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1.
Biling (Camb Engl) ; 26(5): 942-958, 2023 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-37954660

RESUMO

Code-switching, switching between different languages within the same conversation, is a prominent feature in bilingual communication. This study aimed to elucidate to what extent the linguistic abilities and age of dual-language-learning preschoolers influence the frequency and purposes of code-switching (compensatory, to bridge linguistic gaps; preferential, to express content as fluently as possible; pragmatic, to phrase something appropriately for the situation). Parental code-switching ratings of 101 German/French-Turkish/Italian dual-language learners aged 32-78 months were analyzed. Generalized linear mixed models revealed positive but no negative effects of societal- and heritage-language skills on children's code-switching frequencies independent of switching purposes and with no evidence of age effects. Hence, code-switching across the preschool age mainly reflects high linguistic competences. Models with linguistically and psychometrically parallelized language scores indicated a strong switching tendency toward the societal language when proficiency in both languages is high and away from the societal language when language proficiencies are low.

2.
Nat Commun ; 9(1): 2890, 2018 07 23.
Artigo em Inglês | MEDLINE | ID: mdl-30038302

RESUMO

Many properties of solids result from the fact that in a periodic crystal structure, electronic wave functions are delocalized over many lattice sites. Electrons should become increasingly localized when a strong electric field is applied. So far, this Wannier-Stark regime has been reached only in artificial superlattices. Here we show that extremely transient bias over the few-femtosecond period of phase-stable mid-infrared pulses may localize electrons even in a bulk semiconductor like GaAs. The complicated band structure of a three-dimensional crystal leads to a strong blurring of field-dependent steps in the Wannier-Stark ladder. Only the central step emerges strongly in interband electro-absorption because its energetic position is dictated by the electronic structure at an atomic level and therefore insensitive to the external bias. In this way, we demonstrate an extreme state of matter with potential applications due to e.g., its giant optical non-linearity or extremely high chemical reactivity.

3.
Int J Dent Hyg ; 14(1): 4-14, 2016 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-25619863

RESUMO

OBJECTIVES: Air polishing devices are used as an alternative to traditional instrumentation of the root surface. The objective of the systematic review was to analyse patient perception, that is pain and discomfort during treatment with air polishing devices in periodontal therapy. METHODS: The electronic databases MEDLINE, EMBASE and the Cochrane library were screened for studies published through 18th November 2013. Patient perception served as primary outcome. RESULTS: Of the 1266 abstracts screened, nine studies reporting data on patient perception using a visual analogue scale or a patient interview were included in the analysis. Different air polishing powders consisting of sodium bicarbonate, glycine or erythritol were used. Reported discomfort of non-surgical periodontal therapy was consistently equal or lower when air polishing powders consisting of glycine or erythritol were applied compared with root surface instrumentation using hand instruments or ultrasonic devices. CONCLUSION: Air polishing with powders consisting of glycine seems to be associated with less discomfort during non-surgical periodontal therapy, that is supra- and subgingival air polishing.


Assuntos
Polimento Dentário/métodos , Satisfação do Paciente , Periodontia/métodos , Glicina , Humanos , Pós , Raiz Dentária
4.
Int J Dent Hyg ; 14(1): 15-28, 2016 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-25690301

RESUMO

OBJECTIVES: Air-polishing devices are used for the instrumentation of the root surface. Their potential of harm to the hard and/or soft tissues needs to be considered during periodontal treatment. The objective of this systematic review was to analyse the effects of air polishing devices on oral tissues. METHODS: The electronic databases MEDLINE, EMBASE and the Cochrane Library were screened for studies published through 18 November 2013. The surface modifications on human cementum, dentine or gingiva after the instrumentation were considered as outcomes. RESULTS: Of the 1266 abstracts screened, 17 studies were included in the analysis. Different air polishing powders consisting of sodium bicarbonate, calcium carbonate, pumice or glycine were used in different ex vivo or in vitro settings. Thirteen publications reported data on the effects of air polishing devices on cementum and dentine. Hard tissue modifications, including defect depths and volume, caused by sodium bicarbonate or calcium carbonate powders were significantly greater compared to powders consisting of glycine. The soft tissue modifications using different modes of instrumentation were assessed in four publications. The data demonstrate less potential of harm to the gingiva after spraying with glycine powder compared to sodium bicarbonate powder or instrumentation with curettes. CONCLUSION: Glycine powder air polishing may be safely applied to human root surfaces and gingivae.


