Polymorphism in the 3′-untranslated region of TNFα mRNA impairs binding of the post-transcriptional regulatory protein HuR to TNFα mRNA

Tumor necrosis factor α (TNFα) acts as a beneficial mediator in the process of host defence. In recent years major interest has focused on the AU-rich elements (AREs) present in the 3′-untranslated region (3′-UTR) of TNFα mRNA as this region plays a pivotal role in post-transcriptional control of TNFα production. Certain stimuli, such as lipopolysaccharides, a component of the Gram-negative bacterial cell wall, have the ability to relinquish the translational suppression of TNFα mRNA imposed by these AREs in macrophages, thereby enabling the efficient production of the TNFα. In this study we show that the polymorphism (GAU trinucleotide insertional mutation) present in the regulatory 3′-UTR of TNFα mRNA of NZW mice results in the hindered binding of RNA-binding proteins, thereby leading to a significantly reduced production of TNFα protein. We also show that the binding of macrophage proteins to the main ARE is also decreased by another trinucleotide (CAU) insertion in the TNFα 3′-UTR. One of the proteins affected by the GAU trinucleotide insertional mutation was identified as HuR, a nucleo-cytoplasmic shuttling protein previously shown to play a prominent role in the stability and translatability of mRNA containing AREs. Since binding of this protein most likely modulates the stability, translational efficiency and transport of TNFα mRNA, these results suggest that mutations in the ARE of TNFα mRNA decrease the production of TNFα protein in macrophages by hindering the binding of HuR to the ARE.

Electrospray ionization mass spectrometry analysis of changes in phospholipids in RBL-2H3 mastocytoma cells during degranulation

Biological membranes contain an extraordinary diversity of lipids. Phospholipids function as major structural elements of cellular membranes, and analysis of changes in the highly heterogeneous mixtures of lipids found in eukaryotic cells is central to understanding the complex functions in which lipids participate. Phospholipase-catalyzed hydrolysis of phospholipids often follows cell surface receptor activation. Recently, we demonstrated that granule fusion is initiated by addition of exogenous, nonmammalian phospholipases to permeabilized mast cells. To pursue this finding, we use positive and negative mode Fourier-transform ion cyclotron resonance mass spectrometry (FTICR-MS) to measure changes in the glycerophospholipid composition of total lipid extracts of intact and permeabilized RBL-2H3 (mucosal mast cell line) cells. The low energy of the electrospray ionization results in efficient production of molecular ions of phospholipids uncomplicated by further fragmentation, and changes were observed that eluded conventional detection methods. From these analyses we have spectrally resolved more than 130 glycerophospholipids and determined changes initiated by introduction of exogenous phospholipase C, phospholipase D, or phospholipase A2. These exogenous phospholipases have a preference for phosphatidylcholine with long polyunsaturated alkyl chains as substrates and, when added to permeabilized mast cells, produce multiple species of mono- and polyunsaturated diacylglycerols, phosphatidic acids, and lysophosphatidylcholines, respectively. The patterns of changes of these lipids provide an extraordinarily rich source of data for evaluating the effects of specific lipid species generated during cellular processes, such as exocytosis.

Transcription elongation factor of respiratory syncytial virus, a nonsegmented negative-strand RNA virus.

RNA synthesis by the paramyxovirus respiratory syncytial virus, a ubiquitous human pathogen, was found to be more complex than previously appreciated for the nonsegmented negative-strand RNA viruses. Intracellular RNA replication of a plasmid-encoded "minigenome" analog of viral genomic RNA was directed by coexpression of the N, P, and L proteins. But, under these conditions, the greater part of mRNA synthesis terminated prematurely. This difference in processivity between the replicase and the transcriptase was unanticipated because the two enzymes ostensively shared the same protein subunits and template. Coexpression of the M2 gene at a low level of input plasmid resulted in the efficient production of full-length mRNA and, in the case of a dicistronic minigenome, sequential transcription. At a higher level, coexpression of the M2 gene inhibited transcription and RNA replication. The M2 mRNA contains two overlapping translational open reading frames (ORFs), which were segregated for further analysis. Expression of the upstream ORF1, which encoded the previously described 22-kDa M2 protein, was associated with transcription elongation. A model involving this protein in the balance between transcription and replication is proposed. ORF2, which lacks an assigned protein, was associated with inhibition of RNA synthesis. We propose that this activity renders nucleocapsids synthetically quiescent prior to incorporation into virions.

