co-monomer. These core particles were then shelled in a seeded emulsion polymerization reaction, employing styrene, divinylbenzene and methacrylic acid, to generate orthogonally functionalized core-shell microspheres which were internally labeled via the core thiol moieties through reaction with a thiol reactive dye (DY630-maleimide). Following internal labeling, bioconjugation of green fluorescent protein (GFP) to their carboxyl-functionalized surfaces was successfully accomplished using standard coupling protocols. The resultant dual-labeled microspheres were visualized by both of the fully resolvable fluorescence emissions of their cores (DY630) and shells (GFP). In vitro cellular uptake of these microspheres by HeLa cells was demonstrated conventionally by fluorescence-based flow cytometry, whilst MTT assays demonstrated that 92% of HeLa cells remained viable after uptake. Due to their size and surface functionalities, these far-red-labeled microspheres are ideal candidates for in vitro, cellular delivery of proteins, as described in the accompanying paper.