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.

On non-linear dynamics and a periodic control design applied to the potential of membrane action

The Fitzhugh-Nagumo (fn) mathematical model characterizes the action potential of the membrane. The dynamics of the Fitzhugh-Nagumo model have been extensively studied both with a view to their biological implications and as a test bed for numerical methods, which can be applied to more complex models. This paper deals with the dynamics in the (FH) model. Here, the dynamics are analyzed, qualitatively, through the stability diagrams to the action potential of the membrane. Furthermore, we also analyze quantitatively the problem through the evaluation of Floquet multipliers. Finally, the nonlinear periodic problem is controlled, based on the Chebyshev polynomial expansion, the Picard iterative method and on Lyapunov-Floquet transformation (L-F transformation).

A comparative analysis of the chemical nature of defensive secretions of Gonyleptidae (Arachnida : Opiliones : Laniatores)

The arachnids of the order Opiliones (harvestmen) produce substances used in defense. In the present paper, we analyzed 22 species of Gonyleptidae to explore the use of defensive substances in taxonomy and evolutionary biology. Thirty-seven different compounds were detected, 18 of which were preliminarily identified. These compounds were mapped onto a phylogenetic tree showing the relationships within the Gonyleptidae. Data from Cosmetidae were used as an outgroup. Five ketones and six alkyl phenols were reported for the first time in harvestmen. (c) 2005 Elsevier Ltd. All rights reserved.

Essential oil from leaves of Cryptocarya mandioccana Meisner (Lauraceae): Composition and intraspecific chemical variability

The composition of the essential oil from leaves of Cryptocarya mandioccana has been determined by chromatographic fractionation and GC-FID, GC-MS and C-13 NMR analyses, yielding the identification of 64 compounds with predominance of isomeric sesquiterpenes with molecular weights of 204. The main components of the oil obtained by hydrodistillation were beta-caryophyllene, spathulenol, caryophyllene oxide, delta-cadinene, germacrene D, benzaldehyde and bicyclogermacrene. However, the oil obtained by steam distillation contained higher levels of sesquiterpene hydrocarbons, with predominance of P-caryophyllene (C), germacrene D (G) and bicyclogermacrene (B), and was considered to be more representative of the composition of the oil in its natural state. The intraspecific chemical variability of the essential oil obtained by steam distillation was evaluated within populations of trees growing at three separate locations in the state of São Paulo, Brazil. Three distinct chemical groups could be characterised due to differences in the relative percentages of the three main sesquiterpenes from essential oil: CGB [relative contents of C (14-34%), G (5-28%), B (8-15%)], BCG [B (17-34%), C (9-24%), G (12-25%)] and GCB [G (22-42%), C (4-17%), B (7-15%)]. Individuals from groups CGB and BCG were found to be more frequent at south locations while group GCB is predominant in north location. (c) 2006 Elsevier Ltd. All rights reserved.

Chemical composition of flavonoids and styrylpyrones and the genetic variability of isozymes in natural populations of Cryptocarya mandioccana Meisner (Lauraceae)

The chemical composition of leaves of 57 trees of Cryptocarya mandioccana from three populations of southeastern Brazil was investigated through HPLC, assaying six flavonoids, seven styrylpyrones, and seven unidentified compounds. From 51 of the former trees, genotypes were obtained from 40 polymorphic loci of 19 isozymes. Cluster analyses of the phytochemical and genetical variation revealed that trees exhibited four chemotypes and five clusters from isozymes, respectively. Discriminant analyses from selected variables of the isozymic and chemical data sets were performed, respectively, in relation to the four chemotypes and the five isozyme clusters. The classification of individuals presented respective error estimates of 9.16% and 13.57%, indicating that the genetic data could explain the clusters from chernotypes and vice versa at acceptable error levels. Linear regressions with Dummy variable showed significant association of locus Skdh-2 with quercetin-3-O-beta-D-glucopyranoside and cryptofolione, indicating that its alleles would be responsible for the chemotype variation between individuals. (c) 2006 Elsevier Ltd. All rights reserved.

