Isolation of mineralizing Nestin+ Nkx6.1+ vascular muscular cells from the adult human spinal cord

Background: The adult central nervous system (CNS) contains different populations of immature cells that could possibly be used to repair brain and spinal cord lesions. The diversity and the properties of these cells in the human adult CNS remain to be fully explored. We previously isolated Nestin(+) Sox2(+) neural multipotential cells from the adult human spinal cord using the neurosphere method (i.e. non adherent conditions and defined medium). -- Results: Here we report the isolation and long term propagation of another population of Nestin(+) cells from this tissue using adherent culture conditions and serum. QPCR and immunofluorescence indicated that these cells had mesenchymal features as evidenced by the expression of Snai2 and Twist1 and lack of expression of neural markers such as Sox2, Olig2 or GFAP. Indeed, these cells expressed markers typical of smooth muscle vascular cells such as Calponin, Caldesmone and Acta2 (Smooth muscle actin). These cells could not differentiate into chondrocytes, adipocytes, neuronal and glial cells, however they readily mineralized when placed in osteogenic conditions. Further characterization allowed us to identify the Nkx6.1 transcription factor as a marker for these cells. Nkx6.1 was expressed in vivo by CNS vascular muscular cells located in the parenchyma and the meninges. -- Conclusion: Smooth muscle cells expressing Nestin and Nkx6.1 is the main cell population derived from culturing human spinal cord cells in adherent conditions with serum. Mineralization of these cells in vitro could represent a valuable model for studying calcifications of CNS vessels which are observed in pathological situations or as part of the normal aging. In addition, long term propagation of these cells will allow the study of their interaction with other CNS cells and their implication in scar formation during spinal cord injury.

Importancia de la marca paraguas y el made in en la competitividad internacional del granito

[ES] España, al igual que Italia, líder tradicional del sector de granito, ha visto como China, India o Brasil, han escalado posiciones en el ranking de producción y exportación mundial de granito. En un contexto globalizado, es necesario posicionarse frente a estos competidores en los mercados internacionales, y dado que estamos ante un producto genérico que cumple unas condiciones adecuadas de precio y calidad, una forma de identificarlo y diferenciarlo es aportándole valor mediante la creación de una marca. En el trabajo se analiza la utilidad de una estrategia basada en el made in para el caso de Galicia, núcleo fundamental de la industria en España, alternativa que resulta interesante para las empresas consultadas pero cuya puesta en práctica exige la colaboración entre éstas y las instituciones.

Multiscale Method for Turbulent-Flow Computations

A main method of predicting turbulent flows is to solve LES equations, which was called traditional LES method. The traditional LES method solves the motions of large eddies of size larger than filtering scale An while modeling unresolved scales less than Delta_n. Hughes et al argued that many shortcomings of the traditional LES approaches were associated with their inabilities to successfully differentiate between large and small scales. One may guess that a priori scale-separation would be better, because it can predict scale-interaction well compared with posteriori scale-separation. To this end, a multi-scale method was suggested to perform scale-separation computation. The primary contents of the multiscale method are l) A space average is used to differentiate scale. 2) The basic equations include the large scale equations and fluctuation equations. 3) The large-scale equations and fluctuation equations are coupled through turbulent stress terms. We use the multiscale equations of n=2, i.e., the large and small scale (LSS) equations, to simulate 3-D evolutions of a channel flow and a planar mixing layer flow Some interesting results are given.

Huntingdonshire Regional College - The Teacher/Learner Switch

It is said the best ideas are often the simplest ones. At Huntingdonshire Regional College, Ken McKerral, an Advanced Practitioner in E-Learning has developed a very simple idea to engage students and improve teaching and learning. Ken has named his method "The Teacher/Learner Switch". It is a process that uses a unique colour code system, to help differentiate learning outcomes and deliver ownership of time to the students to enrich learning experience.

Uso de células iPS en terapia regenerativa renal: obtención de células iPS mediante vector no integrativo y diferenciación hacia podocitos.

