Comparison of innate immune responses towards rhinovirus infection of primary nasal and bronchial epithelial cells.

BACKGROUND AND OBJECTIVE Rhinoviruses (RV) replicate in both upper and lower airway epithelial cells. We evaluated the possibility of using nasal epithelial cells (NEC) as surrogate of bronchial epithelial cells (BEC) for RV pathogenesis cell culture studies. METHODS We used primary paired NEC and BEC cultures established from healthy subjects and compared the replication of RV belonging to the major (RV16) and minor (RV1B) group, and the cellular antiviral and proinflammatory cytokine responses towards these viruses. We related antiviral and pro-inflammatory responses of NEC isolated from CF and COPD patients with those of BEC. RESULTS RV16 replication and major group surface receptor (ICAM-1) expression were higher in healthy NEC compared with BEC (P < 0.05); RV1B replication and minor group surface receptor (LDLR) expression were similar. Healthy NEC and BEC produced similar levels of IFN-β and IFN-λ2/3 upon RV infection or after simulation with poly(IC). IL-8 production was similar between healthy NEC and BEC. IL-6 release at baseline (P < 0.01) and upon infection with RV16 (P < 0.05) and poly(IC) stimulation (P < 0.05) was higher in NEC. RV1B viral load in NEC was related to RV1B viral load in BEC (r = 0.49, P = 0.01). There was a good correlation of IFN levels between NEC and BEC (r = 0.66, P = 0.0004 after RV1B infection). IL-8 production in NEC was related to IL-8 production in BEC (r = 0.48, P = 0.02 after RV1B infection). CONCLUSION NEC are a suitable alternative cellular system to BEC to study the pathophysiology of RV infections and particularly to investigate IFN responses induced by RV infection.

Ser88 phosphorylation regulates dynein light chain 1 (DLC1) function in the mouse mammary gland

Dynein light chain 1 (DLC1) is a highly conserved and ubiquitously expressed protein which might have critical cellular function as total loss of DLC1 caused Drosophila embryonic death. Despite many proteins and RNAs interaction with it identified, DLC1's function(s) and regulation are largely unknown. Recently, DLC1 was identified as a physiological substrate of P21-activate kinase 1(Pak1) kinase from a human mammary cDNA library in a yeast-2-hybridization screening assay. Studies in primary human tumors and cell culture implicated that DLC1 could promote mammary cancerous phenotypes, and more importantly, Ser88 phosphorylation of DLC1by Pak1 kinase was found to be essential for DLC1's tumorigenic activities. Based on the above tissue culture studies, we hypothesized that Ser88 phosphorylation regulates DLC1. ^ To test this hypothesis, we generated two transgenic mouse models: MMTV-DLC1 and MMTV-DLC1-S88A mice with mammary specific expression of the DLC1 and DLC1-S88A cDNAs. Both of the transgenic mice mammary glands showed rare tumor incidence which indicated DLC1 alone may not be sufficient for tumorigenesis in vivo. However, these mice showed a significant alteration of mammary development. Mammary glands from the MMTV-DLC1 mice had hyperbranching and alveolar hyperplasia, with elevated cell proliferation. Intriguingly, these phenotypes were not seen in the mammary glands from the MMTV-S88A mice. Furthermore, while MMTV-DLC1 glands were normal during involution, MMTV-S88A mice showed accelerated mammary involution with increase apoptosis and altered expression of involution-associated genes. Further analysis of the MMTV-S88A glands showed they had increased steady state level of Bim protein which might be responsible for the early involution. Finally, our in vitro data showed that Ser88 phosphorylation abolished DLC1 dimer and consequently might disturb its interaction with Bim and destabilize Bim. ^ Collectively, our findings provided in vivo evidence that Ser88 phosphorylation of DLC1 can regulate DLC1's function. In addition, Ser88 phosphorylation might be critical for DLC1 dimer-monomer transition. ^

