A LINE2 repetitive DNA sequence from the cichlid fish, Oreochromis niloticus: sequence analysis and chromosomal distribution

We report the cloning and characterization of a long interspersed nucleotide element (LINE) fi-om a cichlid fish, Oreochromis niloticus, and show the distribution of this element, called CiLINE2 for cichlid LINE2, in the chromosomes of this species. The identification of an open reading frame in CiLINE2 with amino acid sequence similarity to reverse transcriptases encoded by LINE-like elements in Caenorhabditis elegans, Platemys spixii, Schistosoma mansoni, Gallus gallus (CRI), Drosophila melanogaster (I factor), and Homo sapiens (LINE2), as well as the structure of the element, suggest it is a member of this family of non-long terminal repeat-containing retrotransposons. Search of a DNA sequence database identified sequences similar to CiLINE2 in four other fish species (Haplotaxodon microlepis, Oreochromis mossambicus, Pseudotropheus zebra, and Fugu rubripes). Southern blot hybridization experiments revealed the presence of sequences similar to CiLINE2 in all Tilapiini species analyzed from the genera Oreochromis, Tilapia, and Sarotherodon, and gave an estimated copy number of about 5500 for the haploid genome of O. niloticus. Fluorescent in situ hybridization showed that CiLINE2 sequences were organized in small clusters dispersed over all chromosomes of O. niloticus, with a higher concentration near chromosome ends. Furthermore the long arm of chromosome 1 was strikingly enriched with this sequence. The distribution of LINE2-related elements might underlie the difference in chromosome banding patterns observed between cold-blooded vertebrates and mammals.

Camponeses Brazil's Peasant Movement in Historical Perspective (1946-2004)

Comparison of the histories of three leading peasant organizations in the Pontal do Paranapanema region of Brazil-the Partido Comunista do Brasil (PCB) from 1945 to 1964, the Confederacao Nacional de Trabalhadores na Agricultura (CONTAG) from 1964 to 1984, and the Movimento dos Trabalhadores Rurais Sem-Terra (MST) from 1984 to 2004-suggests that continuity is as important as change in understanding Brazilian peasant movements. The MST has been considered a "new social movement" in that it has eschewed partisan politics, incorporated families as members rather than just male heads of household, had a national scope and a participatory decision-making structure, and been attuned to the international struggle over globalization. Placing it in historical perspective makes it clear, however, that this is not the first time that militants have organized around the concept of peasants as a political identity; that while the representation of peasants in the leadership of contemporary rural labor organizations may be greater than in the past, earlier peasant leaders also struggled on behalf of their class; that earlier peasant organizations had, if not a national presence, a substantial presence in the agricultural states of the time; and that attempts at international organization to unite peasant struggles around the globe are not entirely new. This is not to deny the innovative features of contemporary movements but to suggest that the investigation of past achievements will contribute to a fuller appreciation of these movements' conditions and prospects.

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.

RESTRUCTURING of SYSTEMS and LOGISTICS of MOVEMENT and ITS IMPACT on URBAN and REGIONAL PLANNING PAULISTA

Report some of the changes in production and consumption occurring in the state of São Paulo. through the restructuring in motion systems, logistics and standards and taxation, as well as the impacts on urban spaces through new economic dynamics, imposed by the demands of corporate, is the purpose of this article. The decentralization of production and consumption towards the interior was made possible by the combination of hierarchical and ordered some basic elements such as technological innovations (ways and means of transport) and organizational (logistics, standards and taxation) which optimized the flow territorial state São Paulo. It is noteworthy, therefore: 1) the improvement of logistics as a strategy, planning and management of transport, storage and communications (including the granting of public services to private), 2) the technological improvement and expansion of motion systems (infrastructure, means of transport) and 3) the systems of rules and regulations through taxation and deregulation affect the circulatory system of a given space. Thus, both systems aims to disentangle the economic flows (goods, services, information, capital and people) and provide a more fluid territorial. The impacts on the State of São Paulo, mainly through its economic dynamics, revert positively and negatively, by changing the way one thinks and performs planning.

