Use of lymphocyte cultures for BrdU replication banding patterns in anuran species (Amphibia)

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

Chromosome banding in three species of Hypsiboas (Hylidae, Hylinae), with special reference to a new case of B-chromosome in anuran frogs and to the reduction of the diploid number of 2n=24 to 2n=22 in the genus

The chromosomes of hylids Hypsiboas albopunctatus, H. raniceps, and H. crepitans from Brazil were analyzed with standard and differential staining techniques. The former species presented 2n = 22 and 2n = 23 karyotypes, the odd diploid number is due to the presence of an extra element interpreted as B chromosome. Although morphologically very similar to the small-sized chromosomes of the A complement, the B was promptly recognized, even under standard staining, on the basis of some characteristics that are usually attributed to this particular class of chromosomes. The two other species have 2n = 24, which is the chromosome number usually found in the species of Hypsiboas karyotyped so far. This means that 2n = 22 is a deviant diploid number, resulted from a structural rearrangement, altering the chromosome number of 2n = 24 to 2n = 22. Based on new chromosome data, some possibilities were evaluated for the origin of B chromosome in Hypsiboas albopunctatus, as well as the karyotypic evolution in the genus, leading to the reduction in the diploid number of 2n = 24 to 2n = 22.

Comparative cytogenetic studies of eleven species of the Tropidurus torquatus group (Sauria, Tropiduridae), with banding patterns

The chromosomes of 173 specimens representing eleven species of the Tropidurus torquatus group, from 33 localities in Brazil, were analysed after Giemsa staining, C-banding, NORs, and replication banding techniques. A karyotype with 2n = 36, including 12 macrochromosomes and 24 microchromosomes (12 M + 24 m), and sex determination of the XY:XX type were found in Tropidurus cocorobensis, T. erythrocephalus, T. etheridgei, T. hispidus, T. hygomi, T. montanus, T. mucujensis, T. oreadicus, and T. torquatus. The two other species, T. itambere and T. psammonastes, presented 2n = 36 (12 M + 23 m) karyotype only in females while males had 2n = 35 (12 M + 23 m), due to the sex determination of the X(1)X(2)Y:X(1)X(1)X(2)X(2) type. Other interspecific differences as well as some intraspecific variation regarding the NORs and C-banding patterns have been observed, mainly in the microchromosome set. on the contrary, the macrochromosomes were highly conservative. Although consistent karyotypic diversity occurred in the torquatus group, the cytogenetic data obtained up to now did not allow us to clarify the phylogenetic relationships of the species. Nevertheless, the geographical distribution of the distinct cytotypes in T. hispidus and T. torquatus suggested that more than one species might be involved in each case.

Cytogenetic analyses of eight species in the genus Leptodactylus Fitzinger, 1843 (Amphibia, Anura, Leptodactylidae), including a new diploid number and a karyotype with multiple translocations

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Comparative cytogenetic analysis on four tree frog species (Anura, Hylidae, Hylinae) from Brazil

A comparative cytogenetic analysis was carried out on four Hylinae tree frogs from Brazil (Aparasphenodon brunoi, Corthomantis greeningi, Osteocephalus langsdorffi and Scinax fuscovarius) using Giemsa staining, BrdU replication banding, Ag-NOR staining, C-banding, DAPI and CMA(3) fluorochrome staining, and fluorescence in situ hybridization (FISH) with an rDNA probe. All the species share closely similar 2n = 24 karyotypes, almost indistinguishable by standard staining. The technique of BrdU incorporation allowed the identification of each pair of homologs and the establishment of extensive homeology for the great majority of the chromosomes, mainly of A. brunoi, C greeningi, and O. langsdorffii. Despite highly conserved replication banding patterns, the use of the other banding techniques disclosed some minor differsences, which reinforces the importance of extensive cytogenctic analyses for the karyotypic characterization of Anuran species. The present cytogenctic data confirm the closer proximity of A. brunoi, C greeningi, and O. langsdorfjii, whereas S. fuscovarius is phylogenetically more distant. Copyright (C) 2003 S. Karger AG, Basel