Design, Synthesis and Development of Transporter Targeting Agents for Image-guided Therapy and Drug Delivery

The purpose of this study was to design, synthesize and develop novel transporter targeting agents for image-guided therapy and drug delivery. Two novel agents, N4-guanine (N4amG) and glycopeptide (GP) were synthesized for tumor cell proliferation assessment and cancer theranostic platform, respectively. N4amG and GP were synthesized and radiolabeled with 99mTc and 68Ga. The chemical and radiochemical purities as well as radiochemical stabilities of radiolabeled N4amG and GP were tested. In vitro stability assessment showed both 99mTc-N4amG and 99mTc-GP were stable up to 6 hours, whereas 68Ga-GP was stable up to 2 hours. Cell culture studies confirmed radiolabeled N4amG and GP could penetrate the cell membrane through nucleoside transporters and amino acid transporters, respectively. Up to 40% of intracellular 99mTc-N4amG and 99mTc-GP was found within cell nucleus following 2 hours of incubation. Flow cytometry analysis revealed 99mTc-N4amG was a cell cycle S phase-specific agent. There was a significant difference of the uptake of 99mTc-GP between pre- and post- paclitaxel-treated cells, which suggests that 99mTc-GP may be useful in chemotherapy treatment monitoring. Moreover, radiolabeled N4amG and GP were tested in vivo using tumor-bearing animal models. 99mTc-N4amG showed an increase in tumor-to-muscle count density ratios up to 5 at 4 hour imaging. Both 99mTc-labeled agents showed decreased tumor uptake after paclitaxel treatment. Immunohistochemistry analysis demonstrated that the uptake of 99mTc-N4amG was correlated with Ki-67 expression. Both 99mTc-N4amG and 99mTc-GP could differentiate between tumor and inflammation in animal studies. Furthermore, 68Ga-GP was compared to 18F-FDG in rabbit PET imaging studies. 68Ga-GP had lower tumor standardized uptake values (SUV), but similar uptake dynamics, and different biodistribution compared with 18F-FDG. Finally, to demonstrate that GP can be a potential drug carrier for cancer theranostics, several drugs, including doxorubicin, were selected to be conjugated to GP. Imaging studies demonstrated that tumor uptake of GP-drug conjugates was increased as a function of time. GP-doxorubicin (GP-DOX) showed a slow-release pattern in in vitro cytotoxicity assay and exhibited anti-cancer efficacy with reduced toxicity in in vivo tumor growth delay study. In conclusion, both N4amG and GP are transporter-based targeting agents. Radiolabeled N4amG can be used for tumor cell proliferation assessment. GP is a potential agent for image-guided therapy and drug delivery.

De Sastres académicos : los estudios culturales como moda-lidad sin objeto

En el presente artículo analizamos las condiciones de aparición y los postulados fundamentales de esta escuela, a partir de las rupturas teóricas y políticas que tuvieron lugar en el seno del marxismo inglés por la década de fines de los años cincuenta. Además, nos preguntamos si podemos olvidar la herencia de la teoría crítica de la cultura a partir de la hegemonía de unos estudios culturales que cada vez se alejan más de una perspectiva de sospecha sobre el sistema social para adoptar una actitud académica complaciente y conservadora.

Balance y perspectivas predominantes en la historia de la cultura escrita : Una aproximación

El artículo reflexiona sobre la obra de los autores que son y serán referentes en los estudios históricos sobre la cultura escrita. No obstante, no pretende abarcarlos en su totalidad. En esta medida, se hace énfasis en las perspectivas teóricas y metodológicas de aquellos que se destacan en el campo. Realizar balances que vislumbren la complejidad de la historia de la cultura escrita como tema de investigación es una tarea obligada y necesaria para historiadores, literatos, sociólogos, antropólogos y bibliotecólogos, entre otros, que se inician en el estudio alrededor de este tema en el contexto latinoamericano y cuyos trabajos monográficos comienzan ya a integrar una corriente histórica en la región

