Solid Lipid Nanoparticles As Nucleic Acid Delivery System: Properties And Molecular Mechanisms.

Solid lipid nanoparticles (SLNs) have been proposed in the 1990s as appropriate drug delivery systems, and ever since they have been applied in a wide variety of cosmetic and pharmaceutical applications. In addition, SLNs are considered suitable alternatives as carriers in gene delivery. Although important advances have been made in this particular field, fundamental knowledge of the underlying mechanisms of SLN-mediated gene delivery is conspicuously lacking, an imperative requirement in efforts aimed at further improving their efficiency. Here, we address recent advances in the use of SLNs as platform for delivery of nucleic acids as therapeutic agents. In addition, we will discuss available technology for conveniently producing SLNs. In particular, we will focus on underlying molecular mechanisms by which SLNs and nucleic acids assemble into complexes and how the nucleic acid cargo may be released intracellularly. In discussing underlying mechanisms, we will, when appropriate, refer to analogous studies carried out with systems based on cationic lipids and polymers, that have proven useful in the assessment of structure-function relationships. Finally, we will give suggestions for improving SLN-based gene delivery systems, by pointing to alternative methods for SLNplex assembly, focusing on the realization of a sustained nucleic acid release.

Audiogram Comparison Of Workers From Five Professional Categories.

Introduction. Noise is a major cause of health disorders in workers and has unique importance in the auditory analysis of people exposed to it. The purpose of this study is to evaluate the arithmetic mean of the auditory thresholds at frequencies of 3, 4, and 6 kHz of workers from five professional categories exposed to occupational noise. Methods. We propose a retrospective cross-sectional cohort study to analyze 2.140 audiograms from seven companies having five sectors of activity: one footwear company, one beverage company, two ceramics companies, two metallurgical companies, and two transport companies. Results. When we compared two categories, we noticed a significant difference only for cargo carriers in comparison to the remaining categories. In all activity sectors, the left ear presented the worst values, except for the footwear professionals (P > 0.05). We observed an association between the noise exposure time and the reduction of audiometric values for both ears. Significant differences existed for cargo carriers in relation to other groups. This evidence may be attributed to different forms of exposure. A slow and progressive deterioration appeared as the exposure time increased.

Efeito da adição de cinza da casca de arroz em misturas cimento-casca de arroz

The rice husk and its ash are abundant and renewable and can be used to obtain alternative building materials. An increase in the consumption of such waste could help minimize the environmental problems from their improper disposal. This study aimed to evaluate the use of ashes as a cargo mineral (filler). However, the rice husk chemically interferes in the conduct of the based cement mixtures. Thus, different mixes cement-rice husk with and without the addition of ash were evaluated in order to highlight the influence of its components (husk; ash), which could otherwise be excluded or be underestimated. Cylindrical samples (test of simple compression and traction by diametrical compression) and samples extracted from manufactured pressed board (test of bending and parallel compression to the surface), were used to evaluate the behavior of different mixtures of components (rice hush; RHA - rice husk ahs). The results of the mechanical tests showed, in general, there is not a statistical difference between the mixtures, which are associated with the chemical suppressive effect of the rice husk ash. The mixture of rice husk of 10 mm, with an addition of 35% of the rice husk ash, is notable for allowing the highest consumption of rice husk and rice husk ash, to reduce 25% the consumption of cement and to allow the storage (without emissions to the atmosphere), around 1.9 ton of CO2 per ton of cement consumed, thus contributing to the reduction of CO2 emissions, which can stimulate rural constructions under an ecological point of view.

Índices de preço para o transporte de cargas: o caso da soja

Esta pesquisa objetivou analisar metodologias para elaboração de índices de preços para o transporte de cargas. O estudo das principais fórmulas da Teoria Econômica culminou com a conclusão que os Índices de Fisher e Walsh são aqueles capazes de atender ao maior número de preceitos lógicos, estatísticos e econômicos. Em seguida, surgem os índices geométricos de Törnqvist, Vartia e Theil. Os Índices de Laspeyres e Paasche, apesar de apresentarem algumas limitações, acabam sendo amplamente utilizados, graças a maior capacidade de operacionalização. A pesquisa apresentou um estudo de caso para o transporte da soja em grão. Foram realizados quatro tratamentos. Obteve-se a variação acumulada no nível geral de preços para o transporte rodoviário de soja no Brasil, no período entre fevereiro de 1998 e março de 2002. De acordo com os resultados, essa variação acumulada teria sido de 76%.

