Differentially expressed genes in giant cell tumor of bone

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

Renomalization of the NN interaction at NNLO: Uncoupled peripheral waves

We apply the subtractive renormalization method to the nucleon-nucleon interaction at Next-to-Next-to-Leading order (NNLO). Here we show the results for some uncoupled peripheral waves.

Renormalizing the NN interaction with multiple subtractions

The aim of this work is to show how to renormalize the nucleon-nucleon interaction at next-to-next-to-leading order using a. systematic subtractive renormalization approach with multiple subtractions. As an example, we calculate the phase shifts for the partial waves with total angular momentum J = 2. The intermediate driving terms at each recursive step as well as the renormalized T-matrix are also shown. We conclude that our method is reliable for singular potentials such as the two-pion exchange and derivative contact interactions.

Electrochemical genosensors for the detection of Bonamia parasite. Selection of single strand-DNA (ssDNA) probes by simulation of the secondary structure folding

A post-PCR nucleic acid work by comparing experimental data, from electrochemical genosensors, and bioinformatics data, derived from the simulation of the secondary structure folding and prediction of hybridisation reaction, was carried out in order to rationalize the selection of ssDNA probes for the detection of two Bonamia species, B. exitiosa and B. ostreae, parasites of Ostrea edulis.Six ssDNA probes (from 11 to 25 bases in length, 2 thiolated and 4 biotinylated) were selected within different regions of B. ostreae and B. exitiosa PCR amplicons (300 and 304 bases, respectively) with the aim to discriminate between these parasite species. ssDNA amplicons and probes were analyzed separately using the "Mfold Web Server" simulating the secondary structure folding behaviour. The hybridisation of amplicon-probe was predicted by means of "Dinamelt Web Server". The results were evaluated considering the number of hydrogen bonds broken and formed in the simulated folding and hybridisation process, variance in gaps for each sequence and number of available bases. In the experimental part, thermally denatured PCR products were captured at the sensor interface via sandwich hybridisation with surface-tethered probes (thiolated probes) and biotinylated signalling probes. A convergence between analytical signals and simulated results was observed, indicating the possibility to use bioinformatic data for ssDNA probes selection to be incorporated in genosensors. (C) 2011 Elsevier B.V. All rights reserved.

SEX-CHROMOSOME POLYMORPHISM AND HETEROGAMETIC MALES REVEALED BY 2 CLONED DNA PROBES IN THE ZW ZZ FISH LEPORINUS-ELONGATUS

In order to study the divergence of teleost sex chromosomes, subtractive cloning was carried out between genomic DNA of males and females of the rainbow trout (XX/XY) and of Leporinus elongatus (ZW/ZZ). Inserts cloned in a plasmid vector were individually tested on Southern blots of DNA of males and females for sex specificity. No sex-specific insert was obtained from trout, but two out of ten inserts cloned from L. elongatus showed sex-specific patterns in this species: one corresponds to a sequence present on both Z and W chromosomes, while the other is W specific. Sequences of these two inserts show neither clear homology with other known sequences, nor an open reading frame. They cross-hybridize with the genomic DNA of Leporinus friderici, but without sex-specific patterns. Twenty-four L. elongatus adults were sexed by gonadal observation, chromosomed examination and Southern hybridization with one or the other insert. Ten males and 11 females had chromosomes and hybridization patterns typical of their sex. One ZW female was recognized as a male with the W-specific probe. This was also the case for two unusual ZW males, one having a male hybridization pattern with the other probe. These three atypical individuals may result from single genetic exchanges between four regions of the Z and the W, giving rise to three atypical W chromosomes. Finding males with such atypical heterochromosomes in a female heterogametic species may indicate that a gradual transition occurs between the heterogametic systems.

Surface modification of Ti dental implants by Nd: YVO4 laser irradiation

Surface modifications have been applied in endosteal bone devices in order to improve the osseointegration through direct contact between neoformed bone and the implant without an intervening soft tissue layer. Surface characteristics of titanium implants have been modified by addictive methods, such as metallic titanium, titanium oxide and hydroxyapatite powder plasma spray, as well as by subtractive methods, such as acid etching, acid etching associated with sandblasting by either AlO2 or TiO2, and recently by laser ablation. Surface modification for dental and medical implants can be obtained by using laser irradiation technique where its parameters like repetition rate, pulse energy, scanning speed and fluency must be taken into accounting to the appropriate surface topography. Surfaces of commercially pure Ti (cpTi) were modified by laser Nd:YVO4 in nine different parameters configurations, all under normal atmosphere. The samples were characterized by SEM and XRD refined by Rietveld method. The crystalline phases alpha Ti, beta Ti, Ti6O, Ti3O and TiO were formed by the melting and fast cooling processes during irradiation. The resulting phases on the irradiated surface were correlated with the laser beam parameters: the aim of the present work was to control titanium oxides formations in order to improve implants osseointegration by using a laser irradiation technique which is of great importance to biomaterial devices due to being a clean and reproducible process. (c) 2007 Elsevier B.V. All rights reserved.

