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1.
Public Health ; 140: 235-243, 2016 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-27233353

RESUMO

OBJECTIVES: There have been increasing calls for more research on interventions to successfully translate evidence-based knowledge into improved health policy and practices. This paper reports on an exploratory study of knowledge translation interventions conducted with participants of global health meetings held in Bangladesh in 2012 and in South Africa in 2013. We measured stakeholders' uptake of evidence-based knowledge in terms of their translation of this knowledge into actions around public health policy and practice. The research sought to determine whether participants shared and used knowledge from the meetings to improve health policy and practices in their settings and the factors influencing sharing and use. STUDY DESIGN: An exploratory study employed quantitative and qualitative methods of online surveys and in-depth interviews to collect data from all meeting participants. METHODS: All participants in the Bangladesh and South Africa meetings were invited to complete an online survey during the meetings and over the following six weeks. Of 411 participants in the 2012 Bangladesh meeting, 148 participants from 22 countries completed the survey. Eleven of these respondents (from eight countries) were interviewed. Of the 436 participants in the 2013 South Africa meeting, 126 respondents from 33 countries completed an online survey; none of these respondents were interviewed. RESULTS: The analysis revealed that most respondents used new knowledge to advocate for policy change (2012: 65.5%; 2013: 67.5%) or improve service quality (2012: 60.1%; 2013: 70.6%). The type of knowledge that respondents most commonly shared was clinical or scientific information (2012: 79.1%; 2013: 66.7%) and country-specific information (2012: 73.0%; 2013: 71.4%). Most 2012 respondents shared knowledge because they thought it would be useful to a co-worker or colleague (79.7%). DISCUSSION: Findings on knowledge use and sharing suggest that most respondents saw themselves as knowledge brokers or intermediaries in a position to influence the translation of knowledge into action in health policy and practices in their countries. Results suggest that supporting knowledge brokers working in a local and regional context to spur change, as described in the paper, has the potential to improve health outcomes. Further research is needed to isolate specific interventions and their knowledge translation outcomes.


Assuntos
Congressos como Assunto , Saúde Global , Disseminação de Informação , Papel Profissional , Pesquisa Translacional Biomédica , Feminino , Política de Saúde , Humanos , Saúde do Lactente , Recém-Nascido , Relações Interprofissionais , Saúde Materna , Serviços de Saúde Materno-Infantil , Gravidez , Melhoria de Qualidade , Inquéritos e Questionários
2.
Gene Ther ; 22(3): 227-36, 2015 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-25567537

RESUMO

Eradication of human immunodeficiency virus-1 (HIV-1) from an infected individual requires a means of inducing production of virus from latently infected cells and stimulating an immune response against the infected cells. We report the development of lentiviral vectors that transduce dendritic cells (DCs) to both induce production of virus from latently infected cells and stimulate antigen-specific cytotoxic T lymphocytes (CTLs). The vectors package Vpx, a lentiviral accessory protein that counteracts the SAMHD1-mediated block to DC transduction, allowing for long-term expression of vector-encoded proteins. The vectors encode influenza or HIV-1-derived epitopes fused via a self-cleaving peptide to CD40L that releases the peptide into the endoplasmic reticulum for entry into the antigen presentation pathway. Expression of CD40L caused transduced DCs to mature and produce Th1-skewing cytokines. The DCs presented antigen to CD8 T cells, enhancing antigen-specific CTLs. Coculture of the transduced DCs with latently infected cells induced high-level virus production, an effect that was mediated by tumor necrosis factor alpha. The ability of a DC vaccine to reactivate latent HIV-1 and stimulate an adaptive immune response provide a means to reduce the size of the latent reservoir in patients. This strategy can also be applied to develop DC vaccines for other diseases.


Assuntos
Vacinas contra a AIDS/imunologia , Infecções por HIV/metabolismo , HIV-1/fisiologia , Subpopulações de Linfócitos T/imunologia , Linfócitos T Citotóxicos/imunologia , Vacinas contra a AIDS/genética , Antígenos CD40/metabolismo , Células Cultivadas , Células Dendríticas/imunologia , Células Dendríticas/metabolismo , Vetores Genéticos/metabolismo , Humanos , Lentivirus/genética , Proteínas Virais Reguladoras e Acessórias/genética , Proteínas Virais Reguladoras e Acessórias/metabolismo , Latência Viral
3.
Calcif Tissue Int ; 93(3): 241-8, 2013 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-23756612

