Vascular smooth muscle cell phenotypic modulation in culture is associated with reorganisation of contractile and cytoskeletal proteins

Smooth muscle cells (SMC) exhibit a functional plasticity, modulating from the mature phenotype in which the primary function is contraction, to a less differentiated state with increased capacities for motility, protein synthesis, and proliferation. The present study determined, using Western analysis, double-label immunofluorescence and confocal microscopy, whether changes in phenotypic expression of rabbit aortic SMC in culture could be correlated with alterations in expression and distribution of structural proteins. Contractile state SMC (days 1 and 3 of primary culture) showed distinct sorting of proteins into subcellular domains, consistent with the theory that the SMC structural machinery is compartmentalised within the cell. Proteins specialised for contraction (alpha -SM actin, SM-MHC, and calponin) were highly expressed in these cells and concentrated in the upper central region of the cell. Vimentin was confined to the body of the cell, providing support for the contractile apparatus but not co-localising with it. In line with its role in cell attachment and motility, beta -NM actin was localised to the cell periphery and basal cortex. The dense body protein alpha -actinin was concentrated at the cell periphery, possibly stabilising both contractile and motile apparatus. Vinculin-containing focal adhesions were well developed, indicating the cells' strong adhesion to substrate. In synthetic state SMC (passages 2-3 of culture), there was decreased expression of contractile and adhesion (vinculin) proteins with a concomitant increase in cytoskeletal proteins (beta -non-muscle [NM] actin and vimentin). These quantitative changes in structural proteins were associated with dramatic chan-es in their distribution. The distinct compartmentalisation of structural proteins observed in contractile state SMC was no longer obvious, with proteins more evenly distributed throughout die cytoplasm to accommodate altered cell function. Thus, SMC phenotypic modulation involves not only quantitative changes in contractile and cytoskeletal proteins, but also reorganisation of these proteins. Since the cytoskeleton acts as a spatial regulator of intracellular signalling, reorganisation of the cytoskeleton may lead to realignment of signalling molecules, which, in turn, may mediate the changes in function associated with SMC phenotypic modulation. (C) 2001 Wiley-Liss, Inc.

Chimeric human papilloma virus-simian/human immunodeficiency virus virus-like-particle vaccines: Immunogenicity and protective efficacy in macaques

Vaccines to efficiently block or limit sexual transmission of both HIV and human papilloma virus (HPV) are urgently needed. Chimeric virus-like-particle (VLP) vaccines consisting of both multimerized HPV L1 proteins and fragments of SIV gag p27, HIV-1 tat, and HIV-1 rev proteins (HPV-SHIV VLPs) were constructed and administered to macaques both systemically and mucosally. An additional group of macaques first received a priming vaccination with DNA vaccines expressing the same SIV and HIV-1 antigens prior to chimeric HPV-SHIV VLP boosting vaccinations. Although HPV L1 antibodies were induced in all immunized macaques, weak antibody or T cell responses to the chimeric SHIV antigens were detected only in animals receiving the DNA prime/HPV-SHIV VLP boost vaccine regimen. Significant but partial protection from a virulent mucosal SHIV challenge was also detected only in the prime/boosted macaques and not in animals receiving the HPV-SHIV VLP vaccines alone, with three of five prime/boosted animals retaining some CD4+ T cells following challenge. Thus, although some immunogenicity and partial protection was observed in non-human primates receiving both DNA and chimeric HPV-SHIV VLP vaccines, significant improvements in vaccine design are required before we can confidently proceed with this approach to clinical trials. (C) 2002 Elsevier Science (USA).