[ES]En las sociedades modernas existe una creciente preocupación por el aumento de la incidencia de la enfermedad renal crónica. Debido a la deficiencia de donantes de órganos y al elevado coste del tratamiento de diálisis, existe la necesidad de desarrollar nuevos tratamientos para estos pacientes. La medicina regenerativa basada en la aplicación de células iPS es una opción prometedora para el tratamiento de esta enfermedad. Sin embargo, la falta de conocimientos sobre el estado pluripotencial de las células y sobre su proceso de diferenciación, así como las limitaciones derivadas del propio procedimiento de reprogramación, impiden su aplicación clínica en un futuro inmediato. Para que se convierta en realidad, numerosas investigaciones se están llevando a cabo con el objetivo de mejorar el procedimiento y hacerlo adecuado para su aplicación clínica. En este trabajo se propone un método que permitiría obtener células iPS a partir de células mesangiales mediante la transfección con un vector no integrativo, el virus Sendai, portador de los genes Oct3/4, Sox2, Klf4 y c-Myc. Al tratarse de un vector no integrativo, se minimizaría el efecto del proceso de reprogramación sobre la estabilidad del genoma celular. Además, en este proyecto se estudiará la capacidad de las células iPS obtenidas para diferenciarse en células progenitoras de podocitos que puedan ser aplicadas específicamente en terapias regenerativas para enfermos renales crónicos.

Dynamics of Streptococcus pneumoniae Serotypes Causing Acute Otitis Media Isolated from Children with Spontaneous Middle-Ear Drainage over a 12-Year Period (1999–2010) in a Region of Northern Spain

10 p.

NMDA modulates oligodendrocyte differentiation of subventricular zone cells through PKC activation

7 p.

Texture measurement on gutted cod during storage in ice using a hand-held instrument

Fish muscle as food is to be seen as highly perishable. In unfrozen fish, freshness is considered the most important quality attribute. It is well known that there are several biochemical changes that can affect dramatically the texture of fish muscle. Immediately after death the fish texture is soft and elastic. In connection with rigor mortis the fish texture changes markedly. It becomes harder during rigor and after its resolution it becomes softer. This softness increases due to proteolysis during further storage at refrigerated conditions. Texture is a very important indicator for evaluating the quality of fish. Barroso et al. (1997) have recently reviewed mechanical methods in use for texture measurements on fresh fish. Further reviews on texture measurement performed on fish muscle were recently published underlining the importance of texture as quality attribute (Hyldig et al 2001, Coppes et al. 2002). The position along the fish can influence the results and was investigated by several authors (Sigurgis-ladottir et. al. 1999). Different methods have been compared for their ability to differentiate between recently killed salmon and salmon stored on ice for up to 24 days (Veland et al. 1999).

Qualitätsvergleich zwischen panierten Filetportionen aus einfach- und doppelgefrorenen TK-Blöcken

Single and double frozen fillet blocks of Alaska pollack and cod both commercially processed of unknown shelf life were further processed to breaded battered portions. The quality of these fillet portions were compared using sensory (QDA), physical and chemical methods. It was difficult to differentiate between SF and DF fillets by sensory method because of the absence of differences in flavour attributes. While no differences could to be found in the texture of cod fillets, in Alaska pollack fillets some texture attributes were significantly different. These differences could not be verified by instrumental texture measurement. In all cases the lightness was different between SF and DF fillets. Probably, after having fixed L* values for SF fillets of commercially important fish species as limit this could be employed in the future to differentiate between single and double frozen products. Due to the unknown shelf life it is difficult to evaluate the results. Therefore, the investigation of the influence of double freezing on the quality needs a special sample preparation. The use of randomly taken commercially processed samples seems not to be useful.

Fischartbestimmung von Caviar durch Protein- und DNA-Analyse

Deutscher Caviar, made from roe of lumpfish or capelin, gives species specific patterns in protein electrophoresis. The same techniques can be used to differentiate caviar from salmon and trout. The differentiation of sturgeon caviar (beluga, osietra, sevruga) is possible by isoelectric focusing, but not by SDS-PAGE. PCR-based methods of DNA-analysis for identification of the origin of sturgeon caviar are under development.

