Generation of a high-titer retroviral vector capable of expressing high levels of the human beta-globin gene.

Retrovirus-mediated gene transfer into hematopoietic cells may provide a means of treating both inherited and acquired diseases involving hematopoietic cells. Implementation of this approach for disorders resulting from mutations affecting the beta-globin gene (e.g., beta-thalassemia and sickle cell anemia), however, has been hampered by the inability to generate recombinant viruses able to efficiently and faithfully transmit the necessary sequences for appropriate gene expression. We have addressed this problem by carefully examining the interactions between retroviral and beta-globin gene sequences which affect vector transmission, stability, and expression. First, we examined the transmission properties of a large number of different recombinant proviral genomes which vary both in the precise nature of vector, beta-globin structural gene, and locus control region (LCR) core sequences incorporated and in the placement and orientation of those sequences. Through this analysis, we identified one specific vector, termed M beta 6L, which carries both the human beta-globin gene and core elements HS2, HS3, and HS4 from the LCR and faithfully transmits recombinant proviral sequences to cells with titers greater than 10(6) per ml. Populations of murine erythroleukemia (MEL) cells transduced by this virus expressed levels of human beta-globin transcript which, on a per gene copy basis, were 78% of the levels detected in an MEL-derived cell line, Hu11, which carries human chromosome 11, the site of the beta-globin locus. Analysis of individual transduced MEL cell clones, however, indicated that, while expression was detected in every clone tested (n = 17), the levels of human beta-globin treatment varied between 4% and 146% of the levels in Hu11. This clonal variation in expression levels suggests that small beta-globin LCR sequences may not provide for as strict chromosomal position-independent expression of beta-globin as previously suspected, at least in the context of retrovirus-mediated gene transfer.

Desenvolvimento e avaliação de métodos de extração e separação cromatográfica em colunas monolíticas e superficialmente porosas para determinação de herbicidas triazínicos em solos e águas

O uso de pesticidas levou ao aumento da produtividade e qualidade dos produtos agrícolas, porém o seu uso acarreta na intoxicação dos seres vivos pela ingestão gradativa de seus resíduos que contaminam o solo, a água e os alimentos. Dessa forma, há a necessidade do monitoramento constante de suas concentrações nos compartimentos ambientais. Para isto, busca-se o desenvolvimento de métodos de extração e enriquecimento de forma rápida, com baixo custo, gerando um baixo volume de resíduos, contribuindo com a química verde. Dentre estes métodos destacam-se a extração por banho de ultrassom e a extração por ponto nuvem. Após o procedimento de extração, o extrato obtido pode ser analisado por técnicas de Cromatografia a Líquido de Alta Eficiência (HPLC) e a Cromatografia por Injeção Sequencial (SIC), empregando fases estacionárias modernas, tais como as monolíticas e as partículas superficialmente porosas. O emprego de SIC com coluna monolítica (C18, 50 x 4,6 mm) e empacotada com partículas superficialmente porosas (C18, 30 x 4,6 mm, tamanho de partícula 2,7 µm) foi estudado para separação de simazina (SIM) e atrazina (ATR), e seus metabólitos, desetilatrazina (DEA), desisopropilatrazina (DIA) e hidroxiatrazina (HAT). A separação foi obtida por eluição passo-a-passo, com fases móveis compostas de acetonitrila (ACN) e tampão Acetato de Amônio/Ácido acético (NH4Ac/HAc) 2,5 mM pH 4,2. A separação na coluna monolítica foi realizada com duas fases móveis: MP1= 15:85 (v v-1) ACN:NH4Ac/HAc e MP2= 35:65 (v v-1) ACN:NH4Ac/HAc a uma vazão de 35 µL s-1. A separação na coluna com partículas superficialmente porosas foi efetivada com as fases móveis MP1= 13:87 (v v-1) ACN: NH4Ac/HAc e MP2= 35:65 (v v-1) ACN:NH4Ac/HAc à vazão de 8 µL s-1. A extração por banho de ultrassom em solo fortificado com os herbicidas (100 e 1000 µg kg-1) resultou em recuperações entre 42 e 160%. A separação de DEA, DIA, HAT, SIM e ATR empregando HPLC foi obtida por um gradiente linear de 13 a 35% para a coluna monolítica e de 10 a 35% ACN na coluna com partículas superficialmente porosas, sendo a fase aquosa constituída por tampão NH4Ac/HAc 2,5 mM pH 4,2. Em ambas as colunas a vazão foi de 1,5 mL min-1 e o tempo de análise 15 min. A extração por banho de ultrassom das amostras de solo com presença de ATR, fortificadas com concentrações de 250 a 1000 µg kg-1, proporcionou recuperações entre 40 e 86%. A presença de ATR foi confirmada por espectrometria de massas. Foram realizados estudos de fortificação com ATR e SIM em amostras de água empregando a extração por ponto nuvem com o surfactante Triton-X114. A separação empregando HPLC foi obtida por um gradiente linear de 13 a 90% de ACN para a coluna monolítica e de 10 a 90% de ACN para a coluna empacotada, sempre em tampão NH4Ac/HAc 2,5 mM pH 4,2. Em ambas as colunas a vazão foi de 1,5 mL min-1 e o tempo de análise 16 min. Fortificações entre 1 e 50 µg L-1 resultaram em recuperações entre 65 e 132%.