A novel technique to produce polygenic resistance to anthracnose in Stylosanthes capitata

An improved cultivar, based on 17 genotypes of S. capitata and six of S.macrocephala, was developed at the wEmbrapa Beef Cattle Research Center, Campo Grande, Brazil. The aim was to create durable, race non-specific resistance to anthracnose controlled by polygenic factors. A mass hybridisation technique was employed to produce a high degree of genetic diversity and sizeable quantities of seed of hybrid-derived progenies of Brazilian and Venezuelan genotypes of S. capitata. Outcrossing resulted in a significant improvement in the forage production of progeny of Venezuelan accessions. The multicross was evaluated in multilocational trials, each representing a large tract of country in the Cerrados ecosystem along a north-south transect from lat.6degrees S to 20degrees S. The genetic shift that occurred in S. capitata was a key element in the formation of the new cultivar. It is a complex mixture of two species, and a recombination of much desired forage traits of Brazilian x Venezuelan genotypes, high forage and seed yields coupled with anthracnose resistance. The new cultivar with its diverse genetic make-up has a wide application in the acid-soil savannas of tropical America. It was released by Embrapa for the Cerrados in 2000.

Cytogenetical and cytotaxonomical analysis of some Brazilian species of Eleocharis (Cyperaceae)

Karyotype analysis of 21 samples of 11 species of Eleocharis ( Cyperaceae) from 10 localities in Brazil, showed the presence of chromosomes without primary constrictions and parallel movement of chromatids at metaphase-anaphase transition. Only the terminal nucleolar constrictions ( satellites) were visualised. The chromosome numbers varied from 2n=6 in E. subarticulata to 2n=54 in E. acutangula, but the chromosome basic number x=5 was confirmed. Generally, C-CMA(3)(+) bands appear mostly in the extremities of the chromosomes, associated to NOR, and interstitial C-CMA(3) bands were found only in E. geniculata and E. acutangula. C-DAPI(+) bands were not found. Fluorescence in situ hybridisation ( FISH) with the 45S rDNA probe was performed in five species. The results showed from four to eight hybridisation signals, always terminal. The analysed species include representatives of the following three subgenera of Eleocharis that occur in Brazil: Limnochloa, Scirpidium and Eleocharis. Species from the subgenus Limnochloa have small and numerous chromosomes. The remaining species, belonging to subgenera Eleocharis and Scirpidium, possess fewer and larger chromosomes. In subgenus Eleocharis, karyotypes of the section Eleocharis were differentiated by symploidy, agmatoploidy and polyploidy, whereas species of the section Eleogenus were all polyploids. Polyploidy seems to be the most frequent event in the karyotype differentiation in Eleocharis, but changes in the chromosome size and repetitive DNA sites were also observed.

Gene expression in placentation of farm animals: An overview of gene function during development

Eutherian mammals share a common ancestor that evolved into two main placental types, i.e., hemotrophic (e.g., human and mouse) and histiotrophic (e.g., farm animals), which differ in invasiveness. Pregnancies initiated with assisted reproductive techniques (ART) in farm animals are at increased risk of failure; these losses were associated with placental defects, perhaps due to altered gene expression. Developmentally regulated genes in the placenta seem highly phylogenetically conserved, whereas those expressed later in pregnancy are more species-specific. To elucidate differences between hemotrophic and epitheliochorial placentae, gene expression data were compiled from microarray studies of bovine placental tissues at various stages of pregnancy. Moreover, an in silico subtractive library was constructed based on homology of bovine genes to the database of zebrafish - a nonplacental vertebrate. In addition, the list of placental preferentially expressed genes for the human and mouse were collected using bioinformatics tools (Tissue-specific Gene Expression and Regulation [TiGER] - for humans, and tissue-specific genes database (TiSGeD) - for mice and humans). Humans, mice, and cattle shared 93 genes expressed in their placentae. Most of these were related to immune function (based on analysis of gene ontology). Cattle and women shared expression of 23 genes, mostly related to hormonal activity, whereas mice and women shared 16 genes (primarily sexual differentiation and glycoprotein biology). Because the number of genes expressed by the placentae of both cattle and mice were similar (based on cluster analysis), we concluded that both cattle and mice were suitable models to study the biology of the human placenta. (C) 2011 Elsevier B.V. All rights reserved.

