Evidence of ecotypic differentiation between populations of the tree species Parapiptadenia rigida due to flooding

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Strain differentiation of Trichophyton rubrum by random amplification of polymorphic DNA (RAPD)

Trichophyton rubrum é um importante agente causal de dermatomicose. Os métodos de tipagem molecular têm sido recentemente desenvolvidos para responder questões sobre epidemiologia e auxiliar no esclarecimento de recidivas, após o tratamento. As seqüências aleatórias 1- (5'-d[GGTGCGGGAA]-3') e 6- (5'-d[CCCGTCAGCA]-3') foram usadas para tipagem molecular deste fungo por RAPD produzindo variabilidade intraespecífica. Cinco padrões foram observados entre os 10 isolados de T. rubrum, com ambas as seqüências. Foi concluído que a análise por RAPD pode ser utilizada para estudos epidemiológicos.

Origin and differentiation of a sex chromosome system in Parodon hilarii (Pisces, Parodontidae). Satellite DNA, G- and C-banding

A satellite DNA sequence of Parodon hilarii ( named pPh2004) was isolated, cloned and sequenced. This satellite DNA is composed of 200 bp, 60% AT rich. In situ hybridization ( FISH) results revealed that the satellite DNA pPh2004 is located in the terminal regions of several chromosomes, forming highly evident blocks in some and punctual marks in others. The comparison between the FISH and C-banding results showed that the location of this satellite DNA coincides with that of most terminal heterochromatins. However, some regions are only marked by FISH whereas other regions are only marked by C-banding. The possible existence of more than one satellite DNA family could explain these partial differences. The in situ hybridization with the satellite DNA and the G- and C-bandings confirmed the presence of a sex chromosome system of the ZZ/ZW type in P. hilarii, as well as the correct identification of the Z chromosome in the karyotype. This chromosome displays a segment of terminal heterochromatin in the long arm, similar to the segment observed in the short arm of the W chromosome, also showing a G- banding pattern similar to that of the short arm and part of the long arm of the W chromosome. A hypothesis on the origin of the W chromosome from an ancestral chromosome similar to the Z chromosome is presented.

Strain differentiation of Trichophyton rubrum by randomly amplified polymorphic DNA and analysis of rDNA nontranscribed spacer

Trichophyton rubrum is the most common pathogen causing dermatophytosis. Molecular strain-typing methods have recently been developed to tackle epidemiological questions and the problem of relapse following treatment. A total of 67 strains of T rubrum were screened for genetic variation by randomly amplified polymorphic DNA (RAPD) analysis, with two primers, 5'-d[GGTGCGGGAA]-3' and 5'-d[CCCGTCAGCA]-3', as well as by subrepeat element analysis of the nontranscribed spacer of rDNA, using the repetitive subelements TRS-1 and TRS-2. A total of 12 individual patterns were recognized with the first primer and 11 with the second. Phylogenetic analysis of the RAPID products showed a high degree of similarity (> 90 %) among the epidemiologically related clinical isolates, while the other strains possessed 60% similarity. Specific amplification of TRS-1 produced three strain-characteristic banding patterns (PCR types); simple patterns representing one copy of TRS-1 and two copies of TRS-2 accounted for around 85 % of all isolates. It is concluded that molecular analysis has important implications for epidemiological studies, and RAPID analysis is especially suitable for molecular typing in T. rubrum.

In situ application of a cellulose bag and an ion exchanger for differentiation of labile and inert metal species in aquatic systems

This work involved the development and application of a new analytical procedure for in-situ characterization of the lability of metal species in aquatic systems by using a system equipped with a diffusion membrane and cellulose organomodified with p-aminobenzoic acid groups (DM-Cell-PAB). To this end, the DM-Cell-PAB system was prepared by adding cellulose organomodified with p-aminobenzoic acid groups (Cell-PAB) to pre-purified cellulose bags. After the DM-Cell-PAB system was sealed, it was examined in the laboratory. The in-situ application involved immersing the DM-Cell-PAB system in two different rivers, enabling us to study the relative lability of metal species (Cu, Cd, Fe, Mn, and Ni) as a function of time and quantity of exchanger. The procedure is simple and opens up a new perspective for understanding environmental phenomena relating to the complexation, transport, stability, and lability of metal species in aquatic systems rich in organic matter.

High levels of genetic differentiation and selfing in the Brazilian cerrado fruit tree Dipteryx alata Vog. (Fabaceae)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Differentiation of epithelial cells in the urinary tract

Uroplakins, cytokeratins and the apical plasma membrane were studied in the epithelia of mouse urinary tract. In the simple epithelium covering the inner medulla of the renal pelvis, no uroplakins or cytokeratin 20 were detected and cells had microvilli on their apical surface. The epithelium covering the inner band of the outer medulla became pseudostratified, with the upper layer consisting of large cells with stalks connecting them to the basal lamina. Uroplakins and cytokeratin 20 were not expressed in these cells. However, some superficial cells appeared without connections to the basal lamina; these cells expressed uroplakins Ia, Ib, II and III and cytokeratin 20, they contained sparse small uroplakin-positive cytoplasmic vesicles and their apical surface showed both microvilli and ridges. Cytokeratin 20 was seen as dots in the cytoplasm. This epithelium therefore showed partial urothelial differentiation. The epithelium covering the outer band of the outer medulla gradually changed from a two-layered to a three-layered urothelium with typical umbrella cells that contained all four uroplakins. Cytokeratin 20 was organized into a complex network. The epithelium possessed an asymmetric unit membrane at the apical cell surface and fusiform vesicles. Umbrella cells were also observed in the ureter and urinary bladder. In males and females, the urothelium ended in the bladder neck and was continued by a non-keratinized stratified epithelium in the urethra in which no urothelial cell differentiation markers were detected. We thus show here the expression, distribution and organization of specific proteins associated with the various cell types in the urinary tract epithelium.