Falling short of expectations? Stress-testing the European banking system. CEPS Policy Brief No. 315, 15 January 2014

Before the ECB takes over responsibility for overseeing Europe’s largest banks, as foreseen in the establishment of a eurozone banking union, it plans to conduct an Asset Quality Review (AQR) throughout the coming year, which will identify the capital shortfalls of these banks. This study finds that a comprehensive and decisive AQR will most likely reveal a substantial lack of capital in many peripheral and core European banks. The authors provide estimates of the capital shortfalls of banks that will be stress-tested under the AQR using publicly available data and a series of shortfall measures. Their analysis identifies which banks will most likely need capital, where a public back stop is likely to be needed and, since many countries are already highly leveraged, where an EU-wide backstop might be necessary.

10Be records of sediment cores from high northern latitudes

The 10Be records of four sediment cores forming a transect from the Norwegian Sea at 70°N (core 23059) via the Fram Strait (core 23235) to the Arctic Ocean at 86°N (cores 1533 and 1524) were measured at a high depth resolution. Although the material in all the cores was controlled by different sedimentological regimes, the 10Be records of these cores were superimposed by glacial/interglacial changes in the sedimentary environment. Core sections with high 10Be concentrations ( >1 * 10**9 at/g) are related to interglacial stages and core sections with low10Be concentrations ( <0.5 * 10**9 at/g) are related to glacial stages. Climatic transitions (e.g., Termination II, 5/6) are marked by drastic changes in the 10Be concentrations of up to one order of magnitude. The average 10Be concentrations for each climatic stage show an inverse relationship to their corresponding sedimentation rates, indicating that the 10Be records are the result of dilution with more or less terrigenous ice-rafted material. However, there are strong changes in the 10Be fluxes (e.g., Termination II) into the sediments which may also account for the observed oscillations. Most likely, both processes affected the 10Be records equally, amplifying the contrast between lower (glacials) and higher (interglacials) 10Be concentrations. The sharp contrast of high and low 10Be concentrations at climatic stage boundaries are an independent proxy for climatic and sedimentary change in the Nordic Seas and can be applied for stratigraphic dating (10Be stratigraphy) of sediment cores from the northern North Atlantic and the Arctic Ocean.