The 1952 cotton goal : 16 million bales from 28 million acres.

"February 1952."

Characterisation of a harvesting step for monoclonal antibodies from mammalian cell culture

The focus of this research was defined by a poorly characterised filtration train employed to clarify culture broth containing monoclonal antibodies secreted by GS-NSO cells: the filtration train blinded unpredictably and the ability of the positively charged filters to adsorb DNA from process material was unknown. To direct the development of an assay to quantify the ability of depth filters to adsorb DNA, the molecular weight of DNA from a large-scale, fed-batch, mammalian cell culture vessel was evaluated as process material passed through the initial stages of the purification scheme. High molecular weight DNA was substantially cleared from the broth after passage through a disc stack centrifuge and the remaining low molecular weight DNA was largely unaffected by passage through a series of depth filters and a sterilising grade membrane. Removal of high molecular weight DNA was shown to be coupled with clarification of the process stream. The DNA from cell culture supernatant showed a pattern of internucleosomal cleavage of chromatin when fractionated by electrophoresis but the presence of both necrotic and apoptotic cells throughout the fermentation meant that the origin of the fragmented DNA could not be unequivocally determined. An intercalating fluorochrome, PicoGreen, was elected for development of a suitable DNA assay because of its ability to respond to low molecular weight DNA. It was assessed for its ability to determine the concentration of DNA in clarified mammalian cell culture broths containing pertinent monoclonal antibodies. Fluorescent signal suppression was ameliorated by sample dilution or by performing the assay above the pI of secreted IgG. The source of fluorescence in clarified culture broth was validated by incubation with RNase A and DNase I. At least 89.0 % of fluorescence was attributable to nucleic acid and pre-digestion with RNase A was shown to be a requirement for successful quantification of DNA in such samples. Application of the fluorescence based assay resulted in characterisation of the physical parameters governing adsorption of DNA by various positively charged depth filters and membranes in test solutions and the DNA adsorption profile of the manufacturing scale filtration train. Buffers that reduced or neutralised the depth filter or membrane charge, and those that impeded hydrophobic interactions were shown to affect their operational capacity, demonstrating that DNA was adsorbed by a combination of electrostatic and hydrophobic interactions. Production-scale centrifugation of harvest broth containing therapeutic protein resulted in the reduction of total DNA in the process stream from 79.8 μg m1-1 to 9.3 μg m1-1 whereas the concentration of DNA in the supernatant of pre-and post-filtration samples had only marginally reduced DNA content: from 6.3 to 6.0 μg m1-1 respectively. Hence the filtration train was shown to ineffective in DNA removal. Historically, blinding of the depth filters had been unpredictable with data such as numbers of viable cells, non-viable cells, product titre, or process shape (batch, fed-batch, or draw and fill) failing to inform on the durability of depth filters in the harvest step. To investigate this, key fouling contaminants were identified by challenging depth filters with the same mass of one of the following: viable healthy cells, cells that had died by the process of apoptosis, and cells that had died through the process of necrosis. The pressure increase across a Cuno Zeta Plus 10SP depth filter was 2.8 and 16.5 times more sensitive to debris from apoptotic and necrotic cells respectively, when compared to viable cells. The condition of DNA released into the culture broth was assessed. Necrotic cells released predominantly high molecular weight DNA in contrast to apoptotic cells which released chiefly low molecular weight DNA. The blinding of the filters was found to be largely unaffected by variations in the particle size distribution of material in, and viscosity of, solutions with which they were challenged. The exceptional response of the depth filters to necrotic cells may suggest the cause of previously noted unpredictable filter blinding whereby a number of necrotic cells have a more significant impact on the life of a depth filter than a similar number of viable or apoptotic cells. In a final set of experiments the pressure drop caused by non-viable necrotic culture broths which had been treated with DNase I or benzonase was found to be smaller when compared to untreated broths: the abilities of the enzyme treated cultures to foul the depth filter were reduced by 70.4% and 75.4% respectively indicating the importance of DNA in the blinding of the depth filter studied.