RESUMO

Bisphosphonate (BP)-related osteonecrosis of the jaw (BRONJ) presents with necrotic bone in the mouth in the setting of BP exposure. It has been studied in cancer patients taking high-dose BP, but BRONJ has also been noted in patients taking lower-dose BP for osteoporosis. The purpose of this study was to characterize the phenotypes and outcomes in a large series of patients with osteoporosis and BRONJ in the setting of BP exposure. We conducted a retrospective case series. The sample was composed of subjects with BRONJ and osteoporosis. Subjects with a history of BP treatment for myeloma or metastatic cancer to the bones were excluded. Descriptive statistics were computed for the study variables. Ninety-one cases of BRONJ met the inclusion criteria. Subjects had a median age of 71 years and were predominantly female (94.5 %). The median time of BP exposure was 60 months (range 2-120). Most subjects were treated with alendronate (82.4 %). The mandible was involved more frequently (58.2 %) than the maxilla (37.3 %). Subjects commonly (65.9 %), but not universally, reported pain. For subjects with treatment outcome data (n = 0), most reported improvement (80.0 %). Although BRONJ is an uncommon condition, the absolute number of cases is fairly large due to the very large number of patients taking BPs for osteoporosis. The findings of this study confirm that BRONJ primarily affects the mandible, a substantial minority present without pain, and patients typically improve with treatment.


Assuntos
Osteonecrose da Arcada Osseodentária Associada a Difosfonatos/diagnóstico , Difosfonatos/efeitos adversos , Osteoporose/complicações , Osteoporose/tratamento farmacológico , Idoso , Idoso de 80 Anos ou mais , Osteonecrose da Arcada Osseodentária Associada a Difosfonatos/epidemiologia , Estudos de Coortes , Feminino , Humanos , Masculino , Mandíbula/efeitos dos fármacos , Pessoa de Meia-Idade , Necrose , Fenótipo , Estudos Retrospectivos , Fatores de Risco , Resultado do Tratamento
4.
Nat Genet ; 23(4): 457-61, 1999 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-10581035

RESUMO

Locus control regions (LCRs) are gene regulatory elements in mammals that can overcome the highly repressive effects normally associated with heterochromatic transgene locations (for example the centromere) in mice. Deletion of essential LCR sequences renders the cognate gene susceptible to this form of repression, so a proportion of the cells from transgenic mice that would normally express the transgene are silenced-a phenomenon known as position effect variegation (PEV). We show here that PEV can also occur when the transgene is non-centromeric and that the extent of variegation can be developmentally regulated. Furthermore, by overexpressing a mammalian homologue (M31) of Drosophila melanogaster heterochromatin protein 1 (HP1; refs 7,8) in transgenic mouse lines that exhibit PEV, it is possible to modify the proportion of cells that silence the transgene in a dose-dependent manner. Thus, we show M31 overexpression to have two contrasting effects which are dependent on chromosomal context: (i) it enhanced PEV in those lines with centromeric or pericentromeric transgene locations; and (ii) it suppressed PEV when the transgene was non-centromeric. Our results indicate that components or modifiers of heterochromatin may have a chromosomal-context-dependent role in gene silencing and activation decisions in mammals.


Assuntos
Proteínas Cromossômicas não Histona/genética , Sequência de Aminoácidos , Animais , Antígenos CD2/genética , Homólogo 5 da Proteína Cromobox , Drosophila melanogaster/genética , Feminino , Expressão Gênica , Heterocromatina/genética , Humanos , Região de Controle de Locus Gênico , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos CBA , Camundongos Transgênicos , Dados de Sequência Molecular , Fenótipo , Linfócitos T/imunologia
5.
J Exp Med ; 188(6): 1125-33, 1998 Sep 21.
Artigo em Inglês | MEDLINE | ID: mdl-9743531

RESUMO

T cell development and selection in the thymus are shaped by the induction of apoptosis. However, a direct role in T cell development and selection for any of the molecules known to regulate apoptosis has remained controversial. We have studied the effect of bax and bcl-2 transgenes in recombination activation gene 1-deficient (RAG-1(-/-)) mice transgenic for the major histocompatibility complex class I-restricted F5 T cell receptor. Overexpression of a bax transgene in the thymus seriously impairs the production of mature T cells, whereas bcl-2 overexpression greatly promotes it. The effect of bax and bcl-2 overexpression on antigen-induced negative selection was studied using fetal thymic organ cultures. This analysis showed that Bcl-2 strongly inhibits negative selection, whereas Bax does not affect it. Our data directly show that Bcl-2 family members have specific roles in T cell selection and also lend support to the hypothesis that Bax and Bcl-2 can antagonize each other's action in a certain apoptosis pathway while in another they can be functionally nonreciprocal.


