Allozyme variability in an invasive drosophilid,Zaprionus indianus (Diptera: Drosophilidae): comparison of a recently introduced Brazilian population with Old World populations

Colonizing species often go through genetic bottlenecks when new territories are invaded. The South American continent has been recently colonized by a generalist African drosophilid, Zaprionus indianus, which has become an agricultural pest in Brazil in the last five years. In this paper we used allozyme electrophoresis to estimate levels of genetic differentiation of Z. indianus collected from sites 4 300 km apart in Brazil. We also compared the level of polymorphism of the Brazilian populations with that found in laboratory strains from Africa and Asia, to verify if a significant decrease in gene variability has taken place during the invasion process. The populations were polymorphic for three out of the 11 loci investigated. Genetic distances and F_ST indices among Brazilian populations were small and generally non significant, suggesting a colonization from one single propagule followed by a rapid demographic expansion. Ancestral and old populations from Africa and Asia were slightly more heterozygous than those from Brazil. Compared to other drosophilids, Z. indianus appears to be characterized by a low proportion (25%) of polymorphic loci. We suggest that the propagule introduced to Brazil had a sufficient size to carry almost all the polymorphism from the (unknown) origin population, although not the precise allelic frequencies.     

Inflammation in disseminated lesions: an analysis of CD4+, CD20+, CD68+, CD31+ and vW+ cells in non-ulcerated lesions of disseminated leishmaniasis

Disseminated leishmaniasis (DL) differs from other clinical forms of the disease due to the presence of many non-ulcerated lesions (papules and nodules) in non-contiguous areas of the body. We describe the histopathology of DL non-ulcerated lesions and the presence of CD4-, CD20-, CD68-, CD31- and von Willebrand factor (vW)-positive cells in the inflamed area. We analysed eighteen biopsies from non-ulcerated lesions and quantified the inflamed areas and the expression of CD4, CD20, CD68, CD31 and vW using Image-Pro software (Media Cybernetics). Diffuse lymphoplasmacytic perivascular infiltrates were found in dermal skin. Inflammation was observed in 3-73% of the total biopsy area and showed a significant linear correlation with the number of vW⁺ vessels. The most common cells were CD68⁺ macrophages, CD20⁺ B-cells and CD4⁺ T-cells. A significant linear correlation between CD4⁺ and CD20⁺ cells and the size of the inflamed area was also found. Our findings show chronic inflammation in all DL non-ulcerated lesions predominantly formed by macrophages, plasmacytes and T and B-cells. As the inflamed area expanded, the number of granulomas and extent of the vascular framework increased. Thus, we demonstrate that vessels may have an important role in the clinical evolution of DL lesions.     

Floral polymorphism in Chamaecrista flexuosa (Fabaceae-Caesalpinioideae): a possible case of atypical enantiostyly?

Background and Aims

Reciprocal herkogamy, including enantiostyly and heterostyly, involves reciprocity in the relative positions of the sexual elements within the flower. Such systems result in morphologically and, since pollen is deposited on and captured from different parts of the pollinator, functionally distinct floral forms. Deviations from the basic pattern may modify the functionality of these mechanisms. For heterostylous species, such deviations are generally related to environmental disturbances, pollination services and/or reduced numbers of one floral morph. Deviations for enantiostylous species have not yet been reported. This study aims to investigate enantiostyly in Chamaecrista flexuosa, in particular the presence of deviations from the standard form, in an area of coastal vegetation in north-east Brazil.

Methods

Observations and investigations of floral biology, the reproductive system, pollinator behaviour, floral morphology and morphometry were performed.

Key Results

In C. flexuosa flowers, anthers of different size but similar function are grouped. The flowers were self-compatible and set fruits after every treatment, except in the spontaneous self-pollination experiment, thereby indicating their dependence on pollen vectors. The flowers were pollinated by bees, especially Xylocopa cearensis and X. grisencens. Pollen is deposited and captured from the ventral portion of the pollinator''s body. Variations in the spatial arrangement of floral elements allowed for the identification of floral morphs based on both morphological and functional criteria. Using morphological criteria, morphologically right (MR) and morphologically left (ML) floral morphs were identified. Three floral morphs were identified using functional criteria: functionally right (FR), functionally central (FC) and functionally left (FL). Combinations of morphologically and functionally defined morphs did not occur in equal proportions. There was a reduced frequency of the MR–FR combination.

Conclusions

The results indicate the occurrence of an atypical enantiostyly in C. flexuosa. This seems to improve reproductive success by increasing the efficiency of pollen deposition and capture.     