Assuntos
Polimento Dentário , Bicarbonato de Sódio , Raiz Dentária , Cemento Dentário , Dentina , Humanos , Pós
5.
Gynecol Obstet Fertil ; 43(4): 329-31, 2015 Apr.
Artigo em Francês | MEDLINE | ID: mdl-25817179

RESUMO

OBJECTIVES: Endometrial cancer is the most common gynecological cancer in France. The therapeutic management is based on preoperative staging. The recommended imaging examination remains the MRI. This is to evaluate ultrasound and MRI in the staging for localized cancers. METHODS: This is a retrospective observational study, conducted from July 2012 to July 2014, at the University Hospital of Nancy, on all patients care for endometrial cancer stage I, who underwent a pelvic ultrasound and MRI for the assessment of myometrial infiltration. RESULTS: Twenty-nine patients were included with a mean age of 69 years and a BMI of 30 kg/m(2). Using ultrasound, we have a sensitivity of 58%, a specificity of 100%, a positive predictive value (PPV) of 100%, a negative predictive value (NPV) of 70% and an accuracy of 75%. Using MRI, we have a sensitivity of 83%, a specificity of 100%, a PPV of 83%, a VPN of 88%, and an accuracy of 86%. CONCLUSION: Transvaginal sonography should be performed before post-menopausal bleeding. It remains possible in the staging of localized cancers.


Assuntos
Neoplasias do Endométrio/diagnóstico por imagem , Neoplasias do Endométrio/patologia , Imageamento por Ressonância Magnética , Estadiamento de Neoplasias/métodos , Idoso , Idoso de 80 Anos ou mais , Feminino , França , Humanos , Pessoa de Meia-Idade , Estudos Retrospectivos , Sensibilidade e Especificidade
6.
J Gynecol Obstet Biol Reprod (Paris) ; 44(7): 679-82, 2015 Sep.
Artigo em Francês | MEDLINE | ID: mdl-25721346

RESUMO

Granulomatous mastitis is a rare disease, often associated with Corynebacterium infection. It raises the problem of diagnosis of breast tumor with a fast evolution and inflammatory character. We report two cases of granulomatous mastitis with Corynebacterium. It concerns the clinical and radiological description, followed by the therapeutic alternatives and future of the patients. The clinical presentation is variable. The treatment consists in a surgical procedure of resection. The medical treatment based of corticosteroids also proves efficient. The association between Corynebacterium presence and this pathology seems frequent and needs a specific bacteriological search.


Assuntos
Infecções por Corynebacterium , Corynebacterium/patogenicidade , Mastite Granulomatosa , Adulto , Infecções por Corynebacterium/complicações , Infecções por Corynebacterium/microbiologia , Infecções por Corynebacterium/terapia , Feminino , Mastite Granulomatosa/etiologia , Mastite Granulomatosa/microbiologia , Mastite Granulomatosa/terapia , Humanos
7.
Vasa ; 39(2): 133-9, 2010 May.
Artigo em Inglês | MEDLINE | ID: mdl-20464668

RESUMO

BACKGROUND: Vascular access patency is of vital importance for patients requiring haemodialysis. This analysis validates potential risk factors and benefits in patients undergoing vascular access procedures. PATIENTS AND METHODS: Vascular access procedures performed over a two-year period were retrospectively analysed. Clinical data and concomitant medication were retrieved from files as were surgical data following a standardized data capture sheet. Outcome parameters were primary (PP) and secondary patency (SP) as well as freedom from repeated revascularization. Minimal follow-up with functioning access was 679 days. RESULTS: During the observation period, 244 patients (mean age 62.2 +/- 0.9 years, 60.7 % male patients, 36.1 % pre-emptive, 31.1 % late referral) underwent vascular accesses procedures. PP and SP were 35.6 % and 45.6 %, respectively, at 540 days. Presence of diabetes mellitus was associated with decreased PP (OR: 0.6, 95 %-CI: 0.3 - 1.0) and SP (OR: 0.4, 95 %-CI: 0.2 - 0.7), whereas female gender was associated with lower SP (OR: 0.6, 95 %-CI: 0.3 - 0.9) and freedom from repeated revascularization rates (OR: 0.6, 95 %-CI: 0.3 - 1.0). In contrast, presence of hyperparathyreoidism was associated with higher SP (OR: 1.7, 95 %-CI: 1.0 - 3.0) and freedom from repeated revascularization (OR: 1.7, 95 %-CI: 1.0 - 3.0) rates. CONCLUSIONS: Haemodialysis access performs worst in patients with diabetes mellitus and in women. The benefit of hyperparathyroidism should be interpreted as hypothesis generating.


