Medusins: a new class of antimicrobial peptides from the skin secretions of phyllomedusine frogs

Natural drug discovery represents an area of research with vast potential. The investigation into the use of naturally-occurring peptides as potential therapeutic agents provides a new “chemical space” for the procurement of drug leads. Intensive and systematic studies on the broad-spectrum antimicrobial peptides found in amphibian skin secretions are of particular interest in the quest for new antibiotics to treat multiple drug-resistant bacterial infections. Here we report the molecular cloning of the biosynthetic precursor-encoding cDNAs and respective mature peptides representing a novel group of antimicrobial peptides from the skin secretions of representative species of phyllomedusine leaf frogs: the Central American red-eyed leaf frog (Agalychnis callidryas), the South American orange-legged leaf frog (Phyllomedusa hypochondrialis) and the Giant Mexican leaf frog, (Pachymedusa dacnicolor). Each novel peptide possessed the highly-conserved sequence, LGMIPL/VAISAISA/SLSKLamide, and each exhibited activity against the Gram-positive bacterium, Staphylococcus aureus and the yeast, Candida albicans, but all were devoid of haemolytic effects at concentrations up to and including the MICs for both organisms. The novel peptide group were named medusins, derived from the name of the hylid frog sub-family, Phyllomedusinae, to which all species investigated belong. These data clearly demonstrate that comparative studies of the skin secretions of phyllomedusine frogs can continue to produce novel peptides that have the potential to be leads in the development of new and effective antimicrobials.

Bradykinin-related peptides (BRPs) from skin secretions of three genera of phyllomedusine leaf frogs and their comparative pharmacological effects on mammalian smooth muscles

While bradykinin has been identified in the skin secretions from several species of amphibian, bradykinin-related peptides (BRPs) are more common constituents. These peptides display a plethora of primary structural variations from the type peptide which include single or multiple amino acid substitutions, N- and/or C-terminal extensions and post-translational modifications such as proline hydroxylation and tyrosine sulfation. Such modified peptides have been reported in species from many families, including Bombinatoridae, Hylidae and Ranidae. The spectrum of these peptides in a given species is thought to be reflective of its predator profile from different vertebrate taxa. Here we report the isolation of BRPs and parallel molecular cloning of their respective biosynthetic precursor-encoding cDNAs from the skin secretions of the Mexican leaf frog (Pachymedusa dacnicolor), the Central American red-eyed leaf frog (Agalychnis callidryas) and the South American orange-legged leaf frog (Phyllomedusa hypochondrialis). Additionally, the eight different BRPs identified were chemically synthesized and screened for bioactivity using four different mammalian smooth muscle preparations and their effects and rank potencies were found to be radically different in these with some acting preferentially through bradykinin B1-type receptors and others through B2-type receptors.

Ranakinestatin-PPF from the Skin Secretion of the Fukien Gold-Striped Pond Frog, Pelophylax plancyi fukienensis: A Prototype of a Novel Class of Bradykinin B2 Receptor Antagonist Peptide from Ranid Frogs

The defensive skin secretions of many amphibians are a rich source of bradykinins and bradykinin-related peptides (BRPs). Members of this peptide group are also common components of reptile and arthropod venoms due to their multiple biological functions that include induction of pain, effects on many smooth muscle types, and lowering systemic blood pressure. While most BRPs are bradykinin receptor agonists, some have curiously been found to be exquisite antagonists, such as the maximakinin gene-related peptide, kinestatin—a specific bradykinin B₂-receptor antagonist from the skin of the giant fire-bellied toad, Bombina maxima. Here, we describe the identification, structural and functional characterization of a heptadecapeptide (DYTIRTRLHQGLSRKIV), named ranakinestatin-PPF, from the skin of the Chinese ranid frog, Pelophylax plancyi fukienensis, representing a prototype of a novel class of bradykinin B₂-receptor specific antagonist. Using a preconstricted preparation of rat tail arterial smooth muscle, a single dose of 10⁻⁶ M of the peptide effectively inhibited the dose-dependent relaxation effect of bradykinin between 10⁻¹¹ M and 10⁻⁵ M and subsequently, this effect was pharmacologically-characterized using specific bradykinin B₁- (desArg-HOE140) and B₂-receptor (HOE140) antagonists; the data from which demonstrated that the antagonism of the novel peptide was mediated through B₂-receptors. Ranakinestatin—PPF—thus represents a prototype of an amphibian skin peptide family that functions as a bradykinin B₂-receptor antagonist herein demonstrated using mammalian vascular smooth muscle.