Correlates of leaf optical properties in tropical forest sun and extreme-shade plants

Thirteens hade-adaptedr ain forest species were comparedw ith twelve sun-adaptedt ropical forest species for correlates to leaf optical properties (described previously in Amer. J. Bot. 73: 1100-1108). The two samples were similar in absorptance of quanta for photosynthesis, but the shade-adaptedt axa: 1) had significantlyl ower specificl eaf weights,i ndicatinga more metabolically efficient production of surface for quantum capture; 2) synthesized less chlorophyll per unit area; and 3) used less chlorophyll for capturing the same quanta for photosynthesis. The anatomical features that best correlate with this increased efficiency are palisade cell shape and chloroplast distribution. Palisade cells with more equal dimensions have more chloroplasts on their abaxial surfaces. This dense layer of chloroplasts maximizes the light capture efficiency limited by sieve effects. The more columnar palisade cells of sun-adapted taxa allow light to pass through the central vacuoles and spaces between cells, making chloroplasts less efficient in energy capture, but allowing light to reach chloroplasts in the spongy mesophyll. Pioneer species may be an exception to these two groups of species. Three pioneer taxa included in this study have columnar palisade cells that are extremely narrow and packed closely together. This layer allows little penetration of light, but exposure of the leaf undersurface may provide illumination of spongy mesophyll chloroplasts in these plants.

Gestão do sistema de produção e práticas enxutas adotadas nas indústrias paraibanas: um estudo multicaso

The evolution of the practices and strategies of manufacturing management, over the years, has made many companies realign their production systems in order to raise their competitiveness and operational performance. However, in most cases these changes are made in a heterogeneous manner, which ends up leaving the production system without a defined goal, which may end up damaging the managerial strategies of the organization as a whole. Thus, some organizations seek to use techniques and/or successful production practices used by other companies, believing can be able to reproduce the same results. An efficient production system must be fully planned and appropriate to the strategic objectives of the organization. Thus, this paper aims to identify the manufacturing management strategies adopted in paraibanas industries, as well as identify the lean practices used by them. Thus, a qualitative study was conducted, using as methodological basis the multicase study. Were made: direct observations, semi-structured interviews and questionnaires applied to those responsible by the production sector of the participating companies. As a result, it was possible to identify the type of manufacturing management system adopted by companies. Where it was detected that Company A uses a system of Modern Mass Production with focus on productivity and low cost and the Company B is using Lean Manufacturing system focused on quality and diversity. In the two organizations was possible to realize the application of lean practices where the Company what does not use the LM, possessed lean practices in standard extremely mature of utilization.

Värdeflödesanalys och Data Envelopment Analysis : En fallstudie för att värdera effektiviteten hos ett logistikföretag

Companies face new challenges almost every day. In order to stay competitive, it is important that companies strive for continuous development and improvement. By describing companies through their processes it is possible to get a clear overview of the entire operation, which can contribute, to a well-established overall understanding of the company. This is a case study based on Stort AB which is a small logistics company specialized in international transportation and logistics solutions. The purpose of this study is to perform value stream mapping in order to create a more efficient production process and propose possible improvements in order to reduce processing time. After performing value stream mapping, data envelopment analysis is used to calculate how lean Stort AB is today and how lean the company can become by implementing the proposed improvements. The results show that the production process can improve efficiency by minimizing waste produced by a bad workplace layout and over-processing. The authors suggested solution is to introduce standardized processes and invest in technical instruments in order to automate the process to reduce process time. According to data envelopment analysis the business is 41 percent lean at present and may soon become 55 percent lean and finally reach an optimum 100 percent lean mode if the process is automated.