Multiple nucleolus organizer regions in Leptodactylus mystacinus (Amphibia, Anura) and comments on its systematic position in the L fuscus group based on cytogenetic and molecular analyses

Specimens of Leptodactylus mystacinus from Brazil were karyotyped with conventional and differential staining. The 2n = 22 karyotype is similar to that found for the majority of the Leptodactylus, the karyotypic conservatism also confirmed by the similarity of the replication banding patterns with those previously described. L. mystacinus has a small amount of C-banded heterochromatin, located mainly at the centromeres, although telomeric or interstitial bands have also been noticed. With DA/CMA(3) some chromosome regions showed slightly bright fluorescence, and with DA/DAPI, no particular AT-rich repetitive region was observed. Silver staining showed an extensive inter- and intraindividual variation in the number and position of Ag-positive regions, in 1p, 4p, 8p, 8q, and 11p. Nevertheless, FISH using rDNA probes confirmed only the signals on the short arms of chromosomes 4 and 8 as true NORs. The remaining silver stained regions are probably due to the heterochromatin with some affinity to the Ag-staining. Phylogenetic analysis based on partial cytochrome b sequence revealed that L. mystacinus forms a basal branch, so that the presence of multiple NORs in pairs 4 and 8 in this species indicates an autapomorphy.

R- and G-band patterns in Astyanax scabripinnis paranae (Pisces, Characiformes, Characidae)

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

Cariótipo de seis espécies de Bokermannohyla dos grupos de B. circumdata e B. pseudopseudis (Anura, Hylidae)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Possible interspecific origin of the B chromosome of Hypsiboas albopunctatus (Spix, 1824) (Anura, Hylidae), revealed by microdissection, chromosome painting, and reverse hybridisation

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

Autism spectrum disorder in a girl with a de novo x;19 balanced translocation

Balanced X-autosome translocations are rare, and female carriers are a clinically heterogeneous group of patients, with phenotypically normal women, history of recurrent miscarriage, gonadal dysfunction, X-linked disorders or congenital abnormalities, and/or developmental delay. We investigated a patient with a de novo X;19 translocation. The six-year-old girl has been evaluated due to hyperactivity, social interaction impairment, stereotypic and repetitive use of language with echolalia, failure to follow parents/caretakers orders, inconsolable outbursts, and persistent preoccupation with parts of objects. The girl has normal cognitive function. Her measurements are within normal range, and no other abnormalities were found during physical, neurological, or dysmorphological examinations. Conventional cytogenetic analysis showed a de novo balanced translocation, with the karyotype 46,X,t(X;19)(p21.2;q13.4). Replication banding showed a clear preference for inactivation of the normal X chromosome. The translocation was confirmed by FISH and Spectral Karyotyping (SKY). Although abnormal phenotypes associated with de novo balanced chromosomal rearrangements may be the result of disruption of a gene at one of the breakpoints, submicroscopic deletion or duplication, or a position effect, X; autosomal translocations are associated with additional unique risk factors including X-linked disorders, functional autosomal monosomy, or functional X chromosome disomy resulting from the complex X-inactivation process.

Pericentric inversion events in karyotypic distinction of Brazilian lizards of genus Phyllopezus (Squamata, Gekkonidae) detected by chromosomal banding patterns