Distúrbios psíquicos menores e condições de trabalho em motoristas de caminhão

OBJETIVO: Estimar a prevalência de distúrbios psíquicos menores e identificar estressores associados entre motoristas de caminhão. MÉTODOS: Estudo transversal conduzido com 460 motoristas de caminhão de uma transportadora de cargas das regiões Sul e Sudeste do Brasil, em 2007. Os trabalhadores preencheram questionário com dados sociodemográficos, estilos de vida e condições de trabalho. As variáveis independentes foram condições de trabalho, incluindo estressores ocupacionais, satisfação e demanda-controle no trabalho. O desfecho avaliado foi a ocorrência de distúrbios psíquicos menores. Foram realizadas análises de regressão logística univariada e múltipla. RESULTADOS: A prevalência de distúrbios psíquicos menores foi de 6,1 por cento. Os estressores mais citados foram congestionamentos, controle de rastreamento e jornada extensa de trabalho. A alta demanda no trabalho, o baixo apoio social e a jornada extensa diária referidos pelos motoristas estiveram associados aos distúrbios psíquicos menores. CONCLUSÕES: O trabalho em jornadas extensas foi associado à ocorrência de distúrbios psíquicos menores, tanto na análise das condições gerais de trabalho quanto como fator referido como estressor pelos motoristas. A regulamentação da jornada de trabalho com limitação de horas de trabalho diário é, portanto, uma medida necessária para a redução da chance de desenvolvimento de distúrbios psíquicos menores em motoristas

Mudanças no âmbito da testagem anti-HIV no Brasil entre 1998 e 2005

OBJETIVO: Analisar mudanças na realização de teste anti-HIV, as razões alegadas entre as pessoas que foram ou não testadas e o recebimento de aconselhamento. MÉTODOS: Estudos transversais conduzidos com homens e mulheres de 16 a 65 anos, com amostras representativas do Brasil urbano em 1998 (n=3.600) e 2005 (n=5.040). Características sociodemográficas, sexuais, reprodutivas e de experiências de vida e saúde foram consideradas na análise. A avaliação das possíveis diferenças nas distribuições das variáveis baseou-se nos testes qui-quadrado de Pearson e F design-based (±<5%). RESULTADOS: Em 1998, 20,2% dos entrevistados haviam realizado o teste e 33,6% em 2005. Foram testadas 60% das mulheres na faixa 25-34 anos, mas as que iniciaram a vida sexual antes dos 16 anos e reportaram quatro ou mais parceiros sexuais nos cinco anos anteriores à entrevista foram menos testadas. Não se observou aumento significativo da testagem entre homens, exceto para os de 55-65 anos, renda per capita entre 1-3 e 5-10 salários mínimos, aposentados, protestantes históricos e adeptos de cultos afro-brasileiros, moradores da região Norte/Nordeste e os que declararam parceria homo/bissexual ou não tiveram relações sexuais nos cinco anos anteriores à entrevista. Não aumentou a freqüência de testagem entre pessoas auto-avaliadas como sob alto risco para o HIV. Entre as mulheres, a freqüência de testagem pré-natal aumentou e a testagem por trabalho diminuiu entre os homens. Em 2005, metade dos testados não recebeu orientação antes ou após o teste. CONCLUSÕES: Houve expansão desigual na testagem, atingindo principalmente mulheres em idade reprodutiva, adultas e pessoas com melhores condições sociais. A testagem parece estar aumentando no País sem a devida atenção à decisão autônoma das pessoas e sem o provimento de maior e melhor oferta de aconselhamento

Structural basis of recognition of monopartite and bipartite nuclear localization sequences by mammalian importin-alpha

Importin-alpha is the nuclear import receptor that recognizes cargo proteins which contain classical monopartite and bipartite nuclear localization sequences (NLSs), and facilitates their transport into the nucleus. To determine the structural basis of the recognition of the two classes of NLSs by mammalian importin-alpha, we co-crystallized an N-terminally truncated mouse receptor protein with peptides corresponding to the monopartite NLS from the simian virus 40 (SV40) large T-antigen, and the bipartite NLS from nucleoplasmin. We show that the monopartite SV40 large T-antigen NLS binds to two binding sites on the receptor, similar to what was observed in yeast importin-alpha. The nucleoplasmin NLS-importin-alpha complex shows, for the first time, the mode of binding of bipartite NLSs to the receptor. The two basic clusters in the NLS occupy the two binding sites used by the monopartite NLS, while the sequence linking the two basic clusters is poorly ordered, consistent with its tolerance to mutations. The structures explain the structural basis for binding of diverse NLSs to the sole receptor protein. (C) 2000 Academic Press.

