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Made available in DSpace on 2020-12-10T20:09:16Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-01-01 Ludwig Institute for Cancer Research U.S. National Cancer Institute Canadian Institutes of Health Research Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Canadian Institutes of Health Research [BESC--M] Fond de la recherche en sante du Quebec Introduction. Persistent human papillomavirus (HPV) type 16 infection is the main causal agent of cervical cancer. Most HPV infections clear spontaneously within 1-2 years. Although not all infected women develop detectable HPV antibodies, about 60-70% seroconvert and retain their antibodies at low levels. Aim. We investigated if cervical HPV16 DNA positivity was associated with HPV16 seroreactivity measured with two different antigen formulations. We assessed if associations were influenced by co-infection with other HPV types and HPV16 viral load. Methodology. We used baseline data for women participating in the Ludwig-McGill cohort, a longitudinal investigation of the natural history of HPV infection and cervical neoplasia. The study enrolled 2462 Brazilian women from 1993 to 1997 (pre-vaccination). ELISA assays were based on L1-only or L1+L2 virus-like particles (VLPs). Seroreactivity was expressed as normalized absorbance ratios. HPV genotyping and viral load were evaluated by PCR protocols. Pearson's r was used to measure correlations between interval-scaled variables. Serological accuracy in HPV16 DNA detection was assessed using receiver operating characteristic (ROC) curves. We analysed the association between HPV DNA positivity and HPV16 seroreactivity by linear regression. Results. Correlations between L1+L2 and L1-only VLPs for detection of HPV16 were poor (r=0.43 and 0.44 for dilutions 1 :10 and 1:50, respectively). The protocol with the best accuracy was L1+L2 VLPs at serum dilution 1: 10 (ROC area=0.73, 95% CI: 0.65-0.85). HPV16 DNA positivity was correlated with HPV16 seroreactivity and was not influenced by co-infection or viral load. To a lesser degree, HPV16 seroreactivity was correlated with infection by other Alpha-9 papillomavirus species. Conclusion. HPV16 DNA positivity and HPV16 seroreactivity are strongly correlated. L1+L2 VLPs perform better than L1-only VLPs for detecting IgG antibodies to HPV16 in women infected with HPV16 or other Alpha-9 HPV species. This study advances our understanding of humoral immune responses against HPV16 by providing insights about the influence of VLP antigen composition to measure humoral immune response against naturally acquired HPV infection. Univ Montreal, Sch Publ Hlth, Dept Social & Prevent Med, Montreal, PQ, Canada Univ Montreal, St Justine Hosp Res Ctr, Montreal, PQ, Canada Univ Estadual Paulista, Inst Biosci, Dept Microbiol & Immunol, Botucatu, SP, Brazil Ludwig Inst Canc Res, Sao Paulo, Brazil Univ Sao Paulo, Inst Canc Estado Sao Paulo, Sao Paulo, Brazil Univ Sao Paulo, Fac Med, Dept Radiol & Oncol, Sao Paulo, Brazil McGill Univ, Div Canc Epidemiol, Montreal, PQ, Canada Univ Estadual Paulista, Inst Biosci, Dept Microbiol & Immunol, Botucatu, SP, Brazil U.S. National Cancer Institute: CA70269 Canadian Institutes of Health Research: MOP-49396 Canadian Institutes of Health Research: CRN-83320 FAPESP: 1999/10790-0 FAPESP: 1994/05415-1 FAPESP: 1997/07172-7