Assuntos
Derivação Arteriovenosa Cirúrgica , Oclusão de Enxerto Vascular/etiologia , Falência Renal Crônica/terapia , Diálise Renal , Grau de Desobstrução Vascular , Derivação Arteriovenosa Cirúrgica/efeitos adversos , Implante de Prótese Vascular/efeitos adversos , Distribuição de Qui-Quadrado , Complicações do Diabetes/etiologia , Feminino , Oclusão de Enxerto Vascular/cirurgia , Humanos , Hiperparatireoidismo/complicações , Estimativa de Kaplan-Meier , Modelos Logísticos , Masculino , Pessoa de Meia-Idade , Razão de Chances , Modelos de Riscos Proporcionais , Reoperação , Estudos Retrospectivos , Medição de Risco , Fatores de Risco , Fatores Sexuais , Fatores de Tempo , Resultado do Tratamento
9.
Aquat Toxicol ; 80(2): 101-8, 2006 Nov 16.
Artigo em Inglês | MEDLINE | ID: mdl-16963132

RESUMO

The zebrafish (Danio rerio) is a sensitive non-mammalian model used for studying polycyclic aromatic hydrocarbon (PAH)-induced chemical carcinogenesis. The susceptibility of zebrafish to PAH-induced carcinogenesis may be related to the ability of the zebrafish P450s to bioactivate these procarcinogens. As a part of our overall effort to identify the various P450 enzymes that are involved in the activation and detoxification of PAHs in zebrafish, therefore, we have examined the ability of recombinant zebrafish CYP1A (zCYP1A) expressed in yeast to metabolize BaP in vitro. Comparison studies also were conducted with liver microsomes from beta-naphthoflavone (BNF)-treated rainbow trout (Oncorhynchus mykiss). Results demonstrated that the trout liver microsomes were almost twice as active as zCYP1A in oxidizing BaP, with Vmax values of 1.7 and 0.94 nmol/min/nmol P450 for trout and zebrafish preparations, respectively. Like trout CYP1A1, cDNA-expressed zCYP1A was found to oxidize BaP to phenols, quinones and diols (BaP-7,8-diol and BaP-9,10-diol) in the presence of exogenous human microsomal epoxide hydrolase (hEH). BaP-7,8-diol is the precursor of the ultimate carcinogen, BaP-7,8-diol-9,10-epoxide (BaPDE). The ability of zCYP1A to bioactivate BaP was confirmed by the formation of DNA adducts when calf thymus DNA was added to the incubation mixture. BaP-DNA binding was enhanced by the addition of hEH to the incubation mixture. HPLC analysis of the [33P]-postlabeled DNA adducts showed the formation of at least four adducts mediated by both zCYP1A and trout liver microsomes, and one of these adducts co-migrated with BaPDE-dG in HPLC analysis. The addition of hEH to the incubation mixture decreased the formation of BaPDE-dG by zCYP1A and by trout liver microsomes while increasing the formation of an unidentified DNA adduct in the case of zCYP1A. zCYP1A also mediated the binding of BaP to protein, providing further evidence that this enzyme is capable of oxidizing BaP to reactive metabolites that bind to macromolecules. It thus appears that zCYP1A may play an important role in BaP-induced carcinogenesis in the zebrafish model by catalyzing the sequential formation of the ultimate diol epoxide carcinogenic metabolite of BaP.


Assuntos
Benzo(a)pireno/metabolismo , Sistema Enzimático do Citocromo P-450/metabolismo , Microssomos Hepáticos/metabolismo , Oncorhynchus mykiss/metabolismo , Proteínas Recombinantes/metabolismo , Proteínas de Peixe-Zebra/metabolismo , Animais , Isótopos de Carbono/análise , DNA/metabolismo , Microssomos Hepáticos/efeitos dos fármacos , Isótopos de Fósforo/análise , Ligação Proteica/efeitos dos fármacos , Saccharomyces cerevisiae
10.
Comp Biochem Physiol C Toxicol Pharmacol ; 140(2): 207-19, 2005 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-15907766

