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1.
Opt Express ; 30(6): 9035-9052, 2022 Mar 14.
Artigo em Inglês | MEDLINE | ID: mdl-35299342

RESUMO

Brightness is one of the most important perceptual correlates of color appearance models (CAMs) when self-luminous stimuli are targeted. However, the vast majority of existing CAMs adopt the presence of a uniform background surrounding the stimulus, which severely limits their practical application in lighting. In this paper, a study on the brightness perception of a neutral circular stimulus surrounded by a non-uniform background consisting of a neutral ring-shaped luminous area and a dark surround is presented. The ring-shaped luminous area is presented with 3 thicknesses (0.33 cm, 0.67 cm and 1.00 cm), at 4 angular distances to the edge of the central stimulus (1.2°, 6.4°, 11.3° and 16.1°) and at 3 luminance levels (90 cd/m2, 335 cd/m2, 1200 cd/m2). In line with the literature, the results of the visual matching experiments show that the perceived brightness decreases in presence of a ring and the effect is maximal at the highest luminance of the ring, for the largest thickness and at the closest distance. Based on the observed results, an image-based model inspired by the physiology of the retina is proposed. The model includes the calculation of cone-fundamental weighted spectral radiance, scattering in the eye, cone compression and receptive field post-receptor organization. The wide receptive field assures an adaptive shift determined by both the adaptation to the stimulus and to the background. It is shown that the model performs well in predicting the matching experiments, including the impact of the thickness, the distance and the intensity of the ring, showing its potential to become the basic framework of a Lighting Appearance Model.


Assuntos
Luz , Visão Ocular , Iluminação , Retina/fisiologia , Percepção Visual/fisiologia
2.
Opt Express ; 27(10): 14423-14436, 2019 May 13.
Artigo em Inglês | MEDLINE | ID: mdl-31163892

RESUMO

Recently, a color appearance model, CAM18sl, has been published. The model can predict the color perception (hue, colorfulness, brightness) of self-luminous stimuli seen against neutral backgrounds varying in luminance. In this paper, the applicability of CAM18sl to situations different from the one for which it was originally developed is explored. The brightness perception predicted by this model (expressed in "bright") is compared with the outcome of the CIE Unified Glare Rating (UGR) for luminaires, the limiting luminance values for Variable Message Signs as indicated in the European standard for outdoor situations, the visual gloss (VG) formula to characterize the gloss of objects, the CIE grey-scale calculation for self-luminous devices, and the predicted brightness value of illuminated objects according to CIECAM02. Although the application domains mentioned above are very distinct, it appears that the predicted brightness perception of the CAM18sl model correlates well with the outcome of the particular and dedicated metrics. It seems that, for brightness perception, CAM18sl can be considered as a general model applicable for a wide range of applications. This could lead to a reduction of a number of application-specific metrics.

3.
J Opt Soc Am A Opt Image Sci Vis ; 35(12): 2000-2009, 2018 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-30645289

RESUMO

A model for brightness and hue perception of self-luminous stimuli surrounded by a self-luminous achromatic background has been developed based on a series of visual experiments. In the model, only the absolute spectral radiance values of the stimulus and background are used as input. Normalized cone excitations are calculated using the 10° Commission Internationale de l'Éclairage (CIE) 2006 cone fundamentals. A von Kries chromatic adaptation transform applied in the CIE 2006 cone space is adopted, and luminance compression and adaptation due to the self-luminous background are included by using a Michaelis-Menten function. Model parameters are determined by fitting the model to the experimental visual data obtained for brightness, hue, and the amount of color versus neutral. The model is validated with additional experimental data. An absolute brightness scale expressed in "bright" is proposed.