Structure of p53 binding to the BAX response element reveals DNA unwinding and compression to accommodate base-pair insertion

The p53 core domain binds to response elements (REs) that contain two continuous half-sites as a cooperative tetramer, but how p53 recognizes discontinuous REs is not well understood. Here we describe the crystal structure of the p53 core domain bound to a naturally occurring RE located at the promoter of the Bcl-2-associated X protein (BAX) gene, which contains a one base-pair insertion between the two half-sites. Surprisingly, p53 forms a tetramer on the BAX-RE that is nearly identical to what has been reported on other REs with a 0-bp spacer. Each p53 dimer of the tetramer binds in register to a half-site and maintains the same protein–DNA interactions as previously observed, and the two dimers retain all the protein–protein contacts without undergoing rotation or translation. To accommodate the additional base pair, the DNA is deformed and partially disordered around the spacer region, resulting in an apparent unwinding and compression, such that the interactions between the dimers are maintained. Furthermore, DNA deformation within the p53-bound BAX-RE is confirmed in solution by site-directed spin labeling measurements. Our results provide a structural insight into the mechanism by which p53 binds to discontinuous sites with one base-pair spacer.     

103 Probing DNA shape and methylation state on a genomic scale with DNase I

DNA binding proteins find their cognate sequences within genomic DNA through recognition of specific chemical and structural features. Here, we demonstrate that high-resolution DNase I cleavage profiles can provide detailed information about the shape and chemical modification status of genomic DNA. Analyzing millions of DNA-backbone hydrolysis events on naked genomic DNA, we show that the intrinsic rate of cleavage by DNase I closely tracks the width of the minor groove. Integration of these DNase I cleavage data with bisulfite sequencing data for the same cell type genome reveals that the cleavage directly adjacent to CpG dinucleotides is enhanced at least eight-fold by cytosine methylation. This phenomenon we show is attributable to methylation-induced narrowing of the minor groove. Furthermore, we demonstrate that it enables simultaneous mapping of DNase I hypersensitivity and regional DNA methylation levels using dense in vivo cleavage data. Taken together, our results suggest a general mechanism through which CpG methylation can modulate protein–DNA interaction strength via the remodeling of DNA shape.     

Semichronic Inhibition of Glutathione Reductase Promotes Oxidative Damage to Proteins and Induces both Transcription and Translation of Tyrosine Hydroxylase in the Nigrostriatal System

We have evaluated the effect of N,N-bis (2-chloroethyl)-N-nitrosourea (BCNU), an inhibitor of glutathione reductase (GR), on the oxidative status along with the integrity of the nigrostriatal dopaminergic system of the rat. The oxidative status was studied by the quantification of carbonyl groups coupled to protein homogenates. Moreover, the specific oxidations in glial fibrillary acidic protein (GFAP) and neurofilament-200 (NF-200) were also measured. The results show that oxidative damage in proteins in the nigrostriatal system is confined to the striatum. Specific carbonyl groups coupled to native NF-200 and GFAP were also increased. These changes were accompanied by reactive astrocytosis in striatum but not in substantia nigra. In substantia nigra, decreased levels of dopamine (DA) and 3,4-dihydroxyphenylacetic acid (DOPAC) were observed following BCNU treatment. In contrast, DA levels were increased in the striatum along with an overall decrease in the ratios of DA metabolites to DA. We also studied the mRNA levels for tyrosine hydroxylase (TH) and the dopamine transporter (DAT) by in situ hybridization. TH mRNA but not DAT mRNA was significantly induced in substantia nigra following BCNU treatment, which was consistent with significant elevations in TH enzyme amount and activity and unchanged DA uptake in striatum. All these results support the DA free radical hypothesis and the key role of the striatal glutathione system in protecting the striatal system against oxidative stress.     

Esterase Polymorphism and the Analysis of Genetic Diversity and Structure in Cactus Populations Descended from Cereus peruvianus Plants Regenerated In Vitro

The genetic structure of Cereus peruvianus populations descended from cultivated plants (F(1) populations) and from plants regenerated in vitro (R(1) populations) was analyzed using α- and β-esterase isozymes in native PAGE. The estimated proportion of polymorphic loci was higher (50%) in the R(1) populations than the F(1) populations (42.85%). The mean observed (0.5599) and expected (0.5620) heterozygosity in R(1) descendents was also higher than the rates in F(1) descendents (H (o)?=?0.4142; H (e)?=?0.4977). A low level of population differentiation was detected in R(1) descendents (F (st)?=?0.05). In contrast, population differentiation was high in F(1) descendents (0.2583). Esterase analysis using PAGE showed that artificial selection by silvicultural management provides high genetic diversity and a large genetic basis for C.?peruvianus, whereas in vitro selection from callus tissue culture involves an increase of heterozygosity levels in descendents from somaclones and a low level of interpopulational divergence.