Made available in DSpace on 2015-03-18T15:55:51Z (GMT). No. of bitstreams: 0 Previous issue date: 2014-11-18Bitstream added on 2015-03-18T16:28:32Z : No. of bitstreams: 1 WOS000345710200001.pdf: 196329 bytes, checksum: 8012259309a44b7de854b5975e589f51 (MD5) Ludwig Institute for Cancer Research U.S. National Cancer Institute Canadian Institutes of Health Research Background: Immunity plays an important role in controlling human papillomavirus (HPV) infection and associated lesions. Unlike infections caused by other viruses, natural HPV infection does not always result in a protective antibody response. Therefore, HPV antibodies are also considered markers of cumulative exposure. The aim of this study was to identify determinants of HPV16 seroreactivity at enrollment among women from the Ludwig-McGill cohort, a natural history study of HPV infection and risk of cervical neoplasia.Methods: HPV16 serology was assessed by ELISA for L1 and L2 capsid antigens, while HPV typing and viral load measurements were performed by PCR-based methods. The associations were analyzed by unconditional logistic regression.Results: Of 2049 subjects, 425 (20.7%) were strongly seropositive for HPV16. In multivariate analysis, seroreactivity was positively correlated with age, lifetime number of sexual partners, frequency of sex, and HPV16 viral load, and negatively associated with duration of smoking.Conclusions: HPV16 seroreactivity is determined by factors that reflect viral exposure. Ludwig Inst Canc Res, Sao Paulo, Brazil Univ Fed Fluminense, Dept Immunobiol, Niteroi, RJ, Brazil Brazilian Natl Canc Inst, Program Cellular Biol, Rio De Janeiro, Brazil McGill Univ, Dept Oncol, Montreal, PQ, Canada Univ Estadual Paulista, Inst Biosci, Dept Microbiol & Immunol, Botucatu, SP, Brazil Inst Gynaecol Canc Res, Sao Paulo, Brazil Univ Montreal, CHUM, Div Gynecol Oncol, Montreal, PQ, Canada McGill Univ, Dept Epidemiol Biostat & Occupat Hlth, Montreal, PQ, Canada ICESP, Sao Paulo Canc Inst, Mol Biol Lab, Sao Paulo, Brazil Univ Sao Paulo, Sch Med, Dept Radiol & Oncol, Sao Paulo, Brazil Univ Sao Paulo, Sch Med, Sao Paulo, Brazil Santa Casa Sao Paulo, Sao Paulo, Brazil Univ Estadual Paulista, Inst Biosci, Dept Microbiol & Immunol, Botucatu, SP, Brazil U.S. National Cancer InstituteCA70269 Canadian Institutes of Health Research49396 Canadian Institutes of Health Research83320

Submitted by Guilherme Lemeszenski (guilherme@nead.unesp.br) on 2014-02-26T17:19:33Z No. of bitstreams: 1 WOS000222434200023.pdf: 392582 bytes, checksum: eb50adacff00ab5cc37867f0de853492 (MD5) Made available in DSpace on 2014-02-26T17:19:33Z (GMT). No. of bitstreams: 1 WOS000222434200023.pdf: 392582 bytes, checksum: eb50adacff00ab5cc37867f0de853492 (MD5) Previous issue date: 2004-07-01 Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-20T13:50:12Z No. of bitstreams: 1 WOS000222434200023.pdf: 392582 bytes, checksum: eb50adacff00ab5cc37867f0de853492 (MD5) Made available in DSpace on 2014-05-20T13:50:12Z (GMT). No. of bitstreams: 1 WOS000222434200023.pdf: 392582 bytes, checksum: eb50adacff00ab5cc37867f0de853492 (MD5) Previous issue date: 2004-07-01 We report the results of a transcript finishing initiative, undertaken for the purpose of identifying and characterizing novel human transcripts, in which RT-PCR was used to bridge gaps between paired EST Clusters, mapped against the genomic sequence. Each pair of EST Clusters selected for experimental validation was designated a transcript finishing unit (TFU). A total of 489 TFUs were selected for validation, and an overall efficiency of 43.1% was achieved. We generated a total of 59,975 bp of transcribed sequences organized into 432 exons, contributing to the definition of the structure of 211 human transcripts. The structure of several transcripts