University Summer Quarter Chorus and High School Music Intensive Orchestra, July 11, 1940

Concert program for Summer Quarter Music, July 11, 1940

Summer School Band/Summer School Chorus, July 18, 1939

Concert program for Summer School Band/Summer School Chorus, July 18, 1939

The University Singers and The University Evening Chorus March 1, 1966

Concert program for The University Singers and the University Evening Chorus March 1, 1966

Music Score - The Drunkard's Home, As it was Temperance Song and As it is Chorus

A music score called "The Drunkard's Home" by Frank Howard. The front cover of the score has an illustration of two different experiences. One side of the illustration appears to be a family sitting by the fire as the patriarch reads the newspaper and the matriarch tends to one of the children. The other side of the illustration shows children on the floor in torn clothing as a female is slumped over a table and two men stumble in the door.

Tree frogs.

La selva es el hogar de más de la mitad de los animales del mundo desde diminutos insectos a los grandes mamíferos pretende enseñar a los niños el ciclo de vida de las ranas arbóreas y la forma en que interactúan con su entorno, sus hábitos , su estilo de vida y las amenazas a los que se enfrenta por la actuación de los hombres al modificar las condiciones ambientales. La obra contiene un mapa de distribución de su hábitat, glosario y bibliografía.

Frogs and toads.

Aprobada por expertos en educación la obra trata temas de la vida real. Texto con frases sencillas y repetición de palabras para motivar al niño a desarrollar su interés por la lectura y la curiosidad por el mundo en que vive.Tiene glosario alfabético.

Frogs.

Escrito para responder las preguntas que los niños pequeños hacen sobre las ranas. Proporciona comparaciones visuales, pasatiempos, chistes, acertijos y rompecabezas. El texto tiene dos tamaños de letra y dos niveles de dificultad para los lectores más expertos.

The matrix-tolerance hypothesis: an empirical test with frogs in the Atlantic Forest

The matrix-tolerance hypothesis suggests that the most abundant species in the inter-habitat matrix would be less vulnerable to their habitat fragmentation. This model was tested with leaf-litter frogs in the Atlantic Forest where the fragmentation process is older and more severe than in the Amazon, where the model was first developed. Frog abundance data from the agricultural matrix, forest fragments and continuous forest localities were used. We found an expected negative correlation between the abundance of frogs in the matrix and their vulnerability to fragmentation, however, results varied with fragment size and species traits. Smaller fragments exhibited stronger matrix-vulnerability correlation than intermediate fragments, while no significant relation was observed for large fragments. Moreover, some species that avoid the matrix were not sensitive to a decrease in the patch size, and the opposite was also true, indicating significant differences with that expected from the model. Most of the species that use the matrix were forest species with aquatic larvae development, but those species do not necessarily respond to fragmentation or fragment size, and thus affect more intensively the strengthen of the expected relationship. Therefore, the main relationship expected by the matrix-tolerance hypothesis was observed in the Atlantic Forest; however we noted that the prediction of this hypothesis can be substantially affected by the size of the fragments, and by species traits. We propose that matrix-tolerance model should be broadened to become a more effective model, including other patch characteristics, particularly fragment size, and individual species traits (e. g., reproductive mode and habitat preference).

Are leaf-litter frogs and lizards affected by edge effects due to forest fragmentation in Brazilian Atlantic forest?

FAPESP Fundacao de Amparo a Pesquisa do Estado de Sao Paulo

Do green and golden bell frogs (Litoria aurea) occupy habitats with fungicidal properties?

The Green and Golden Bell frog Litoria aureo is in major decline in Australia, where its distribution is now confined mainly to the east coast of New South Wales (NSW). Infection by the newly emerged amphibian fungal pathogen Batrachochytrium dendrobatidis has been identified as one of the main threats affecting L. aurea. Surprisingly, some of the sites in NSW sustaining the largest populations of this species are industrial and urban habitats that are often disturbed and polluted, which could protect L. aurea from chytrid infection if pollution had fungicidal capacity.The aim of this study was to characterise the trace metal concentration of several L. aurea breeding sites in the Sydney and IIlawarra regions of NSW and to evaluate the fungicidal efficacy of the main "'ace metals identified. Selected L. aurea sites were sampled throughout the breeding season (September to February) to establish the concentration of trace metals in both surface sediment and waters. Physico-chemical parameters including pH and salinity were also measured. Of the trace metals identified, copper and zinc were consistently elevated across sites. Over 50% of sites exceeded the National Sediment Quality Guideline for both copper and zinc concentration, and over 90% of sites exceeded the National Water Quality Guideline for these metals. Consequently, we evaluated their effect on the growth and survival of a laboratory culture of B. dendrobatidis,These tests were performed in media containing dissolved metal concentrations of 0.02 - 0.65 mgL^-1 Cu and 0.24 - 5.0 mgL^-1 Zn. Growth rates were inferred by total fungal density in liquid culture (based on spectral absorbance measurements), final dry weight, and the density of zoospores in fungal cultures grown for 28 days. Both copper and zinc were found to reduce the growth and proliferation of B. dendrobatidis, but in a non-linear manner. This suggests that L. aurea may be gaining some protection from B. dendrobatidis infection at several of the sites examined.