Utilization of coal combustion residues in glass manufacturing process

The focus of paper is to asses and evaluate new utilisation method of coals combustion resides in glass manufacturing process. Mathematical model of glass manufacturing material balance was used to find favourable proportion of normally used batch materials and coal ash. It was found that possible to substitute up to 20 % of batch with coal ash. On the world glass production scale there is a potential to save 8,4 million tons of silica sand, 6 million tons of dolomite, 3 million tons of clay and 0,2 million tons of lime borate. Furthermore, potential to utilize 2 % of coal combustion products with suggested method.

Avaliação da crioinjúria em ovócitos imaturos e maturados in vitro e produção de embriões a partir de ovócitos bovinos vitrificados em Open Pulled Straw (OPS) e Cryotop

Dissertação (mestrado)—Universidade de Brasília, Faculdade de Ciências Médicas, Programa de Pós-Graduação em Ciências Médicas, 2012.

A reestruturação logística dos serviços no Exército : Estudo de caso: O fardamento.

O Exército tem vindo a sofrer inúmeras reestruturações em virtude das revisões de que foi alvo o Conceito Estratégico de Defesa Nacional (2013). A atual orgânica, aprovada pela Resolução do Conselho de Ministro n.º 26/2013, de 19 abril, estabeleceu como linhas de ação prioritárias (de entre outras) a adaptação e racionalização de estruturas e a rentabilização de meios e capacidades, por forma a obter ganhos de eficiência, economias de escala e inovação de procedimentos a curto, médio e longo prazo. Com o intuito de adaptar, racionalizar e rentabilizar as estruturas edificadas e as capacidades logísticas, desenvolveu-se o projeto “Defesa 2020”, que visa implementar um modelo sustentável potenciador da racionalização da supracitada despesa pública e incrementador da eficiência na utilização dos recursos. Hodiernamente, as entidades não primam somente pela adoção de estratégias a nível de melhorias de eficiência económica, mas também a nível do acompanhamento da evolução tecnológica. Neste contexto, o presente trabalho tem como objeto de estudo a nova orgânica do Exército e os novos procedimentos do Sistema Logístico na gestão do Fardamento, com o fim de verificar as eventuais oportunidades de melhoria que os mesmos poderão ser alvo. Decorrente do expresso acima, derivam os objetivos deste que se consubstanciam na viabilidade e melhorias possíveis de aplicar aos serviços Logísticos do Exército na gestão do Fardamento, com a génese no estudo benchmarking dos serviços Logísticos do Ejército de Tierra (Espanha) por força da sua proximidade geográfica e da sua maior maturidade. Para o estudo da problemática levantada e para a concretização dos objetivos citados, realizou-se uma recolha de dados que teve como base o método hipotético-dedutivo. Os resultados obtidos nesta investigação demonstram que a gestão do fardamento no Exército Português se encontra numa fase embrionária, o que permite a adoção de melhorias que possibilitem um desempenho mais favorável e funcionamento mais eficaz, deparando-se com conspectos que podem ser melhorados, nomeadamente o planeamento de rede de transportes, a acessibilidade de vendas aos militares e a partilha de informação de artigos disponíveis e suas características. Conclui-se que é possível aperfeiçoar procedimentos no Sistema Logístico tendo em conta a eficiência e eficácia necessária à organização, no meio onde se insere.

Gestión de la cadena de suministro de Bimbo

El presente trabajo es un informe final sobre los procesos de producción, logística y distribución de la empresa Bimbo. El informe está conformado por aspectos teóricos y prácticos, de esta manera, brinda información clara y concreta sobre los antecedentes más importantes de esta organización y su contribución a la productividad del país. La información presentada ha sido recolectada mediante una revisión teórica y una visita a sus instalaciones en Veracruz, México. De esta manera, se han estudiado sus procesos y logística de producción y cómo contribuyen en la eficiencia y calidad de la empresa; así mismo, son fundamentales para diagnosticar los aspectos a mejorar de la entidad. Por otro lado, gracias a la visita, se logró desarrollar habilidades administrativas que hasta el momento solo estaban en teoría. Llegar a un contexto real y con problemáticas evidentes permitió consolidar el aprendizaje que se obtuvo durante años en la carrera para plasmarlos en la labor. Así mismo, el detallar las instalaciones de Bimbo permitió alcanzar una mayor perspectiva con respecto a los aspectos importantes dentro del panorama organizacional; tales como la producción, la gestión y mejora de los procesos de la compañía. Esto se logró realizando labores correspondientes a la inspección, observación, revisión de logística y el análisis de los procesos productivos. El trabajo práctico indudablemente sirvió para acrecentar los conocimientos obtenidos en la universidad.

