A logística como ferramenta eficaz no transporte urbano de passageiros: o caso da empresa de transporte metropolitano Belém Rio

Este estudo trata a questão da Logística na operacionalização do transporte público por ônibus da Região Metropolitana de Belém - RMB, particularmente, da concessionária Empresa Belém Rio, que atua na Capital do Pará. O foco principal foi perceber se existem indicadores de desempenho eficaz na operacionalização mais racional e equilibrada desse modal de transporte. A pesquisa foi conduzida por uma revisão bibliográfica de estudos da Logística e suas operações, como também da utilização dessa ferramenta de gestão no transporte, seguido de uma apresentação do enfoque evolutivo da RMB, com especial atenção a questão do transporte de passageiros. A metodologia foi preparada para atender à pesquisa de campo e ao estudo descritivo. Consistiu basicamente na determinação de variáveis extraídas de indicadores que tiveram por base o conceito de Logística. O questionário foi estruturado com base nessas vertentes e aplicados à uma população de 44 sujeitos, funcionários da referida empresa. Além desse procedimento foi realizada uma entrevista com a gerência executiva, na intenção não de tratamento, mas de buscar informações que pudessem subsidiar a análise. Os dados coletados no instrumento metodológico foram tratados e sistematizados em gráficos e tabelas por ferramentas estatísticas. Na análise dos dados quantificados, foi utilizada a literatura para cruzamento de informações, além do que o uso de algumas falas de respondentes. Dentro dos resultados obtidos, encontram-se observações que podem subsidiar algumas tomadas de decisão para a melhoria de desempenho da citada empresa. Além do que, disponibilizadas percepções que poderão ser utilizadas por outros estudos.

Spliceosomal factors are recognized by Chagasic sera

We investigated the possibility that Chagas' patients develop an autoimmune response to human UsnRNPs or Sm epitopes. Using purified human UsnRNPs we detected anti-human UsnRNPs antibodies in sera from patients with Chagas' disease. The antibodies were also detected using peptide-ELISA containing the Sm-motif 1 domain, showing that 61% (31/51) of the Chagas sera contained antibodies against the Sm-motif 1. Our preliminary results obtained by immunoprecipitation using Chagas chronically infected Balb/C sera and HeLa nuclear extracts showed that some autoantibodies could also be found during the course of the disease. Groups of 20 mice tone-month-old) were infected i.p. with Y strain 30 bloodstream trypomastigotes and killed at 60 days post-infection and sera were used for immunoprecipitation analysis as mentioned, These antibodies cross-reacted with U2 snRNP in HeLa nuclear extract and revealed many different bands in T. cruzi nuclear extract. These results suggest that the autoantibodies may have a role in the pathogenesis of the disease,

Presence of autoantibodies against small nuclear ribonucleoprotein epitopes in Chagas' patients' sera

We investigated the possibility that Chagas' patients develop an autoimmune response to human UsnRNPs (small nuclear ribonucleoprotein) or Sm epitopes. Using purified human UsnRNPs, we detected anti-human UsnRNPs antibodies in sera from patients suffering from Chagas' disease. The antibodies it-ere also detected using peptide enzyme-linked immunosorbent assays containing the Sm-motif 1 domain. The latter technique showed that 61% (31/51) of the Chagas' patients' sera contained antibodies against Sm-motif I. The detection of anti-UsnRNPs autoantibodies in Chagas patients' sera strongly encourages further studies using animal models to determine how these autoantibodies appear.

The contribution of 700,000 ORF sequence tags to the definition of the human transcriptome

open reading frame expressed sequences tags (ORESTES) differ from conventional ESTs by providing sequence data from the central protein coding portion of transcripts. We generated a total of 696,745 ORESTES sequences from 24 human tissues and used a subset of the data that correspond to a set of 15,095 full-length mRNAs as a means of assessing the efficiency of the strategy and its potential contribution to the definition of the human transcriptome. We estimate that ORESTES sampled over 80% of all highly and moderately expressed, and between 40% and 50% of rarely expressed, human genes. In our most thoroughly sequenced tissue, the breast, the 130,000 ORESTES generated are derived from transcripts from an estimated 70% of all genes expressed in that tissue, with an equally efficient representation of both highly and poorly expressed genes. In this respect, we find that the capacity of the ORESTES strategy both for gene discovery and shotgun transcript sequence generation significantly exceeds that of conventional ESTs. The distribution of ORESTES is such that many human transcripts are now represented by a scaffold of partial sequences distributed along the length of each gene product. The experimental joining of the scaffold components, by reverse transcription-PCR, represents a direct route to transcript finishing that may represent a useful alternative to full-length cDNA cloning.