Biostratigraphic events close to the Tortonian-Messinian boundary at ODP Hole 107-654A (Table 1)

Correlations of biostratigraphic datums to the geomagnetic reversal time scale (GRTS) at Leg 107 sites provide a means of correlating these datums to sections outside the Mediterranean. Unfortunately, poor recovery and core deformation due to rotary drilling at Sites 651, 652, and 654 severely hampered efforts to acquire detailed magnetostratigraphies and biostratigraphies. However, many biostratigraphic markers could be correlated to the GRTS, including those close to the Miocene/Pliocene and Tortonian/Messinian boundaries. These boundaries are interpreted to occur in Chrons 3r and 3B, respectively (chron nomenclature after Cox, 1982). Comparison of the correlation of Plio-Pleistocene calcareous plankton biostratigraphic events to the GRTS in the Mediterranean and in the open oceans indicates that many events are broadly synchronous between the two environments. The outstanding exception is the first occurrence of Globorotalia margaritae which is delayed in the Mediterranean by about 1 m.y.

(Table 1) Abundances of Discoaster species in ODP Hole 130-806C

Members of the calcareous nannofossil genus Discoaster have been used extensively to subdivide Tertiary deep-sea sediments into biostratigraphic zones or subzones (e.g., Martini, 1971; Bukry, 1973). Haq and Lohmann (1976) mapped biogeographic migrations of this group through time and over latitude. They suggested that expansions and contractions of Discoaster-dominated assemblages across latitudes reflect sea-surface temperature changes. Subsequently, late Pliocene Discoaster species were counted at closely spaced sample intervals from various Atlantic sites (Backman et al., 1986; Backman and Pestiaux, 1987; Chepstow-Lusty et al., 1989, 1991), and Indian Ocean as well as Pacific Ocean sites (Chepstow-Lusty, 1990). In addition to the biostratigraphic information revealing positions and the precision by which the different late Pliocene Discoaster species can be determined, these studies also demonstrated that discoasters strongly fluctuate in abundance as a function of time. These abundance variations occur in equatorial as well as temperate temperature regimes, and show periodicities that reflect orbital frequencies. Chepstow-Lusty et al. (1989, 1991) also suggested that the oscillating abundances partly represent productivity pressure, because discoasters tend to show low abundances under high productivity conditions and vice versa. In the Pacific Ocean, counts showing late Pliocene Discoaster abundances exist from three sites, namely Ocean Drilling Program (ODP) Site 677 in the eastern equatorial upwelling region, Core V28-179 from the central equatorial region, and Core V32-127 from the mid-latitude Hess Rise. The two Vema cores are condensed and show sedimentation rates below 0.5 cm/1000 yr, thus offering a poorly resolved stratigraphy. Hole 806C from the Ontong Java Plateau provided an opportunity to establish a highly resolved Discoaster record from the western extreme of the equatorial Pacific under an environmental setting that differed from ODP Site 677 by being less influenced by intense upwelling. The Discoaster counting technique is described by Backman and Shackleton (1983).

Paleotemperature reconstructions of the Atlantic Sector of the Southern Ocean

Diatom assemblages from ODP Leg 177 sites 1093, 1094 and core PS2089-2, from the present Antarctic sea ice free zone and close to the Polar Front, were analyzed in order to reconstruct the climate development around the Mid-Brunhes Event 400 000 yr ago, as reflected by summer sea surface temperature (SSST) and sea ice distribution. Dense sample spacing allows a mean temporal resolution during Marine Isotope Stage (MIS) 11 (423-362 ka) of 300-400 yr. SSST values were estimated from diatom assemblages using a transfer function technique. The distribution pattern of sea ice diatoms indicates that the present-day ice free Antarctic Zone was seasonally covered by sea ice during the cold MIS 12 and MIS 10. These glacial periods are characterized by sea ice fluctuations with a periodicity of 3 and 1.85 kyr, suggesting the occurrence of Dansgaard-Oeschger-style millennial-scale oscillations in the Atlantic sector of the Southern Ocean during the glacial stages MIS 12 and MIS 10. Termination V (MIS 12/11) is characterized by a distinct temperature increase of 4-6°C, intersected especially at the southern site 1094 and core PS2089-2 by two distinct cooling events reminiscent of the Younger Dryas, which are associated with a northward shift of the winter sea ice edge in the Antarctic Zone. The SSST record is characterized by distinct temperature intervals bounded by stepwise, rapid changes. Maximum temperatures were reached during Termination V and the early MIS 11, exceeding modern values by 2°C over a period of 8 kyr. This pattern indicates a very early response of the Southern Ocean to global climate on Milankovitch-driven climate variability. The SSST optimum is marked by millennial-scale temperature oscillations with an amplitude of ca. 1°C and periodicities of ca. 1.85 and 1.47 kyr, probably reflecting changes in the ocean circulation system. The SSSTs during the MIS 11 temperature optimum do not exceed values obtained from other interglacial optima such as the early periods of MIS 5 or MIS 1 from the Antarctic Zone. However, the total duration of the warmest period was distinctly longer than observed from other interglacials. The comparison of the South Atlantic climate record with a high-resolution record from ODP Leg 162, site 980from the North Atlantic shows a strong conformity in the climate development during the studied time interval.

