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1.
J Ment Health ; 28(5): 467-474, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-28719230

RESUMO

Background: While the onset of many mental health problems occurs in adolescence, these problems are severely undertreated in this age group. To inform early intervention for adolescents, we investigated the effect of gender and education type on perception of barriers to help seeking, mental health literacy, and the awareness and use of mental health services. Method: A web-based survey using vignettes, open-ended and multiple choice items was administered to upper secondary school students in two counties in Norway. Results: The survey was completed by 1249 students (88% response rate) with an average age of 17.6 years and 56% were female. Compared to males, the females were better in identifying psychological problems of anxiety and trauma, awareness of mental health services (p < 0.001) and perceived more barriers for seeking help (cost and waiting time; p < 0.001). For use of all mental health services, the effect of education type was greater than the effect of gender. Conclusion: For adolescents, gender appears to play a significant, but not exclusive, role in the inclination to seek professional help for mental health problems. We hypothesise that the observed gender difference in use of services is related to the gender difference in awareness of referral pathway services and the influence of parents in help-seeking process.


Assuntos
Comportamento de Busca de Ajuda , Transtornos Mentais/psicologia , Transtornos Mentais/terapia , Aceitação pelo Paciente de Cuidados de Saúde/psicologia , Adolescente , Feminino , Acessibilidade aos Serviços de Saúde , Humanos , Masculino , Psicologia do Adolescente , Fatores Sexuais
2.
Vet Res ; 48(1): 52, 2017 09 19.
Artigo em Inglês | MEDLINE | ID: mdl-28927447

RESUMO

Breeding towards genetic resistance to prion disease is effective in eliminating scrapie. In sheep, classical forms of scrapie have been eradicated almost completely in several countries by breeding programs using a prion protein (PrP) gene (PRNP) amino acid polymorphism. For goats, field and experimental studies have provided evidence for several amino acid polymorphisms that are associated with resistance to scrapie, but only limited data are available concerning the susceptibility of caprine PRNP genotypes to BSE. In this study, goat kids representing five PRNP genotypes based on three polymorphisms (M142, Q211 and K222 and the wild type I142, R211 and Q222) were orally challenged with bovine or goat BSE. Wild type goats were killed with clinical signs between 24-28 months post inoculation (mpi) to both challenges, and goats with genotype R/Q211 succumbed between 29-36 mpi. I/M142 goats developed clinical signs at 44-45 mpi and M/M142 goats remained healthy until euthanasia at 48 mpi. None of the Q/K222 goats showed definite clinical signs. Taken together the highest attack ratios were seen in wild type and R/Q211 goats, and the lowest in I/M142, M/M142 and Q/K222. In all genotype groups, one or more goats remained healthy within the incubation period in both challenges and without detectable PrP deposition in the tissues. Our data show that both the K222 and M142 polymorphisms lengthen the incubation period significantly compared to wild type animals, but only K222 was associated with a significant increase in resistance to BSE infection after oral exposure to both BSE sources.


Assuntos
Resistência à Doença/genética , Encefalopatia Espongiforme Bovina/prevenção & controle , Doenças das Cabras/prevenção & controle , Príons/efeitos adversos , Animais , Cruzamento , Bovinos , Códon/genética , Encefalopatia Espongiforme Bovina/genética , Feminino , Doenças das Cabras/genética , Doenças das Cabras/patologia , Cabras , Masculino , Proteínas Priônicas
3.
Water Res ; 125: 180-190, 2017 11 15.
Artigo em Inglês | MEDLINE | ID: mdl-28854385

RESUMO

Real time control (RTC) is increasingly seen as a viable method to optimise the functioning of wastewater systems. Model exercises and case studies reported in literature claim a positive impact of RTC based on results without uncertainty analysis and flawed evaluation periods. This paper describes two integrated RTC strategies at the wastewater treatment plant (WWTP) Eindhoven, the Netherlands, that aim to improve the use of the available tanks at the WWTP and storage in the contributing catchments to reduce the impact on the receiving water. For the first time it is demonstrated that a significant improvement can be achieved through the application of RTC in practice. The Storm Tank Control is evaluated based on measurements and reduces the number of storm water settling tank discharges by 44% and the discharged volume by an estimated 33%, decreasing dissolved oxygen depletion in the river. The Primary Clarifier Control is evaluated based on model simulations. The maximum event NH4 concentration in the effluent reduced on average 19% for large events, while the load reduced 20%. For all 31 events the reductions are 11 and 4% respectively. Reductions are significant taking uncertainties into account, while using representative evaluation periods.