RESUMO

A full-length zebrafish (Danio rerio) cytochrome P450 (CYP) 2K6 cDNA, was obtained (GenBank accession No. AF283813) through polymerase chain reaction cloning using degenerated primers based on a consensus CYP2 sequence and the heme-binding domain. This first CYP2K family member cloned from zebrafish had 1861 bp which contained 27 bp of 5'-untranslated region (5'-UTR), an open reading frame (ORF) of 1518 bp, and a 300 bp 3'-UTR with a poly A tail. The deduced 506 amino acid sequence of CYP2K6 had 63%, 62% and 59% identity with rainbow trout CYP2K1, CYP2K4 and CYP2K3, respectively; and 45%, 42%, and 42% identity with rabbit CYP2C1, human CYP2C19 and mouse CYP2C39, respectively. CYP2K6 mapped to 107.49cR on LG3 using the LN54 radiation hybrid panel. Its mRNA was detected at 5 days post-fertilization and in the adult liver and ovary among nine tissues examined. The ORF, including the 27 bp of the 5'-UTR, was cloned into pFastBac donor vector and then transferred into the baculovirus genome (bacmid DNA) in DH10Bac competent cells. The recombinant bacmid DNA was used to infect Spodoptera frugiperda insect cells to express the CYP2K6 protein (Bv-2K6). As its ortholog, rainbow trout Bv-2K1 [Yang, Y.H., Miranda, C.L., Henderson, M.C., Wang-Buhler, J.-L., Buhler, D.R., 2000. Heterologous expression of CYP2K1 and identification of the expressed protein (Bv-2K1) as lauric acid (omega-1)-hydroxylase and aflatoxin B1 exo-epoxidase. Drug Metab. Disp. 28,1279-83.], Bv-2K6 also catalyzed the conversion of aflatoxin B1 (AFB1) to its exo-8,9-epoxide as assessed by the trapping of a glutathione (GSH) adduct in the presence of a specific mouse alpha class glutathione S-transferase. The identity of the AFB1-GSH adduct was verified by liquid chromatography-mass spectrometry (LC-MS) and mass spectrometry-mass spectrometry (MS-MS) analysis. Although rainbow trout Bv-2K1 was capable of oxidizing lauric acid, zebrafish Bv-2K6 protein showed no activity against this substrate.


Assuntos
Aflatoxina B1/metabolismo , Hidrocarboneto de Aril Hidroxilases/genética , Sequência de Aminoácidos , Animais , Hidrocarboneto de Aril Hidroxilases/biossíntese , Baculoviridae , Sequência de Bases , Biotransformação , Cromatografia Líquida de Alta Pressão , Cromatografia Líquida , Clonagem Molecular , Citocromo P-450 CYP4A/metabolismo , Família 2 do Citocromo P450 , Embrião não Mamífero , Proteínas de Peixes/genética , Biblioteca Gênica , Espectrometria de Massas , Dados de Sequência Molecular , Reação em Cadeia da Polimerase , Proteínas Recombinantes/metabolismo , Alinhamento de Sequência , Spodoptera , Esteroide Hidroxilases/genética , Distribuição Tecidual , Peixe-Zebra/crescimento & desenvolvimento
11.
Bioinformatics ; 21(7): 880-8, 2005 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-15539453

RESUMO

MOTIVATION: Annotation of operons in a bacterial genome is an important step in determining an organism's transcriptional regulatory program. While extensive studies of operon structure have been carried out in a few species such as Escherichia coli, fewer resources exist to inform operon prediction in newly sequenced genomes. In particular, many extant operon finders require a large body of training examples to learn the properties of operons in the target organism. For newly sequenced genomes, such examples are generally not available; moreover, a model of operons trained on one species may not reflect the properties of other, distantly related organisms. We encountered these issues in the course of predicting operons in the genome of Bacteroides thetaiotaomicron (B.theta), a common anaerobe that is a prominent component of the normal adult human intestinal microbial community. RESULTS: We describe an operon predictor designed to work without extensive training data. We rely on a small set of a priori assumptions about the properties of the genome being annotated that permit estimation of the probability that two adjacent genes lie in a common operon. Predictions integrate several sources of information, including intergenic distance, common functional annotation and a novel formulation of conserved gene order. We validate our predictor both on the known operons of E.coli and on the genome of B.theta, using expression data to evaluate our predictions in the latter.