Assuntos
Proteínas Proto-Oncogênicas c-bcl-2/fisiologia , Proteínas Proto-Oncogênicas/fisiologia , Subpopulações de Linfócitos T/citologia , Animais , Apoptose/efeitos dos fármacos , Apoptose/genética , Apoptose/imunologia , Diferenciação Celular/efeitos dos fármacos , Diferenciação Celular/genética , Diferenciação Celular/imunologia , Separação Celular , Feto , Citometria de Fluxo , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Camundongos Transgênicos , Técnicas de Cultura de Órgãos , Proteínas Proto-Oncogênicas/genética , Proteínas Proto-Oncogênicas c-bcl-2/genética , Subpopulações de Linfócitos T/efeitos dos fármacos , Subpopulações de Linfócitos T/fisiologia , Timo/citologia , Timo/efeitos dos fármacos , Timo/fisiologia , Transgenes/imunologia , Proteína X Associada a bcl-2
6.
J Exp Med ; 189(3): 575-86, 1999 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-9927519

RESUMO

Bad is a distant relative of Bcl-2 and acts to promote cell death. Here, we show that Bad expression levels are greatly increased in thymocytes during apoptosis. We generated bad transgenic mice to study the action of upregulated Bad expression on T cell apoptosis. The T cells from these mice are highly sensitive to apoptotic stimuli, including anti-CD95. The numbers of T cells are greatly depleted and the processes of T cell development and selection are perturbed. We show that the proapoptotic function of Bad in primary T cells is regulated by Akt kinase and that Bad overexpression enhances both cell cycle progression and interleukin 2 production after T cell activation. These data suggest that Bad can act as a key regulator of T cell apoptosis and that this is a consequence of its upregulation after exposure to death stimuli.


Assuntos
Apoptose , Proteínas de Transporte/biossíntese , Proteínas Serina-Treonina Quinases , Linfócitos T/imunologia , Timo/imunologia , Animais , Complexo CD3/metabolismo , Ciclo Celular , Dexametasona/farmacologia , Raios gama/efeitos adversos , Proteínas de Homeodomínio/genética , Interleucina-12/biossíntese , Camundongos , Camundongos Transgênicos , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Quinases/metabolismo , Proteínas Proto-Oncogênicas/metabolismo , Proteínas Proto-Oncogênicas c-akt , Transdução de Sinais , Timo/citologia , Regulação para Cima , Proteína de Morte Celular Associada a bcl , Receptor fas/imunologia
7.
Animal ; 14(2): 409-417, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31354111

RESUMO

Lameness is an important economic problem in the dairy sector, resulting in production loss and reduced welfare of dairy cows. Given the modern-day expansion of dairy herds, a tool to automatically detect lameness in real-time can therefore create added value for the farmer. The challenge in developing camera-based tools is that one system has to work for all the animals on the farm despite each animal having its own individual lameness response. Individualising these systems based on animal-level historical data is a way to achieve accurate monitoring on farm scale. The goal of this study is to optimise a lameness monitoring algorithm based on back posture values derived from a camera for individual cows by tuning the deviation thresholds and the quantity of the historical data being used. Back posture values from a sample of 209 Holstein Friesian cows in a large herd of over 2000 cows were collected during 15 months on a commercial dairy farm in Sweden. A historical data set of back posture values was generated for each cow to calculate an individual healthy reference per cow. For a gold standard reference, manual scoring of lameness based on the Sprecher scale was carried out weekly by a single skilled observer during the final 6 weeks of data collection. Using an individual threshold, deviations from the healthy reference were identified with a specificity of 82.3%, a sensitivity of 79%, an accuracy of 82%, and a precision of 36.1% when the length of the healthy reference window was not limited. When the length of the healthy reference window was varied between 30 and 250 days, it was observed that algorithm performance was maximised with a reference window of 200 days. This paper presents a high-performing lameness detection system and demonstrates the importance of the historical window length for healthy reference calculation in order to ensure the use of meaningful historical data in deviation detection algorithms.


Assuntos
Algoritmos , Doenças dos Bovinos/diagnóstico , Indústria de Laticínios/métodos , Coxeadura Animal/diagnóstico , Animais , Bovinos , Fazendas , Feminino , Marcha , Postura , Sensibilidade e Especificidade , Suécia
8.
Science ; 170(3954): 181-3, 1970 Oct 09.
Artigo em Inglês | MEDLINE | ID: mdl-5456611

RESUMO

Crude extracts of tentacles of two polychaetous annelids completely inhibit growth of Erlich ascites tumor in 60 to 100 percent of treated mice. Dialyzed extracts of one of these annelids, Lanice conchilega, show activity in the retentate after pronase digestion, suggesting that antitumor activity is associated with a nonprotein component of the crude tentacle extract.


Assuntos
Anelídeos , Antineoplásicos/uso terapêutico , Carcinoma de Ehrlich/tratamento farmacológico , Extratos de Tecidos/uso terapêutico , Animais , Antineoplásicos/toxicidade , Bioensaio , Diálise , Feminino , Havaí , Biologia Marinha , Camundongos , Tamanho do Órgão , Baço/anatomia & histologia , Extratos de Tecidos/toxicidade
9.
Science ; 197(4305): 784-6, 1977 Aug 19.
Artigo em Inglês | MEDLINE | ID: mdl-887922

RESUMO

In tree shrews (Tupaia glis) reared with one eye closed, Y-cells were almost entirely absent in the binocular segment of the lateral geniculate laminae receiving input from the deprived eye. Y-cells were found in the monocular segment of these laminae, and in the binocular segment of the laminae with input from the normal eye. X-cells were present in both the deprived and normal laminae and appeared unaffected by the deprivation. A number of abnormal cells were also found, and these were located primarily in the binocular segment where Y-cells were absent.