Function of microRNA-146a and NF-κB in physiologic and pathologic hematopoiesis

During inflammation and infection, hematopoietic stem and progenitor cells (HSPCs) are stimulated to proliferate and differentiate into mature immune cells, especially of the myeloid lineage. MicroRNA-146a (miR-146a) is a critical negative regulator of inflammation. Deletion of the gene encoding miR-146a—expressed in all blood cell types—produces effects that appear as dysregulated inflammatory hematopoiesis, leading to a decline in the number and quality of hematopoietic stem cells (HSCs), excessive myeloproliferation, and, ultimately, to exhaustion of the HSCs and hematopoietic neoplasms. Six-week-old deleted mice are normal, with no effect on cell numbers, but by 4 months bone marrow hypercellularity can be seen, and by 8 months marrow exhaustion is becoming evident. The ability of HSCs to replenish the entire hematopoietic repertoire in a myelo-ablated mouse also declines precipitously as miR-146a-deficient mice age. In the absence of miR-146a, LPS-mediated serial inflammatory stimulation accelerates the effects of aging. This chronic inflammatory stress on HSCs in deleted mice involves a molecular axis consisting of upregulation of the signaling protein TRAF6 leading to excessive activity of the transcription factor NF-κB and overproduction of the cytokine IL-6. At the cellular level, transplant studies show that the defects are attributable to both an intrinsic problem in the miR-146a-deficient HSCs and extrinsic effects of miR-146a-deficient lymphocytes and non-hematopoietic cells. This study has identified a microRNA, miR-146a, to be a critical regulator of HSC homeostasis during chronic inflammatory challenge in mice and has provided a molecular connection between chronic inflammation and the development of bone marrow failure and myeloproliferative neoplasms. This may have implications for human hematopoietic malignancies, such as myelodysplastic syndrome, which frequently displays downregulated miR-146a expression.

Molecular neurogenetics of eye development in Drosophila

The compound eye of Drosophila melanogaster begins to differentiate during the late third larval instar in the eye-antennal imaginal disc. A wave of morphogenesis crosses the disc from posterior to anterior, leaving behind precisely patterned clusters of photoreceptor cells and accessory cells that will constitute the adult ommatidia of the retina. By the analysis of genetically mosaic eyes, it appears that any cell in the eye disc can adopt the characteristics of any one of the different cell types found in the mature eye, including photoreceptor cells and non-neuronal accessory cells such as cone cells. Therefore, cells within the prospective retinal epithelium assume different fates presumably via information present in the environment. The sevenless^+ (sev^+) gene appears to play a role in the expression of one of the possible fates, since the mutant phenotype is the lack of one of the pattern elements, namely, photoreceptor cell R7. The sev^+ gene product had been shown to be required during development of the eye, and had also been shown in genetic mosaics to be autonomous to presumptive R7. As a means of better understanding the pathway instructing the differentiation R7, the gene and its protein product were characterized.

The sev+ gene was cloned by P-element transposon tagging, and was found to encode an 8.2 kb transcript expressed in developing eye discs and adult heads. By raising monoclonal antibodies (MAbs) against a sev^+- β-galactosidase fusion protein, the expression of the protein in the eye disc was localized by immuno-electronmicroscopy. The protein localizes to the apical cell membranes and microvilli of cells in the eye disc epithelium. It appears during development at a time coincident with the initial formation of clusters, and in all the developing photoreceptors and accessory cone cells at a time prior to the overt differentiation of R7. This result is consistent with the pluripotency of cells in the eye disc. Its localization in the membranes suggests that it may receive information directing the development of R7. Its localization in the apical membranes and microvilli is away from the bulk of the cell contacts, which have been cited as a likely regions for information presentation and processing. Biochemical characterization of the sev^+ protein will be necessary to describe further its role in development.