Chromosome mapping of repetitive sequences in Anostomidae species: Implications for genomic and sex chromosome evolution

Background: Members of the Anostomidae family provide an interesting model system for the study of the influence of repetitive elements on genome composition, mainly because they possess numerous heterochromatic segments and a peculiar system of female heterogamety that is restricted to a few species of the Leporinus genus. The aim of this study was to isolate and identify important new repetitive DNA elements in Anostomidae through restriction enzyme digestion, followed by cloning, characterisation and chromosome mapping of this fragment. To identify repetitive elements in other Leporinus species and expand on studies of repetitive elements in Anostomidae, hybridisation experiments were also performed using previously described probes of LeSpeI repetitive elements. Results: The 628-base pair (bp) LeSpeII fragment was hybridised to metaphase cells of L. elongatus individuals as well as those of L. macrocephalus, L. obtusidens, L. striatus, L. lacustris, L. friderici, Schizodon borellii and S. isognathus. In L. elongatus, both male and female cells contained small clusters of LeSpeII repetitive elements dispersed on all of the chromosomes, with enrichment near most of the terminal portions of the chromosomes. In the female sex chromosomes of L. elongatus (Z2,Z2/W1W 2), however, this repeated element was absent. In the remaining species, a dispersed pattern of hybridisation was observed on all chromosomes irrespective of whether or not they were sex chromosomes. The repetitive element LeSpeI produced positive hybridisations signals only in L. elongatus, L. macrocephalus and L. obtusidens, i.e., species with differentiated sex chromosomes. In the remaining species, the LeSpeI element did not produce hybridisation signals. Conclusions: Results are discussed in terms of the effects of repetitive sequences on the differentiation of the Anostomidae genome, especially with respect to sex chromosome evolution. LeSpeII showed hybridisation patterns typical of Long Interspersed Elements (LINEs). The differential distribution of this element may be linked to sex chromosome differentiation in L. elongatus species. The relationship between sex chromosome specificity and the LeSpeI element is confirmed in the species L. elongatus, L. macrocephalus and L. obtusidens. © 2012 da Silva et al.; licensee BioMed Central Ltd.

Cytogenetic analysis in Thoracocharax stellatus (Kner, 1858) (Characiformes, Gasteropelecidae) from Paraguay river basin, Mato Grosso, Brazil

Thoracocharax stellatus (Characiformes, Gasteropelecidae) is a small Neotropical species of fish, widely distributed in several rivers of South America. Evidence for karyotype heteromorphysm in populations from different geographical regions has been reported for this species. In this way, populations of T. stellatus from the Paraguay River basin were cytogenetically characterized and the results were compared with other studies performed in the same species but from different basins. The results showed a diploid number of 2n = 54 for T. stellatus, with chromosomes arranged in 6 metacentric (m), 6 submetacentric (sm), 2 subtelocentric (st) and 40 acrocentric (a), for both sexes, with a simple Nucleolus Organiser Region (NOR) system reported by the techniques of silver nitrate impregnation and fluorescent in situ hybridisation (FISH) using 18S rDNA sequences as probe. The distribution of constitutive heterochromatin, observed by the C-band technique and Chromomycin A3 staining showed great similarity among the analyzed populations and consists mainly of discrete blocks in the pericentromeric and telomeric regions of most chromosomes. The presence of female heterogamety was alsoobserved indicating a ZZ/ZW system with W chromosome almost totally heterochromatic. The results also show cytogenetic diversity of the group and are useful to understand the mechanisms of karyotype evolution of the family. © Edson Lourenço da Silva et al.

Crustal growth in the 3.4-2.7Ga São José de Campestre Massif, Borborema Province, NE Brazil