Assuntos
Algoritmos , Inteligência Artificial , Bacteroides/genética , Mapeamento Cromossômico/métodos , Escherichia coli/genética , Óperon/genética , Reconhecimento Automatizado de Padrão/métodos , Genoma Bacteriano , Modelos Genéticos , Modelos Estatísticos , Software
12.
Pac Symp Biocomput ; : 5-16, 2004.
Artigo em Inglês | MEDLINE | ID: mdl-14992488

RESUMO

We propose an assay to detect and quantify alternative splicing simultaneously for numerous genes in a pool of cellular mRNA. The assay exploits polymerase colonies, a recently developed method for sampling and amplifying large numbers of individual transcript molecules into discrete spots on a gel. The proposed assay combines the advantages of microarrays for transcript quantitation with the sensitivity and precision of methods based on counting single transcript molecules. Given a collection of spots s(i), each containing an unknown splice variant of some known gene G(i), we design a series of hybridizations to short oligonucleotide probes to determine in parallel which exons of G(i) are present in every spot s(i). We give algorithms to minimize the cost of such designs.


Assuntos
Processamento Alternativo , Biologia Computacional , Técnicas Genéticas , Éxons , Etiquetas de Sequências Expressas , Perfilação da Expressão Gênica , Análise de Sequência com Séries de Oligonucleotídeos , RNA Mensageiro/genética
13.
Bioinformatics ; 19 Suppl 1: i122-9, 2003.
Artigo em Inglês | MEDLINE | ID: mdl-12855448

RESUMO

MOTIVATION: Comparing two protein databases is a fundamental task in biosequence annotation. Given two databases, one must find all pairs of proteins that align with high score under a biologically meaningful substitution score matrix, such as a BLOSUM matrix (Henikoff and Henikoff, 1992). Distance-based approaches to this problem map each peptide in the database to a point in a metric space, such that peptides aligning with higher scores are mapped to closer points. Many techniques exist to discover close pairs of points in a metric space efficiently, but the challenge in applying this work to proteomic comparison is to find a distance mapping that accurately encodes all the distinctions among residue pairs made by a proteomic score matrix. Buhler (2002) proposed one such mapping but found that it led to a relatively inefficient algorithm for protein-protein comparison. RESULTS: This work proposes a new distance mapping for peptides under the BLOSUM matrices that permits more efficient similarity search. We first propose a new distance function on peptides derived from a given score matrix. We then show how to map peptides to bit vectors such that the distance between any two peptides is closely approximated by the Hamming distance (i.e. number of mismatches) between their corresponding bit vectors. We combine these two results with the LSH-ALL-PAIRS-SIM algorithm of Buhler (2002) to produce an improved distance-based algorithm for proteomic comparison. An initial implementation of the improved algorithm exhibits sensitivity within 5% of that of the original LSH-ALL-PAIRS-SIM, while running up to eight times faster.


Assuntos
Algoritmos , Bases de Dados de Proteínas , Proteínas/química , Proteoma/química , Alinhamento de Sequência/métodos , Análise de Sequência de Proteína/métodos , Sequência de Aminoácidos , Sequência Conservada , Dados de Sequência Molecular , Proteínas/classificação , Proteoma/classificação , Homologia de Sequência de Aminoácidos
14.
Vasa ; 32(1): 10-3, 2003 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-12677758

RESUMO

BACKGROUND: Proliferation and migration of vascular smooth muscle cells (SMCs) mark the key processes in the development of bypass graft disease and during neointima formation in restenosis after angioplasty. Growth factors are potent SMC mitogens as they are involved in SMC proliferation and in extracellular matrix (ECM) synthesis. Based on these premises, we examined the effect of the proliferation inhibitor rapamycin in human SMC culture and in a rabbit vascular injury model. MATERIALS AND METHODS: Injection of rapamycin or its vehicle was performed with an infusion-balloon catheter directly into the vessel wall during vascular injury. The intima/media ratio was determined histologically whereas the protein expression was analysed using the powerful two-dimensional gel electrophoresis (2D page) technique. Inhibition of proliferation after rapamycin application was estimated in a human SMC culture for time and dose dependent effects. RESULTS: Rapamycin treatment resulted in a significant reduction of intima media ratio compared to vehicle treated animals after three weeks (0.65 +/- 0.1 vs. 1.2 +/- 0.2 intima-media-ratio, p < 0.05). 2D electrophoresis analysis proved increased ECM synthesis following angioplasty (i.e., lamin, vimentin) in vehicle treated animals. Local rapamycin administration resulted in profound reduction of ECM synthesis after vascular injury. In in-vitro experiments exposure of cultured human SMCs to rapamycin resulted in a significant and dose-dependent (1 nm-100 nm) reduction of human smooth muscle cell proliferation measured by cell counting. CONCLUSION: These above mentioned results suggest that protein synthesis in addition to reduction of cellular proliferation plays an important role following vascular injury, since application of rapamycin resulted in the reduction of SMC proliferation and ECM-synthesis.