reported here was confirmed during the course of this project, through the generation of their corresponding full-length cDNA sequences. Nevertheless, for 21% of the validated TFUs, a full-length cDNA sequence is not yet available in public databases, and the structure of 69.2% of these TFUs was not correctly predicted by computer programs. The TF strategy provides a significant contribution to the definition of the complete catalog of human genes and transcripts, because it appears to be particularly useful for identification of low abundance transcripts expressed in a restricted Set of tissues as well as for the delineation of gene boundaries and alternatively spliced isoforms. Univ São Paulo, Inst Quim, BR-05513970 São Paulo, Brazil Ludwig Inst Canc Res, BR-01509010 São Paulo, Brazil Univ São Paulo, Dept Epidemiol, Sch Publ Hlth, BR-01246904 São Paulo, SP, Brazil Univ Ribeirao Preto, Dept Quim Informat Bioinformat, BR-14096380 Ribeirao Preto, Brazil Univ São Paulo, Fac Med Ribeirao Preto, Centro Terapia Celular, Hemocentro, BR-14051140 Ribeirao Preto, SP, Brazil Univ São Paulo, Fac Med Ribeirao Preto, Dept Clin Med, BR-14051140 Ribeirao Preto, SP, Brazil Inst Ludwig, Lab Biol Computacional, BR-01509010 São Paulo, SP, Brazil Univ São Paulo, Lab Genet Cardiol Mol, Inst Coracao, BR-05403000 São Paulo, SP, Brazil Univ Fed São Paulo, Bioinform Lab, Hlth Informatics Dept, BR-04039032 São Paulo, Brazil Univ Estad Campinas, Dept Clin Med, Fac Ciências Med, BR-13083970 Campinas, SP, Brazil Univ Fed São Paulo, Rheumatol Div, BR-04113001 São Paulo, Brazil Univ São Paulo, Dept Histol Embriol, Inst Ciências Biomed, BR-05508900 São Paulo, Brazil Univ Fed São Paulo, Dept Microbiol Immunol Parasitol, BR-04023062 São Paulo, Brazil Univ São Paulo, Lab Cellular Mol Endocrinol, Sch Med, BR-01246903 São Paulo, Brazil Univ Fed São Paulo, Dept Med, Disciplina Endocrinol, Lab Endocrinol Mol, BR-04039002 São Paulo, Brazil Univ São Paulo, Inst Ciências Biomed, Lab Transferencia Genica, BR-05508900 São Paulo, Brazil Univ São Paulo, Fac Med Ribeirao Preto, Dept Biol Celular Mol Bioagentes Patogenicas, BR-14049900 Ribeirao Preto, SP, Brazil Univ Estadual Paulista, Lab Biol Mol, Hemocentro, Fac Med, BR-18618970 Botucatu, SP, Brazil Univ Fed Sao Carlos, Dept Genet Evol, BR-13565905 Sao Carlos, SP, Brazil Univ São Paulo, Fac Ciências Farmaceuticas Ribeirao Preto, BR-14040903 Ribeirao Preto, SP, Brazil Univ São Paulo, Fac Filosofia Ciências Letras Ribeirao Preto, Ribeirao Preto, SP, Brazil Univ São Paulo, Dept Patol, Fac Med Vet Zootecnia, BR-05508000 São Paulo, Brazil Univ Estadual Campinas, Dept Genet Med, Fac Ciências Med, BR-13081970 Campinas, SP, Brazil Univ São Paulo, Fac Med Ribeirao Preto, Dept Genet, BR-14040900 Ribeirao Preto, SP, Brazil Univ São Paulo, Dept Bioquim, Inst Quim, BR-05599970 São Paulo, Brazil Univ São Paulo, Dept Neurol, Fac Med, BR-01246903 São Paulo, Brazil Univ Estadual Campinas, Lab Genoma Funcional, Centro Biol Mol Engenharia Genet, BR-13083970 Campinas, SP, Brazil Univ Estadual Campinas, Dept Genet Evol, Inst Biol, Campinas, SP, Brazil Univ São Paulo, Dept Radiol, Disciplina Oncol, Fac Med, BR-01246903 São Paulo, Brazil Univ Vale Paraiba, Inst Pesquisa Desenvolvimento, BR-12244000 Sao Jose Dos Campos, SP, Brazil Univ São Paulo, Inst Biociencias, Centro Estudios Genoma Humano, Dept Biol, BR-055508900 São Paulo, Brazil Univ Fed São Paulo, Lab Mol Gynecol, Dept Gynecol, BR-04039001 São Paulo, Brazil São Paulo State Univ, Sch Pharmacol, Dept Biol Sci, BR-14801902 Araraquara, SP, Brazil Univ São Paulo, Discipl Genet, Fac Odontol, BR-14040900 