Route of administration of chimeric BPV1VLP determines the character of the induced immune responses

To examine the mucosal immune response to papillomavirus virus-like particles (PV-VLP), mice were immunized with VLP intrarectally (i.r.), intravaginally (i.va.) or intramuscularly (i.m.) without adjuvant. PV-VLP were assembled with chimeric BPV-1 L1 proteins incorporating sequence from HIV-1 gp 120, either the V3 loop or a shorter peptide incorporating a known CTL epitope (HIVP18I10). Antibody specific for BPV-1 VLP and P18 peptide was detected in serum following i.m., but not i.r. or i.va. immunization. Denatured VLP induced a much reduced immune response when compared with native VLP, Immune responses following mucosal administration of VLP were generally weaker than following systemic administration. VLP specific IgA was higher in intestine washes following i.r. than i.va. immunization, and higher in vaginal washes following i.m. than i.r. or i.va. immunization. No differences in specific antibody responses were seen between animals immunized with BPV-1 P18 VLP or with BPV-1 V3 VLP. Cytotoxic T lymphocyte precursors specific for the P18 CTL epitope were recovered from the spleen following i.m., i.va. or i.r. immunization with P18 VLP, and were similarly detected in Peyer's patches following i.m. or i.r. immunization. Thus, mucosal or systemic immunization with PV VLP induces mucosal CTL responses and this may be important for vaccines for mucosal infection with human papillomaviruses and for other viruses.

Investigations into a power-combining structure using a reflect array of dual-feed aperture-coupled microstrip patch antennas

This paper details an investigation of a power combiner that uses a reflect array of dual-feed aperture-coupled microstrip patch antennas and a corporate-fed dual-polarized array as a signal distributing/combining device. In this configuration, elements of the reflect array receive a linearly polarized wave and retransmit it with an orthogonal polarization using variable-length sections of microstrip lines connecting receive and transmit ports. By applying appropriate lengths of these delay lines, the array focuses the transmitted wave onto the feed array. The operation of the combiner is investigated for a small-size circular reflect array for the cases of -3 dB, -6 dB and -10 dB edge illumination by the 2 x 2-element dual-polarized array.

Design, development, and testing of X-band amplifying reflectarrays

The design and development of two X-band amplifying reflectarrays is presented. The arrays use dual-polarized aperture coupled patch antennas with FET transistors and phasing circuits to amplify a microwave signal and to radiate it in a chosen direction. Two cases are considered, one when a reflectarray converts a spherical wave due to a feed horn into a plane wave radiated into a boresight direction, and two, when the reflectarray converts a spherical wave due to a dual-polarized four-element feed array into a co-focal spherical wave. This amplified signal is received in an orthogonal port of the feed array so that the entire structure acts as a spatial power combiner. The two amplifying arrays are tested in the near-field zone for phase distribution over their apertures to achieve the required beam formation. Alternatively, their radiation patterns or gains are investigated.

High-frequency climatic oscillations recorded in a Holocene coral reef at Leizhou Peninsula, South China sea

A detailed study of the Goniopora reef profile at Dengloujiao, Xuwen County, Leizhou Peninsula, the northern coast of the South China Sea suggests that a series of high-frequency, large-amplitude and abrupt cold events occurred during the Holocene Hypsithermal, an unusual phenomenon termed Leizhou Events in this paper. This period (corresponding to C-14 age of 6.2 -6.7 kaBP or calendar age of 6.7-7.2 kaBP), when the climatic conditions were ideal for coral. reefs to develop, can be divided into at least nine stages. Each stage (or called a climate optimum), lasting about 20 to 50 a, was terminated by an abrupt cold nap and (or) a sea-level lowering event in winter, leading to widespread emergence and death of the Goniopora corals, and growth discontinuities on the coral surface. Such a cyclic process resulted in the creation of a > 4m thick Goniopora reef flat. During this period, the crust subsided periodically but the sea level was rising. The reef profile provides valuable archives for the study of decadal-scale mid-Holocene climatic oscillations in the tropical area of South China. Our results provide new evidence for high-frequency climate instability in the Holocene Hypsithermal, and challenge the traditional understanding of Holocene climate.

Archetype change in professional organizations: Survey evidence from large law firms

This paper examines the proposition that the traditional archetype of the professional partnership is said to have changed into a more 'business-like' entity, the managed professional business. It broadens the restricted case sample base on which much of the evidence has been adduced, by developing a survey questionnaire through which 197 large British law firms were sampled. Change, consistent with the notion of a more commercially oriented and consciously managed organization, is concentrated in the market-facing area of the firm but coexists with areas of continuity in the governance of the firm and its strategic management. The findings reveal a more managerial form of organization in which the core elements of the traditional form of professional organization have not been transformed. These results contest the assertion of either transformational or sedimented change found in other, case-based research and suggest that archetype change needs theoretically to be distinguished from the general phenomenon of greater managerialism within the professional service firm.