4.
ESMO Open ; 9(2): 102235, 2024 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-38320429

RESUMO

BACKGROUND: The use of circulating tumor DNA (ctDNA) concentration for metastatic cancer surveillance is promising, but uncertainty remains about cut-offs with clinical validity. MATERIALS AND METHODS: This observational study recruited 136 subjects with advanced metastatic breast cancer (irrespective of ERBB2/hormone receptor status) for sequencing of their primary tumor in search for PIK3CA hotspot variants amenable for monitoring by droplet digital PCR (ddPCR). The study analyzed 341 on-treatment samples from 19 patients with PIK3CA variants H1047R or E545K enrolled for long-term (median 85 weeks, range 13-125 weeks), frequent (every 3-5 weeks, median of 14 time points per subject, range 2-29) blood sampling for ctDNA quantification by ddPCR, orthogonally validated by deep sequencing. The diagnostic accuracy of ctDNA versus cancer antigen 15-3 (CA15-3) concentrations to predict disease progression within 12 weeks was investigated using receiver operating characteristic (ROC) analysis. Likelihood ratios were used for rational selection of ctDNA result intervals. RESULTS: ctDNA [area under the ROC curve (AUC) 0.848, 95% confidence interval (CI) 0.791-0.895] showed superior diagnostic performance than CA15-3 (AUC 0.670, 95% CI 0.601-0.735, P < 0.001) to predict clinical progression within 12 weeks. ctDNA levels below 10 mutant allele copies/ml had high negative predictive value (88%), while levels above 100 copies/ml detected 64% of progressions 10 weeks earlier versus standard of care. Logistic regression analysis indicated complementary value of ctDNA and the presence of two consecutive CA15-3 rises, resulting in a model with 86% (95% CI 74% to 93%) positive predictive value and a clinically meaningful result in 89% of blood draws. CONCLUSIONS: Intensive ctDNA quantification improves metastatic breast cancer surveillance and enables individualized risk-based scheduling of clinical care.


Assuntos
Neoplasias da Mama , DNA Tumoral Circulante , Humanos , Feminino , DNA Tumoral Circulante/genética , Neoplasias da Mama/tratamento farmacológico , Biomarcadores Tumorais/genética , Progressão da Doença , Classe I de Fosfatidilinositol 3-Quinases/genética
5.
Clin Radiol ; 68(3): 317-22, 2013 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-22959851

RESUMO

Different conditions that may lead to enlarged nerves or nerve roots include hereditary motor and sensory neuropathy (HMSN), neurofibromatosis (NF) type 1, chronic inflammatory demyelinating polyradiculoneuropathy (CIDP), and intraneural perineurioma. Differential diagnosis of hypertrophic mono- and polyradiculopathies remains challenging but is important because of different treatments and prognosis. Magnetic resonance imaging (MRI) can identify the hypertrophic nerve segments and guide a fascicular biopsy. A fascicular biopsy will often be necessary for precise diagnosis.


Assuntos
Imageamento por Ressonância Magnética/métodos , Mononeuropatias/diagnóstico , Polineuropatias/diagnóstico , Biópsia , Meios de Contraste , Diagnóstico Diferencial , Humanos , Mononeuropatias/patologia , Polineuropatias/patologia
6.
B-ENT ; 8(2): 127-9, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-22896932

RESUMO

We report a case of Pott's puffy tumour, a subperiosteal abscess of the frontal bone associated with an underlying frontal osteomyelitis, in a 5-year-old boy. Ultrasonography played a crucial role in the diagnosis of our patient, suggesting the presence of a Pott's puffy tumour with epidural abscess by showing a subperiosteal abscess associated with erosion of the frontal bone. Subsequently, the diagnosis of Pott's puffy tumour with epidural abscess was confirmed by contrast-enhanced CT scanning. Prompt neurosurgical intervention with drainage of abscesses and debridement of bone sequestrate, together with prolonged antibiotic therapy, significantly contributes to a favorable outcome.


Assuntos
Tumor de Pott/diagnóstico por imagem , Pré-Escolar , Humanos , Imageamento Tridimensional , Masculino , Tumor de Pott/cirurgia , Tomografia Computadorizada por Raios X , Ultrassonografia
7.
J Anim Physiol Anim Nutr (Berl) ; 95(2): 198-205, 2011 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-20796081