Assuntos
Corpos Geniculados/citologia , Musaranhos/anatomia & histologia , Vias Visuais/citologia , Percepção Visual , Animais , Corpos Geniculados/crescimento & desenvolvimento , Vias Visuais/crescimento & desenvolvimento
10.
Science ; 196(4289): 538-40, 1977 Apr 29.
Artigo em Inglês | MEDLINE | ID: mdl-403608

RESUMO

Chloroform extracts of several seaweeds, of the family Oscillatoriaceae, from Enewetak Atoll, Marshall Islands, display activity against P-388 lymphocytic mouse leukemia. A P-388 active compound, debromoaplysiatoxin, has been isolated from Lyngbya gracilis and characterized. This compound also has dermonecrotic activity and may be the dermatitis-producing substance in L. majuscula, the causative agent of "swimmers' itch" outbreaks in Hawaiian waters.


Assuntos
Antineoplásicos/uso terapêutico , Cianobactérias , Leucemia Experimental/tratamento farmacológico , Leucemia Linfoide/tratamento farmacológico , Extratos Vegetais/uso terapêutico , Toxinas Biológicas/uso terapêutico , Animais , Cianobactérias/análise , Dermatite de Contato/etiologia , Camundongos , Toxinas Biológicas/isolamento & purificação
11.
Animal ; 13(12): 3009-3017, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31516101

RESUMO

Economic pressures continue to mount on modern-day livestock farmers, forcing them to increase herds sizes in order to be commercially viable. The natural consequence of this is to drive the farmer and the animal further apart. However, closer attention to the animal not only positively impacts animal welfare and health but can also increase the capacity of the farmer to achieve a more sustainable production. State-of-the-art precision livestock farming (PLF) technology is one such means of bringing the animals closer to the farmer in the facing of expanding systems. Contrary to some current opinions, it can offer an alternative philosophy to 'farming by numbers'. This review addresses the key technology-oriented approaches to monitor animals and demonstrates how image and sound analyses can be used to build 'digital representations' of animals by giving an overview of some of the core concepts of PLF tool development and value discovery during PLF implementation. The key to developing such a representation is by measuring important behaviours and events in the livestock buildings. The application of image and sound can realise more advanced applications and has enormous potential in the industry. In the end, the importance lies in the accuracy of the developed PLF applications in the commercial farming system as this will also make the farmer embrace the technological development and ensure progress within the PLF field in favour of the livestock animals and their well-being.


Assuntos
Criação de Animais Domésticos/métodos , Fazendeiros , Fazendas/economia , Gado , Criação de Animais Domésticos/economia , Bem-Estar do Animal/normas , Animais , Fazendas/organização & administração , Humanos
12.
Atmos Meas Tech ; 12(11): 6079-6089, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-32514321

RESUMO

In this work, a new commercially available, laser-based, and ultra-portable formaldehyde (HCHO) gas sensor is characterized, and its usefulness for monitoring HCHO mixing ratios in both indoor and outdoor environments is assessed. Stepped calibrations and intercomparison with well-established laser-induced fluorescence (LIF) instrumentation allow a performance evaluation of the absorption-based, mid-infrared HCHO sensor from Aeris Technologies, Inc. The Aeris sensor displays linear behavior (R2 > 0.940) when compared with LIF instruments from Harvard and NASA Goddard. A non-linear least-squares fitting algorithm developed independently of the sensor's manufacturer to fit the sensor's raw absorption data during post-processing further improves instrument performance. The 3σ limit of detection (LOD) for 2, 15, and 60 min integration times are 2190, 690, and 420 pptv HCHO, respectively, for mixing ratios reported in real-time, though the LOD improves to 1800, 570, and 300 pptv HCHO, respectively, during post-processing. Moreover, the accuracy of the sensor was found to be ±(10% + 0.3) ppbv when compared against LIF instrumentation sampling ambient air. This sub-ppbv precision and level of accuracy are sufficient for most HCHO levels measured in indoor and outdoor environments. While the compact Aeris sensor is currently not a replacement for the most sensitive research-grade instrumentation available, its usefulness for monitoring HCHO is clearly demonstrated.