Assuntos
Divisão Celular/efeitos dos fármacos , Displasia Fibromuscular/patologia , Músculo Liso Vascular/efeitos dos fármacos , Sirolimo/farmacologia , Túnica Média/efeitos dos fármacos , Angioplastia com Balão , Animais , Movimento Celular/efeitos dos fármacos , Células Cultivadas/efeitos dos fármacos , Células Cultivadas/patologia , Relação Dose-Resposta a Droga , Eletroforese em Gel Bidimensional , Humanos , Masculino , Proteínas Musculares/biossíntese , Músculo Liso Vascular/patologia , Coelhos , Túnica Média/patologia
16.
Toxicol Appl Pharmacol ; 177(2): 94-102, 2001 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-11740908

RESUMO

In mammals the cytochrome P450 3A (CYP3A) subfamily isoforms are primarily expressed in liver and intestines with lesser amounts found in other tissues. The aim of this study was to examine the cellular localization and the expression pattern of CYP3A27 in the gastrointestinal tract (GI tract) of a freshwater teleost species, the rainbow trout (Oncorhynchus mykiss), a fish model used extensively for toxicological and carcinogenesis research. Using an avidin biotinylated enzyme complex and 3,3'-diaminobenzidine staining, strong cytoplasmic immunohistochemical staining was observed for CYP3A27 protein in hepatocytes and in enterocytes of the intestinal ceca and the proximal descending intestine when probed with a polyclonal antibody raised against rainbow trout P450 LMC5, a CYP3A protein. The intensity of epithelial staining decreased distally along the GI tract with faint staining observed in the epithelial cells examined near the vent. Western blot analysis was supportive of the immunohistochemistry results. Northern blot analysis also demonstrated that CYP3A27 mRNA was expressed along the entire GI tract. The major area of CYP3A27 mRNA expression was in the intestinal ceca, followed by the proximal descending intestine, at levels that were about three- to five-fold and two- to four-fold, respectively, greater than seen in the liver of the fish studied. Monooxygenase activities of intestinal ceca microsomes against testosterone and progesterone confirmed the presence of active CYP3A enzyme in this tissue. These results suggest that the intestine of rainbow trout may possesses substantial capacity for first-pass metabolism of xenobiotics by CYP3A27, which makes it an excellent model in which to study the consequence of such metabolism.


Assuntos
Hidrocarboneto de Aril Hidroxilases , Sistema Enzimático do Citocromo P-450/metabolismo , Sistema Digestório/enzimologia , Oncorhynchus mykiss/metabolismo , Oxirredutases N-Desmetilantes/metabolismo , Animais , Northern Blotting , Western Blotting , Ceco/enzimologia , Citocromo P-450 CYP3A , Sistema Enzimático do Citocromo P-450/biossíntese , Sistema Enzimático do Citocromo P-450/genética , Sistema Digestório/metabolismo , Feminino , Regulação Enzimológica da Expressão Gênica , Hidroxilação , Imuno-Histoquímica , Isoenzimas/biossíntese , Isoenzimas/genética , Isoenzimas/metabolismo , Masculino , Microssomos Hepáticos/enzimologia , Oxirredutases N-Desmetilantes/biossíntese , Oxirredutases N-Desmetilantes/genética , Progesterona/metabolismo , RNA Mensageiro/biossíntese , RNA Mensageiro/genética , Testosterona/metabolismo
17.
Bioinformatics ; 17(5): 419-28, 2001 May.
Artigo em Inglês | MEDLINE | ID: mdl-11331236