Antibodies to yeast Sm motif 1 cross-react with human Sm core polypeptides

Two regions common to all UsnRNP core polypeptides have been described: Sm motif 1 and Sm motif 2. Rabbits were immunized with a 22 amino-acid peptide containing one segment of Sm motif 1 (YRGTLVSTDNYFNLQL-NEAEEF, corresponding to residues 11-32) from yeast F protein. After immunization, the rabbit sera contained antibodies that not only reacted specifically with the peptide from yeast F protein but also cross-reacted with Sm polypeptides from mammals; that is, with purified human U1snRNPs. The results suggest that the peptide used and human Sm polypeptides contain a common feature recognized by the polyclonal antibodies. A large collection of human systemic lupus erythematosus sera was assayed using the yeast peptide as an antigen source. Seventy per cent of systemic lupus erythematosus sera contain an antibody specificity that cross-reacts with the yeast peptide.

Trypanosoma cruzi: establishment of permeable cells for RNA processing analysis with drugs

Pre-mRNA maturation in trypanosomatids occurs through a process called trans-splicing which involves excision of introns and union of exons in two independent transcripts. For the first time, we present the standardization of Trypanosoma cruzi permeable cells (Y strain) as a model for trans-splicing study of mRNAs in trypanosomes, following by RNase protection reaction, which localizes the SL exon and intron. This trans-splicing reaction in vitro was also used to analyze the influence of NFOH-121, a nitrofurazone-derivative, on this mechanism. The results suggested that the prodrug affects the RNA processing in these parasites, but the trans-splicing reaction still occurred.

Cloning and molecular characterization of Trypanosoma cruzi U2, U4, U5, and U6 small nuclear RNAs

Small nuclear RNAs (snRNAs) are important factors in the functioning of eukaryotic cells that form several small complexes with proteins; these ribonucleoprotein particles (U snRNPs) have an essential role in the pre-mRNA processing, particularly in splicing, catalyzed by spliceosomes, large RNA-protein complexes composed of various snRNPs. Even though they are well defined in mammals, snRNPs are still not totally characterized in certain trypanosomatids as Trypanosoma cruzi. For this reason we subjected snRNAs (U2, U4, U5, and U6) from T. cruzi epimastigotes to molecular characterization by polymerase chain reaction (PCR) and reverse transcription-PCR. These amplified sequences were cloned, sequenced, and compared with those other of trypanosomatids. Among these snRNAs, U5 was less conserved and U6 the most conserved. Their respective secondary structures were predicted and compared with known T. brucei structures. In addition, the copy number of each snRNA in the T. cruzi genome was characterized by Southern blotting.

Comparison of the genomes of two Xanthomonas pathogens with differing host specificities

The genus Xanthomonas is a diverse and economically important group of bacterial phytopathogens, belonging to the gamma-subdivision of the Proteobacteria. Xanthomonas axonopodis pv. citri (Xac) causes citrus canker, which affects most commercial citrus cultivars, resulting in significant losses worldwide. Symptoms include canker lesions, leading to abscission of fruit and leaves and general tree decline(1). Xanthomonas campestris pv. campestris (Xcc) causes black rot, which affects crucifers such as Brassica and Arabidopsis. Symptoms include marginal leaf chlorosis and darkening of vascular tissue, accompanied by extensive wilting and necrosis(2). Xanthomonas campestris pv. campestris is grown commercially to produce the exopolysaccharide xanthan gum, which is used as a viscosifying and stabilizing agent in many industries(3). Here we report and compare the complete genome sequences of Xac and Xcc. Their distinct disease phenotypes and host ranges belie a high degree of similarity at the genomic level. More than 80% of genes are shared, and gene order is conserved along most of their respective chromosomes. We identified several groups of strain-specific genes, and on the basis of these groups we propose mechanisms that may explain the differing host specificities and pathogenic processes.

