Elements of the granular gland peptidome and transcriptome persist in air-dried skin of the South American orange-legged leaf frog, Phyllomedusa hypocondrialis.

The defensive strategy of amphibians against predator attack relies heavily on the secretion of noxious/toxic chemical cocktails from specialized skin granular glands. Bioactive peptides constitute a major component of secretions in many species and the most complex are produced by neotropical leaf frogs of the sub-family Phyllomedusinae. We recently reported that these skin secretions contain elements of both the granular gland peptidome and transcriptome and that polyadenylated mRNAs constituting the latter are protected from degradation by interactions with endogenous amphipathic peptides. This thus permits parallel amino acid sequencing of peptides and nucleic acid sequencing of cloned precursor transcripts from single lyophilized samples of secretion. Here we report that the protection afforded is sufficiently robust to permit transcriptome studies by cloning of full-length polyadenylated peptide precursor encoding mRNAs from libraries constructed using ambient temperature air-dried skin from recently deceased specimens as source material. The technique was sufficiently sensitive to permit the identification of cDNAs encoding antimicrobial peptides constituted by six different isoforms of phylloseptin and two dermaseptins. Also, for the first time, establishment of the nucleic acid and amino acid sequence of the precursor encoding the phyllomedusine frog skin bradykinin-related peptide, phyllokinin, from cloned cDNA, was achieved. These data unequivocally demonstrate that the granular gland transcriptome persists in air-dried amphibian skin—a finding that may have fundamental implications in the study of archived materials but also in the wider field of molecular biology.

Adsorption and photocatalytic bleaching of acid orange 7 on P25 titania

The observed adsorption of acid orange 7, AO7(-), on P25 titania over a range of pH values (pH 2-8) gives a good fit to data generated using a charge distribution, multisite complexation, i.e. CD-MUSIC, model, modified for aggregated dye adsorption. For this system the model predicts that both the apparent dark Langmuir adsorption constant. K-L, and the number of adsorption sites, n(o), increase with decreasing pH, and are negligible above pH 6. At pH 2 the CD-MUSIC model predicts the fraction of singly co-ordinated sites occupied by the dye,f(AO7), is ca. 32% under the in situ monitoring experimental conditions used in this work to study the photocatalytic bleaching of AO7(-) under UV light illumination ([TiO2] = 20 mgdm(-3); [AO7(-)](total) = 4.86 x 10(-5) M). Although AO7(-) adsorption on P25 titania is insignificant above pH 6 and increases almost linearly and markedly below this pH, the measured initial rate of bleaching of AO7(-), photocatalysed by titania using UV appears to only increase modestly (

FACTORS AFFECTING THE KINETICS OF METHYL-ORANGE REDUCTION PHOTOSENSITIZED BY COLLOIDAL CDS

The kinetics of photoreduction of methyl orange by ascorbic acid sensitized by colloidal CdS has been studied. Different experimental factors such a [O2], pH and temperature, as well as the presence of potential competitive species like MV2+ and Cd2+ have been taken as variables in this study. O2 and Cd2+ clearly inhibit the photoreduction but the presence of MV2+ increases the reaction rate. The pH greatly influences the kinetics and temperature (T) has little effect. The results are interpreted using a reaction scheme proposed in earlier papers where dispersions of crystalline CdS were used as the photocatalyst and EDTA as the hole scavenger.

METHYL-ORANGE AS A PROBE OF PHOTOELECTROCHEMICAL REACTIONS SENSITIZED BY COLLOIDAL CDS

The kinetics of photoreduction of methyl orange by ethylenediaminetetraacetic acid (EDTA) sensitized by colloidal CdS are reported as a function of [methyl orange], [O2] and [EDTA]. The results are interpreted using a reaction scheme which was proposed in an earlier paper for the same reaction sensitized by a powdered dispersion of highly crystalline CdS. An analysis of the results for the CdS colloid based on this reaction scheme shows that the rate of dye reduction by photogenerated electrons is approximately 50 times greater than the rate of oxygen reduction and the rate of scavenging of the photogenerated holes is approximately 7000 times greater than the rate of recombination. These findings are discussed in the light of similar observations reported for powdered CdS.

Acridine orange leukocyte fluorography in mice

Simultaneous non-invasive visualization of blood vessels and nerves in patients can be obtained in the eye. The retinal vasculature is a target of many retinopathies. Inflammation, readily manifest by leukocyte adhesion to the endothelial lining, is a key pathophysiological mechanism of many retinopathies, making it a valuable and ubiquitous target for disease research. Leukocyte fluorography has been extensively used in the past twenty years; however, fluorescent markers, visualization techniques, and recording methods have differed between studies. The lack of detailed protocol papers regarding leukocyte fluorography, coupled with lack of uniformity between studies, has led to a paucity of standards for leukocyte transit (velocity, adherence, extravasation) in the retina. Here, we give a detailed description of a convenient method using acridine orange (AO) and a commercially available scanning laser ophthalmoscope (SLO, HRA-OCT Spectralis) to view leukocyte behavior in the mouse retina. Normal mice are compared to mice with acute and chronic inflammation. This method can be readily adopted in many research labs.