Balance y perspectivas predominantes en la historia de la cultura escrita : Una aproximación

El artículo reflexiona sobre la obra de los autores que son y serán referentes en los estudios históricos sobre la cultura escrita. No obstante, no pretende abarcarlos en su totalidad. En esta medida, se hace énfasis en las perspectivas teóricas y metodológicas de aquellos que se destacan en el campo. Realizar balances que vislumbren la complejidad de la historia de la cultura escrita como tema de investigación es una tarea obligada y necesaria para historiadores, literatos, sociólogos, antropólogos y bibliotecólogos, entre otros, que se inician en el estudio alrededor de este tema en el contexto latinoamericano y cuyos trabajos monográficos comienzan ya a integrar una corriente histórica en la región

Balance y perspectivas predominantes en la historia de la cultura escrita : Una aproximación

El artículo reflexiona sobre la obra de los autores que son y serán referentes en los estudios históricos sobre la cultura escrita. No obstante, no pretende abarcarlos en su totalidad. En esta medida, se hace énfasis en las perspectivas teóricas y metodológicas de aquellos que se destacan en el campo. Realizar balances que vislumbren la complejidad de la historia de la cultura escrita como tema de investigación es una tarea obligada y necesaria para historiadores, literatos, sociólogos, antropólogos y bibliotecólogos, entre otros, que se inician en el estudio alrededor de este tema en el contexto latinoamericano y cuyos trabajos monográficos comienzan ya a integrar una corriente histórica en la región

Seasonal changes in the D/H ratio of fatty acids of pelagic microorganisms

Culture studies of microorganisms have shown that the hydrogen isotopic composition of fatty acids depends on their metabolism, but there are only few environmental studies available to confirm this observation. Here we studied the seasonal variability of the deuterium/hydrogen (D/H) ratio of fatty acids in the coastal Dutch North Sea and compared this with the diversity of the phyto- and bacterioplankton. Over the year, the stable hydrogen isotopic fractionation factor epsilon between fatty acids and water ranged between -172 per mil and -237 per mil, the algal-derived polyunsaturated fatty acid nC20:5 being the most D-depleted and nC18:0 the least D-depleted fatty acid. The D-depleted nC20:5 is in agreement with culture studies, which indicates that photoautotrophic microorganisms produce fatty acids which are significantly depleted in D relative to water. The epsilon-lipid/water of all fatty acids showed a transient shift towards increased fractionation during the spring phytoplankton bloom, indicated by increasing chlorophyll a concentrations and relative abundance of the nC20:5 PUFA, suggesting increased contributions of photoautotrophy. Time periods with decreased fractionation (less negative epsilon-lipid/water values) can be explained by an increased contribution by heterotrophy to the fatty acid pool. Our results show that the hydrogen isotopic composition of fatty acids is a useful tool to assess the community metabolism of coastal plankton.

CO2 and vitamin B12 interactions determine bioactive trace metal requirements of a subarctic Pacific diatom

Phytoplankton growth can be limited by numerous inorganic nutrients and organic growth factors. Using the subarctic diatom Attheya sp. in culture studies, we examined how the availability of vitamin B(12) and carbon dioxide partial pressure (pCO(2)) influences growth rate, primary productivity, cellular iron (Fe), cobalt (Co), zinc (Zn) and cadmium (Cd) quotas, and the net use efficiencies (NUEs) of these bioactive trace metals (mol C fixed per mol cellular trace metal per day). Under B(12)-replete conditions, cells grown at high pCO(2) had lower Fe, Zn and Cd quotas, and used those trace metals more efficiently in comparison with cells grown at low pCO(2). At high pCO(2), B(12)-limited cells had ~50% lower specific growth and carbon fixation rates, and used Fe ~15-fold less efficiently, and Zn and Cd ~3-fold less efficiently, in comparison with B(12)-replete cells. The observed higher Fe, Zn and Cd NUE under high pCO(2)/B(12)-replete conditions are consistent with predicted downregulation of carbon-concentrating mechanisms. Co quotas of B(12)-replete cells were 5- to 14-fold higher in comparison with B(12)-limited cells, suggesting that >80% of cellular Co of B(12)-limited cells was likely from B(12). Our results demonstrate that CO(2) and vitamin B(12) interactively influence growth, carbon fixation, trace metal requirements and trace metal NUE of this diatom. This suggests the need to consider complex feedback interactions between multiple environmental factors for this biogeochemically critical group of phytoplankton in the last glacial maximum as well as the current and future changing ocean.