Assuntos
Eliminação de Resíduos Líquidos/métodos , Amônia/análise , Países Baixos , Rios , Eliminação de Resíduos Líquidos/instrumentação
4.
J Anim Breed Genet ; 133(5): 366-74, 2016 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-26991480

RESUMO

Scrapie is a neurodegenerative disease occurring in goats and sheep. Several haplotypes of the prion protein increase resistance to scrapie infection and may be used in selective breeding to help eradicate scrapie. In this study, frequencies of the allelic variants of the PrP gene are determined for six goat breeds in the Netherlands. Overall frequencies in Dutch goats were determined from 768 brain tissue samples in 2005, 766 in 2008 and 300 in 2012, derived from random sampling for the national scrapie surveillance without knowledge of the breed. Breed specific frequencies were determined in the winter 2013/2014 by sampling 300 breeding animals from the main breeders of the different breeds. Detailed analysis of the scrapie-resistant K222 haplotype was carried out in 2014 for 220 Dutch Toggenburger goats and in 2015 for 942 goats from the Saanen derived White Goat breed. Nine haplotypes were identified in the Dutch breeds. Frequencies for non-wild type haplotypes were generally low. Exception was the K222 haplotype in the Dutch Toggenburger (29%) and the S146 haplotype in the Nubian and Boer breeds (respectively 7 and 31%). The frequency of the K222 haplotype in the Toggenburger was higher than for any other breed reported in literature, while for the White Goat breed it was with 3.1% similar to frequencies of other Saanen or Saanen derived breeds. Further evidence was found for the existence of two M142 haplotypes, M142 /S240 and M142 /P240 . Breeds vary in haplotype frequencies but frequencies of resistant genotypes are generally low and consequently selective breeding for scrapie resistance can only be slow but will benefit from animals identified in this study. The unexpectedly high frequency of the K222 haplotype in the Dutch Toggenburger underlines the need for conservation of rare breeds in order to conserve genetic diversity rare or absent in other breeds.


Assuntos
Frequência do Gene , Variação Genética , Cabras/classificação , Cabras/genética , Proteínas Priônicas/genética , Animais , Haplótipos , Países Baixos , Linhagem
6.
Vet Rec ; 178(7): 168, 2016 Feb 13.
Artigo em Inglês | MEDLINE | ID: mdl-26755614

RESUMO

Scrapie in goats is a transmissible, fatal prion disease, which is endemic in the British goat population. The recent success in defining caprine PRNP gene variants that provide resistance to experimental and natural classical scrapie has prompted the authors to conduct a survey of PRNP genotypes in 10 goat breeds and 52 herds to find goats with the resistant K222 allele. They report here the frequencies in 1236 tested animals of the resistance-associated K222 and several other alleles by breed and herd. Eight animals were found to be heterozygous QK222 goats (0.64 per cent genotype frequency, 95 per cent CI 0.28 to 1.27 per cent) but no homozygous KK222 goats were detected. The K222 allele was found in Saanen, Toggenburg and Anglo-Nubian goats. The fact that only a few goats with the K222 allele have been identified does not preclude the possibility to design and implement successful breeding programmes at national level.