MHC class II expression is regulated in dendritic cells independently of invariant chain degradation

We have investigated the mechanisms that control MHC class II (MHC II) expression in immature and activated dendritic cells (DC) grown from spleen and bone marrow precursors. Degradation of the MHC II chaperone invariant chain (li), acquisition of peptide cargo by MHC II, and delivery of MHC II-peptide complexes to the cell surface proceeded similarly in both immature and activated DC. However, immature DC reendocytosed and then degraded the MHC II-peptide complexes much faster than the activated DC. MHC II expression in DC is therefore not controlled by the activity of the protease(s) that degrade Ii, but by the rate of endocytosis of peptide-loaded MHC II. Late after activation, DC downregulated MHC II synthesis both in vitro and in vivo.

The trans-membrane protein p25 forms highly specialized domains that regulate membrane composition and dynamics

Trans-membrane proteins of the p24 family are abundant, oligomeric proteins predominantly found in cis-Golgi membranes. They are not easily studied in vivo and their functions are controversial. We found that p25 can be targeted to the plasma membrane after inactivation of its canonical KKXX motif (KK to SS, p25SS), and that p25SS causes the co-transport of other p24 proteins beyond the Golgi complex, indicating that wild-type p25 plays a crucial role in retaining p24 proteins in cis-Golgi membranes. We then made use of these observations to study the intrinsic properties of these proteins, when present in a different membrane context. At the cell surface, the p25SS mutant segregates away from both the transferrin receptor and markers of lipid rafts, which are enriched in cholesterol and glycosphingolipids. This suggests that p25SS localizes to, or contributes to form, specialized membrane domains, presumably corresponding to oligomers of p25SS and other p24 proteins. Once at the cell surface, p25SS is endocytosed, together with other p24 proteins, and eventually accumulates in late endosomes, where it remains confined to well-defined membrane regions visible by electron microscopy. We find that this p25SS accumulation causes a concomitant accumulation of cholesterol in late endosomes, and an inhibition of their motility - two processes that are functionally linked. Yet, the p25SS-rich regions themselves seem to-exclude not only Lamp1 but also accumulated cholesterol. One may envision that p25SS accumulation, by excluding cholesterol from oligomers, eventually overloads neighboring late endosomal membranes with cholesterol beyond their capacity (see Discussion). In any case, our data show that p25 and presumably other p24 proteins are endowed with the intrinsic capacity to form highly specialized domains that control membrane composition and dynamics. We propose that p25 and other p24 proteins control the fidelity of membrane transport by maintaining cholesterol-poor membranes in the Golgi complex.

GAIP participates in budding of membrane carriers at the trans-Golgi network

Galpha interacting protein (GAIP) is a regulator of G protein signaling protein that associates dynamically with vesicles and has been implicated in membrane trafficking, although its specific role is not yet known. Using an in vitro budding assay, we show that GAIP is recruited to a specific population of trans-Golgi network-derived vesicles and that these are distinct from coatomer or clathrin-coated vesicles. A truncation mutant (NT-GAIP) encoding only the N-terminal half of GAIP is recruited to trans -Golgi network membranes during the formation of vesicle carriers. Overexpression of NT-GAIP induces the formation of long, coated tubules, which are stabilized by microtubules. Results from the budding assay and from imaging in live cells show that these tubules remain attached to the Golgi stack rather than being released as carrier vesicles. NT-GAIP expression blocks membrane budding and results in the accumulation of tubular carrier intermediates. NT-GAIP-decorated tubules are competent to load vesicular stomatitis virus protein G-green fluorescent protein as post-Golgi, exocytic cargo and in cells expressing NT-GAIP there is reduced surface delivery of vesicular stomatitis virus protein G-green fluorescent protein. We conclude that GAIP functions as an essential part of the membrane budding machinery for a subset of post-Golgi exocytic carriers derived from the trans-Golgi network.

