A novel adeno-associated virus serotype (AAV)-8 capsid mutant improves human coagulation factor IX expression in vivo

Recombinant AAV-8 vectors have shown significant promise for hepatic gene therapy of hemophilia B. However, the theme of AAV vector dose dependent immunotoxicity seen with AAV2 vectors earlier seem to re-emerge with AAV8 vectors as well. It is therefore important to develop novel AAV8 vectors that provide enhanced gene expression at significantly less vector doses. We hypothesized that AAV8 during its intracellular trafficking, are targeted for destruction in the cytoplasm by the host-cellular kinase/ubiquitination/proteasomal degradation machinery and modification of specific serine/threonine kinase or ubiquitination targets on AAV8 capsid (Fig.1A) may improve its transduction efficiency. To test this, point mutations at specific serine (S)/threonine (T) > alanine (A) or lysine (K)>arginine (R) residues were generated on AAV8 capsid. scAAV8-EGFP vectors containing the wild-type (WT) and each one of the 5 S/T/K-mutant(S276A, S501A, S671A, T251A and K137R) capsids were evaluated for their liver transduction efficiency at a dose of 5 X 1010 vgs/ animal in C57BL/6 mice in vivo. The best performing mutant was found to be the K137R vector in terms of either the gene expression (46-fold) or the vector copy numbers in the hepatocytes (22-fold) compared to WT-AAV8 (Fig.1B). The K137R-AAV8 vector that showed significantly decreased ubiquitination of the viral capsid had reduced activation of markers of innate immune response [IL-6, IL-12, tumor necrosis factor α, Kupffer cells and TLR-9]. In addition, animals injected with the K137R mutant also demonstrated decreased (2-fold) levels of cross-neutralizing antibodies when compared to animals that received the WT-AAV8 vector. To study further the utility of the novel AAV8-K137R mutant in a therapeutic setting, we delivered human coagulation factor IX (h.FIX) under the control of liver specific promoters (LP1 or hAAT) at two different doses (2.5x10^10 and 1x10^11 vgs per mouse) in 8-12 weeks old male C57BL/6 mice. As can be seen in Fig.1C/D, the circulating levels of h.FIX were higher in all the K137R-AAV8 treated groups as compared to the WT-AAV8 treated groups either at 2 weeks (62% vs 37% for hAAT constructs and 47% vs 21% for LP1 constructs) or 4 weeks (78% vs 56% for hAAT constructs and 64% vs 30% for LP1 constructs) post hepatic gene transfer. These studies demonstrate the feasibility of the use of this novel vector for potential gene therapy of hemophilia B.

Bioestímulo e bioaumento na remediação de solo contaminado com óleo lubrificante usado escala piloto

O óleo lubrificante mineral é amplamente utilizado no cenário mundial no funcionamento de máquinas e motores. No entanto, o ciclo de vida deste petro-derivado resulta na geração de um resíduo (óleo lubrificante usado), o qual é nocivo ao meio ambiente quando não descartado adequadamente ou reciclado. No Brasil, apesar das normas que tratam especificamente do armazenamento, recolhimento e destino de óleo lubrificante usado, grande parte do mesmo ainda é despejado diretamente no meio ambiente, sem qualquer tratamento, sendo de grande importância estudos que visem o entendimento dos processos e o desenvolvimento de tecnologias de remediação de áreas contaminadas por esse resíduo. O objetivo geral do presente trabalho foi conduzir estudos de tratabilidade de solo arenoso contaminado experimentalmente com 5% (m m-1 seco) de óleo lubrificante usado, através de duas diferentes estratégias de biorremediação: bioestímulo e bioaumento. Foram conduzidos dois experimentos. No primeiro, foi avaliada a atividade microbiana aeróbia na biodegradação do OLU através do método respirométrico de Bartha. No segundo, foram montados três biorreatores de fase sólida simulando biopilhas estáticas com aeração forçada, cada um contendo 125 kg de solo e 5% (m m-1 seco) de óleo lubrificante automotivo usado, os quais receberam como tratamento: bioestímulo por ajuste de pH e umidade (BIOSca); bioestímulo por ajuste de pH e umidade associado ao bioaumento com a adição de composto maduro (BIOA1ca) ; e bioestímulo por ajuste de pH e umidade associado ao bioaumento com a adição de composto jovem (BIOA2ca). Foram também montados três biorreatores de bancada simulando biopilhas estáticas sem aeração forçada, cada um contendo 3 kg de solo e 5% (m m-1) do mesmo contaminante, sendo que o primeiro continha solo sem contaminação - CONTsa, o segundo, solo contaminado com ajuste de pH BIOSsa e o terceiro, solo contaminado com adição de 0,3% de azida sódica - ABIOsa. Os tratamentos foram avaliados pela remoção de hidrocarbonetos totais de petróleo (HTPs) e após 120 dias de experimento obteve-se remoções de HTPs de 84,75%, 99,99% e 99,99%, com BIOS, BIOA1 e BIOA2, respectivamente, demonstrando que a estratégia de bioestímulo associada ao bioaumento foram promissoras na remediação do solo contaminado pelo óleo lubrificante usado. Os tratamentos que receberam composto (BIOA1 e BIOA2) não apresentaram diferenças quanto à remoção de HTPs, evidenciando que a fase de maturação dos compostos não apresentou influência na eficiência do processo. No entanto, verificou-se uma eficiência nos tratamentos que receberam composto quando comparado ao tratamento sem adição de composto