Flowering synchronization in hybrid rice parental lines.

The cultivation of hybrid rice is a technology that allows for an increase in grain yield of 30% relative to the grain yield of conventional cultivars. However, the main challenge for this technology is related to seed production, which has high production costs and low seed yields. Therefore, agronomic techniques that could enhance flowering synchrony of parental lines in the field are essential for an efficient production system of hybrid rice seeds. The objective of this work was to study the effects of sowing depth, plant density and fertilization with nitrogen or phosphorus as potential techniques to increase the pollen availability in the field and, consequently, the flowering synchrony between parental lines in the production of hybrid rice seeds. The experiments were conducted during two growing seasons in the Central Region of Brazil. All of the experiments were conducted as a randomized complete block in a split plot scheme; however, the experiment with P fertilization had a factorial design. Our research allow inferring that nitrogen fertilization technique applied to the soil or foliar at the time of panicle differentiation does not affect the time of onset of flowering of rice varieties INTA Puitá CL and L106R, which are potential R lines for the production of hybrid rice. Agronomic techniques of variation in sowing depth, seeding rate and the phosphate fertilization affect the time of onset of flowering from 10 to 19 degree-days, which could represent two days in the crop cycle, for the line L106R. Such techniques constitute potential alternatives for use in hybrid rice seed production systems and could be applied in alternated blocks of R lines in the field to obtain longer periods of pollen availability in the field.

Technical and scale efficiency of cassava production system in Delta State, Nigeria: an application of Two-Stage DEA approach

The present study examines the level of pure technical and scale efficiencies of cassava production system including its sub-processes (that is production and processing stages) of 278 cassava farmers/processors from three regions of Delta State, Nigeria by applying Two-Stage Data Envelopment Analysis (DEA) approach. Results reveal that pure technical efficiency (PTE) is significantly lower at the production stage 0.41 vs 0.55 for the processing stage, but scale efficiency (SE) is high at both stages (0.84 and 0.87), implying that productivity can be improved substantially by reallocation of resources and adjusting operation size. The socio-economic determinants exert differential impacts on PTE and SE at each stage. Overall, education, experience and main occupation as farmer significantly improve SE while subsistence pressure reduces it. Extension contact significantly improves SE at the processing stage but reduces PTE and SE overall. Inverse size-PTE and size-SE relationships exist in cassava production system. In other words, large/medium farms are technically and scale inefficient. Gender gap exists in performance. Male farmers are technically efficient at processing stage but scale inefficient overall. Farmers in northern region are technically efficient. Investments in education, extension services and infrastructure are suggested as policy options to improve the cassava sector in Nigeria.

Scale and scope economies and the efficient vertical and horizontal configuration of the water industry:a survey of the literature

This paper surveys the literature on scale and scope economies in the water and sewerage industry. The magnitude of scale and scope economies determines the cost efficient configuration of any industry. In the case of a regulated sector, reliable estimates of these economies are relevant to inform reform proposals that promote vertical (un)bundling and mergers. The empirical evidence allows some general conclusions. First, there is considerable evidence for the existence of vertical scope economies between upstream water production and distribution. Second, there is only mixed evidence on the existence of (dis)economies of scope between water and sewerage activities. Third, economies of scale exist up to certain output level, and diseconomies of scale arise if the company increases its size beyond this level. However, the optimal scale of utilities also appears to vary considerably between countries. Finally, we briefly consider the implications of our findings for water pricing and point to several directions for necessary future empirical research on the measurement of these economies, and explaining their cross country variation.