The Archean (3.45-2.70Ga) rocks of the São José do Campestre Massif (SJCM) in the Borborema Province (NE Brazil) make up a small area (~6000km2) and are composed of granitoids and metasupracrustal rocks that define a complex magmatic and deformational history. The massif provides the opportunity to study mantle- and crustal-derived magmas generated since the Palaeoarchean. The orthogneisses of the SJCM are composed of: (1) tonalite to granodiorite with diorite enclaves (Bom Jesus gneiss, 3412±8Ma; TDM Nd model ages from 4.1 to 3.5Ga and negative epsilon Nd values); (2) biotite and ferroan-diopside monzogranite (Presidente Juscelino complex, 3356±21Ma and 3251±44Ma; TDM model ages range from 4.1 to 3.4Ga and epsilon Nd values that are slightly positive to negative); (3) hornblende tonalite to granodiorite (Brejinho complex, 3333±77Ma and 3187±8Ma; dominantly positive epsilon Nd values and TDM ages from 3.6 to 3.2Ga); (4) biotite monzogranite (São Pedro do Potengi gneiss, 3120±22Ma; TDM =3.5Ga; negative epsilon Nd value); (5) ferroan-diopside-grossular anorthosite and metagabbro (Senador Elói de Souza complex, 3033±3Ma); and (6) quartz diorite to syenogranite (São José do Campestre complex; 2685±9Ma and 2655±4Ma; negative epsilon Nd values and TDM ages from 3.9 to 3.3Ga). The orthogneisses are subalkaline to faintly alkaline, magnesian to ferroan, M- and I-type granitoids that follow either the K-enrichment or the trondhjemite trends. Each group has a subset with REE characteristics similar to Archean TTG and another that is analogous to Phanerozoic granitoids. They have negative Ta-Nb and Ti anomalies and have trace element contents of granitoids from subduction zones. Geochemical and Nd isotope data suggest that subducted oceanic crust and a depleted and metasomatised mantle wedge both acted as the magma sources. We propose a convergent tectonic model in which hybridisation of the upper mantle occurs through interactions with adakitic or trondhjemitic melts and recycling of earlier crust. The results imply that both the subducted oceanic crust and the mantle wedge played major roles in continent formation throughout successive episodes of arc accretion in Palaeo- and Mesoarchean times. The Archean rocks of the SJCM shares some similarities with the Pilbara, Kaapvaal, West African, and São Francisco cratons. However, the most reliable comparisons with the SJCM are with the neighbouring basement of the Nigeria and Cameroon shields. © 2012 Elsevier B.V.

Cytogenetic analysis of Phyllomedusa distincta Lutz, 1950 (2n = 2x = 26), P. tetraploidea Pombal and Haddad, 1992 (2n = 4x = 52), and their natural triploid hybrids (2n = 3x = 39) (Anura, Hylidae, Phyllomedusinae)

Background: Natural polyploidy has played an important role during the speciation and evolution of vertebrates, including anurans, with more than 55 described cases. The species of the Phyllomedusa burmeisteri group are mostly characterized by having 26 chromosomes, but a karyotype with 52 chromosomes was described in P. tetraploidea. This species was found in sintopy with P. distincta in two localities of São Paulo State (Brazil), where triploid animals also occur, as consequence of natural hybridisation. We analyse the chromosomes of P. distincta, P. tetraploidea, and their triploid hybrids, to enlighten the origin of polyploidy and to obtain some evidence on diploidisation of tetraploid karyotype.Results: Phyllomedusa distincta was 2n = 2x = 26, whereas P. tetraploidea was 2n = 4x = 52, and the hybrid individuals was 2n = 3x = 39. In meiotic phases, bivalents were observed in the diploid males, whereas both bivalents and tetravalents were observed in the tetraploid males. Univalents, bivalents or trivalents; metaphase II cells carrying variable number of chromosomes; and spermatids were detected in the testis preparations of the triploid males, indicating that the triploids were not completely sterile. In natural and experimental conditions, the triploids cross with the parental species, producing abnormal egg clutches and tadpoles with malformations. The embryos and tadpoles exhibited intraindividual karyotype variability and all of the metaphases contained abnormal constitutions. Multiple NORs, detected by Ag-impregnation and FISH with an rDNA probe, were observed on chromosome 1 in the three karyotypic forms; and, additionally, on chromosome 9 in the diploids, mostly on chromosome 8 in the tetraploids, and on both chromosome 8 and 9 in the triploids. Nevertheless, NOR-bearing chromosome 9 was detected in the tetraploids, and chromosome 9 carried active or inactive NORs in the triploids. C-banding, base-specific fluorochrome stainings with CMA3 and DAPI, FISH with a telomeric probe, and BrdU incorporation in DNA showed nearly equivalent patterns in the karyotypes of P. distincta, P. tetraploidea, and the triploid hybrids.Conclusions: All the used cytogenetic techniques have provided strong evidence that the process of diploidisation, an essential step for stabilising the selective advantages produced by polyploidisation, is under way in distinct quartets of the tetraploid karyotype. © 2013 Gruber et al.; licensee BioMed Central Ltd.

Identificação e caracterização de possiveis marcadores moleculares em carcinoma renal de células claras

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Expressão gênica diferencial em resposta aos efeitos da proteína anti-inflamatória Anexina A1 nas células epiteliais pigmentadas da retina humana

Pós-graduação em Genética - IBILCE