Value chain analysis for sea cucumber in the Philippines

This study examined the sea cucumber industry in the Philippines through the value chain lens. The intent was to identify effective pathways for the successful introduction of sandfish culture as livelihood support for coastal communities. Value chain analysis is a high-resolution analytical tool that enables industry examination at a detailed level. Previous industry assessments have provided a general picture of the sea cucumber industry in the country. The present study builds on the earlier work and supplies additional details for a better understanding of the industry's status and problems, especially their implications for the Australian Center for International Agricultural Research (ACIAR) funded sandfish project "Culture of sandfish (Holothuria scabra) in Asia- Pacific" (FIS/2003/059). (PDF contains 54 pages)

A study of the proposed Diamond Creek Reservoir

Politically the Colorado river is an interstate as well as an international stream. Physically the basin divides itself distinctly into three sections. The upper section from head waters to the mouth of San Juan comprises about 40 percent of the total of the basin and affords about 87 percent of the total runoff, or an average of about 15 000 000 acre feet per annum. High mountains and cold weather are found in this section. The middle section from the mouth of San Juan to the mouth of the Williams comprises about 35 percent of the total area of the basin and supplies about 7 percent of the annual runoff. Narrow canyons and mild weather prevail in this section. The lower third of the basin is composed of mainly hot arid plains of low altitude. It comprises some 25 percent of the total area of the basin and furnishes about 6 percent of the average annual runoff.

The proposed Diamond Creek reservoir is located in the middle section and is wholly within the boundary of Arizona. The site is at the mouth of Diamond Creek and is only 16 m. from Beach Spring, a station on the Santa Fe railroad. It is solely a power project with a limited storage capacity. The dam which creats the reservoir is of the gravity type to be constructed across the river. The walls and foundation are of granite. For a dam of 290 feet in height, the back water will be about 25 m. up the river.

The power house will be placed right below the dam perpendicular to the axis of the river. It is entirely a concrete structure. The power installation would consist of eighteen 37 500 H.P. vertical, variable head turbines, directly connected to 28 000 kwa. 110 000 v. 3 phase, 60 cycle generators with necessary switching and auxiliary apparatus. Each unit is to be fed by a separate penstock wholly embedded into the masonry.

Concerning the power market, the main electric transmission lines would extend to Prescott, Phoenix, Mesa, Florence etc. The mining regions of the mountains of Arizona would be the most adequate market. The demand of power in the above named places might not be large at present. It will, from the observation of the writer, rapidly increase with the wonderful advancement of all kinds of industrial development.

All these things being comparatively feasible, there is one difficult problem: that is the silt. At the Diamond Creek dam site the average annual silt discharge is about 82 650 acre feet. The geographical conditions, however, will not permit silt deposites right in the reservoir. So this design will be made under the assumption given in Section 4.

The silt condition and the change of lower course of the Colorado are much like those of the Yellow River in China. But one thing is different. On the Colorado most of the canyon walls are of granite, while those on the Yellow are of alluvial loess: so it is very hard, if not impossible, to get a favorable dam site on the lower part. As a visitor to this country, I should like to see the full development of the Colorado: but how about THE YELLOW!