Molecular Cloning of a Novel Tryptophyllin Peptide from the Skin of the Orange-Legged Monkey Frog, Phyllomedusa hypochondrialis

Tryptophyllins are a group of small (4–14 amino acids), heterogenous peptides, mostly from the skins of hylid frogs from the genera, Phyllomedusa and Litoria. To date, more than forty TPHs have been discovered in species from these two genera. Here, we describe the identification of a novel tryptophyllin type 3 peptide, PhT-3, from the extracts of skin of the orange-legged monkey frog, Phyllomedusa hypochondrialis, and molecular cloning of its precursor-encoding cDNA from a cDNA library constructed from the same skin sample. Full primary structural characterization was achieved using a combination of direct Edman degradation, mass spectrometry and deduction from cloned skin-derived cDNA. The open-reading frame of the precursor cDNA was found to consist of 63 amino acid residues. The mature peptide arising from this precursor contains a post-translationally modified N-terminal pyroglutamate (pGlu) residue, formed from acid-mediated cyclization of an N-terminal Gln (Q) residue, and with the structure: pGlu-Asp-Lys-Pro-Phe-Trp-Pro-Pro-Pro-Ile-Tyr-Pro-Met. Pharmacological assessment of a synthetic replicate of this peptide on phenylephrine preconstricted rat tail artery segments, revealed a reduction in relaxation induced by bradykinin. PhT-3 was also found to mediate antiproliferative effects on human prostate cancer cell lines.

The majority-minority conundrum in Northern Ireland::An Orange Order perspective

Researchers have argued that, depending on the framing of the Northern Ireland conflict, each group could either be a minority or a majority relative to the other. This complicates macrosocial explanations of the conflict which make specific predictions on the basis of minority or majority positions. The present paper argues that this conundrum may have arisen from the inherent variability in microidentity processes that do not fit easily with macroexplanations. In this paper the rhetoric of relative group position is analysed in political speeches delivered by leading members of an influential Protestant institution in Northern Ireland. It is apparent that minority and majority claims are not fixed but are
flexibly used to achieve local rhetorical goals. Furthermore, the speeches differ before and after the Good Friday Agreement, with a reactionary “hegemonic” Unionist position giving way to a “majority-rights power sharing” argument and a “pseudo-minority” status giving way to a “disempowered minority” argument. These results suggest a view of the Northern Ireland conflict as a struggle for “symbolic power,” i.e., the ability to flexibly define the intergroup situation to the ingroup’s advantage.

Quantum yield measurements for the photocatalytic oxidation of Acid Orange 7 (AO7) and reduction of 2,6-dichloroindophenol (DCIP) on transparent TiO2 films of various thickness

This work comprises the photoactivity assessment of transparent sol–gel TiO2 coatings of various thickness using two test systems. The initial rates of both photocatalytic reactions, namely the oxidative bleaching of Acid Orange 7 (AO7) and the reductive bleaching of 2,6-dichlorindophenol (DCIP) increase linearly with increasing titania film thickness as well as with increasing absorbed light flux. The latter work revealed quantum yields (QY) of 0.19% and 92% for the AO7 and DCIP test system, respectively. The low QY for the AO7 oxidation is due to the combination of a slow irreversible reduction of oxygen and also for the oxidation of AO7, thus favouring the high efficiency for electron–hole recombination that is typical for aqueous organic pollutants. In contrast, the very high QY for the photocatalysed reduction of DCIP is due to the presence of a vast excess of glycerol which traps the photogenerated holes efficiently and so allow time for the slower reduction of dye to take place. Furthermore, the oxidation of glycerol results in the generation of highly reducing R-hydroxyalkyl radicals that are able to also reduce DCIP. As a consequence of this ‘current doubling’ effect, the observed QY (92%) is much higher than the apparent theoretical value of 50%.

The effect of pH on the photonic efficiency of the destruction of methyl orange under controlled periodic illumination with UV-LED sources

The nature of photon interaction and reaction pH can have significant impacts on semiconductor photocatalysis. This paper describes the effect of pH on the photonic efficiency of photocatalytic reactions in the aqueous phase using TiO2 catalysts. The reactor was irradiated using periodic illumination with UV-LEDs through control of the illumination duty cycle (γ) through a series of light and dark times (Ton/Toff). Photonic efficiencies for methyl orange degradation were found to be comparable at high γ irrespective of pH. At lower γ, pH effects on photonic efficiency were very distinct across acidic, neutral and alkaline pH indicating an effect of complementary parameters. The results suggest photonic efficiency is greatest as illumination time, Ton approaches interfacial electron-transfer characteristic time which is within the range of this study or charge-carrier lifetimes upon extrapolation and also when electrostatic attraction between surface-trapped holes, {TiIVOH}ads+ and substrate molecules is strongest.

Effect of controlled periodic-based illumination on the photonic efficiency of photocatalytic degradation of methyl orange

The use of controlled periodic illumination with UV LEDs for enhancing photonic efficiency of photocatalytic decomposition processes in water has been investigated using methyl orange as a model compound. The impact of the length of light and dark time periods (T _ON/T _OFF times) on photodegradation and photonic efficiency using a UV LED-illuminated photoreactor has been studied. The results have shown an inverse dependency of the photonic efficiency on duty cycle and a very little effect on T _ON or T _OFF time periods, indicating no effect of rate-limiting steps through mass diffusion or adsorption/desorption in the reaction. For this reactor, the photonic efficiency under controlled periodic illumination (CPI) matches to that of continuous illumination, for the same average UV light intensities. Furthermore, under CPI conditions, the photonic efficiency is inversely related to the average UV light intensity in the reactor, in the millisecond time regime. This is the first study that has investigated the effect of controlled periodic illumination using ultra band gap UV LED light sources in the photocatalytic destruction of dye compounds using titanium dioxide. The results not only enhance the understanding of the effect of periodic illumination on photocatalytic processes but also provide a greater insight to the potential of these light sources in photocatalytic reactions.