Assuntos
Alelos , Doenças das Cabras/genética , Príons/genética , Scrapie/genética , Animais , Genótipo , Cabras , Polimorfismo Genético , Inquéritos e Questionários , Reino Unido
7.
J Virol ; 90(2): 805-12, 2016 01 15.
Artigo em Inglês | MEDLINE | ID: mdl-26512080

RESUMO

UNLABELLED: Susceptibility or resistance to prion infection in humans and animals depends on single prion protein (PrP) amino acid substitutions in the host, but the agent's modulating role has not been well investigated. Compared to disease incubation times in wild-type homozygous ARQ/ARQ (where each triplet represents the amino acids at codons 136, 154, and 171, respectively) sheep, scrapie susceptibility is reduced to near resistance in ARR/ARR animals while it is strongly enhanced in VRQ/VRQ carriers. Heterozygous ARR/VRQ animals exhibit delayed incubation periods. In bovine spongiform encephalopathy (BSE) infection, the polymorphism effect is quite different although the ARR allotype remains the least susceptible. In this study, PrP allotype composition in protease-resistant prion protein (PrP(res)) from brain of heterozygous ARR/VRQ scrapie-infected sheep was compared with that of BSE-infected sheep with a similar genotype. A triplex Western blotting technique was used to estimate the two allotype PrP fractions in PrP(res) material from BSE-infected ARR/VRQ sheep. PrP(res) in BSE contained equimolar amounts of VRQ- and ARR-PrP, which contrasts with the excess (>95%) VRQ-PrP fraction found in PrP in scrapie. This is evidence that transmissible spongiform encephalopathy (TSE) agent properties alone, perhaps structural aspects of prions (such as PrP amino acid sequence variants and PrP conformational state), determine the polymorphic dependence of the PrP(res) accumulation process in prion formation as well as the disease-associated phenotypic expressions in the host. IMPORTANCE: Transmissible spongiform encephalopathies (TSEs) are fatal neurodegenerative and transmissible diseases caused by prions. Amino acid sequence variants of the prion protein (PrP) determine transmissibility in the hosts, as has been shown for classical scrapie in sheep. Each individual produces a separate PrP molecule from its two PrP gene copies. Heterozygous scrapie-infected sheep that produce two PrP variants associated with opposite scrapie susceptibilities (136V-PrP variant, high; 171R-PrP variant, very low) contain in their prion material over 95% of the 136V PrP variant. However, when these sheep are infected with prions from cattle (bovine spongiform encephalopathy [BSE]), both PrP variants occur in equal ratios. This shows that the infecting prion type determines the accumulating PrP variant ratio in the heterozygous host. While the host's PrP is considered a determining factor, these results emphasize that prion structure plays a role during host infection and that PrP variant involvement in prions of heterozygous carriers is a critical field for understanding prion formation.


Assuntos
Predisposição Genética para Doença , Príons/metabolismo , Scrapie/genética , Alelos , Animais , Heterozigoto , Período de Incubação de Doenças Infecciosas , Príons/genética , Ovinos , Fatores de Tempo
8.
Water Res ; 88: 245-256, 2016 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-26512802

RESUMO

Gully pots are essential assets designed to relief the downstream system by trapping solids and attached pollutants suspended in runoff. This study applied a methodology to develop a quantitative gully pot sedimentation and blockage model. To this end, sediment bed level time series from 300 gully pots, spanning 15 months, were collected. A generalised linear mixed modelling (GLMM) approach was applied to model and quantify the accumulation of solids in gully pots and to identify relevant physical and catchment properties that influence the complex trapping processes. Results show that the retaining efficiency decreases as sediment bed levels increase. Two typical silting evolutions were identified. Approximately 5% of all gully pots experienced progressive silting, eventually resulting in a blockage. The other gully pots show stabilising sediment bed levels. The depth of the sand trap, elapsed time since cleaning and the road type were identified to be the main properties discriminating progressive accumulation from stabilising sediment bed levels. Furthermore, sediment bed levels exhibit no residual spatial correlation, indicating that the vulnerability to a blockage is reduced as adjacent gully pots provide a form of redundancy. The findings may aid to improve maintenance strategies in order to safeguard the performance of gully pots.