Design de jornal impresso: a relação entre formato e usabilidade

A presente pesquisa Design de jornal impresso: a relação entre formato e usabilidade tem como principal objetivo reforçar a importância do envolvimento do usuário no design de produtos, através da compreensão da influência que o formato definido pelo tamanho das páginas e organização dos cadernos exerce sobre a usabilidade do jornal impresso. São apresentados os aspectos tecnológicos e econômicos relacionados a esse produto e verificado a influência exercida por eles sobre o formato. É feito um mapeamento sobre os principais aspectos do design de jornais e sobre os diferentes formatos de jornais, suas origens e sua relevância para o design de jornais. Busca-se mapear o formato dos jornais em diferentes mercados e os possíveis impactos das mudanças de formato na circulação, no conteúdo e na publicidade dos jornais impressos. Faz-se uma revisão do panorama atual dos jornais eletrônicos e suas possíveis interferências nos hábitos de leitura. Revisa-se a bibliografia referente à usabilidade, concentrando-se na metodologia de design de documentos e sua aplicabilidade para testes de usabilidade em jornais impressos. Por fim, descreve-se a pesquisa de campo através de entrevistas com especialistas em design de jornais, de observações sistemáticas em gráficas de jornal e sobre o mercado de jornais impressos da cidade do Rio de Janeiro e de testes de usabilidade aplicados a jornais. Conclui-se que, considerando o público participante dos testes profissionais que trabalham com impressão de jornais , apenas o aspecto do formato relacionado à organização dos cadernos parece exercer influência sobre a usabilidade dos jornais.

Innovación, capacidad productiva, formación en el puesto de trabajo y productividad

[ES] Este estudio analiza la relación entre productividad laboral, innovación radical, innovación incremental, tecnología incorporada en maquinaria y equipos, utilización de la capacidad productiva y formación. Los datos utilizados son de empresas españolas, manufactureras y de servicios, procedentes de la Business Environment and enterprise Performance Survey (BEEPS). La técnica empleada para estimar los coeficientes del modelo es la regresión mínimo cuadrática ordinaria, ya que la variable dependiente (productividad laboral) es continua. Los resultados indican que la innovación radical y la formación influyen positiva y significativamente sobre la productividad laboral. La influencia de la tecnología incorporada también es significativa, pero de signo negativo. Finalmente, señalar que las empresas que más exportan y las de mayor tamaño alcanzan mayores niveles de productividad. Los hallazgos encontrados en este estudio tienen implicaciones para los responsables de la política económica. Los decisores políticos españoles deben promover y subsidiar la adquisición de maquinaria y equipos más eficientes para las empresas que alcanzan menores niveles de productividad, así como subsidiar la formación de trabajadores cualificados en el manejo de estos nuevos equipos. Además, también deben promover y subsidiar el desarrollo de actividades de elevado contenido en I+D para las empresas que consiguen elevados niveles de productividad, a fin de que incrementen su desempeño en innovación radical. La promoción y subsidiación de programas de formación relacionados con la I+D también resulta esencial en este tipo de empresas, máxime en un escenario caracterizado por un cambio tecnológico intenso y que se produce con gran rapidez.

Estudo sobre Produção mais Limpa (P+L) aplicado a um serviço de referência de uma Instituição de Ensino e Pesquisa - Rio de Janeiro (RJ).

O objetivo deste trabalho foi avaliar a potencialidade de implantação da Produção Mais Limpa (P+L), através do estudo de caso em Laboratório Biomédico de Referência em uma instituição pública de ensino e pesquisa localizada no Rio de Janeiro. Esta investigação é exploratória e analítica, utilizando-se como instrumentos a revisão bibliográfica e documental, a observação direta e a entrevista com aplicação de questionários voltados aos responsáveis pela área ambiental do laboratório pesquisado. A análise foi realizada confrontando-se os dados levantados com as recomendações da metodologia de P+L, identificando-se as lacunas e oportunidades para a melhoria dos serviços e processos de trabalho. O laboratório possui instalações modernas, organização, sistemas de avaliação da matéria-prima e insumos usados, além do gerenciamento dos resíduos. Contudo, nem todos os procedimentos são validados ou estão adequados às normas. Em geral, problemas em laboratórios dizem respeito ao uso excessivo de substâncias perigosas e ao manejo inadequado de resíduos, o qual pode ser contornado com a P+L, tendo como enfoque a prevenção da poluição e a minimização na fonte geradora. A redução do consumo de materiais e insumos, além da implantação de mudanças nos processos de trabalho, podem diminuir os custos financeiros e os impactos ambientais, como foi demonstrado no estudo. Para a melhoria da gestão dos laboratórios, recomenda-se a continuidade na aquisição, manutenção de equipamentos e infraestrutura. É importante a divulgação de informações ambientais e treinamento permanente para funcionários e alunos. A Sustentabilidade Ambiental só pode ser alcançada quando for bem entendida e absorvida por todos, sendo a alta administração das instituições a maior responsável para liderar esse processo. Para estudos futuros, propõe-se melhor definição e ampliação dos indicadores para o monitoramento e aprimoramento da gestão ambiental. Complementarmente, indicam-se estudos sobre a aquisição de conceitos pelos atores sobre a P+L e como eles podem contribuir com a Sustentabilidade Ambiental e a melhoria no ambiente de trabalho. Espera-se que esta pesquisa auxilie com o aperfeiçoamento da gestão no laboratório estudado e em instituições similares que a venham implantar a P+L.

