Combinatorial synthesis and directed evolution applied to the production of alpha-helix forming antimicrobial peptides analogues

Antimicrobial peptides (AMPs) are effector molecules of innate immune systems found in different groups of organisms, including microorganisms, plants, insects, amphibians and humans. These peptides exhibit several structural motifs but the most abundant AMPs assume an amphipathic alpha-helical structure. The alpha-helix forming antimicrobial peptides are excellent candidates for protein engineering leading to an optimization of their biological activity and target specificity. Nowadays several approaches are available and this review deals with the use of combinatorial synthesis and directed evolution in order to provide a high-throughput source of antimicrobial peptides analogues with enhanced lytic activity and specificity.

Physiology of temperature regulation: Comparative aspects

Few environmental factors have a larger influence on animal energetics than temperature, a fact that makes thermoregulation a very important process for survival. In general, endothermic species, i.e., mammals and birds, maintain a constant body temperature (Tb) in fluctuating environmental temperatures using autonomic and behavioural mechanisms. Most of the knowledge on thermoregulatory physiology has emerged from studies using mammalian species, particularly rats. However, studies with all vertebrate groups are essential for a more complete understanding of the mechanisms involved in the regulation of Tb. Ectothermic vertebrates-fish, amphibians and reptiles-thermoregulate essentially by behavioural mechanisms. With few exceptions, both endotherms and ectotherms develop fever (a regulated increase in Tb) in response to exogenous pyrogens, and regulated hypothermia (anapyrexia) in response to hypoxia. This review focuses on the mechanisms, particularly neuromediators and regions in the central nervous system, involved in thermoregulation in vertebrates, in conditions of euthermia, fever and anapyrexia. (c) 2006 Elsevier B.V. All rights reserved.

Short-term movement and retreat sites of Leptodactylus labyrinthicus (Anura : Leptodactylidae) during the breeding season: A spool-and-line tracking study

Capture/recapture studies significantly increase our knowledge of the natural history of anuran amphibians. Many different methods have been employed in these studies, but a number of new techniques still require experimental validation. During two reproductive seasons in a Cerrado remnant in southeastern Brazil, we investigated the movement patterns and habitat use of the pepper frog, Leptodactylus labyrinthicus, using a spool-and-line device. This low-cost device did not appear to interfere with the activities of the frogs and allowed for constant monitoring, showing precise routes of movement and great predictability of relocations. Both sexes were active at night. During the day, males and females made use of retreat sites under vegetation or in burrows constructed by small- and medium-sized mammals. Males and females did not use standardized routes; there were no significant differences between their movements, and movements were not correlated with body size or environmental conditions. Individuals are able to move further than 100 m per day, a characteristic that may enable this species to colonize or recolonize open areas.

A REVIEW OF THE GENUS MEGAELOSIA (ANURA, LEPTODACTYLIDAE) WITH A DESCRIPTION OF A NEW SPECIES

A new species of Megaelosia is described from the Atlantic Forest in the northern part of São Paulo, southeastern Brazil. Based on examination of topotypic specimens of Megaelosia massarti, this species is removed from the synonymy of M. goeldii. Tadpoles of M. lutzae and M. massarti are described and natural history observations of M. massarti and M. goeldii are reported. Diagnoses, measurements, figures, distributional data, and a key for the species of Megaelosia are provided.

A new island species of Scinax (Anura; Hylidae) from southeastern Brazil

A new species of hylid frog, from the genus Scinax, is described from Ilha de Porcos Pequena at the southern coast of São Paulo State, southeastern Brazil. The new species belongs to the Scinax perpusillus species group and is diagnosed by the following set of characters: moderate-size (males 16.2-18.8 turn SVL, female 18.8-20.6 mm SVL); canthus rostralis distinct and well defined; V-shaped depression between nostrils; eyes protruding and prominent, glandular skin surface of legs. This new species is found only on Ilha de Porcos Pequena, an island of approximately 24 ha and, therefore, is threatened because of restricted range size and susceptibility to habitat modification.

Anatomy of the oral cavity of hylid larvae from the genera Aplastodiscus, Bokermannohyla, and Hypsiboas (Amphibia, anura): Description and systematic implications

Internal larval oral anatomy was used to explore morphological diversity and its contribution to the systematics of the genera Aplastodiscus, Bokerinannohyla, and Hypsiboas, belonging to the tribe Cophomantini. Internal oral morphology was examined for tadpoles of 12 species. All species have a large pair of infralabial papillae on the buccal floor and other papillae on the prelingual region. In Aplastodiscus and Bokerinannohyla, the large infralabial papillae have digitiform secondary projections. The number and arrangement of the buccal floor papillae varies among species, but they are more numerous in Aplastodiscus and Bokerinannohyla. The arrangement of the postnarial papillae is variable, but in Aplastodiscus and Bokerinannohyla, they show a definite, inverted V-shape pattern. The lateral ridge papillae are more complex in larvae of Bokertnannohyla with long digitiform secondary projections. Tadpoles of only Aplastodiscus albofrenatus, Aplastodiscus eugenioi, and Bokerinannohyla luctuosa have papillae on the buccal roof arena and larvae of all species have lateral roof papillae except Hypsiboas albomarginatus and Hypsiboas cinerascens. Larvae of Aplastodiscus, Bokermannohyla, and Hypsiboas presumably share the presence of vacuities anterior to the internal nares; although this character state is clearly synapomorphic within hylids, it is still uncertain whether it is exclusive of these three genera or whether it is present in the other genera of the tribe Cophomantini (Hyloscirtus and Myersiohyla). The inclusion of internal oral anatomy characters, such as the narial vacuities, in systematic studies is certainly valuable because it will provide additional information toward the understanding of phylogenetic relationships.