13.
Theriogenology ; 70(6): 871-9, 2008 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-18440627

RESUMO

This project examined reproductive characteristics of female and male Jackson's hartebeest (Alcelaphus buselaphus jacksoni), a subspecies that originates in Africa and is currently a model for studies of endangered hartebeest subspecies. Progestagen concentrations were measured in fecal samples collected thrice weekly for 1 year from three non-pregnant, adult females, in the Northern Hemisphere (31 degrees 37'N, 81 degrees 09'W). When their ovaries were active, the females exhibited regular luteal cycles with an overall mean (+/-S.D.) cycle length of 21.4+/-4.1 days (n=31 luteal phases). Peak luteal progestagen concentration was 1.73+/-0.63 microg/g, with a nadir concentration of 0.79+/-0.24 microg/g. Cyclic activity ceased from 6 April to 28 June, 7 April to 8 July, and from 18 February to 20 August, for the three females, respectively. During this acyclic period, mean progestagen concentration was 0.90+/-0.23 microg/g. Ejaculates were collected by electroejaculation from seven males throughout all seasons, with mean (+/-S.D.) 40+/-18% motility, 4.1+/-0.19 progressive motility (scale, 0-5), 1373+/-826x10(6) sperm/mL, and 42+/-28% morphologically normal sperm. These data characterized basic reproductive traits for Jackson's hartebeest and established the females as spontaneously ovulating and seasonally polyestrous when housed in the Northern Hemisphere, whereas males produced apparently viable sperm throughout the year.


Assuntos
Antílopes/fisiologia , Reprodução/fisiologia , Estações do Ano , Animais , Antílopes/sangue , Ciclo Estral/sangue , Feminino , Masculino , Progestinas/análise , Progestinas/sangue , Análise do Sêmen , Comportamento Sexual Animal/fisiologia
14.
J Anim Sci ; 95(4): 1512-1520, 2017 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-28464112

RESUMO

Footpad dermatitis and lameness are a major welfare concern in broiler chicken farming. In general, footpad lesions are linked to poor environmental conditions. Ulcers that arise from advanced lesions can negatively affect the gait of the birds, with effects on the animal welfare, including, in the worst cases, inability to reach the feed or water. In this study, the degree of footpad dermatitis and lameness was manually scored on 4 broiler farms across Europe, as part of an EU-wide welfare assessment program. The welfare of the chickens was assessed 3 times per production cycle (at wk 3, 4, and 5), scoring footpad dermatitis, lameness, and litter quality. In the same broiler farms, variables such as air temperature and relative humidity were automatically measured over the same period. These variables were combined into a widely accepted thermal comfort index and associated to upper and lower thresholds, which made it possible to quantify the percentage of time the birds spent out of the thermal comfort zone (POOC). The data was analyzed by combining data from the welfare assessments with environmental data collected by the automated monitoring systems. Considering the comparison between POOC classes, the highest probabilities of footpad dermatitis and lameness were obtained when POOC values exceeded the 70% threshold. Therefore, the analysis showed that footpad dermatitis and lameness were more frequent when the flock was exposed to poor environmental conditions for prolonged periods ( < 0.001). Since environmental conditions can be continuously measured, and the risk factor for footpad dermatitis and lameness increases with poor environmental conditions, there is the possibility to develop a detection and control system of severe lesions.


Assuntos
Dermatite/veterinária , Coxeadura Animal/epidemiologia , Doenças das Aves Domésticas/epidemiologia , Criação de Animais Domésticos , Bem-Estar do Animal , Animais , Causalidade , Galinhas , Dermatite/epidemiologia , Meio Ambiente , Europa (Continente) , Umidade , Fatores de Risco , Temperatura
15.
Cancer Res ; 50(17): 5301-7, 1990 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-2386938