Molecular and cytogenetic analysis of the telomeric (TTAGGG)n repetitive sequences in the Nile tilapia, Oreochromis niloticus (Teleostei: Cichlidae)

The majority of chromosomes in Oreochromis niloticus, as with most fish karyotyped to date, cannot be individually identified owing to their small size. As a first step in establishing a physical map for this important aquaculture species of tilapia we have analyzed the location of the vertebrate telomeric repeat sequence, (TTAGGG)n, in O. niloticus. Southern blot hybridization analysis and a Bal31 sensitivity assay confirm that the vertebrate telomeric repeat is indeed present at O. niloticus chromosomal ends with repeat tracts extending for 4-10 kb on chromosomal ends in erythrocytes. Fluorescent in situ hybridization revealed that (TTAGGG)n is found not only at telomeres, but also at two interstitial loci on chromosome 1. These data support the hypothesis that chromosome 1, which is significantly larger than all the other chromosomes in the karyotype, was produced by the fusion of three chromosomes and explain the overall reduction of chromosomal number from the ancestral teleost karyotype of 2n=48 to 2n=44 observed in tilapia.

Short interspersed repetitive elements (SINEs) from the cichlid fish, Oreochromis niloticus, and their chromosomal localization by fluorescent in situ hybridization

In the present study, we describe the cloning and characterization of a new SINE-like element from O. niloticus (ROn-2) and show the distribution of this SINE and a previously isolated SINE, ROn-1, in the chromosomes of O. niloticus. The ROn-2 element is 359 base pairs (bp) in length, contains short direct terminal repeats, a tRNA-related region similar to tRNA Val and tRNA Arg, a tRNA-unrelated region, and a poly-A tail. Analysis of the chromosomal distribution of ROn-1 and ROn-2 by fluorescent in situ hybridization showed that both SINE sequences are present in all chromosomes of tilapia, and organized in small clusters. The only exception was a large cluster of ROn-1 repeats found in the middle of the long arm of chromosome 1. In view of our data we discuss the hypothesis that the absence of large clusters of SINE sequences and the structural composition of these sequences may explain the absence of base-specific fluorochrome bands in the chromosomes of tilapia.

Electromyographic fatigue threshold of erector spinae muscle induced by a muscular endurance test in health men

Purpose: To identify the electromyographic fatigue threshold in the erector spinae muscle. Methods: Eight 19 to 24-year-old male volunteers participated in this study, in which surface electrodes were used, as well as a biological signals acquisition module (Lynx) with a sampling frequency of 1000Hz, a 1000 times gain, a 20Hz high pass filter and a 500Hz low pass filter. The test consisted of repeated isometric contractions of the erector spinae muscle in a 45° hip flexion posture, with 30%, 40%, 50% and 60% of the maximum voluntary isometric contraction. Results: A positive correlation of the RMS (root mean square) value as a function of time was found for most of the subjects with 40% (N = 6), 50% (N = 7) and 60% (N = 8) loads of the maximum voluntary isometric contraction. Conclusions: It was concluded, from this study, that the proposed protocol provides evidence, through the electromyographic signal, of the development of fatigue in the erector spinae muscle with loads of 40%, 50% and 60% of the maximum voluntary isometric contraction. The protocol also allows the electromyographic fatigue threshold and its probable applicability in the diagnosis of this phenomenon during repetitive activities to be determined.

Kinesiographic study of complete denture movement related to mucosa displacement in edentulous patients.

The mucosa that covers the residual ridges of edentulous patients may present some distortion or displacement when occlusal loading is applied in complete dentures. This distortion and movement of the denture can result in acceleration of residual ridge resorption and loss of retention and stability. The aim of this study was to analyze the pattern of upper complete denture movement related to underlying mucosa displacement. A sample of 10 complete denture wearers was randomly selected, which had acceptable upper and lower dentures and normal volume and resilience of residual ridges. The kinesiographic instrument K6-I Diagnostic System was used to measure denture movements, according to the method proposed by Maeda et al.7, 1984. Denture movements were measured under the following experimental conditions: (A) 3 maximum voluntary clenching cycles and (B) unilateral chewing for 20 seconds. The results showed that under physiological load, oral mucosa distortion has two distinct phases: a fast initial displacement as load is applied and a slower and incomplete recovery when load is removed. Intermittent loading such as chewing progressively reduces the magnitude of the denture displacement and the recovery of the mucosa is gradually more incomplete.