RESUMO

MOTIVATION: Comparison of multimegabase genomic DNA sequences is a popular technique for finding and annotating conserved genome features. Performing such comparisons entails finding many short local alignments between sequences up to tens of megabases in length. To process such long sequences efficiently, existing algorithms find alignments by expanding around short runs of matching bases with no substitutions or other differences. Unfortunately, exact matches that are short enough to occur often in significant alignments also occur frequently by chance in the background sequence. Thus, these algorithms must trade off between efficiency and sensitivity to features without long exact matches. RESULTS: We introduce a new algorithm, LSH-ALL-PAIRS, to find ungapped local alignments in genomic sequence with up to a specified fraction of substitutions. The length and substitution rate of these alignments can be chosen so that they appear frequently in significant similarities yet still remain rare in the background sequence. The algorithm finds ungapped alignments efficiently using a randomized search technique, locality-sensitive hashing. We have found LSH-ALL-PAIRS to be both efficient and sensitive for finding local similarities with as little as 63% identity in mammalian genomic sequences up to tens of megabases in length


Assuntos
Algoritmos , Alinhamento de Sequência/estatística & dados numéricos , Animais , Cromossomos Humanos Par 22/genética , Biologia Computacional , DNA/genética , Bases de Dados Factuais , Globinas/genética , Humanos , Região de Controle de Locus Gênico , Camundongos , Receptores de Antígenos de Linfócitos T alfa-beta/genética , Software
18.
Science ; 292(5518): 929-34, 2001 May 04.
Artigo em Inglês | MEDLINE | ID: mdl-11340206

RESUMO

We demonstrate an integrated approach to build, test, and refine a model of a cellular pathway, in which perturbations to critical pathway components are analyzed using DNA microarrays, quantitative proteomics, and databases of known physical interactions. Using this approach, we identify 997 messenger RNAs responding to 20 systematic perturbations of the yeast galactose-utilization pathway, provide evidence that approximately 15 of 289 detected proteins are regulated posttranscriptionally, and identify explicit physical interactions governing the cellular response to each perturbation. We refine the model through further iterations of perturbation and global measurements, suggesting hypotheses about the regulation of galactose utilization and physical interactions between this and a variety of other metabolic pathways.


Assuntos
Galactose/metabolismo , Perfilação da Expressão Gênica , Genoma Fúngico , Proteoma , Proteínas de Saccharomyces cerevisiae , Saccharomyces cerevisiae/metabolismo , Biologia Computacional , Meios de Cultura , Bases de Dados Factuais , Proteínas Fúngicas/metabolismo , Galactosefosfatos/metabolismo , Regulação Fúngica da Expressão Gênica , Modelos Biológicos , Modelos Genéticos , Proteínas de Transporte de Monossacarídeos/metabolismo , Mutação , Análise de Sequência com Séries de Oligonucleotídeos , RNA Fúngico/genética , RNA Fúngico/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Saccharomyces cerevisiae/genética
19.
J Biochem Mol Toxicol ; 15(5): 243-55, 2001.
Artigo em Inglês | MEDLINE | ID: mdl-11835621

RESUMO

A 2,037 bp CYP1A1 cDNA (GenBank AF072899) was cloned through screening of a lambdaZipLox cDNA library constructed from the liver of a leaping mullet (Liza saliens) fish captured from Izmir Bay on the Aegean coast of Turkey using rainbow trout CYP1A1 cDNA as a probe. This clone has a 130 bp 5'-flanking region, a 1,563 bp open reading frame (ORF) encoding a 521-amino acid protein (58,972 Da), and a 344 bp 3'-untranslated region without a poly (A) tail. Alignment of the deduced amino acids of CYP1A1 cDNAs showed 58% and 69-96% identities with human and 12 other fish species, respectively. Southern blot analysis suggested that this CYP1A1 cDNA was from a single-copy gene. Based on the comparison with CYP1A1 genes reported for fish and mammals, the leaping mullet CYP1A1 gene is probably split into 7 exons. The intron insertion sites were predicted. Alignment of the CYP1A1 cDNA encoded amino acids from 13 fish and 7 mammalian species disclosed differences in highly conserved amino acids between aquatic and land vertebrates. The possible associated secondary structure; conserved motifs and substrate-binding sites were discussed. The phylogenetic relationships of CYP1A1s among 13 fish species were analyzed by a distance method.