Modelización y simulación numérica de sistemas geotérmicos planos

En el presente proyecto se estudiará la conveniencia de utilizar simulación numérica para evaluar, diseñar y mejorar el diseño de instalaciones geotérmicas planas. Para ello, se modelizará el conjunto de terreno y sistema geotérmico mediante el acoplamiento de un modelo tridimensional (terreno) y un modelo unidimensional, formado por la red de tubería geotérmica, que estará dispuesta en un plano cuya orientación dependerá de las aplicaciones. Para la simulación numérica se utilizará un software de análisis y resolución por elementos finitos tipo comercial. Finalmente, se definirán unos parámetros geométricos de diseño y, teniendo en cuenta las propiedades térmicas de cada material, se realizarán estudios paramétricos para determinar la influencia de cada grupo de parámetros. Ello requerirá la realización de cientos de simulaciones numéricas que permitirán optimizar el rendimiento de sistemas geotérmicos planos tanto horizontales (soleras), como verticales (muros pantalla). ABSTRACT In this project it is going to be studied the convenience of numerical simulation as to evaluate, design and improve the design of geothermal planar installations. With this purpose, the terrain and the geothermal system will be modeled by coupling a three-dimensional model (terrain) and a one-dimensional model formed by the geothermal piping network disposed in a plane with an orientation which will depend on the applications. For the numerical simulation commercial software of analyzing and resolving by finite elements will be used. Finally, geometric design parameters will be defined, and taking into account the thermal properties of each material, parametric studies will be performed to determine the influence of each group of parameters. This will require the completion of hundreds of numerical simulations which will enhance the performance of both horizontal (slabs) and vertical (diaphragm walls) geothermal systems.

HIV type-1 infection of the cotton rat (Sigmodon fulviventer and S. hispidus)

Cotton rats (Sigmodon hispidus and S. fulviventer) are susceptible to many viruses that infect humans (e.g., poliovirus, respiratory syncytial virus, influenza virus, adenovirus, and parainfluenza virus) and have been influential in developing therapeutic clinical intervention strategies for many viral infections of man. This study set out to determine whether cotton rats are susceptible to infection with HIV type 1 (HIV-1). Results indicate that HIV-1 does infect the cotton rat and S. fulviventer is more susceptible than S. hispidus. The virus was passaged from animal to animal for a total of three serial passages; but HIV replicated poorly in vivo, was only detectable as proviral DNA, and never exceeded one provirus per 1.8 × 105 cotton rat peripheral blood mononuclear cells. Infection induced a distinct and characteristic anti-HIV antibody response that, in some animals, included neutralizing antibodies, recognized all of the major HIV-1 antigens and the antibodies lasted out to 52 wk post-infection. Neonate S. fulviventer were not more susceptible to infection than adults. In vitro culture studies produced indirect evidence of viral replication by detection of viral gag gene RNA in reverse transcriptase–PCR assays on viral culture supernatants. Collectively, these results indicate that HIV-1 can replicate in a nontransgenic rodent and that this system may have potential as an animal model for HIV-1 infection if viral replication rates can be improved in vivo.