A New Species of Proceratophrys (Anura: Cycloramphidae) from Midwestern Brazil

We describe adult morphology, advertisement call, and natural history aspects of a new frog species from riparian habitats alongside intermittent headwater streams in southwestern Cerrado, the second largest biome in Brazil. Presently known from three localities in Mato Grosso State, the new species belongs to the Proceratophrys cristiceps group. It is characterized mainly by small size (adult snout vent length approximately 46 mm), snout rounded in dorsal view and obtuse in lateral view, absence of flared lips, dorsal skin granular, and absence of both postocular swellings and prominent palpebral appendages. The advertisement call of the new species consists of a single note with 19-25 pulses. Calls are repeated at mean intervals of 0.7 sec. Mean dominant frequency is 1,250.2 Hz, with ascendant modulation.

The neuranatomical basis of central control of cardiorespiratory interactions in vertebrates

It seems that a dual location for vagal preganglionic neurones (VPNs) has important functional correlates in all vertebrates. This may be particularly the case with the central control exerted over the heart by cardiac VPNs (CVPNs). About 30 % of VPNs but up to 70 % of CVPNs are in the nucleus ambiguus (NA) of mammals. There is a similar proportional representation of VPNs between the major vagal nuclei in amphibians and turtles but in fish and crocodilians; the proportion of VPNs in the NA is closer to 10% and in some lizards and birds it is about 5%. However, the CVPNs are distributed unequally between these nuclei so that 45 % of the CVPNs are located in the NA of the dogfish, and about 30% in the NA of Xenopus and the duck. This topographical separation of CVPNs seems to be of importance in the central control of the heart. Cells in one location may show respiration-related activity (e.g those in the dorsal vagal nucleus (DVN) of dogfish and in the NA of mammals) while cells in the other locations do not. Their different activities and separate functions will be determined by their different afferent inputs from the periphery or from elsewhere in the CNS, which in turn will relate to their central topography. Thus, CVPNs in the NA of mammals receive inhibitory inputs from neighbouring inspiratory neurones, causing respiratory sinus arrythmia (RSA), and the CVPNs in the DVN of the dogfish may generate cardiorespiratory synchrony (CRS).

Natural History of the Lutz's Frog Cycloramphus lutzorum Heyer, 1983 (Anura: Cycloramphidae) in the Brazilian Atlantic Forest: Description of the Advertisement Call, Tadpole, and Karyotype

We describe the advertisement call, tadpole, karyotype, and additional information on the natural history of Cycloramphus lutzorum from southern Brazil. Sonograms were generated from digitally recorded calls. Tadpoles were collected in the field for description in the lab, and an adult was collected for karyotyping. Data on seasonal activity were gathered monthly from November 2005 to November 2007. All tadpoles (N = 21), juveniles (N = 18), and adults (N = 52) were found exclusively in streams. Reproduction, as identified by calling frogs, occurred from July through November. Frogs call all day long, but mostly at dusk, from rock crevices inside the stream edges near the splash zone. The call is short and loud, with 11 pulsed notes, of 491-641 ms, with a dominant frequency of 0.98-1.39 kHz. We describe the exotrophic and semiterrestrial tadpoles, always found in constantly humid vertical rock walls in the stream. Tadpoles of C. lutzorum are recognized by differences in labial tooth row formula, eye diameter, body shape, position of nares, and development of tail. Like congeneric species, the karyotype of C. lutzorum comprises 26 metacentric and submetacentric chromosomes. Cycloramphus lutzorum is restricted to and adapted for living in fast flowing streams, many of which are threatened by deforestation, pollution, and habitat loss. Therefore, we recommend the status of C. lutzorum be changed from its current "Data Deficient" to "Near Threatened (NT)" in the IUCN species red list.

A comparative study of the extracutaneous pigmentary system in three anuran amphibian species evaluated during the breeding season.

The present investigation examined the extracutaneous pigmentation pattern of three species of anuran amphibians, Dendropsophus nanus, Physalaemus cuvieri, and Rhinella schneideri, during the course of their breeding seasons. Pigmentation intensity in the different organs was graded on a 4-category scale, in which category 0 refers to organs without pigment and category 3 refers to intensely pigmented organs, with 2 intermediate stages of progressively stronger pigmentation. Rhinella schneideri showed testicular pigmentation, with intra-specific variation (categories 0 and 1). In P. cuvieri the pigmentation in the lungs varied in time: all the animals showed pigmentation (category 1) at the beginning of the breeding season, and as the season progressed the absence of pigments became the most common pattern. In the liver of the first animals collected, the pigment intensity was high (category 2) with many iridophores present, but in the last specimens collected no iridophores were found. The variation in D. nanus occurred in the kidneys, where animals collected at the beginning of the season did not show pigmentation. Renal veins displayed few melanocytes (category 1); animals collected at the end of the season showed more pigmentation in the kidneys (category 2), whereas in the renal veins the intensity remained the same. The changes observed in the extracutaneous pigment system in some organs, during the reproductive period, may be due to physiological alterations or may represent a species-specific characteristic.

The Archipelago of Fernando de Noronha: An Intriguing Malformed Toad Hotspot in South America

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)