Assuntos
Aminoácidos/metabolismo , Citocromo P-450 CYP1A1/genética , Fígado/metabolismo , Smegmamorpha/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Northern Blotting , Southern Blotting , Clonagem Molecular , Citocromo P-450 CYP1A1/química , Citocromo P-450 CYP1A1/metabolismo , DNA Complementar , Humanos , Dados de Sequência Molecular , Filogenia , Estrutura Secundária de Proteína , Homologia de Sequência de Aminoácidos
20.
Toxicol Appl Pharmacol ; 168(2): 91-101, 2000 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-11032764

RESUMO

There is growing concern that exposure to chemicals in the environment can disrupt the endocrine systems of wildlife and humans, causing reproductive problems or other adverse effects. The expression of many cytochrome P450s (CYPs) is under hormonal control, hence, levels of these enzymes can be affected by exposure to endocrine-disrupting chemicals. Previous research has reported that treatment of fish and other animals with the estrogenic and androgenic hormones 17beta-estradiol (E2) and testosterone (T) alters the P450 content or enzyme activities in the treated animals. However, the results of many of these studies are either incomplete or in disagreement and in most cases the effect on specific P450 forms has not been determined. Therefore, to better understand the effects of gonadal hormones on the expression of P450s and their associated enzyme activities, it was of interest to undertake a comprehensive investigation of the transcriptional and translational expression of three constitutive hepatic P450s in the rainbow trout (Oncorhynchus mykiss) following hormone exposure. Accordingly, juvenile trout were injected intraperitoneally with propylene glycol vehicle and the most active estrogenic and androgenic hormones E2 (3 mg/kg) or T (3 mg/kg) on days 1, 4, 7, 13, and 15 and euthanized on day 19. After treatment with E2, hepatic microsomes showed significantly lower levels (percentage of control) in total P450 contents (52%), lauric acid hydroxylase (32%), and 6beta-progesterone hydroxylase activities (27%), [(3)H]aflatoxin-DNA binding (31%), and the protein levels of individual cytochrome P450s (CYPs) LMC1 (CYP2M1), LMC2, (CYP2K1), and LMC5 (CYP3A27) (average for three isoforms a reduction to 29% of control values) with only minor differences between sexes. Treatment with T had either no effect or resulted in small increases in total P450 in males (42%), in lauric acid hydroxylase in females (24%), and in 6beta-progesterone hydroxylase activity in males (21%). Biological variabilities among fish were high and a polymorphic or new LMC2-like form was detected at about 52 kDa in some liver microsomal samples after exposure of fish to either hormone. Female liver RNAs were analyzed through Northern blots and an average decrease of 94% in CYP2 M1, CYP2K1, and CYP3A27 mRNA levels occurred in the E2-treated trout. In livers from T-treated trout, the changes of mRNA levels of CYP2M1 and CYP3A27 were negligible, but CYP2K1 mRNA level decreased by about 60%. Additional CYP2K1 cDNA hybridizable mRNAs were seen in some fish as faint bands at about 2.8 kb for both hormone treatments. Results of this study, therefore, indicated that E2 down-regulated while T produced small but variable effects on the hepatic mRNA/protein levels of CYP2K1, CYP2M1, and CYP3A27 in juvenile rainbow trout. This study, therefore, suggests that exposure of fish and other wildlife to environmental endocrine disruptors, especially estrogen mimics, can adversely affect a number of physiological processes through mechanisms involving altered levels of expression of specific P450 isozymes.


Assuntos
Hidrocarboneto de Aril Hidroxilases , Sistema Enzimático do Citocromo P-450/metabolismo , Estradiol/toxicidade , Proteínas de Peixes , Hormônios Esteroides Gonadais/toxicidade , Microssomos Hepáticos/efeitos dos fármacos , Oncorhynchus mykiss/metabolismo , RNA Mensageiro/metabolismo , Testosterona/toxicidade , Aflatoxina B1/biossíntese , Aflatoxina B1/metabolismo , Aflatoxina B1/toxicidade , Animais , Northern Blotting , Western Blotting , Catálise/efeitos dos fármacos , Sistema Enzimático do Citocromo P-450/biossíntese , Sistema Enzimático do Citocromo P-450/genética , Família 2 do Citocromo P450 , DNA/metabolismo , Adutos de DNA/biossíntese , Eletroforese em Gel de Poliacrilamida , Feminino , Isoenzimas/biossíntese , Isoenzimas/genética , Isoenzimas/metabolismo , Masculino , Microssomos Hepáticos/enzimologia , Microssomos Hepáticos/metabolismo , Oxigenases de Função Mista/biossíntese , Oxigenases de Função Mista/genética , Oxigenases de Função Mista/metabolismo , Biossíntese de Proteínas/efeitos dos fármacos , RNA Mensageiro/genética , Esteroide Hidroxilases/biossíntese , Esteroide Hidroxilases/genética , Esteroide Hidroxilases/metabolismo , Transcrição Gênica/efeitos dos fármacos
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