Proceedings of the Ocean Drilling Program.

Vols. for 176- consist of print manual containing summary report accompanied by CD-ROMs in pocket, containing (in 2 or more CDs) PDF version of the full-text of the report along with log and core data.

Modeling the age of tropical moist forest fragments in heavily-cleared lowland landscapes of Colombia

Increasingly, large areas of native tropical forests are being transformed into a mosaic of human dominated land uses with scattered mature remnants and secondary forests. In general, at the end of the land clearing process, the landscape will have two forest components: a stable component of surviving mature forests, and a dynamic component of secondary forests of different ages. As the proportion of mature forests continues to decline, secondary forests play an increasing role in the conservation and restoration of biodiversity. This paper aims to predict and explain spatial and temporal patterns in the age of remnant mature and secondary forests in lowland Colombian landscapes. We analyse the age distributions of forest fragments, using detailed temporal land cover data derived from aerial photographs. Ordinal logistic regression analysis was applied to model the spatial dynamics of mature and secondary forest patches. In particular, the effect of soil fertility, accessibility and auto-correlated neighbourhood terms on forest age and time of isolation of remnant patches was assessed. In heavily transformed landscapes, forests account for approximately 8% of the total landscape area, of which three quarters are comprised of secondary forests. Secondary forest growth adjacent to mature forest patches increases mean patch size and core area, and therefore plays an important ecological role in maintaining landscape structure. The regression models show that forest age is positively associated with the amount of neighbouring forest, and negatively associated with the amount of neighbouring secondary vegetation, so the older the forest is the less secondary vegetation there is adjacent to it. Accessibility and soil fertility also have a negative but variable influence on the age of forest remnants. The probability of future clearing if current conditions hold is higher for regenerated than mature forests. The challenge of biodiversity conservation and restoration in dynamic and spatially heterogeneous landscape mosaics composed of mature and secondary forests is discussed. (c) 2004 Elsevier B.V. All rights reserved.

Quantifying aphasic people's social lives in the context of non-aphasic peers

Background: Developing the knowledge base on the impact of aphasia on people's social lives has become increasingly important in recent times to further our understanding of the broad consequences of communication disability and thus provide appropriate services. Past research clearly indicates that relationships and social activities with family members and others undergo change with the onset of aphasia in an individual, however more evidence of a quantitative nature would be beneficial. Aims: The current research furthers our knowledge by quantifying chronically aphasic older people's regular social contacts and social activities, and places them in context by comparing them with healthy older people of similar age and education. Methods & Procedures: A total of 30 aphasic participants aged 57 to 88 years, and 71 non-aphasic controls aged 62 to 98 years were interviewed by a speech and language therapist using self-report measures of Social Network Analysis (Antonucci & Akiyama, 1987) and Social Activities Checklist (Cruice, 2001, in Worrall & Hickson, 2003). Demographic information was also collected. Descriptive statistics are presented and independent samples t tests were used to examine differences between the groups. Outcomes & Results: Participants with primarily mild to moderate aphasic impairment reported a considerable range of social contacts (5-51) and social activities (8-18). Many significant differences were evident between the two groups' social contacts and activities. On average, aphasic participants had nine fewer social contacts (mainly friend'' relationships) and three fewer social activities (mainly leisure'' activities) than their non-aphasic peers. The majority of controls were satisfied with their social activities, whereas the majority of aphasic participants were not and wanted to be doing more. There were some general similarities between the groups, in terms of range of social contacts, overall pattern of social relationships, and core social activities. Conclusions: Older people with chronic aphasia had significantly fewer social contacts and social activities than their peers. People with aphasia expressed a desire to increase the social activity of their lives. Given the importance of leisure activity and relationships with friends as well as family for positive well-being, speech and language therapists may direct their rehabilitation efforts towards two areas: (1) conversational partner programmes training friends to maintain these relationships; and (2) encouraging and supporting aphasic clients in leisure activities of their choice.