Alternatively spliced products of the human kinesin light chain 1 (KNS2) gene

Conventional kinesin is a microtubule-based molecular motor involved in the transport of membranous and non-membranous cargoes. The kinesin holoenzyme exists as a heterotetramer, consisting of two heavy chain and two light chain subunits. It is thought that one function of the light chains is to interact with the cargo. Alternative splicing of kinesin light chain pre-mRNA has been observed in lower organisms, although evidence for alternative splicing of the human gene has not been reported. We have identified 19 variants of the human KNS2 gene (KLC1) that are generated by alternative splicing of downstream exons, but calculate that KNS2 has the potential to produce 285919 spliceforms. Corresponding spliceforms of the mouse KLC1 gene were also identified. The alternative exons are all located 3' of exon 12 and the novel spliceforms produce both alternative carboxy termini and alternative 3' untranslated regions. The observation of multiple light chain isoforms is consistent with their proposed role in specific cargo attachment.

Dimensional X: a dificuldade na obten????o da informa????o em meio a problemas na gest??o de pessoas

Ant??nio foi convidado para assumir um cargo em que deveria gerenciar os sistemas de informa????o de um grande minist??rio. Com a miss??o inicial de fornecer uma informa????o sobre a for??a de trabalho, percebeu que havia problemas que iam muito al??m da simples utiliza????o desses sistemas, como a pol??tica de capacita????o de pessoal, a rotatividade de servidores, a falta de integra????o entre as ??reas e outros problemas na ??rea de gest??o de pessoas

G??nero, ra??a e espa??os de poder: o caso de Maria Ant??nia e Geraldo

O caso relata uma situa????o de promo????o em ??rg??o p??blico na qual dois funcion??rios ??? uma mulher negra e um homem branco - se mostram como candidatos ao cargo. S??o apresentados os perfis e habilidades profissionais dos dois servidores e, ao final, a decis??o tomada pelo Secret??rio, chefe de ambos. O estudo suscita discuss??es sobre pr??ticas discriminat??rias nos espa??os de poder das institui????es p??blicas e o papel do dirigente para combater ou difundir tais pr??ticas. Pode ser aplicado em cursos sobre lideran??a, gest??o de pessoas e ??tica e servi??o p??blico

Gentileza ou ass??dio: conflitos de g??nero e de ra??a nas rela????es de trabalho

Com o afastamento do chefe do setor por quest??es m??dicas, Carla, que j?? exercia eventualmente a chefia da equipe, tornou-se a substituta do cargo, s?? n??o contava com o ass??dio de Ricardo, funcion??rio querido por todos. As investidas foram ficando cada vez mais agressivas e Carla n??o sabia mais como lidar com a situa????o. Come??ou a ter problemas com a sua equipe e sofrer difama????es por parte de Ricardo. O cen??rio encontrado pelo chefe ao voltar da licen??a foi de uma equipe desestabilizada. O caso trata do tema de ass??dio moral, agregado ?? quest??o racial e de g??nero, e suscita discuss??o sobre como um dirigente deve atuar para evitar a pr??tica dos ass??dios moral e sexual na equipe que gerencia, bem como para remediar situa????es j?? consolidadas. Pode ser aplicado em cursos sobre lideran??a, gest??o de pessoas e ??tica e servi??o p??blico

H?? um g??nero mais adequado para determinada fun????o?

Carolina foi aprovada em concurso p??blico para cargo t??cnico de hidrot??cnico e assinou contrato de experi??ncia por 90 dias. Contudo, durante a realiza????o das atividades inerentes ao cargo, n??o recebeu nenhuma orienta????o do seu supervisor. No momento da avalia????o de desempenho, foi mal avaliada tanto pelo supervisor quanto por sua chefia, que desde o in??cio demonstrava que o cargo n??o era apropriado para mulheres. Por conta dessa avalia????o, Carolina buscou ajuda na Ouvidoria do ??rg??o, mas acabou sendo demitida. Optou, ent??o, por recorrer ?? Justi??a. O caso trata de situa????o na qual ?? colocada em xeque a quest??o de g??nero para determinada fun????o no servi??o p??blico. O estudo pode ser aplicado em cursos com enfoque em quest??es vinculadas ?? diversidade de g??nero, ??s posturas das chefias que lidam com avalia????o de desempenho, bem como ao papel da Ouvidoria