Identification of human chromosome 22 transcribed sequences with ORF expressed sequence tags

Transcribed sequences in the human genome can be identified with confidence only by alignment with sequences derived from cDNAs synthesized from naturally occurring mRNAs. We constructed a set of 250,000 cDNAs that represent partial expressed gene sequences and that are biased toward the central coding regions of the resulting transcripts. They are termed ORF expressed sequence tags (ORESTES). The 250,000 ORESTEs were assembled into 81,429 contigs. of these, 1,181 (1.45%) were found to match sequences in chromosome 22 with at least one ORESTES contig for 162 (65.6%) of the 247 known genes, for 67 (44.6%) of the 150 related genes, and for 45 of the 148 (30.4%) EST-predicted genes on this chromosome. Using a set of stringent criteria to validate our sequences, we identified a further 219 previously unannotated transcribed sequences on chromosome 22. of these, 171 were in fact also defined by EST or full length cDNA sequences available in GenBank but not utilized in the initial annotation of the first human chromosome sequence. Thus despite representing less than 15% of all expressed human sequences in the public databases at the time of the present analysis, ORESTEs sequences defined 48 transcribed sequences on chromosome 22 not defined by other sequences. All of the transcribed sequences defined by ORESTEs coincided with DNA regions predicted as encoding exons by GENSCAN.

Trypanosoma cruzi: identification and characterization of a novel ribosomal protein L27 (TcrL27) that cross-reacts with an affinity-purified anti-Sm antibody

Small nuclear ribonucleoproteins (snRNPs)are involved in trans-splicing processing of pre-mRNA in Trypanosoma cruzi. To clone T. cruzi snRNPs we screened an epimastigote cDNA library with a purified antibody raised against the Sm-binding site of a yeast sequence. A clone was obtained containing a 507 bp-insert with an ORF of 399 bp and coding for a protein of 133 amino acids. Sequence analysis revealed high identity with the L27 ribosomal proteins from different species including: Canis familiaris, Homo sapiens, Schizosaccharomyces pombe and Saccharomyces cerevisiae. This protein has not been previously described in the literature and seems to be a new ribosomal protein in T. cruzi and was given the code TcrL27. To express this recombinant T. cruzi L27 ribosomal protein in E. coli, the insert was subcloned into the pET32a vector and a 26 kDa recombinant protein was purified. Immunoblotting studies demonstrated that this purified recombinant protein was recognized by the same anti-Sm serum used in the library screening as well as by chagasic and systemic lupus erythemathosus (SLE) sera. Our results suggest that the T. cruzi L27 ribosomal protein may be involved in autoimmunity of Chagas disease.

Influence of management and environmental factors on mating success of Africanized queen honey bees

Environmental factors and management techniques were evaluated in São Paulo, Brazil, for enhanced production of Africanized queen honey bees. Queens were reared by the Doolittle method; 12 breeder, 6 cell-builder, and 36 3-frame nucleus colonies were used. Nine groups of four virgin Africanized queen honey bees were subjected to the following treatments: queens were either 1-2, 3-4 or 5-6 days old and were released into mating nuclei containing either capped brood, uncapped brood or no brood. This was repeated sixteen times between August 1990 and August 1992. Seven repetitions occurred during nectar flow periods and nine repetitions occurred during nectar-dearth periods. Overall, 59% of 576 queens were successfully introduced and mated. The best results (93% success) were obtained during nectar flows, with 3- to 4-day-old queens released into nuclei containing capped brood. During nectar dearths the best mating success came from queens introduced into broodless nuclei (63%), the age of the queen did not influence mating success. Mating success decreased when wind velocity increased; this was the only significant meteorological effect found.

The generation and utilization of a cancer-oriented representation of the human transcriptome by using expressed sequence tags

Whereas genome sequencing defines the genetic potential of an organism, transcript sequencing defines the utilization of this potential and links the genome with most areas of biology. To exploit the information within the human genome in the fight against cancer, we have deposited some two million expressed sequence tags (ESTs) from human tumors and their corresponding normal tissues in the public databases. The data currently define approximate to23,500 genes, of which only approximate to1,250 are still represented only by ESTs. Examination of the EST coverage of known cancer-related (CR) genes reveals that <1% do not have corresponding ESTs, indicating that the representation of genes associated with commonly studied tumors is high. The careful recording of the origin of all ESTs we have produced has enabled detailed definition of where the genes they represent are expressed in the human body. More than 100,000 ESTs are available for seven tissues, indicating a surprising variability of gene usage that has led to the discovery of a significant number of genes with restricted expression, and that may thus be therapeutically useful. The ESTs also reveal novel nonsynonymous germline variants (although the one-pass nature of the data necessitates careful validation) and many alternatively spliced transcripts. Although widely exploited by the scientific community, vindicating our totally open source policy, the EST data generated still provide extensive information that remains to be systematically explored, and that may further facilitate progress toward both the understanding and treatment of human cancers.