RESUMO

In this study, the effect of dietary antioxidants on the plasma oxidative status of growing birds fed a diet rich in polyunsaturated fatty acids was investigated. One-day-old broilers were fed for 42 days a diet containing 4% linseed oil and supplemented with single plant extracts rich in antioxidants (natural tocopherols, rosemary, grape seed, green tea, tomato) or a combination of some of these plant extracts, in two different total doses (100 and 200 mg product/kg feed). A diet with synthetic antioxidants with and without α-tocopheryl acetate (200 mg/kg feed) were also included. The plasma oxidative status was evaluated measuring the ferric reducing ability of plasma (FRAP), the superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activity. Lipid peroxidation was measured by thiobarbituric acid-reactive substances (TBARS). No significant effect of the dietary treatments was observed for FRAP as well as for TBARS. However, diet affected GSH-Px activity (p = 0.002) and a trend for an effect on SOD activity was observed (p=0.084). A higher GSH-Px activity was found for 200 mg/kg tomato extract and natural α-tocopherol in relation to the corresponding 100 mg/kg treatment, and the lowest GSH-Px activity was measured for the synthetic antioxidants treatment. The lowest and highest SOD activity were found for the 200. and 100 mg/kg treatment with tomato extract respectively. In conclusion, the oxidative status and lipid oxidation of plasma in broilers was not affected by feeding natural antioxidant extracts at the doses in the present study, but some changes in antioxidant enzyme activities were observed, of which the implication remains to be elucidated.


Assuntos
Galinhas/sangue , Galinhas/metabolismo , Dieta/veterinária , Suplementos Nutricionais , Extratos Vegetais/farmacologia , Ração Animal/análise , Fenômenos Fisiológicos da Nutrição Animal , Animais , Relação Dose-Resposta a Droga , Ferro/química , Ferro/metabolismo , Malondialdeído/sangue , Substâncias Reativas com Ácido Tiobarbitúrico/metabolismo
9.
J Dairy Sci ; 93(2): 495-505, 2010 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-20105521

RESUMO

Milk with an increased content of unsaturated fatty acids was obtained by incorporating 60% of extruded linseed into the concentrate of cows. Two groups of Holstein cows (3 animals/group) were fed a concentrate (control or linseed enriched) together with the same roughage diet (ad libitum). After an adaptation period of 3 wk, evening and morning milk samples were collected every 7 d for 3 wk. Milk was decreamed and anhydrous milk fat (AMF) was isolated from the fat fraction by using the Bureau of Dairy Industries method. The objective of this study was to investigate if the crystallization mechanism of milk fat changed when the content of unsaturated fatty acids was increased. Therefore, the crystallization behavior of a milk fat enriched with unsaturated fatty acids was compared with that of a control milk fat. Nonisothermal crystallization was investigated with differential scanning calorimetry, and 1-step and 2-step isothermal crystallization behaviors were investigated using pulsed nuclear magnetic resonance, differential scanning calorimetry, and x-ray diffraction. A higher content of unsaturated fatty acids in AMF resulted in an increased proportion of low melting triglycerides. These triglycerides lowered the solid fat content profile, particularly at refrigerator temperatures. Furthermore, they induced some changes in the crystallization and melting behaviors of milk fat compared with a control AMF, although no fundamental changes in the crystallization mechanism could be revealed. Even though a lower melting point could be observed for milk fat with a higher content of unsaturated fatty acids, a similar degree of supercooling was needed to initiate crystallization, resulting in a shift in onset temperature of crystallization toward lower temperatures. In addition, slower crystallization kinetics were measured, such as a lower nucleation rate and longer induction times, although crystallization occurred in a similar polymorphic crystal lattice. During melting, a shift in offset temperature toward lower temperatures could be observed for the 3 melting fractions of AMF in addition to a higher proportion of low melting triglycerides. These results demonstrate that a higher content of unsaturated fatty acids has some effect on the crystallization behavior of milk fat. This knowledge could be used to produce dairy products of similar or superior quality compared with conventional products by intervening in the production process of dairy products.


Assuntos
Suplementos Nutricionais , Gorduras/química , Linho , Leite/química , Animais , Bovinos , Cristalização , Dieta/veterinária , Ácidos Graxos/análise , Ácidos Graxos Insaturados , Feminino
10.
Gait Posture ; 29(1): 54-8, 2009 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-18760925