Ribeirao Preto, SP, Brazil Univ Fed São Paulo, Dept Biofisica, BR-04023062 São Paulo, Brazil Univ Estadual Paulista, Dept Biol, Inst Biociencias Letras Ciências Exatas, BR-15054000 Sao Jose do Rio Preto, SP, Brazil Univ Estadual Paulista, Dept Genet, Inst Biociencias, BR-18618000 Botucatu, SP, Brazil Univ São Paulo, Dept Bioquim Imunol, Fac Med Ribeirao Preto, BR-14049900 Ribeirao Preto, SP, Brazil Univ Estadual Campinas, Lab Genet Mol Cancer, Dept Clin Med, Fac Ciências Med, BR-13083970 Campinas, SP, Brazil Univ Estadual Campinas, Dept Patol Clin, Fac Ciências Med, BR-13083970 Campinas, SP, Brazil Univ São Paulo, Setor Vetores Virals, Lab Cardiol Mol, Inst Coracao, BR-15403000 São Paulo, Brazil Univ Estadual Paulista, Lab Biol Mol, Hemocentro, Fac Med, BR-18618970 Botucatu, SP, Brazil São Paulo State Univ, Sch Pharmacol, Dept Biol Sci, BR-14801902 Araraquara, SP, Brazil Univ Estadual Paulista, Dept Biol, Inst Biociencias Letras Ciências Exatas, BR-15054000 Sao Jose do Rio Preto, SP, Brazil Univ Estadual Paulista, Dept Genet, Inst Biociencias, BR-18618000 Botucatu, SP, Brazil

Submitted by Guilherme Lemeszenski (guilherme@nead.unesp.br) on 2014-02-26T17:18:53Z No. of bitstreams: 1 WOS000220363200029.pdf: 614453 bytes, checksum: 0bea3ab6505bdc83189b3db61b9a4c2a (MD5) Made available in DSpace on 2014-02-26T17:18:53Z (GMT). No. of bitstreams: 1 WOS000220363200029.pdf: 614453 bytes, checksum: 0bea3ab6505bdc83189b3db61b9a4c2a (MD5) Previous issue date: 2004-04-01 Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-20T15:29:48Z No. of bitstreams: 1 WOS000220363200029.pdf: 614453 bytes, checksum: 0bea3ab6505bdc83189b3db61b9a4c2a (MD5) Made available in DSpace on 2014-05-20T15:29:48Z (GMT). No. of bitstreams: 1 WOS000220363200029.pdf: 614453 bytes, checksum: 0bea3ab6505bdc83189b3db61b9a4c2a (MD5) Previous issue date: 2004-04-01 Leptospira species colonize a significant proportion of rodent populations worldwide and produce life-threatening infections in accidental hosts, including humans. Complete genome sequencing of Leptospira interrogans serovar Copenhageni and comparative analysis with the available Leptospira interrogans serovar Lai genome reveal that despite overall genetic similarity there are significant structural differences, including a large chromosomal inversion and extensive variation in the number and distribution of insertion sequence elements. Genome sequence analysis elucidates many of the novel aspects of leptospiral physiology relating to energy metabolism, oxygen tolerance, two-component signal transduction systems, and mechanisms of pathogenesis. A broad array of transcriptional regulation proteins and two new families of afimbrial adhesins which contribute to host tissue colonization in the early steps of infection were identified. Differences in genes involved in the biosynthesis of lipopolysaccharide 0 side chains between the Copenhageni and Lai serovars were identified, offering an important starting point for the elucidation of the organism's complex polysaccharide surface antigens. Differences in adhesins and in lipopolysaccharide might be associated with the adaptation of serovars Copenhageni and Lai to different animal hosts. Hundreds of genes encoding surface-exposed lipoproteins and transmembrane outer membrane proteins were identified as candidates for development of vaccines for the prevention of leptospirosis. Univ São Paulo, Ctr Biotechnol, Inst Butantan, BR-05503900 São Paulo, SP, Brazil Univ São Paulo, Inst Quim, São Paulo, Brazil Univ São Paulo, Inst Ciências Biomed, BR-05508 São Paulo, Brazil