Assuntos
Hidrologia/métodos , Modelos Estatísticos , Probabilidade , Distribuição Aleatória , Reprodutibilidade dos Testes
9.
Water Sci Technol ; 69(12): 2397-406, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-24960000

RESUMO

Storm water runoff is a major contributor to the pollution of receiving waters. Storm water characteristics may vary significantly between locations and events. Hence, for each given location, this necessitates a well-designed monitoring campaign prior to selection of an appropriate storm water management strategy. The challenge for the design of a monitoring campaign with a given budget is to balance detailed monitoring at a limited number of locations versus less detailed monitoring at a large number of locations. This paper proposes a methodology for the selection of monitoring locations for storm water quality monitoring, based on (pre-)screening, a quick scan monitoring campaign, and final selection of locations and design of the monitoring setup. The main advantage of the method is the ability to prevent the selection of monitoring locations that turn out to be inappropriate. In addition, in this study, the quick scan resulted in a first useful dataset on storm water quality and a strong indication of illicit connections at one of the monitoring locations.


Assuntos
Monitoramento Ambiental/métodos , Poluentes Químicos da Água/química , Qualidade da Água/normas , Animais , Países Baixos , Fatores de Tempo , Água/química
10.
N Z Vet J ; 62(6): 343-50, 2014 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-24961961

RESUMO

AIMS: To estimate the number of cases of scrapie that would occur in sheep of different prion protein (PrP) genotypes if scrapie was to become established in New Zealand, and to compare the performance of two commercially available, rapid ELISA kits using ovine retro-pharyngeal lymph nodes (RLN) from non-infected and infected sheep of different PrP genotypes. METHODS: Using published data on the distribution of PrP genotypes within the New Zealand sheep flock and the prevalence of cases of scrapie in these genotypes in the United Kingdom, the annual expected number of cases of scrapie per genotype was estimated, should scrapie become established in New Zealand, assuming a total population of 28 million sheep. A non-infected panel of RLN was collected from 737 sheep from New Zealand that had been culled, found in extremis or died. Brain stem samples were also collected from 131 of these sheep. A second panel of infected samples comprised 218 and 117 RLN from confirmed scrapie cases that had originated in Europe and the United States of America, respectively. All samples were screened using two commercial, rapid, transmissible spongiform encephalopathy ELISA kits: Bio-Rad TeSeE ELISA (ELISA-BR), and IDEXX HerdChek BSE-Scrapie AG Test (ELISA-ID). RESULTS: If scrapie became established in New Zealand, an estimated 596 cases would occur per year; of these 234 (39%) and 271 (46%) would be in sheep carrying ARQ/ARQ and ARQ/VRQ PrP genotypes, respectively. For the non-infected samples from New Zealand the diagnostic specificity of both ELISA kits was 100%. When considering all infected samples, the diagnostic sensitivity was 70.4 (95% CI=65.3-75.3)% for ELISA-BR and 91.6 (95% CI=88.2-94.4)% for ELISA-ID. For the ARQ/ARQ genotype (n=195), sensitivity was 66.2% for ELISA-BR and 90.8% for ELISA-ID, and for the ARQ/VRQ genotype (n=107), sensitivity was 81.3% for ELISA-BR and 98.1% for ELISA-ID. CONCLUSIONS: In this study, the ELISA-ID kit demonstrated a higher diagnostic sensitivity for detecting scrapie in samples of RLN from sheep carrying scrapie-susceptible PrP genotypes than the ELISA-BR kit at comparable diagnostic specificity. CLINICAL RELEVANCE: The diagnostic performance of the ELISA-ID kit using ovine RLN merits the consideration of including this assay in the national scrapie surveillance programme in New Zealand.