RESUMO

Rat liver cytosolic sulfotransferase activity forms the highly reactive sulfuric acid ester of N-hydroxy-2-acetylaminofluorene (N-OH-2AAF), an ultimate carcinogen in 2-acetylaminofluorene (2AAF) hepatocarcinogenesis. A previous report demonstrated that 2AAF-induced liver hyperplastic nodules displayed a persistent loss of cytosolic N-OH-2AAF sulfotransferase activity following a hepatocarcinogenesis-producing regimen of 2AAF administration. As an initial step in examining the mechanism responsible for lowering N-OH-2AAF sulfotransferase activity, a monospecific polyclonal antibody to aryl sulfotransferase IV (AST IV) was produced and used in the assessment of AST IV as a candidate enzyme for liver cytosolic N-OH-2AAF sulfotransferase activity. Studies comparing the levels of N-OH-2AAF sulfotransferase activity of highly purified AST IV and rat liver cytosols with corresponding immunochemical analysis of AST IV contents demonstrated that there was sufficient AST IV activity in liver cytosols to indicate that it was the primary enzyme catalyzing cytosolic N-OH-2AAF sulfation. A subsequent immunochemical survey of nine extrahepatic tissues showed no detectable AST IV content and indicated that AST IV expression may be tissue specific. An immunochemical comparison of AST IV levels in control liver cytosols (high in sulfotransferase activity) with cytosols from 2AAF-derived hyperplastic nodules (low in sulfotransferase activity) or liver tumors (no sulfotransferase activity) showed low or no detectable levels, respectively, of AST IV. In addition, an immunochemical analysis of four rat hepatoma cell lines showed they contained no detectable levels of AST IV. These results suggested a strong correlation existed between a decrease in AST IV expression and tumor development. When the liver cytosols of rats taken from early, intermediate, and late stages of 2AAF carcinogenesis were analyzed for the development of a persistent loss of N-OH-2AAF sulfotransferase activity, a parallel loss of cytosolic N-OH-2AAF sulfotransferase activity and AST IV content was observed in rats which had proceeded from a stage of low risk to high risk for liver cancer. These findings indicated that (a) AST IV, a liver-specific enzyme, was the principle enzyme comprising cytosolic N-OH-2AAF sulfotransferase activity and (b) the decrease in sulfotransferase activity in nodules and tumors resulted from a decrease in the level of AST IV expression. Furthermore, it is suggested that a persistent decrease in AST IV expression may reflect a role for AST IV as part of a resistance phenotype in which transforming liver cells are able to escape the cytotoxic effects of highly reactive 2AAF metabolites and progress to cancer.


Assuntos
2-Acetilaminofluoreno/toxicidade , Arilsulfotransferase/metabolismo , Neoplasias Hepáticas Experimentais/enzimologia , Fígado/enzimologia , Animais , Anticorpos , Arilsulfotransferase/isolamento & purificação , Citosol/enzimologia , Eletroforese em Gel de Poliacrilamida , Fígado/efeitos dos fármacos , Fígado/patologia , Neoplasias Hepáticas Experimentais/patologia , Masculino , Peso Molecular , Especificidade de Órgãos , Ratos , Valores de Referência , Sulfotransferases/metabolismo
16.
Cancer Res ; 51(2): 504-9, 1991 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-1670761

RESUMO

Enzymatic sulfation of N-hydroxylated arylamines by mammalian hepatic cytosol sulfotransferases (AST; EC 2.8.2.1) is an important metabolic step which generates ultimate carcinogens. The metabolic activity of AST IV, the putative isozymic form of AST primarily responsible for catalyzing N-hydroxy-2-acetylaminofluorene sulfation, is modulated during 2-acetylaminofluorene (AAF)-induced rat hepatocarcinogenesis. To characterize the molecular mechanisms regulating the differential expression of AST IV, we have assessed polyadenylated mRNA derived from the livers of Sprague-Dawley rats undergoing different stages of AAF hepatocarcinogenesis for general in vitro translation capacity and specific expression of AST IV and albumin. Following 1 and 3 cycles of a cyclical feeding regimen (3 weeks 0.05% AAF, then 1 week basal diet), the mRNA capacity for translation was lowered and the expression of AST IV and albumin was down-regulated about 2-fold each but recovered to normal levels when treated rats were subsequently placed on basal diet for 3 continuous weeks. Cytosolic albumin levels were determined by Western blot analysis to be lowered about 1.5-2-fold. In contrast, however, mRNA from rats on basal diets for 3 weeks subsequent to cycle 5 of the feeding regimen recovered only about 50% of the capacity for AST IV expression, although overall translation capacity and albumin expression returned to normal levels. This pattern of reversible expression, followed by irreversible expression of AST IV at early and late stages of AAF hepatocarcinogenesis, respectively, provides the first evidence correlating the modulation of hepatic mRNA capacity for AST IV expression with differential cytosolic AST IV activity in the AAF hepatocarcinogenesis model. The results further suggest that during early stages in hepatocarcinogenesis modulation of mRNA protein synthesis functions may be a critical factor in AAF-mediated lowering of AST IV expression, while other persistent genetic lesions are likely playing a more significant role at the late stages of the carcinogenic process leading to neoplastic transformation of initiated hepatocytes.


Assuntos
2-Acetilaminofluoreno/toxicidade , Arilsulfotransferase/genética , Neoplasias Hepáticas Experimentais/patologia , Fígado/enzimologia , Biossíntese de Proteínas/efeitos dos fármacos , RNA Mensageiro/genética , Envelhecimento , Animais , Citosol/enzimologia , Feminino , Fígado/efeitos dos fármacos , Fígado/crescimento & desenvolvimento , Fígado/patologia , Masculino , Poli A/genética , Poli A/isolamento & purificação , RNA Mensageiro/efeitos dos fármacos , RNA Mensageiro/isolamento & purificação , Ratos , Ratos Endogâmicos
17.
Cancer Res ; 45(7): 3282-7, 1985 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-2860970