IL-10 Mediates Suppression of the CD8 T Cell IFN-γ Response to a Novel Viral Epitope in a Primed Host

Priming to Ag can inhibit subsequent induction of an immune response to a new epitope incorporated into that Ag, a phenomenon referred to as original antigenic sin. In this study, we show that prior immunity to a virus capsid can inhibit subsequent induction of the IFN-gamma effector T cell response to a novel CD8-restricted antigenic epitope associated with the virus capsid. Inhibition does not involve Ab to the virus capsid, as it is observed in animals lacking B cells. CD8-restricted virus-specific T cell responses are not required, as printing to virus without CTL induction is associated with inhibition. However, IL-10(-/-) mice, in contrast to IL-10(+/+) mice, generate CD8 T cell and Ab responses to novel epitopes incorporated into a virus capsid, even when priming to the capsid has resulted in high titer Ab to the capsid. Furthermore, capsid-primed mice, unable to mount a response to a novel epitope in the capsid protein, are nevertheless able to respond to the same novel epitope delivered independently of the capsid. Thus, inhibition of responsiveness to a novel epitope in a virus-primed animal is a consequence of secretion of IL-10 in response to presented Ag, which inhibits local generation of new CD8 IFN-gamma-secreting effector T cells. Induction of virus- or tumor Ag-specific CD8 effector T cells in the partially Ag-primed host may thus be facilitated by local neutralization of IL-10.

Writing the body of the mother: Narrative moments in Tsushima Yuko, Ariyoshi Sawako and Enchi Fumiko

This discussion argues the transformative potential inherent in the corporeal experience of motherhood as represented in selected textual moments of Japanese narrative. Narratives that address the experiences of the body of the mother are informed and given substance by an intense physicality, and thus have the potential to contest processes of social inscription in addition to suggesting alternative possibilities for all readers, not just those occupying an embodied maternal space. The discussion features brief events from the work of three writers who have written as mothers: Tsushima Y(u)macrko, Ariyoshi Sawako and Enchi Fumiko. In Yama o hashiru onna (1980; translated as Woman Running in the Mountains, 1991), Tsushima Y(u)macrko invites the reader to consider the embodied response to light of Takiko, a young pregnant woman. Emiko, the protagonist of Hishoku (Without Colour, 1967) by Ariyoshi Sawako, is the Japanese wife of an African American and has just given birth to a child. The daughter protagonist in Enchi Fumiko's 'Kami' ('Hair', 1957) operates a hairdressing business that is viable only with her mother's unpaid labour. The narratives are read through a matrix of post-structuralist theories of embodiment, drawing on the work of writers such as Julia Kristeva, Luce Irigaray and Elizabeth Grosz.

Intraempreendedorismo e a inova????o na gest??o p??blica federal

O objetivo deste artigo ?? apresentar tr??s a????es empreendedoras em organiza????es p??blicas premiadas pela ENAP no concurso Inova????o na Gest??o P??blica Federal. A partir da revis??o de literatura e estudo de caso, ser??o confrontados os diversos tipos e modelos de processos de inova????o, para identificar as dificuldades apontadas e as solu????es adotadas nas a????es premiadas. Foram selecionados tr??s casos de iniciativas inovadoras no INSS, apontando a origem, caracter??sticas, amplitude e o modelo do processo. Nosso quadro te??rico de refer??ncia est?? estruturado no sentido de aprofundar a percep????o, bem como nos dar condi????es de comparar alguns estudos e apontar lacunas, se existentes. A pesquisa foi realizada por meio de metodologia qualitativa, explorat??ria e adotando a estrat??gia de investiga????o do estudo de caso. A revis??o de literatura teve como objetivo identificar o que se encontra dispon??vel em livros, artigos, teses e, inclusive, experi??ncias. A conclus??o alcan??ada no artigo ?? a de que, apesar dos alegados entraves e limita????es, o intraempreendedorismo se faz presente nas organiza????es p??blicas e vem crescentemente sendo adotado pelas chefias ou mesmo por funcion??rios sem nenhum tipo de fun????o comissionada, como relatado em um dos casos. A inova????o nas organiza????es p??blicas, representada pelo intraempreendedorismo das experi??ncias premiadas pelo concurso, confirma nosso quadro te??rico na medida em que a????es empreendedoras geradoras de inova????o se fazem presentes nas organiza????es p??blicas, em todos os seus processos e meios.