Assuntos
Ensaio de Imunoadsorção Enzimática/veterinária , Linfonodos/patologia , Kit de Reagentes para Diagnóstico/veterinária , Scrapie/diagnóstico , Animais , Ensaio de Imunoadsorção Enzimática/métodos , Príons/genética , Sensibilidade e Especificidade , Ovinos
11.
Environ Manage ; 53(2): 333-42, 2014 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-24276896

RESUMO

Bioenergy production involves different agents with potentially different objectives, and an agent's decision often has transboundary impacts on other agents along the bioenergy value chain. Understanding and estimating the transboundary impacts is essential to portraying the interactions among the different agents and in the search for the optimal configuration of the bioenergy value chain. We develop an agent-based model to mimic the decision making by feedstock producers and feedstock-to-biofuel conversion plant operators and propose multipliers (i.e., ratios of economic values accruing to different segments and associated agents in the value chain) for assessing the transboundary impacts. Our approach is generic and thus applicable to a variety of bioenergy production systems at different sites and geographic scales. We apply it to the case of producing ethanol using corn stover in Iowa, USA. The results from the case study indicate that stover removal rate is site specific and varies considerably with soil type, as well as other factors, such as stover price and harvesting cost. In addition, ethanol production using corn stover in the study region would have strong positive ripple effects, with the values of multipliers varying with greenhouse gas price and national energy security premium. The relatively high multiplier values suggest that a large portion of the value associated with corn stover ethanol production would accrue to the downstream end of the value chain instead of stover producers.


Assuntos
Agricultura/tendências , Zea mays , Agricultura/economia , Ração Animal , Biocombustíveis , Biomassa , Conservação dos Recursos Naturais , Tomada de Decisões
12.
Water Sci Technol ; 67(11): 2458-66, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-23752377

RESUMO

Sewer systems are costly to construct and even more costly to replace, requiring proper asset management. Sewer asset management relies to a large extent on available information. In sewer systems where pipe corrosion is the dominant failure mechanism, visual inspection by closed circuit television (CCTV) and core sampling are among the methods mostly applied to assess sewer pipe condition. This paper compares visual inspection and drill core analysis in order to enhance further understanding of the limitations and potentials of both methods. Both methods have been applied on a selected sewer reach in the city of The Hague, which was reportedly subject to pipe corrosion. Results show that both methods, visual inspection and core sampling, are associated with large uncertainties and that there is no obvious correlation between results of visual inspection and results of drill core analysis.


Assuntos
Drenagem Sanitária , Análise de Falha de Equipamento/métodos , Eliminação de Resíduos Líquidos/instrumentação , Materiais de Construção , Esgotos
13.
J Environ Manage ; 113: 383-9, 2012 Dec 30.
Artigo em Inglês | MEDLINE | ID: mdl-23088934

RESUMO

Fertilizer use, widely practiced in forest plantation management to stimulate tree growth, contributes to greenhouse gas (GHG) emissions. We explore how accounting for GHG consequences affects optimal fertilizer application rates of commercial forest plantations. A generic model that maximizes the equivalent annual net benefit of timber production and GHG balance is developed and applied to loblolly pine (Pinus taeda L.) plantations in the southern United States. We find that fertilizer use still is a viable practice for managing loblolly pine plantations in the region although fertilizer application rate should be reduced when GHG consequences are valued. A greater reduction in fertilizer application rate is recommended where wood is used for paper production because life cycle GHG emissions of paper products are much higher than those of solid wood or bioenergy products. A higher fertilizer rate should be applied when forest residues are used for the production of bioenergy that offsets GHG emissions from consuming fossil fuels.


Assuntos
Biomassa , Pinus taeda , Fertilizantes , Efeito Estufa
14.
Water Sci Technol ; 66(8): 1614-20, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-22907442

RESUMO

Long-term and high-frequency in-sewer monitoring opens up a broad range of possibilities to study (influences on) water quantity and quality variations. Using data from the Eindhoven wastewater system in The Netherlands both dry weather flow and wet weather flow situations have been studied. For approximately 160 dry weather days mean diurnal variations of flow and pollutant concentrations have been derived. For wet weather situations (≈ 40 storm events) peak load factors have been studied. Generally, peak load factors for all considered pollutant parameters are larger than one. Peak load factors for particulate matter are larger than for dissolved constituents. Also, the smallest catchment area consistently shows the largest mean peak factors and vice versa.