RESUMO

The effects of chemical carcinogenesis to produce premalignant hyperplastic nodules in rat liver on concomitant immune function were studied. Induction of hyperplastic nodules in Fischer rats was accomplished using a combined regimen of diethylnitrosamine, 2-acetylaminofluorene, and partial hepatectomy. Hyperplastic nodules were detected in carcinogen-treated rats from 5 to 23 weeks as confirmed by gross pathology, histopathology, and significantly elevated liver gamma-glutamyltransferase activity. Suppression of natural killer activity of either peritoneal or peripheral blood lymphoid, but not splenic, cells for YAC-1 target cells occurred during 5 to 20 weeks in carcinogen-treated rats. Spleen and blood lymphocyte mitogenic responses to concanavalin A and pokeweed mitogen were also suppressed at most intervals from 8 through 20 weeks. Control groups given individual carcinogen or partial hepatectomy alone or in dual combination were not suppressed in their immune function and failed to develop hyperplastic foci or changes in liver gamma-glutamyltransferase. Our findings indicate that immunosuppression of natural killer and lymphocyte mitogenic functions occurs for a protracted period concurrently with the development of the premalignant hyperplastic state in rat liver. The data suggest a potential role for immune competency during the onset of malignant neoplasia.


Assuntos
Tolerância Imunológica , Células Matadoras Naturais/imunologia , Neoplasias Hepáticas Experimentais/imunologia , Linfócitos/imunologia , Lesões Pré-Cancerosas/imunologia , Animais , Citotoxicidade Imunológica , Hiperplasia , Fígado/patologia , Ativação Linfocitária , Masculino , Ratos , Ratos Endogâmicos F344 , gama-Glutamiltransferase/análise
18.
Cancer Res ; 52(17): 4779-86, 1992 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-1511441

RESUMO

A complementary DNA (cDNA) for rat hepatic aryl sulfotransferase IV (AST IV) was isolated, characterized, and used as a hybridization probe to evaluate the molecular basis for the differential expression of AST IV during 2-acetylaminofluorine (2AAF)-induced hepatocarcinogensis. The AST IV cDNA clone was obtained by immunochemical screening of a male Sprague-Dawley rat liver cDNA library. The AST IV cDNA was found to be 1.3 kilobases long and to encode a fusion protein which was reactive with an antibody to AST IV and enzymatically able to generate the sulfuric acid ester of N-hydroxy-2AAF. Sequence analysis of the AST IV cDNA showed it to be 1127 residues in length and to have essentially complete homology with PST-I cDNA, a previously reported (S. Ozawa, et al., Nucleic Acids Res., 18: 4001, 1990), 1028-base cDNA for an uncharacterized rat liver aryl sulfotransferase. Comparison of the PST-I/AST IV cDNA-deduced amino acid sequence with data from a partial (51%) amino acid sequence analysis of purified AST IV showed complete amino acid homology, confirming the identity of the cDNA and establishing that AST IV was an N-blocked, 291-amino acid protein with a molecular mass of 33,909 daltons. The AST IV cDNA sequence differed from the PST-I cDNA in two principal ways: the 5' end lacked 18 coding bases, and the 3' end contained a 190-base extention in the untranslated region, including a consensus sequence for signalling polyadenylation. Studies of AST IV gene transcript levels showed that the livers of rats fed 2AAF for 3 wk (early stage hepatocarcinogenesis) and hyperplastic nodules from the livers of rats fed 2AAF for 19 wk (intermediate stage hepatocarcinogenesis) displayed transcript levels similar to those of livers from normal rats. This contrasted with the 60 to 70% lower than normal capacity of the mRNA fractions to express AST IV observed during in vitro translation. These results indicated that modulation of AST IV expression at early and intermediate stages of hepatocarcinogenesis involved regulatory mechanisms at the translational level. In contrast, mRNA fractions isolated from some 2AAF-induced liver tumors or from known chemical carcinogen-derived rat hepatoma cell lines showed losses of both AST IV transcript level and in vitro translation capacity, suggesting that regulation at the transcriptional level may become important at late stages of 2AAF-induced hepatocarcinogenesis. These results indicated that the molecular mechanisms for the 2AAF-mediated down regulation of AST IV expression during 2AAF-induced hepatocarcinogenesis involved alterations in regulation at both translational and transcriptional levels.