Self-fertility and polyembryony in South American yellow trumpet trees (Handroanthus chrysotrichus and H. ochraceus, Bignoniaceae): a histological study of postpollination events

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

Synthesis, X-ray Crystal Structure and Theoretical Calculations of Antileishmanial Neolignan Analogues

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Comparative study of ovule and fruit development in species of Hypolytrum and Rhynchospora (Cyperaceae, Poales)

The development of the ovule and of the fruit of Hypolytrum bullatum and H. schraderianum (Mapanioideae) and of Rhynchospora consanguinea and R. rugosa (Cyperoideae) are described. All species share anatropous, bitegmic and crassinucellate ovules, funicular obturator, megagametophyte of the Polygonum type, presence of starch grains in the mature megagametophyte, free-nuclear endosperm, Onagrad-type embryogeny, testal-tegmic seed, and a simple fruit of the achene type. Rhynchospora species have characters typical of the family: micropyle formed by the inner integument alone; 3-4-layered parietal tissue; and hard achene. Hypolytrum species differ in those characters by presenting a slightly zigzag micropyle formed by both integuments connected with the funicular obturator, 5-8-layered parietal tissue, and fibrous-spongy achene. The peculiar formation of the micropyle in Hypolytrum is a feature reported here for the first time in the family. The ontogeny provides evidence for a better understanding of the dispersal unit in Hypolytrum supporting the classification as a true achene, like that of Rhynchospora, which is characteristic of the family.

Biodiversity, Species Interactions and Ecological Networks in a Fragmented World

Biodiversity is organised into complex ecological networks of interacting species in local ecosystems, but our knowledge about the effects of habitat fragmentation on such systems remains limited. We consider the effects of this key driver of both local and global change on both mutualistic and antagonistic systems at different levels of biological organisation and spatiotemporal scales.There is a complex interplay of patterns and processes related to the variation and influence of spatial, temporal and biotic drivers in ecological networks. Species traits (e.g. body size, dispersal ability) play an important role in determining how networks respond to fragment size and isolation, edge shape and permeability, and the quality of the surrounding landscape matrix. Furthermore, the perception of spatial scale (e.g. environmental grain) and temporal effects (time lags, extinction debts) can differ markedly among species, network modules and trophic levels, highlighting the need to develop a more integrated perspective that considers not just nodes, but the structural role and strength of species interactions (e.g. as hubs, spatial couplers and determinants of connectance, nestedness and modularity) in response to habitat fragmentation.Many challenges remain for improving our understanding: the likely importance of specialisation, functional redundancy and trait matching has been largely overlooked. The potentially critical effects of apex consumers, abundant species and supergeneralists on network changes and evolutionary dynamics also need to be addressed in future research. Ultimately, spatial and ecological networks need to be combined to explore the effects of dispersal, colonisation, extinction and habitat fragmentation on network structure and coevolutionary dynamics. Finally, we need to embed network approaches more explicitly within applied ecology in general, because they offer great potential for improving on the current species-based or habitat-centric approaches to our management and conservation of biodiversity in the face of environmental change.

Chromosomal evolution in Gekkonidae. I. Chromosome painting between Gekko and Hemidactylus species reveals phylogenetic relationships within the group