Assuntos
Monitoramento Ambiental/métodos , Esgotos/análise , Poluentes Químicos da Água/análise
15.
Water Sci Technol ; 66(7): 1534-9, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-22864441

RESUMO

This paper assesses the influence of (differential) settlement on sewer system functioning. Based on historical data of the vertical position of sewer invert levels, tilt measurements and in-sewer inspection videos, settlement and settlement related influences are analyzed for a case study area in Amsterdam. The average settlement rate of this system was 4 mm/year. Given this settlement rate, it is shown that settlement significantly influences sewer system functioning within the lifetime of a sewer (system).


Assuntos
Esgotos , Países Baixos
16.
Water Res ; 46(20): 6868-80, 2012 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-22748326

RESUMO

Stormwater runoff is a major contributor to the pollution of receiving waters. This study focuses at characterising stormwater in order to be able to determine the impact of stormwater on receiving waters and to be able to select the most appropriate stormwater handling strategy. The stormwater characterisation is based on determining site mean concentrations (SMCs) and their uncertainties as well as the treatability of stormwater by monitoring specific pollutants concentration levels (TSS, COD, BOD, TKN, TP, Pb, Cu, Zn, E.coli) at three full scale stormwater treatment facilities in Arnhem, the Netherlands. This has resulted in 106 storm events being monitored at the lamella settler, 59 at the high rate sand filter and 132 at the soil filter during the 2 year monitoring period. The stormwater characteristics in Arnhem in terms of SMCs for main pollutants TSS and COD and settling velocities differ from international data. This implies that decisions for stormwater handling made on international literature data will very likely be wrong due to assuming too high concentrations of pollutants and misjudgement of the treatability of stormwater. The removal rates monitored at the full scale treatment facilities are within the expected range, with the soil filter and the sand filter having higher removal rates than the lamella settler. The full scale pilots revealed the importance of incorporating gross solids removal in the design of stormwater treatment facilities, as the gross solids determine operation and maintenance requirements.


Assuntos
Chuva , Eliminação de Resíduos Líquidos , Movimentos da Água , Poluição da Água/prevenção & controle , Monitoramento Ambiental , Filtração/métodos , Dióxido de Silício , Solo , Incerteza , Poluentes da Água
17.
Water Sci Technol ; 66(1): 145-50, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-22678211

RESUMO

Storm water separating manifolds in house connections have been introduced as a cost effective solution to disconnect impervious areas from combined sewers. Such manifolds have been applied by the municipality of Breda, the Netherlands. In order to investigate the performance of the manifolds, a monitoring technique (distributed temperature sensing or DTS) using fiber optic cables has been applied in the sewer system of Breda. This paper describes the application of DTS as a research tool in sewer systems. DTS proves to be a powerful tool to monitor the performance of (parts of) a sewer system in time and space. The research project showed that DTS is capable of monitoring the performance of house connections and identifying locations of inflow of both sewage and storm runoff. The research results show that the performance of storm water separating manifolds varies over time, thus making them unreliable.


Assuntos
Monitoramento Ambiental/métodos , Chuva , Temperatura , Movimentos da Água , Drenagem Sanitária , Habitação , Eliminação de Resíduos Líquidos
18.
Water Sci Technol ; 64(7): 1519-26, 2011.
Artigo em Inglês | MEDLINE | ID: mdl-22179651

RESUMO

This paper introduces an integrated approach for the assessment of receiving water quality and the relative contribution of the urban drainage system to perceived receiving water quality problems. The approach combines mass balances with relatively simple receiving water impact models. The research project has learned that the urban drainage system is only one of the determining factors with respect to receiving urban water quality problems. The morphology of the receiving waters and the non-sewer sources of pollution, such as waterbirds, dogs, or inflow of external surface water might be equally important. This conclusion underlines the necessity to changes today's emission based approach and adopt an integral and immission based approach. The integrated approach is illustrated on a case study in Arnhem, where the receiving water quality remained unsatisfactory even after retrofitting a combined sewer system into a separated sewer system.