Assuntos
Arilsulfotransferase/genética , Neoplasias Hepáticas Experimentais/genética , Neoplasias Hepáticas/genética , 2-Acetilaminofluoreno/toxicidade , Sequência de Aminoácidos , Animais , Sequência de Bases , Clonagem Molecular , DNA/genética , Expressão Gênica , Fígado/enzimologia , Neoplasias Hepáticas/enzimologia , Neoplasias Hepáticas Experimentais/induzido quimicamente , Masculino , Dados de Sequência Molecular , Oligodesoxirribonucleotídeos/química , Lesões Pré-Cancerosas/genética , RNA Mensageiro/genética , RNA Neoplásico/genética , Ratos , Proteínas Recombinantes de Fusão/imunologia , Proteínas Recombinantes de Fusão/metabolismo , Alinhamento de Sequência
19.
J Comp Neurol ; 192(3): 531-47, 1980 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-7419743

RESUMO

Microelectrode recordings were made in the binocular portion of the tree shrew striate cortex to determine how orientation selective cells are distributed topographically in area 17 of this species. Seventy-five percent of the cells sampled were activated well by elongated visual stimuli and were quite selective for stimulus orientation. Ninety-five percent of the orientation-selective cells had orientation tuning ranges (Wilson and Sherman, '76) between +/- 5 degrees and +/- 40 degrees from their optimal orientation. Orientation-selective cells with the same or similar optimal orientations were distributed in cortex in a columnar manner (Hubel and Wiesel, '62), as determined from electrode penetrations nearly normal to the cortical surface. Penetrations parallel to the cortical surface revealed a highly ordered representation of optimal stimulus orientation, generally characterized by sequential changes in optimal orientation with electrode movement across the striate cortex. In addition, relatively consistent differences were observed in the rates and patterns of orientation shift on these penetrations depending on the direction of electrode movement across the cortex. Penetrations parallel to the 17--18 border yielded moderate-to-high rates of orientation change (mean slope = 434 degrees/mm), with the changes generally progressing through a complete clockwise or counterclockwise cycle of 180 degrees or more before a major reversal in the direction of orientation shift was encountered. In contrast, penetrations perpendicular to the border yielded low-to-moderate slopes (mean slope = 239 degrees/mm). On these penetrations a more limited range of optimal orientations (< 180 degrees) was usually encountered, due to frequent reversals in the direction of orientation shift. Also, extended regions (100--200) micrometers long) of constant optimal orientation were observed in these penetrations. The different patterns of orientation change found on these orthogonal penetrations across the striate cortex indicate that the orientation column system in this species is anisotropically organized with respect to the 17--18 border. Further, the regions of constant optimal orientation frequently encountered on penetrations perpendicular to the 17--18 border suggest that the anisotropy is subserved by a system of elongated zones of iso-orientation arranged approximately perpendicular to the 17--18 border.


Assuntos
Orientação/fisiologia , Córtex Visual/fisiologia , Animais , Autorradiografia , Dominância Cerebral/fisiologia , Feminino , Masculino , Neurônios/fisiologia , Tupaiidae , Vias Visuais/fisiologia , Percepção Visual/fisiologia
20.
J Comp Neurol ; 358(3): 401-13, 1995 Jul 31.
Artigo em Inglês | MEDLINE | ID: mdl-7560294

RESUMO

A laminar analysis of the receptive field properties of relay cells in the binocular region of the tree shrew dorsal lateral geniculate nucleus (LGN) found three main subdivisions. Lamina 1 (receiving ipsilateral eye input) and lamina 2 (contralateral) comprise a pair of layers that contain only ON-center neurons. Laminae 4 (contralateral) and 5 (ipsilateral) comprise a pair of layers with mostly OFF-center cells (86%). Laminae 3 and 6 (both contralaterally innervated) also form a distinct pair, although lamina 3 contains a mixture of cells with ON-centers (43%) or OFF-centers (57%), and lamina 6 contains mostly cells with ON-OFF centers and suppressive surrounds (81%). Cells located in the interlaminar zones resembled neurons in laminae 3 and 6. In comparison with the cells in the OFF-center laminae 4 and 5, the ON-center cells in laminae 1 and 2 had smaller, more elliptical receptive field centers with stronger responses to flashed visual stimuli. In addition, cells in the ipsilateral eye laminae 1 and 5 showed a greater change in center diameter, with eccentricity from the area centralis, than cells in the contralateral eye laminae 2 and 4. Principal components analysis using six receptive field properties (latency to optic chiasm stimulation, receptive field center diameter, maintained discharge rate, response onset latency, peak spike density, and phasic-tonic index) suggested that the cells in laminae 3 and 6 and the interlaminar zones are W-like. Principal components analysis of the same receptive field properties in laminae 1, 2, 4, and 5 did not reveal differences clearly related to X-like (parvocellular) and Y-like (magnocellular) categories. Ninety-seven percent of the cells tested for linearity of spatial summation in laminae 1, 2, 4, and 5 were linear. We conclude that the dominant organizational features of the tree shrew LGN are the ON-center, OFF-center, and W pairs of layers that project to different regions within the striate cortex.


Assuntos
Córtex Cerebral/anatomia & histologia , Corpos Geniculados/anatomia & histologia , Células Ganglionares da Retina/ultraestrutura , Tupaiidae/anatomia & histologia , Vias Visuais/anatomia & histologia , Animais , Estimulação Elétrica , Neurônios/ultraestrutura , Campos Visuais
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