RESUMO

The purpose of the current study was to examine spontaneous overground walk-to-run transitions (WRT). For the first time, subjects' WRT was examined during an overground protocol that allowed them to accelerate freely. The overground speed profile prior to reaching the WRT was analysed together with the spatiotemporal characteristics of the actual transition. Nine women (height: 166.4+/-3.5 cm) performed five spontaneous WRT. Speed, step frequency (SF) and step length (SL) of the accelerating walking steps and the transition step were determined. By means of fourth degree polynomials, subjects' spatiotemporal profiles prior to reaching WRT were determined. A step length index (SLI) was used to calculate the contribution of SF and SL to the increase in walking speed. Subjects took on average 5.9+/-0.9 walking steps prior to reaching transition. When speeding up towards the transition to running, subjects chose to accelerate predominantly in the first half of the walking acceleration period, followed by smaller speed increments in the second half. The SLI values indicated that subjects tended to increase walking speed by increasing SL, more than SF, except during the first 20% of the acceleration period. WRT-speed was 2.664+/-0.230 m s(-1), which was higher than in former treadmill studies using a constant acceleration protocol (+/-2.1 m s(-1)). Subjects made a speed jump of 0.417 m s(-1) from the last walking step to the WRT-step. We can conclude that further transition studies studying the interaction between the acceleration and gait transition behaviour are necessary in order to complete the understanding of the transition phenomenon.


Assuntos
Corrida/fisiologia , Caminhada/fisiologia , Aceleração , Adulto , Análise de Variância , Fenômenos Biomecânicos , Feminino , Humanos , Locomoção/fisiologia , Fatores de Tempo
11.
Poult Sci ; 87(8): 1682-8, 2008 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-18648067

RESUMO

Natural tocopherols (TC), rosemary (RO), green tea (GT), grape seed, and tomato extracts were supplemented in single and in combinations at total concentrations of 100 and 200 mg.kg(-1) of feed in a 4% linseed oil-containing diet to investigate the oxidative stability of broiler breast muscle. Supplementation with 300 mg.kg(-)1 of synthetic antioxidants alone and synthetic antioxidants with alpha-tocopheryl acetate at a concentration of 200 mg.kg(-1) (100 IU) feed was used as a control. Fresh patties were prepared and stored under light at 4 degrees C. After freezing for 8 mo and overnight thawing, 3 other patties were prepared and similarly stored under light at 4 degrees C. During display, samples were evaluated for oxidative stability measurements. For lipid oxidation, the treatment with synthetic antioxidants and 200 mg.kg(-1) of alpha-tocopheryl acetate yielded the lowest TBA reactive species (TBARS) values. For TC, grape seed, and tomato extracts, TBARS values for 100 mg.kg(-1) were higher (P < 0.05) than 200 mg.kg(-1) treatments, whereas no differences (P > 0.05) in TBARS values were observed for RO between 100 and 200 mg.kg(-1). In contrast, GT showed higher TBARS values at 200 mg.kg(-1). Administration of combinations of TC, RO, and GT did not reveal synergistic effects but confirmed the increase in TBARS values with increasing doses of GT. No differences (P > 0.05) among the different antioxidant treatments were detected for protein oxidation. The muscle alpha-tocopherol content linearly responded to the feed alpha-tocopherol content and thus there were no indications for a sparing effect on alpha-tocopherol from other antioxidant treatments. In summary, dietary natural antioxidant extracts were less effective than the treatment with synthetic antioxidants combined with alpha-tocopheryl acetate for protecting against oxidation, but there were marked differences between different natural antioxidant extracts.


Assuntos
Antioxidantes/farmacologia , Lipídeos/química , Carne , Proteínas Musculares/química , Músculos Peitorais/efeitos dos fármacos , Extratos Vegetais/farmacologia , alfa-Tocoferol/análogos & derivados , Animais , Galinhas , Masculino , Indústria de Embalagem de Carne/métodos , Oxirredução , Músculos Peitorais/química , Substâncias Reativas com Ácido Tiobarbitúrico/análise , Tocoferóis , alfa-Tocoferol/farmacologia
12.
J Bone Joint Surg Br ; 89(4): 538-41, 2007 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-17463127

RESUMO

A prospective study of serum and urinary ion levels was undertaken in a triathlete who had undergone a metal-on-metal resurfacing arthroplasty of the hip four years previously. The one month study period included the final two weeks of training, the day of the triathlon, and the two weeks immediately post-race. Serum cobalt and chromium levels did not vary significantly throughout this period, including levels recorded on the day after the 11-hour triathlon. Urinary excretion of chromium increased immediately after the race and had returned to pre-race levels six days later. The clinical implications are discussed.