Geckos are a large group of lizards characterized by a rich variety of species, different modes of sex determination and diverse karyotypes. In spite of many unresolved questions on lizards' phylogeny and taxonomy, the karyotypes of most geckos have been studied by conventional cytogenetic methods only. We used flow-sorted chromosome-specific painting probes of Japanese gecko (Gekko japonicus), Mediterranean house gecko (Hemidactylus turcicus) and flat-tailed house gecko (Hemidactylus platyurus) to reveal homologous regions and to study karyotype evolution in seven gecko species (Gekko gecko, G. japonicus, G. ulikovskii, G. vittatus, Hemidactylus frenatus, H. platyurus and H. turcicus). Generally, the karyotypes of geckos were found to be conserved, but we revealed some characteristic rearrangements including both fissions and fusions in Hemidactylus. The karyotype of H. platyurus contained a heteromorphic pair in all female individuals, where one of the homologues had a terminal DAPI-negative and C-positive heterochromatic block that might indicate a putative sex chromosome. Among two male individuals studied, only one carried such a polymorphism, and the second one had none, suggesting a possible ZZ/ZW sex determination in some populations of this species. We found that all Gekko species have retained the putative ancestral karyotype, whilst the fission of the largest ancestral chromosome occurred in the ancestor of modern Hemidactylus species. Three common fissions occurred in the ancestor of Mediterranean house and flat-tailed house geckos, suggesting their sister group relationships. PCR-assisted mapping on flow-sorted chromosome libraries with conserved DMRT1 gene primers in G. japonicus indicates the localization of DMRT1 gene on chromosome 6.

Differentiation of floral color and odor in two fly pollinated species of Metrodorea (Rutaceae) from Brazil

We investigated if differences in morphological characters in two species of Metrodorea (Rutaceae) from Brazilian semideciduous forests correspond to some pollination divergence. M. nigra and M. stipularis are sympatric species, display a similar floral morphology, are protandrous, self-incompatible, their flower periods overlap, and both are pollinated by flies. M. nigra main pollinators are Pseudoptiloleps nigripoda (Muscidae) and Fannia sp. (Fanniidae); M. stipularis major pollinators are Phaenicia eximia (Calliphoridae), Palpada sp. and Ornidia obesa (Syrphidae). The distinct floral odor (disagreeable in M. nigra and sweet in M. stipularis) and color (brownish violet vs. pale yellow) determine the differences on type and number of floral visitors observed. Several species from semideciduous forests initially considered to be pollinated by diverse insects, present flies as main pollinators, stressing the importance of fly pollination in such habitats.

Chromosomal similarities and differences among four Neotropical Elateridae (Conoderini and Pyrophorini) and other related species, with comments on the NOR patterns in Coleoptera

This work deals with the comparative cytogenetic analysis of four Neotropical Elateridae species and reviews the nucleolar organizer region (NOR) patterns on Coleoptera chromosomes, for the first time. The cytogenetic characterization of Conoderus malleatus (Conoderini), Pyrearinus candelarius, Pyrophorus divergens and Pyrophorus punctatissimus (Pyrophorini) was accomplished through the study of mitotic and meiotic cells submitted to standard (Giemsa) and differential staining [silver impregnation and GC-specific chromomycin A 3 (CMA 3) plus AT-specific 4′-6-diamidino-2-phenylindole (DAPI) fluorochromes]. The analysis of spermatogonial cells revealed the diploid numbers: 2n = 17 in C. malleatus and 2n = 15 in P. candelarius, P. divergens and P. punctatissimus. In these species, the X0 type sex-determination system and the acrocentric morphology of almost all chromosomes were observed. The study of meiotic cells of the four species revealed the occurrence of total synapsis between the autosomes, the presence of one terminal or interstitial chiasma in the majority of the bivalents, and the reductional behaviour and regular segregation of all chromosomes. Although the three Pyrophorini species demonstrated many similar karyotypical characteristics, there was one discrepancy, which was noted in the diplotene cells and concerns the number of bivalents with two chiasmata; P. candelarius only presented one bivalent, P. divergens showed two bivalents and P. punctatissimus exhibited up to four bivalents with two chiasmata. Testicular cells impregnated with silver nitrate demonstrated two terminal NORs located on the fourth autosomal pair of the Conoderini species and on the second autosomal pair of the three Pyrophorini representatives. Use of CMA 3/distamycin A (DA)/DAPI staining on the P. candelarius and P. punctatissimus chromosomes revealed that the CMA 3 labelled regions were coincident with the NORs. The main strategies of karyotypical differentiation that have occurred among the four Elateridae species and other related species, and the general trends of the NOR shifts during Coleoptera chromosomal evolution are discussed in this work. © 2007 The Authors.