A general method to design dominant negatives to B-HLHZip proteins that abolish DNA binding

We describe a method to design dominant-negative proteins (D-N) to the basic helix–loop–helix–leucine zipper (B-HLHZip) family of sequence-specific DNA binding transcription factors. The D-Ns specifically heterodimerize with the B-HLHZip dimerization domain of the transcription factors and abolish DNA binding in an equimolar competition. Thermal denaturation studies indicate that a heterodimer between a Myc B-HLHZip domain and a D-N consisting of a 12-amino acid sequence appended onto the Max dimerization domain (A-Max) is −6.3 kcal⋅mol−1 more stable than the Myc:Max heterodimer. One molar equivalent of A-Max can totally abolish the DNA binding activity of a Myc:Max heterodimer. This acidic extension also has been appended onto the dimerization domain of the B-HLHZip protein Mitf, a member of the transcription factor enhancer binding subfamily, to produce A-Mitf. The heterodimer between A-Mitf and the B-HLHZip domain of Mitf is −3.7 kcal⋅mol−1 more stable than the Mitf homodimer. Cell culture studies show that A-Mitf can inhibit Mitf-dependent transactivation both in acidic extension and in a dimerization-dependent manner. A-Max can inhibit Myc-dependent foci formation twice as well as the Max dimerization domain (HLHZip). This strategy of producing D-Ns may be applicable to other B-HLHZip or B-HLH proteins because it provides a method to inhibit the DNA binding of these transcription factors in a dimerization-specific manner.

Comércio, bens de prestígio e insígnias de poder: as agências centro-ocidentais africanas nos relatos de viagem de Henrique de Carvalho em sua expedição à Lunda (1884-1888)

Ao longo da segunda metade do século XIX, a região da África Centro-Ocidental foi palco do processo de intensificação de expedições europeias rumo ao interior do continente que conjugavam interesses econômicos, políticos e científicos. Esta pesquisa busca analisar o papel de relevo ocupado pela cultura material na agenda científica da expedição portuguesa à Lunda chefiada pelo militar português Henrique de Carvalho entre 1884 e 1888. Pretendemos também avaliar as potencialidades que o estudo sobre os objetos apresentam enquanto fontes para a compreensão mais ampla acerca das agências históricas africanas. Para tanto, selecionamos as obras Descripção da Viagem à Mussumba do Muatiânvua (1890-1894) e Ethnographia e História Tradicional dos Povos da Lunda (1890) e, de maneira complementar, o Album da Expedição ao Muatianvua (1887) e o catálogo Collecção Henrique de Carvalho (Expedição à Lunda), editado pela Sociedade de Geografia de Lisboa (1896). Assim, pela ótica da história social, pretendemos investigar como as exigências e predileções centro-africanas por determinados artigos moldaram as relações comerciais travadas nesse período, abordando os processos de incorporação e ressignificação de objetos particularmente, bens de prestígio e insígnias de poder - interpretados como expressões de identidades, códigos sociais e hierarquias políticas no âmbito dessas sociedades e de suas relações com os europeus.

Brewing Behind Barbed Wire: An Archaeology of Saké at Amache

After the Japanese attack at Pearl Harbor on December 7, 1941, approximately 120,000 people of Japanese ancestry living on the west coast of the United States were forcibly removed from their home communities. These people were designated as "evacuees" by the U.S. Government and were incarcerated within a network of federal government facilities the largest of which were internment centers operated by the War Relocation Authority that held mostly U.S. citizens. The Granada Relocation Center (Amache) was the smallest of these internment centers. The presence of saké at Amache indicates that Japanese Americans continued important practices of daily life despite restrictions under confinement. This thesis investigates the practices of saké production and consumption at Amache and examines the importance of these practices in Japanese American daily life. In order to understand these practices, this research draws on multiple lines of evidence. This includes investigations of an assemblage of the material culture associated with saké, research into the history and methods of production and consumption, collection of oral histories, review of archival data, and the application of practice theory. These data provide insight into practices that are not well understood by researchers of Japanese American internment due to their illicit nature. This research endeavors to characterize how saké was produced and used at Amache and provides a way to understand how cultural practices maintain aspects of everyday life in ways that may have little to do with intentional resistance.