Assuntos
Artroplastia de Quadril , Cromo/farmacocinética , Cobalto/farmacocinética , Prótese de Quadril , Esportes , Ciclismo , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Desenho de Prótese , Corrida , Natação
13.
J Hosp Infect ; 64(3): 264-70, 2006 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-16934365

RESUMO

Important improvements have been made in wound care over the last decade. However, few data are available on the influence that these have outside their intended use. This study aimed to clarify the effects of the use of wound cleansers on bacterial contamination of the immediate surroundings. Little evidence was found from either laboratory or clinical settings that wound-derived micro-organisms become airborne during wound cleansing. Bacterial dispersion around wounds may be attributed to general activity rather than wound cleansing. If simple precautions are taken, risks for personnel and patients in hospitals and consultation rooms during wound cleansing can be minimized.


Assuntos
Anti-Infecciosos Locais/uso terapêutico , Úlcera Cutânea/terapia , Infecção dos Ferimentos/prevenção & controle , Ferimentos e Lesões/terapia , Ar , Poluentes Atmosféricos/isolamento & purificação , Bactérias/crescimento & desenvolvimento , Contagem de Colônia Microbiana , Humanos , Transmissão de Doença Infecciosa do Paciente para o Profissional , Mecânica , Modelos Biológicos , Infecção dos Ferimentos/microbiologia , Infecção dos Ferimentos/transmissão
14.
Meat Sci ; 74(3): 476-86, 2006 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-22063052

RESUMO

The effect of dietary oil (linseed or soybean oil) and antioxidant treatment (α-tocopheryl acetate (AT; 40ppm) versus a cocktail (AOC; 200ppm): α-tocopheryl acetate+rosemary+citric acid+gallic acid) on colour, lipid and protein oxidation of fresh and processed pork was investigated. No effect of oil source on different parameters of oxidation was seen. No effect of antioxidant treatment on colour stability of fresh longissimus thoracis (LT) or cooked cured ham (CCH) was observed. For both antioxidant treatments, lipid oxidation in fresh LT and CCH was well controlled during display. However, lipid oxidation increased significantly in pre-frozen uncured cooked meat under aerobic conditions. No unambiguous effect of antioxidant treatment on protein oxidation was observed. There seemed to be no clear link between colour, protein and lipid oxidation. At the dose used in this study, no additional or synergistic effects of the extra components of the AOC on the different oxidation parameters was found.

15.
Int J Food Microbiol ; 100(1-3): 41-53, 2005 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-15854691

RESUMO

The individual-based approach of the lag phase is gaining interest, especially for pathogens that initially contaminate food products in low amounts. In this paper, the effect of temperature (30, 10, 7, 4 and 2 degrees C) and pH (7.4, 6.1, 5.5, 5.0, 4.7 and 4.4) on the individual cell lag phase of Listeria monocytogenes was examined in a factorial design, using OD measurements. Individual lag phases of about 100 individual cells per condition were examined and calculated using a linear extrapolation method. Generation times were calculated out of the slope. The obtained data were analyzed at three different levels: in a first approach, the mean values were calculated for each set of environmental conditions and compared to predictions made by the USDA's Pathogen Modeling Program (PMP) for analogous growth conditions. The PMP predictions of the generation times were in the same order of magnitude as the obtained data, although a persistent underestimation could be observed. The observed individual cell lag data differed from lag phase predictions by PMP. Possible reasons for this discrepancy are discussed. Secondly, histograms of individual lag phase measurements were constructed for the different temperature-pH combinations. In this way, the influence of both factors on the variability of individual lag phases could be estimated. At low stress levels, most individual cells showed a short lag phase resulting in a compression of the histograms at the zero-lag level, while, at high stress levels, the histograms shifted to longer lag phases with a significant increase in variability. Thirdly, 37 different distribution types were fitted to the datasets to reveal the distributions that fitted best the obtained data. The gamma distribution was preferred at moderate stress levels, while the Weibull distribution was chosen for harsher growth conditions.