Laryngeal electromyography in movement disorders: Preliminary data

This study describes preliminary laryngeal electromyography (LEMG) data and botulinum toxin treatment in patients with dysphonia due to movement disorders. Twenty-five patients who had been clinically selected for botulinum toxin administration were examined, 19 with suspected laryngeal dystonia or spasmodic dysphonia (SD), 5 with vocal tremor, and 1 with Gilles de la Tourette syndrome (GTS). LEMG evaluations were performed before botulinum toxin administration using monopolar electrodes. Electromyography was consistent with dystonia in 14 patients and normal in 5, and differences in frequency suggesting essential tremor in 3 and Parkinson tremors in 2. The different LEMG patterns and significant improvement in our patients from botulinum toxin therapy has led us to perform laryngeal electromyography as a routine in UNICAMP movement disorders ambulatory.

Tuberculose e o estudo molecular da sua epidemiologia

Systems that can distinguish epidemiologically-related Mycobacterium tuberculosis strains from unrelated ones are extremely valuable. Molecular biology techniques have allowed a great deal of information to be acquired about the infectious disease tuberculosis (TB) that was very hard or impossible to obtain by conventional epidemiology. A typing method based on bacterial DNA genome differences, known as RFLP (restriction fragment length polymorphism), is widely used to discriminate strains in the epidemiologic study of TB. However, RFLP is laborious and there is a tendency to replace it by other methods. Thus, other DNA sequences have been employed as epidemiological markers, as in Spoligotyping, a fast technique based on PCR followed by differential hybridization of amplified products. The polymorphism observed among different isolates is probably the product of strain-dependent recombination. MIRU (mycobacterial interspersed repetitive unit) typing is a reproducible and fast assay, involving the generation of genotypes based on the study of 12 loci containing VNTRs (variable-number tandem repeats) in strains of the M. tuberculosis complex. It compares strains from different geographic areas and allows the movement of individual lineages to be tracked, as in RFLP. This approach enables a greater number of isolates to be analyzed, leading to the identification of a larger number of foci of transmission within the population and thus to improved ways of slowing the progress of the disease.

Spatial electric load forecasting using a local movement approach

An agent based model for spatial electric load forecasting using a local movement approach for the spatiotemporal allocation of the new loads in the service zone is presented. The density of electrical load for each of the major consumer classes in each sub-zone is used as the current state of the agents. The spatial growth is simulated with a walking agent who starts his path in one of the activity centers of the city and goes to the limits of the city following a radial path depending on the different load levels. A series of update rules are established to simulate the S growth behavior and the complementarity between classes. The results are presented in future load density maps. The tests in a real system from a mid-size city show a high rate of success when compared with other techniques. The most important features of this methodology are the need for few data and the simplicity of the algorithm, allowing for future scalability. © 2009 IEEE.

Knowledge on dental trauma and orthodontic tooth movement held by a group of orthodontists

The objective of this work was to assess the knowledge about orthodontic tooth movement and dental trauma held by a group of orthodontists in specific areas of Brazil. For this purpose, 166 questionnaires with 15 objective questions about this subject were distributed. One hundred and five questionnaires were properly filled and collected after 30 days. It was concluded that, except for avulsion, the knowledge on dental injuries held by the professionals interviewed was considered unsatisfactory, and about 40% of them were not acquainted with the recommendations for the orthodontic movement of traumatized teeth.

Characterization and genome organization of a repetitive element associated with the nucleolus organizer region in leporinuselongatus (anostomidae: Characiformes)

Chromosome mapping and studies of the genomic organization of repetitive DNA sequences provide valuable insights that enhance our evolutionary and structural understanding of these sequences, as well as identifying chromosomal rearrangements and sex determination. This study investigated the occurrence and organization of repetitive DNA sequences in Leporinus elongatus using restriction enzyme digestion and the mapping of sequences by chromosomal fluorescence in situ hybridization (FISH). A 378-bp fragment with a 54.2% GC content was isolated after digestion with the SmaI restriction enzyme. BLASTN search found no similarity with previously described sequences, so this repetitive sequence was named LeSmaI. FISH experiments were conducted using L. elongatus and other Anostomidae species, i.e. L. macrocephalus,L. obtusidens, L. striatus, L. lacustris, L. friderici, Schizodon borellii, S. isognathus, and Abramites hypselonotus which detected signals that were unique to male and female L. elongatus individuals. Double-FISH using LeSmaI and 18S rDNA showed that LeSmaI was located in a nucleolus organizer region (NOR) in the male and female metaphases of L. elongatus. This report also discusses the role of repetitive DNA associated with NORs in the diversification of Anostomidae species karyotypes. Copyright © 2012 S. Karger AG, Basel.

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.