A INFLUÊNCIA DAS TEORIAS ECONÔMICAS NA DECISÃO DE TERCEIRIZAR O DESPACHO ADUANEIRO NO SETOR DE COMÉRCIO ATACADISTA NA CIDADE DE SÃO PAULO

O objetivo central deste trabalho é verificar a influência das teorias econômicas na decisão das empresas de comércio atacadista em terceirizar o despacho aduaneiro na cidade de São Paulo. Para isso, a história da terceirização é apresentada brevemente, seguida do levantamento e estudos sobre terceirização e fundamentação das teorias econômicas que mostram possíveis influências na decisão das empresas em terceirizar o despacho aduaneiro: a Teoria do Custo das Transações, a Teoria da Agência, a Teoria do Resource-Based View (RBV) e a Teoria das Competências Essenciais. Além de contextualizar o comércio exterior brasileiro, levando em conta sua história e representatividade econômica, foram apresentadas as atividades ligadas ao despacho aduaneiro, que podem levar as empresas a terceirizar este serviço na cidade de São Paulo. Após a explanação sobre o tema, por meio de pesquisa empírica valendo-se de questionário de perguntas fechadas, aplicado a uma amostra não probabilística por conveniência, verificou-se a relação existente entre as teorias mencionadas acima e o quanto elas influenciam o processo das empresas importadoras e exportadoras do comércio atacadista para terceirizarem o despacho aduaneiro, foi usado o teste ANOVA e possíveis complementações como o do Teste de Dunnett T3 e do Teste de Tukey que constataram a diferença de respostas entre os respondentes que terceirizam totalmente, parcialmente ou não terceirizam esta atividade, além da diferença entre estes níveis de terceirização verificou-se que todas as teorias econômicas apresentadas neste trabalho possuem alguma influência sobre a terceirização do despacho aduaneiro.