Assuntos
Microbiologia de Alimentos , Concentração de Íons de Hidrogênio , Listeria monocytogenes/crescimento & desenvolvimento , Modelos Biológicos , Temperatura , Contagem de Colônia Microbiana , Cinética , Modelos Estatísticos , Valor Preditivo dos Testes
16.
Gene ; 136(1-2): 215-9, 1993 Dec 22.
Artigo em Inglês | MEDLINE | ID: mdl-8294005

RESUMO

We have constructed a cosmid vector, Tropist3, based on the lambda origin double-cos-site vector Lawrist4, which is designed for efficient cloning and mapping of genomic DNA. Tropist3 contains two cloning sites in addition to the HindIII and BamHI sites present in Lawrist4; a SalI site allows cloning of Sau3AI partial digests following partial filling-in of the ends, and a PmlI site is suitable for blunt-end cloning. Both these strategies reduce the chance of co-cloning two inserts. Tropist3 also contains NotI, PacI, SacII and KpnI sites flanking the cloning region; these allow most inserts to be excised cleanly and mapped by partial digestion followed by hybridization with short vector sequences which lie adjacent to the cloning sites. This will also be useful for recloning inserts into different vectors, or for cosmid sequencing projects.


Assuntos
Mapeamento Cromossômico/métodos , Cosmídeos , Vetores Genéticos , Sequência de Bases , Clonagem Molecular/métodos , DNA , Escherichia coli/genética , Dados de Sequência Molecular , Mycobacterium/genética , Oligodesoxirribonucleotídeos , Mapeamento por Restrição
17.
Antivir Ther ; 6(1): 21-39, 2001 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-11417759

RESUMO

HIV drug resistance is one of the major limitations in the successful treatment of HIV-infected patients using currently available antiretroviral combination therapies. When appropriate, drug susceptibility profiles should be taken into consideration in the choice of a specific combination therapy. Guidelines recommending resistance testing in certain circumstances have been issued. Many clinicians have access to resistance testing and will increasingly use these results in their treatment decisions. In this document, we comment on the different methods available, and the relevant issues relating to the clinical application of these tests. Specifically, the following recommendations can be made: (i) genotypic and phenotypic HIV-1 drug resistance analyses can yield complementary information for the clinician. However, insufficient information currently exists as to which approach is preferable in any particular clinical setting; (ii) when HIV-1 drug resistance testing is required, it is recommended that testing be performed on plasma samples obtained before starting, stopping or changing therapy, on samples that have a viral load above the detection limit of the resistance test; (iii) the panel recommends that genotypic and phenotypic HIV-1 drug resistance testing for clinical purposes be performed in a certified laboratory under strict quality control and quality assurance standards; and (iv) the panel recommends that resistance testing laboratories provide clinicians with resistance reports that include a list of drug-related resistance mutations (genotype) and/or a list of drug-related fold resistance values (phenotype), with interpretations of each by an experienced virologist. The interpretation of genotypic and phenotypic analysis is a complex and developing science, and in order to understand HIV-1 drug resistance reports, communication between the requesting clinician and the expert that interpreted the resistance report is recommended.


Assuntos
HIV-1/efeitos dos fármacos , Testes de Sensibilidade Microbiana , Síndrome da Imunodeficiência Adquirida/tratamento farmacológico , Resistência Microbiana a Medicamentos , Seguimentos , Genótipo , Guias como Assunto , HIV-1/genética , Humanos , Testes de Sensibilidade Microbiana/normas , Fenótipo , Controle de Qualidade
18.
J Immunol Methods ; 177(1-2): 243-50, 1994 Dec 28.
Artigo em Inglês | MEDLINE | ID: mdl-7529809

RESUMO

Production of chimeric and multimeric peptides is of interest for the analysis of topographic relationships between T and B cell stimulatory epitopes. Recombinant DNA technology has certain advantages over conventional chemical peptide synthesis for the production of peptide constructs of large size (more than 40 amino acid residues). We describe a methodology which is versatile and independent of the expression vector used because it only relies on the incorporation of appropriate restriction enzyme sites in oligonucleotides. The method was verified using two 20mer sequences from the 38 kDa antigen of M. tuberculosis. Peptide 201-220, containing an antibody binding linear epitope, has been made immunogenic in vivo when combined with T cell stimulatory peptide 350-369 in a chimeric peptide. The results demonstrate that a distinct orientation of the constituent peptides was essential for achieving optimal immunogenicity.