Cytogenetic investigations based on conventional and differential staining analysis (C-and replication R-banding and Ag-staining) were carried out on eight specimens of Phyllopezus periosus, 17 of P. pollicaris pollicaris, and one of P. pollicaris przewalskii collected from different localities of Brazil. P. periosus and P. p. pollicaris share the same diploid number of 2n = 40 chromosomes, and their karyotypes are very distinctive regarding to the number of biarmed and uniarmed chromosomes. After careful side-by-side comparison of R-banded chromosomes in both taxa, pronounced homology between, at least, eight pairs was revealed. The R-banding patterns allowed us to postulate that karyotype differentiation could be due to pericentric inversion events. P. p. przewalskii (2n = 38) exhibited a very similar karyotype to that found in P. p. pollicaris, except for the presence of one metacentric pair, which probably resulted from a Robertsonian rearrangement. Single and multiple pairs of NOR-bearing chromosomes, showing variation in number and location, were detected among the three forms of Phyllopezus. Similar C-banding patterns were found in P. periosus and P. p. pollicaris. Sex chromosomes were not positively identified.

Chromosome banding patterns of four species of bats, with special reference to a case of X-autosome translocation

The karyotypes of 4 species of bats, Artibeus lituratus (Phyllostomatidae), Pipistrellus pipistrellus (Vespertilionidae), Pteropus alecto and P. giganteus (Pteropodidae), were studied after several banding techniques. For A. lituratus, in which an X-autosome translocation was observed, an analysis of the replication pattern in the rearranged chromosome was also made after BrdU incorporation.

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.

A Management Tool for the Replication of Operating Systems in Wireless Communication Networks

To simplify computer management, various administration systems based on wired connections adopt advanced techniques to manage software configuration. Nevertheless, the strong relation between hardware and software makes for an individualism of that management, besides penalizing computational mobility and ubiquity. All these issues lead to degradation of scalability, flexibility and the facility to install and maintain distributed applications. This article presents an environment for centralized wireless communication network management, named WSE-OS (Wireless Sharing Environment - Operating Systems): a model based on Virtual Desktop Infrastructure (VDI) which associates virtualization techniques and safe remote access systems to create a distributed architecture as a base for a managing system. WSE-OS is capable of accomplishing the replication of operating system images using wireless communication network, besides offering abstraction of hardware to its clients, making the management more flexible and independent of wired connections. Results obtained from this work indicate that WSE-OS allows disseminating, through a single software configuration, the execution of data related to operating system images in client computers. WSE-OS can also be used as a management tool for operating systems in a wireless network.

The genetics of hypertension modifies the renal cell replication response induced by experimental diabetes

To investigate whether the genetics of hypertension modifies renal cell responses in experimental diabetes, we studied the renal cell replication and its regulation by two cyclin-dependent kinase (Cdk) inhibitors, p27(Kip1) and p21(Cip1), in prehypertensive spontaneously hypertensive rats (SUR) and their genetically normotensive counterparts, Wistar Kyoto (WKY) rats, with and without streptozotocin-induced diabetes. In diabetic SIIR, the number of proliferating glomerular (0.6 +/- 0.3 positive cells/50 glomeruli) and tubulointerstitial (2.8 +/- 0.6 positive tubulointerstitial cells/50 grid fields) cells assessed by the bromodeoxyuridine technique was significantly (P = 0.0002) lower than in control SIIR (13.2 +/- 1.7 and 48.6 +/- 9.7, respectively) and control (14.0 +/- 1.8 and 63.9 +/- 10.6) and diabetic (14.3 +/- 3.5 and 66.4 +/- 11.5) WKY rats. Proliferating cell nuclear antigen, another marker of cell proliferation, was significantly reduced in replicating glomerular (P = 0.0002) and tubulointerstitial (P < 0.0001) cells in diabetic SHR. In freshly isolated glomeruli, the level of p27(Kip1) detected by Western blotting was significantly higher In diabetic SIIR than in nondiabetic SHR (1.52 +/- 0.14 vs. 1.00 +/- 0.10% of control, P = 0.014). The expression of p21(Cip1) in isolated glomeruli did not differ among the groups of rats. In conclusion, the response of renal cell replication to diabetes differs markedly between prehypertensive SIIR and their WKY control rats. The decreased glomerular cell proliferation in prehypertensive diabetic SIIR is at least partly mediated by a higher expression of the Cdk inhibitor p27(Kip1).