Univ São Paulo, Inst Biociencias, São Paulo, Brazil Univ São Paulo, Inst Biol, São Paulo, Brazil Univ São Paulo, Piracicaba, Brazil Univ Estadual Paulista, Fac Ciências Agrarias & Vet, Jaboticabal, Brazil Univ Fed Sao Carlos, Ctr Ciências Agrarias, Araras, Brazil Fac Ciências Farmaceut, Ribeirao Preto, Brazil Fac Filosofia Ciências & Letras, Ribeirao Preto, Brazil Univ São Paulo, BR-14049 Ribeirao Preto, Brazil Univ Mogi Cruzes, Fac Ciências Agron, Mogi Cruzes, Brazil Univ Mogi Cruzes, Inst Biociencias, Mogi Cruzes, Brazil Univ Estadual Paulista, Botucatu, SP, Brazil Univ Mogi Cruzes, Nucleo Integrado Biotecnol, Mogi Das Cruzes, Brazil Univ Estadual Paulista, Dept Genet & Evolucao, Campinas, SP, Brazil Univ Estadual Paulista, Inst Computacao, Campinas, SP, Brazil Univ Estadual Paulista, Inst Agron Campinas, Campinas, SP, Brazil Univ Estadual Paulista, Ctr Biol Mol & Engn Genet, Campinas, SP, Brazil Univ Fed Bahia, Serv Immunol, BR-41170290 Salvador, BA, Brazil UEFS, LAPEM, Dept Ciências Biol, Feira de Santana, BA, Brazil Inst Oswaldo Cruz, Dept Bioquim & Biol Mol, BR-20001 Rio de Janeiro, Brazil Univ Fed Pelotas, Ctr Biotechnol, Pelotas, Brazil Univ Fed Rio Grande Norte, Ctr Biociencias, BR-59072970 Natal, RN, Brazil Cornell Univ, Weill Med Coll, Div Int Med & Infect Dis, New York, NY USA Ludwig Inst Canc Res, New York, NY USA Univ Calif Los Angeles, Sch Med, Los Angeles, CA USA Vet Affairs Greater Los Angeles Healthcare Syst, Div Infect Dis, Los Angeles, CA USA Koninklin Inst Tropen, Royal Trop Inst, KIT Biomed Res, Amsterdam, Netherlands Univ Estadual Paulista, Fac Ciências Agrarias & Vet, Jaboticabal, Brazil Univ Estadual Paulista, Botucatu, SP, Brazil Univ Estadual Paulista, Dept Genet & Evolucao, Campinas, SP, Brazil Univ Estadual Paulista, Inst Computacao, Campinas, SP, Brazil Univ Estadual Paulista, Inst Agron Campinas, Campinas, SP, Brazil Univ Estadual Paulista, Ctr Biol Mol & Engn Genet, Campinas, SP, Brazil

Purpose: the physiologic role of the cellular prion protein (PrPc) is unknown. Mice devoid of PrPc develop normally and show only minor deficits. However, electrophysiologic and histologic alterations found in these mice suggest a possible:role fur PrPc in seizure threshold and/or epilepsy.Methods: We tested the sensitivity of PrPc knockout mice to seizures induced by single convulsant or repeated subconvulsant (kindling) doses of pentylenetetrazol (PTZ), and to status epilepticus (SE) induced by kainic acid or pilocarpine.Results. in PTZ kindling, seizure severity progressed faster in the PrPc knockout group, in which 92.8% reached stage 5 or death after 4 days of stimulation, as opposed to 38.4% in wildtype animals: After 10 injections, mortality was 85.7% among knockouts and 15.3% among controls. After a single PTZ injection (60 mg/kg), overall mortality due to seizures was 91% in knockout mice, but only 33% among wild-type animals. Pilocarpine-induced SE (320 mg/kg) caused an 86.7% mortality in knockouts, as opposed to 40% in wild-type animals. Finally, after kainic acid injections (10 mg/kg), 70% of the knockouts developed at least one severe seizure, and 50% showed repetitive seizures, whereas no wild-type animal exhibited observable seizures.Conclusions: Animals lacking cellular prion protein expression are more susceptible to seizures induced by various convulsant agents. This is perhaps the most striking alteration yet found in PrPc-null mice, who at first analysis appeared to be completely normal. A possible role for PrPc in chronic and idiopathic (familial), secondary, or cryptogenic epilepsies in humans remains to be investigated. Univ Fed Rio Grande Sul, ICBS, Dept Bioquim, Ctr Memoria, BR-90035003 Porto Alegre, RS, Brazil UNIFESP, EPM, Dept Expt Neurol, São Paulo, Brazil Ludwig Inst Canc Res, São Paulo, Brazil UNIFESP, EPM, Dept Expt Neurol, São Paulo, Brazil Web of Science