CONCURSOS: É A SOLICITADORIA EQUIVALENTE A DIREITO?

Conclusão: ou seja, num certo concurso que abra para v.g. licenciatura em Direito e/ou “áreas afins”, um licenciado em Solicitadoria não pode ser excluído de ser concorrente, quando um licenciado em Direito é admitido. § Conclusion: that is, in a contest that opens to vg law degree and / or "related fields", a graduate in Solicitors can not be excluded from being competitor, when a degree in law is admitted.

Avaliação da rede de centros de atenção psicossocial: entre a saúde coletiva e a saúde mental

OBJETIVO: Avaliar os modelos assistenciais, de gestão e de formação de trabalhadores de uma rede centros de atenção psicossocial (CAPS). MÉTODOS: Pesquisa avaliativa qualitativa, sustentada pela hermenêutica gadameriana, realizada na cidade de Campinas (SP), em 2006-2007. Os dados foram coletados por meio de 20 grupos focais, em CAPS III, realizados com diferentes grupos de interesse (trabalhadores, gestores municipais, usuários, familiares e gestores locais). Após a transcrição do material gravado de cada grupo, foram construídas narrativas, seguindo o referencial teórico de Ricoeur. Na segunda etapa de grupos focais essas narrativas foram apresentadas aos participantes para contestá-las, corrigi-las e validá-las. Os resultados preliminares foram discutidos em oficinas para elaboração de um guia de boas práticas em CAPS III. RESULTADOS: Foram identificados pontos fortes e fragilidades no que concerne à atenção à crise, articulação com a rede básica, formulação de projetos terapêuticos, gestão e organização em equipes de referência, formação educacional e sofrimento psíquico. CONCLUSÕES: A rede de centros de atenção psicossocial em Campinas destaca-se pela sua originalidade na implantação de seis CAPS III , e pela sua eficácia na continência com usuários e familiares no momento da crise e na reabilitação. A organização por técnico e/ou equipe de referência prevalece, assim como a construção de projetos terapêuticos. A redução das equipes noturnas desponta como principal problema e fonte de estresse para os trabalhadores. A formação dos profissionais se mostrou insuficiente para os desafios enfrentados por esses serviços.

Does diaphragmatic breathing technique really improve its range of motion? An objective assessment

Purpose - To verify the results of a diaphragmatic breathing technique (DBT) on diaphragmatic range of motion in healthy subjects. Methods - A total of 51 healthy subjects (10 male; 41 female), mean age 20 years old and a body mass index (BMI) ranging from 15.6 to 34.9 kg/m2, were enrolled in this study. Diaphragmatic range of motion was assessed by M-mode ultrasound imaging. Measurements were made before and after the DBT implementation in a standard protocol, based on 3 seconds of inspiration starting from a maximum expiration. Differences between assessments were analyzed by descriptive statistics and t-test (p < 0.05). Results - Mean value range of motion before DBT was 55.3 ± 13.4 mm and after DBT was 63.8 ± 13.2 mm showing a significant improvement of 8.5 ± 14.7 mm (p < 0.001). A strong correlation between the slope and the range of motion was found (r = 0.71, p < 0.001). Conclusions - Based on ultrasound measurements, it has been proved that DBT really contributes to a higher diaphragmatic range of motion. Future studies are needed in order to understand the influence of protocol parameters (e.g. inspiration time). Clinical implications - In the contest of evidence-based practice in physiotherapy, it has been showed by objective measurements that DBT improves the diaphragm range of motion, translating into a more efficient ventilatory function and thus can be used in clinical setting. To our knowledge this is the first study to assess the effects of DBT on range of motion of diaphragm muscle with ultrasound imaging.