Assuntos
Antígenos de Bactérias/química , Proteínas Recombinantes de Fusão/biossíntese , Sequência de Aminoácidos , Animais , Formação de Anticorpos , Sequência de Bases , Clonagem Molecular/métodos , Epitopos , Feminino , Ativação Linfocitária , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Dados de Sequência Molecular , Mycobacterium tuberculosis/imunologia , Peptídeos/imunologia
19.
Biochem Pharmacol ; 61(10): 1293-303, 2001 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-11322933

RESUMO

In this study, we investigated the combined effects of EGF and collagen I gel on the phenotype of cultured rat hepatocytes and we focussed our investigations on the regulation of xenobiotic-mediated induction of CYP, cell cycle progression and activation of capases 8 and 3. We found that EGF, added to basal culture medium or phenobarbital (3.2 mM) containing medium, provoked a moderate decrease of CYP1A1 and CYP2B1/2 activities. However, EGF did not exert any inhibitory effect on 3-methylcholantrene (5 microM) and beta-naphtoflavone (25 microM) induction of CYP1A1 activities. In collagen gel sandwich cultures, hepatocytes remained arrested in mid-G1 phase of the cell cycle, even in the presence of EGF. In conventional primary cultures, caspases 8 and 3 were activated at 3 and 5 days after plating respectively. In collagen gel sandwich cultures, we found that neither collagen I nor EGF prevented activation of caspase 8 while collagen I gel inhibited activation of caspase 3, preventing spontaneous apoptosis of cultured rat hepatocytes. In contrast, EGF transiently increased caspase 3 activity at day 1 after plating. Altogether, our data demonstrate that collagen I gel triggers intracellular signals which strongly affect cultured hepatocyte phenotype, leading to a cell cycle arrest in G1 phase and long-term survival through the inhibition of caspase 3 activation and that EGF-free medium improves survival and liver-specific gene expression in hepatocytes maintained in collagen I gel sandwich cultures.


Assuntos
Caspases/metabolismo , Citocromo P-450 CYP1A1/metabolismo , Citocromo P-450 CYP2B1/metabolismo , Fator de Crescimento Epidérmico/farmacologia , Hepatócitos/efeitos dos fármacos , Xenobióticos/farmacologia , Animais , Caspase 3 , Ciclo Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Colágeno/farmacologia , Citocromo P-450 CYP1A1/efeitos dos fármacos , Citocromo P-450 CYP2B1/efeitos dos fármacos , Replicação do DNA/efeitos dos fármacos , Interações Medicamentosas , Ativação Enzimática , Indução Enzimática/efeitos dos fármacos , Hepatócitos/enzimologia , Técnicas In Vitro , Masculino , Ratos , Ratos Sprague-Dawley , Albumina Sérica/metabolismo
20.
Microb Drug Resist ; 6(4): 283-7, 2000.
Artigo em Inglês | MEDLINE | ID: mdl-11272256

RESUMO

Mutations in the pncA gene, encoding pyrazinamidase, are considered the major mechanism of pyrazinamide (PZA) resistance in Mycobacterium tuberculosis, but resistant strains containing the wild-type gene have been described. The correlation of pncA sequence with PZA resistance level was examined for 21 M. tuberculosis clinical isolates. Susceptibility patterns were determined for 100, 300, and 900 microg/ml concentrations of the drug in BACTEC. Insertions and deletions and a substitution in the putative promoter region led to high-level resistance, whereas substitutions within the open reading frame seemed to confer variable levels of resistance. Variable resistance levels and PZase activities were also observed among isolates lacking pncA mutations. The high-level resistance (900 microg/ml) in pncA wild-type isolates highlights the clinical significance of these isolates. These data also suggest that there may still be more than one alternative mechanism leading to PZA resistance in M. tuberculosis isolates.


Assuntos
Amidoidrolases/genética , Amidoidrolases/metabolismo , Antituberculosos/farmacologia , Mycobacterium tuberculosis/efeitos dos fármacos , Pirazinamida/farmacologia , Biomarcadores , Resistência Microbiana a Medicamentos/genética , Humanos , Testes de Sensibilidade Microbiana , Mutação , Mycobacterium tuberculosis/enzimologia , Mycobacterium tuberculosis/genética , Pró-Fármacos/metabolismo , Pirazinamida/análogos & derivados , Pirazinamida/metabolismo
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