The Driftnet Fishery in the Fort Pierce-Port Salerno Area off Southeast Florida

From May through September 1987, observations were made on 38 trips in the driftnet fishery off the Fort Pierce-Port Salerno area off southeast Florida. Of the number and weight of fish landed on observed trips, 91.6 percent consisted of king mackerel, Scomberomorus cavalla, the targeted species. Over 33 species of fishes were observed among the discarded by-catch. The most frequently occurring species in the discards was little tunny, Euthynnus alletteratus, which made up 67.0 percent by number of the discarded by-catch. Total landings for all commercial gear from Saint Lucie and Martin counties (the counties of the study area) increased 516,741 pounds from 1986 to 1987. In 1986, 55 percent of the catch was from handline and 45 percent from driftnet landings. In 1987, 78 percent was from driftnet and 22 percent from handline landings. A comparison of lengths from recreational and commercial landings showed recreationally caught fish to be, on the average, smaller. No marine mammals, birds, or turtles were entangled in the net on observed trips. Data on cost of nets. fuel, and supplies plus the distribution of earnings among the crew were obtained for five driftnet boats.

A guide to the fisheries of Ceylon

This bulletin has been written primarily to serve as a handbook to the Indo-Pacific Fisheries Council at its sessions in Colombo during December 1958. It presents an outline of the fishing industry as found in Ceylon today, in non-technical language so that it may also be understood by laymen. The large gaps in our knowledge are brought out in the section on "resources"; the section on "administration" gives an indication of progress made towards solving the problems of the industry, while sections on "fishing methods" and "utilization of catch" will help us to appreciate the handicaps and difficulties of those concerned with the industry. The Government Administration of the fisheries of Ceylon is being assisted to a considerable extent by the generous gifts of machinery and equipment as well as the loan of skilled technical personnel made by other countries through the FAO and the Colombo Plan. The progress made so far has helped to place the industry in a favorable position for further improvements.

Product scheduling for thermal energy reduction in papermaking industries

Papermaking is considered as an energy-intensive industry partly due to the fact that the machinery and procedures have been designed at the time when energy was both cheap and plentiful. A typical paper machine manufactures a variety of different products (grades) which impose variable per-unit raw material and energy costs to the mill. It is known that during a grade change operation the products are not market-worthy. Therefore, two different production regimes, i.e. steady state and grade transition can be recognised in papermaking practice. Among the costs associated with paper manufacture, the energy cost is 'more variable' due to (usually) day-to-day variations of the energy prices. Moreover, the production of a grade is often constrained by customer delivery time requirements. Given the above constraints and production modes, the product scheduling technique proposed in this paper aims at optimising the sequence of orders in a single machine so that the cost of production (mainly determined by the energy) is minimised. Simulation results obtained from a commercial board machine in the UK confirm the effectiveness of the proposed method. © 2011 IFAC.

盆地地下水资源开发的数值模拟研究－以山西灵石静升盆地为例

Groundwater basin is important for water supply in northern China. The paper took the Jingsheng Basin in Lingshi County, Shanxi Province as a case to study the basin groundwater system by numerical modeling. The hydrogeological characteristics were analysed basing on the field investigation, and a three-dimensional groundwater flow model was established to describe the groundwater flow system in the Jingsheng groundwater basin. The boundary of the model was determined by using geophysics and GIS data, and the lumped parameter model of runoff was used to depict the transform between the surface water and groundwater, and the groundwater dating data was used to calibrate the model. All these methods were used to improve the model. The Software Visual MODFLOW 2000 was applied to set up the numerical groundwater flow model. The groundwater flow pattern in the average year, the high-water year and the low-water year were simulated respectively by the model. Some new cognition to the groundwater movement in Jingsheng Basin was obtained in the paper. The difficult problems were resolved when using the conventional and theoretical analysis to forecast and appraise the exploitation of the groundwater, and supplies the instructional technology base for the reasonable exploitation and optimization collocation. The numerical model will improve evaluation of the basin groundwater resources.