Sialidases (EC 3.2.1.18) are commonly found in viruses, bacteria, fungi, protozoa, and vertebrates, but not in invertebrates. We have previously reported the presence of a new siaIidase activity in the gut of exclusively hematophagous insects of the Triatoma genus, which transmit Chagas' disease (Amino, R., Acosta, A, Morita, O. M., Chioccola, V. L. P., and Schenkman, S. (1995) Glycobiology 5, 625-631). Here we show that this sialidase is present in the salivary gland of Triatoma infestans, and it is released with the saliva during the insect bite. the sialidase was purified to homogeneity (>5000 times) to a specific activity of more than 20 units/mg. It elutes from a gel filtration column with a volume corresponding to the size of 33 kDa, and it migrates as a single 26-kDa band in SDS-polyacrylamide gel electrophoresis, which is unusually smaller when compared with other known sialidases. T. infestans sialidase hydrolyzes preferentially alpha 2-->3-linked sialic acids at pH 4-8, with maximaI activity between pH 5.5 and 6.5, which is compatible with the optimal pH of secreted sialidases. the sialidase is competitively inhibited by 2-deoxy-2,3-dehydro-N-acetyl-neuraminic acid (K-i = 0.075 mM) and differently from many sialidases, with exception of Salmonella typhimurium sialidase, it is inhibited competitively by HEPES (K-i = 15 mM). the fact that T. infestans sialidase is released with the saliva and can hydrolyze sialyl-Lewis(x) blood groups, which are the ligands for selectins, suggests that it might have a role in the blood feeding. Universidade Federal de São Paulo, Escola Paulista Med, Dept Microbiol Imunol & Parasitol, BR-04023062 São Paulo, Brazil Universidade Federal de São Paulo, Escola Paulista Med, Dept Histol, BR-04023062 São Paulo, Brazil Ludwig Inst Canc Res, BR-01509010 São Paulo, Brazil Universidade Federal de São Paulo, Escola Paulista Med, Dept Microbiol Imunol & Parasitol, BR-04023062 São Paulo, Brazil Universidade Federal de São Paulo, Escola Paulista Med, Dept Histol, BR-04023062 São Paulo, Brazil Web of Science

Made available in DSpace on 2013-08-12T18:36:36Z (GMT). No. of bitstreams: 0 Previous issue date: 2010-01-21 Made available in DSpace on 2013-09-30T18:22:18Z (GMT). No. of bitstreams: 0 Previous issue date: 2010-01-21 Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-20T13:32:36Z No. of bitstreams: 0 Made available in DSpace on 2014-05-20T13:32:36Z (GMT). No. of bitstreams: 0 Previous issue date: 2010-01-21 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) AIM: To verify the methylation status of CDH1, DAPK, COX2, hMLH1 and CDKN2A genes and to evaluate their association with Helicobacter pylori (H. pylori)-cagA(+) and Epstein Barr virus (EBV) infections in gastric adenocarcinomas.METHODS: Methylation-specific PCR (MSP) assay was performed in 89 primary gastric carcinomas (intestinal and diffuse types). Microsatellite instability (MSI) analysis was performed using the BAT26 primer set and PCR products were analyzed with the ABI PRISM 3100 Genetic Analyzer using Genescan 3.7 software (Applied Biosystems). Detection of H, pylori and genotyping were performed by PCR, using specific primers for ureaseC and cagA genes. The presence of EBV was assessed by in situ hybridization. Statistical analyses were performed