Editorial

This second issue of Knowledge Management Research & Practice (KMRP) continues the international nature of the first issue, with papers from authors based on four different continents. There are five regular papers, plus the first of what is intended to be an occasional series of 'position papers' from respected figures in the knowledge management field, who have specific issues they wish to raise from a personal standpoint. The first two regular papers are both based on case studies. The first is 'Aggressively pursuing knowledge management over two years: a case study a US government organization' by Jay Liebowitz. Liebowitz is well known to both academics and practictioners as an author on knowledge management and knowledge based systems. Government departments in many Western countries must soon face up to the problems that will occur as the 'baby boomer' generation reaches retirement age over the next decade. This paper describes how one particular US government organization has attempted to address this situation (and others) through the introduction of a knowledge management initiative. The second case study paper is 'Knowledge creation through the synthesizing capability of networked strategic communities: case study on new product development in Japan' by Mitsuru Kodama. This paper looks at the importance of strategic communities - communities that have strategic relevance and support - in knowledge management. Here, the case study organization is Nippon Telegraph and Telephone Corporation (NTT), a Japanese telecommunication firm. The third paper is 'Knowledge management and intellectual capital: an empirical examination of current practice in Australia' by Albert Zhou and Dieter Fink. This paper reports the results of a survey carried out in 2001, exploring the practices relating to knowledge management and intellectual capital in Australia and the relationship between them. The remaining two regular papers are conceptual in nature. The fourth is 'The enterprise knowledge dictionary' by Stuart Galup, Ronald Dattero and Richard Hicks. Galup, Dattero and Hicks propose the concept of an enterprise knowledge dictionary and its associated knowledge management system architecture as offering the appropriate form of information technology to support various different types of knowledge sources, while behaving as a single source from the user's viewpoint. The fifth and final regular paper is 'Community of practice and metacapabilities' by Geri Furlong and Leslie Johnson. This paper looks at the role of communities of practice in learning in organizations. Its emphasis is on metacapabilities - the properties required to learn, develop and apply skills. This discussion takes work on learning and core competences to a higher level. Finally, this issue includes a position paper 'Innovation as an objective of knowledge management. Part I: the landscape of management' by Dave Snowden. Snowden has been highly visible in the knowledge management community thanks to his role as the Director of IBM Global Services' Canolfan Cynefin Centre. He has helped many government and private sector organizations to consider their knowledge management problems and strategies. This, the first of two-part paper, is inspired by the notion of complexity. In it, Snowden calls for what he sees as a 20th century emphasis on designed systems for knowledge management to be consigned to history, and replaced by a 21st century emphasis on emergence. Letters to the editor on this, or any other topic related to knowledge management research and practice, are welcome. We trust that you will find the contributions stimulating, and again invite you to contribute your own paper(s) to future issues of KMRP.

The drying of small drops of particulate slurries

The literature on the evaporation of drops of pure liquids, drops containing solids and droplet sprays has been critically reviewed. An experimental study was undertaken on the drying of suspended drops of pure water and aqueous sodium sulphate decahydrate with concentrations varying from 5 to 54. 1 wt. %. Individual drops were suspended from a glass filament balance in a 26 mm I.D. vertical wind tunnel, designed and constructed to supply hot de-humidified air, to simulate conditions encountered in commercial spray driers. A novel thin film thermocouple was developed to facilitate the simultaneous measurement of drop weight and core temperature. The heat conduction through the thermocouple was reduced because of its unique design; using essentially a single 50μ diameter nickel wire. For pure water drops, the Nusselt number was found to be a function of the Reynolds, Prandtl and Transfer numbers for a temperature range between 19 to 79°C.                  Nu = 2 + 0.19 (1/B)0.24 Re0.5 Pr0.33 Two distinct periods were observed during the drying of aqueous sodium sulphate decahydrate. The first period was characterised by the evaporation from a free liquid surface, whilst drying in the second period was controlled by the crust resistance. Fracturing of the crust occurred randomly but was more frequent at higher concentrations and temperatures. A model was proposed for the drying of slurry drops, based on a receding evaporation interface. The model was solved numerically for the variation of core temperature, drop weight and crust thickness as a function of time. Experimental results were in excellent agreement with the model predictions although at higher temperatures modifications to the model had to be made to accommodate the unusual behaviour of sodium sulphate slurries, i.e. the formation of hydrates.

Are there two distinct populations of cored senile plaques in senile dementia of the Alzheimer type?

The relationship between plaque diameter (PD) and core diameter (CD) was studied in four brains from each of four SDAT brains. The regions studied were parahippocampal gyrus (PHG), hippocampus, frontal and inferior temporal lobes. The largest diameters of 100 cored classical plaques and their cores were measured. CD was positively correlated with PD (Pearson's 'r' 0.4 - 0.95) in all region studied. Significant linear regressions of CD on PD with positive slopes (0.10 - 0.65) were found. Two distinct types of regression were found. Type A had a steep slope and a negative intercept on the ordinate whereas Type B had a shallow slope and a positive intercept. Both types can be found within the same brain but Type A or B predominate in a particular tissue. The data suggest that core development may occur either early or late in the development of the plaque. The two types of plaque may thus have different aetiologies. Such an interpretation is consistent with current ideas of plaque formation.