Structural and ultrastructural aspects of ontogenesis and differentiation of resin secretory cavities in Hymenaea stigonocarpa (Fabaceae-Caesalpinioideae) leaves

Cell lysis in the formation of secretory cavities in plants has been questioned by some authors and considered as result of technical artifacts. To describe the formation of secretory resin cavities in Hymenaea stigonocarpa leaves, leaflet samples at different stages of differentiation were collected, fixed, and processed for light and electron microscopy as per usual methods. The initial cells of secretory resin cavities are protodermal and grow towards the mesophyll ground meristem; these cells then divide producing cell groups that are distinguished by the shape and arrangement of cytoplasm, and density. At the initial stages of differentiation of the secretory cavities, some central cells in these groups show dark cytoplasm and condensed nuclear chromatin. Later, there is cell wall loosening, tonoplast and plasmalemma rupture resulting in cell death. These cells, however, maintain organelle integrity until lysis, when the cell wall degrades and the plasmalemma ruptures, releasing protoplast residues, marked characteristics of programmed cell death. The secretory epithelium remains active until complete leaf expansion when the cavity is filled with resin and the secretory activity ceases. There are no wall residues between central cells in adult cavities. Our results demonstrate lysigeny and the importance of ontogenetic studies in determining the origin of secretory cavities.

Cytogenetic characterization of six species of flatfishes with comments to karyotype differentiation patterns in Pleuronectiformes (Teleostei)

The karyotypes and cytogenetic characteristics of flatfishes species Paralichthys orbignyanus, Paralichthys patagonicus, Citarichthys spilopterus and Etropus crossotus (Paralichthyidae), Bothus ocellatus (Bothidae) and Symphurus tessellatus (Cynoglossidae) were investigated by conventional [Giemsa staining, C-banding, Ag- and chromomycin (CMA(3))-stainings] and molecular [in situ hybridization (ISH)] cytogenetic techniques. The results showed 2n = 46 and FN = 48 (2msm + 46sta) in P. orbignyanus, 2n = 46 and FN = 46 (46sta) in P. patagonicus, 2n = 26 and FN = 44 (18msm + 8sta) in C. spilopterus, 2n = 38 and FN = 64 (26msm + 12sta) in E. crossotus, 2n = 32 and FN = 50 (18msm + 14sta) in B. ocellatus, and 2n = 46 and FN = 62 (46msm + 62sta) in S. tessellatus. All species exhibited weak C-band positive segments in terminal and centromeric positions of some chromosome pairs. Silver staining of the nucleolus organizer regions (Ag-NOR) technique showed a single Ag-NOR-bearing chromosome pair in all species except E. crossotus. All these sites were CMA(3) positive and showed clear ISH signals after probing with a 18S rRNA probe. Etropus crossotus presented until seven chromosomes with Ag-NORs and CMA(3) positively stained segments in five chromosome pairs. Conversely only one chromosome pair was identified with the ISH experiments in this species. The available results show that the fishes of the order Pleuronectiformes experienced a marked chromosome evolution that included reduction in diploid number, mainly due to Robertsonian rearrangements, and several chromosome inversions. (c) 2007 the Authors Journal compilation (c) 2007 the Fisheries Society of the British Isles.

Allozyme differentiation of two populations of the genus Neoplecostomus Eigenmann & Eigenmann, 1888 (Teleostei, Loricariidae) from the upper Paraná River basin, Brazil

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

XX : XY sex chromosome system with X heterochromatinization: an early stage of sex chromosome differentiation in the Neotropic electric eel Eigenmannia virescens

An early stage of sex chromosome differentiation is reported to occur in the electric eel Eigenmannia virescens (Pisces, Sternopygidae) from populations of two tributaries of the Parana river system (Brazil). Cytogenetic studies carried out in the two populations showed that the Mogi-Guacu population is characterized by 2n = 38 chromosomes and undifferentiated sex chromosomes and the Tiete population presents 2n = 38 both for males and females and an XX:XY sex chromosome system. The X-chromosome is acrocentric, easily recognized by the presence of a conspicuous heterochromatin block in its distal portion; the Y-chromosome is probably one of the medium sized acrocentrics present in the male karyotype. BrdU induced R-bands of the two populations did not reveal any difference in the euchromatic regions of the chromosomes. AluI and HaeIII restriction enzyme digestion patterns and chromomycin A3 staining of the X-chromosome are presented. The possible role of heterochromatinization in the evolution of sex chromosomes in fish is discussed.

5S rDNA chromosomal mapping and COI sequence analysis reveal differentiation among distinct populations of a characid fish Serrapinnus notomelas

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