using the chi(2) or Fisher's exact test.RESULTS: The most frequent hypermethylated gene was COX-2 (63.5%) followed by DAPK (55.7%), CDH1 (51%), CDKN2A (36%) and hMLH1 (30.3%). Intestinal and diffuse adenocarcinomas showed different methylation profiles and there was an association between methylation of E-CDH1 and H. pylori-cagA(+) in the intestinal adenocarcinoma type. MSI was correlated with hMLH1 methylation. There was an inverse correlation between DAPK hypermethylation and MSI.CONCLUSION: We found a strong association between CDH1 methylation and H, pylori-cagA+ in intestinal-type gastric cancer, association of MSI and better prognosis and an heterogeneous COX-2 overexpression. (C) 2010 Baishideng. All rights reserved. UNESP São Paulo State Univ, Ctr Blood Transfus, Botucatu Med Sch, BR-18618970 Botucatu, SP, Brazil Universidade Federal do Ceará (UFC), Dept Pathol, BR-60430160 Fortaleza, Ceara, Brazil UNESP São Paulo State Univ, Dept Pathol, Sch Med, BR-18618970 Botucatu, SP, Brazil Ludwig Inst Canc Res, New York, NY 10065 USA Ludwig Inst Canc Res, BR-01323903 São Paulo, Brazil UNESP São Paulo State Univ, Dept Surg Gastroenterol, Sch Med, BR-18618970 Botucatu, SP, Brazil UNESP São Paulo State Univ, Ctr Blood Transfus, Botucatu Med Sch, BR-18618970 Botucatu, SP, Brazil UNESP São Paulo State Univ, Dept Pathol, Sch Med, BR-18618970 Botucatu, SP, Brazil UNESP São Paulo State Univ, Dept Surg Gastroenterol, Sch Med, BR-18618970 Botucatu, SP, Brazil

Theoretical considerations predict that amplification of expressed gene transcripts by reverse transcription-PCR using arbitrarily chosen primers will result in the preferential amplification of the central portion of the transcript. systematic, high-throughput sequencing of such products would result in an expressed sequence tag (EST) database consisting of central, generally coding regions of expressed genes. Such a database would add significant value to existing public EST databases, which consist mostly of sequences derived from the extremities of cDNAs, and facilitate the construction of contigs of transcript sequences. We tested our predictions, creating a database of 10,000 sequences from human breast tumors. The data confirmed the central distribution of the sequences, the significant normalization of the sequence population, the frequent extension of contigs composed of existing human ESTs, and the identification of a series of potentially important homologues of known genes. This approach should make a significant contribution to the early identification of important human genes, the deciphering of the draft human genome sequence currently being compiled, and the shotgun sequencing of the human transcriptome. Ludwig Inst Canc Res, BR-01509010 Sao Paulo, Brazil Univ Sao Paulo, Inst Quim, Dept Bioquim, BR-05599970 Sao Paulo, Brazil Univ Fed Sao Paulo, Escola Paulista Med, BR-04023062 Sao Paulo, Brazil Univ Sao Paulo, Fac Med, Dept Radiol, BR-01296903 Sao Paulo, Brazil Fac Med Ribeirao Preto, Dept Clin Med, BR-14049900 Sao Paulo, Brazil Univ Estadual Campinas, Hemoctr, BR-13089970 Sao Paulo, Brazil Fac Ciencias Farmaceut Ribeirao Preto, Dept Ciencias Farmaceut, BR-14049900 Sao Paulo, Brazil Swiss Inst Bioinformat, CH-1066 Epalinges, Switzerland Swiss Inst Expt Canc Res, CH-1066 Epalinges, Switzerland Ludwig Inst Canc Res, Off Informat Technol, CH-1066 Epalinges, Switzerland UCL, Ludwig Inst Canc Res, London W1P 7LD, England Hosp AC Camargo Fund Antonio Prudente, BR-01509010 Sao Paulo, Brazil Univ Fed Sao Paulo, Escola Paulista Med, BR-04023062 Sao Paulo, Brazil Web of Science