Assuntos
Poluição da Água/prevenção & controle , Qualidade da Água , Animais , Cães , Monitoramento Ambiental/métodos , Países Baixos , Esgotos , Urbanização , Abastecimento de Água/normas
19.
J Virol ; 85(23): 12537-46, 2011 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-21917981

RESUMO

Classical scrapie is a prion disease in sheep and goats. In sheep, susceptibility to disease is genetically influenced by single amino acid substitutions. Genetic breeding programs aimed at enrichment of arginine-171 (171R) prion protein (PrP), the so-called ARR allele, in the sheep population have been demonstrated to be effective in reducing the occurrence of classical scrapie in the field. Understanding the molecular basis for this reduced prevalence would serve the assessment of ARR adaptation. The prion formation mechanism and conversion of PrP from the normal form (PrP(C)) to the scrapie-associated form (PrP(Sc)) could play a key role in this process. Therefore, we investigated whether the ARR allele substantially contributes to scrapie prion formation in naturally infected heterozygous 171Q/R animals. Two methods were applied to brain tissue of 171Q/R heterozygous sheep with natural scrapie to determine the relative amount of the 171R PrP fraction in PrP(res), the proteinase K-resistant PrP(Sc) core. An antibody test differentiating between 171Q and 171R PrP fragments showed that PrP(res) was mostly composed of the 171Q allelotype. Furthermore, using a novel tool for prion research, endoproteinase Lys-C-digested PrP(res) yielded substantial amounts of a nonglycosylated and a monoglycosylated PrP fragment comprising codons 114 to 188. Following two-dimensional gel electrophoresis, only marginal amounts (<9%) of 171R PrP(res) were detected. Enhanced 171R(res) proteolytic susceptibility could be excluded. Thus, these data support a nearly zero contribution of 171R PrP in PrP(res) of 171R/Q field scrapie-infected animals. This is suggestive of a poor adaptation of classical scrapie to this resistance allele under these natural conditions.


Assuntos
Encéfalo/metabolismo , Resistência a Medicamentos , Endopeptidase K/farmacologia , Príons/genética , Príons/metabolismo , Scrapie/metabolismo , Scrapie/patologia , Alelos , Animais , Western Blotting , Encéfalo/patologia , Suscetibilidade a Doenças , Eletroforese em Gel Bidimensional , Citometria de Fluxo , Genótipo , Técnicas Imunoenzimáticas , Ovinos
20.
J Gen Virol ; 92(Pt 1): 222-32, 2011 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-20943889

RESUMO

With increased awareness of the diversity of transmissible spongiform encephalopathy (TSE) strains in the ruminant population, comes an appreciation of the need for improved methods of differential diagnosis. Exposure to bovine spongiform encephalopathy (BSE) has been associated with the human TSE, variant Creutzfeldt-Jakob disease, emphasizing the necessity in distinguishing low-risk TSE types from BSE. TSE type discrimination in ruminants such as cattle, sheep, goats and deer, requires the application of several prion protein (PrP)-specific antibodies in parallel immunochemical tests on brain homogenates or tissue sections from infected animals. This study uses in a single incubation step, three PrP-specific antibodies and fluorescent Alexa dye-labelled anti-mouse Fabs on a Western blot. The usual amount of brain tissue needed is 0.5 mg. This multiplex application of antibodies directed towards three different PrP epitopes enabled differential diagnosis of all established main features of classical scrapie, BSE and Nor98-like scrapie in sheep and goats, as well as the currently known BSE types C, H and L in cattle. Moreover, due to an antibody-dependent dual PrP-banding pattern, for the first time CH1641 scrapie of sheep can be reliably discriminated from the other TSE isolate types in sheep.


Assuntos
Técnicas de Laboratório Clínico/métodos , Doenças Priônicas/diagnóstico , Doenças Priônicas/veterinária , Príons/classificação , Animais , Anticorpos Monoclonais , Western Blotting/métodos , Bovinos , Cervos , Diagnóstico Diferencial , Cabras , Humanos , Sensibilidade e Especificidade , Ovinos
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