Other mutations in Drosophila have eye phenotypes. These were analyzed to find which ones affected the initial patterning of cells in the eye disc, in order to identify other genes, like sev, whose gene products may be involved in generating the pattern. The adult eye phenotypes ranged from severe reduction of the eye, to variable numbers of photoreceptor cells per ommatidium, to sub de defects in the organization of the supporting cells. Developing eye discs from the different strains were screened using a panel of MAbs, which highlight various developmental stages. Two identified matrix elements in and anterior to the furrow, while others identified the developing ommatidia themselves, like the anti-sev MAb. Mutation phenotypes were shown to appear at many stages of development. Some mutations seem to affect the precursor cells, others, the setting up of the pattern, and still others, the maintenance of the pattern. Thus, additional genes have now been identified that may function to support the development of a complex pattern.

Comportamento de bugios (Alouatta clamitans Cabrera, 1940) na Ilha Grande, RJ

Alouatta clamitans é uma espécie endêmica da Mata Atlântica, bioma que vem sendo continuamente reduzido, o que torna de extrema urgência o conhecimento sobre a espécie. No Estado do Rio de Janeiro, sua área de ocorrência abrange a região da Ilha Grande, município de Angra dos Reis. A Ilha Grande possui uma extensa área de preservação, o Parque Estadual da Ilha Grande, que atua na conservação de cerca de 62,5% da sua extensão. O isolamento das espécies em ilhas pode provocar o desenvolvimento de características morfológicas e comportamentais diferentes das espécies do continente. No entanto, não existem trabalhos sistematizados sobre a ecologia e o comportamento da espécie no local. Este estudo objetivou analisar aspectos do comportamento de Alouatta clamitans na Ilha Grande, contribuindo para uma melhor compreensão sobre a biologia da espécie. Durante nove meses foram registrados dados de composição social e comportamento de grupos da espécie através da amostragem por varredura instantânea e todas as ocorrências. Observou-se que o tamanho médio dos grupos foi de cinco indivíduos e a composição social por grupo foi representada por um a dois machos adultos, uma a três fêmeas adultas e imaturos de diferentes classes etárias, com predominância de grupos unimacho. Em média, os grupos eram compostos por 22% de machos adultos, 38% de fêmeas adultas, 4% de machos subadultos, 27% de juvenis e 9% de infantes. O comportamento mais observado foi o repouso (45,2%), seguido da alimentação (28%), movimentação (21,7%) e comportamento social (5,1%), e dentre os comportamentos sociais, o mais exibido foi a vocalização (45,8%), seguido dos comportamentos de catação (33,7%), agonístico (7,9%), brincadeira (5,8%), marcação (4,2%) e comportamento sexual (2,6%). Não foram encontradas diferenças estatisticamente significativas nestas atividades entre os períodos seco e chuvoso. As vocalizações foram predominantemente emitidas por machos e adultos e estiveram relacionadas ao encontro de grupos. O comportamento de catação teve as fêmeas adultas como principais iniciadoras e os machos adultos, principais receptores, sendo realizado durante o comportamento de repouso, após a cópula, após e durante encontro de grupos e após perseguições. Os comportamentos agonísticos tiveram relação com o encontro de grupos em 40% dos registros e em 33,3% destes ocorreu entre fêmeas e pareceu estar associado à disputa por alimento e espaço, mas não houve registros de agressão física. O comportamento de marcação envolveu a utilização da garganta e das costas e esteve relacionado com encontros inter-grupais e com a ocorrência de chuvas. Cinco cópulas foram registradas no período de estudo nos meses de setembro, outubro e fevereiro e tiveram duração menor que um minuto. Nos encontros com primatas de outras espécies, os bugios pareceram neutros em relação aos estímulos. Os dados obtidos sobre a composição dos grupos, padrão de atividades e comportamentos sociais observados na Ilha Grande, de maneira geral, mostraram-se semelhantes aos resultados obtidos em outros trabalhos sobre a espécie e o gênero, de maneira que podemos concluir que os grupos, mesmo residentes em ilha, não demonstraram modificações comportamentais significativas que possam diferenciar-lhes de populações estudadas no continente.

Diagnosis of colorectal cancer using Raman spectroscopy of laser-trapped single living epithelial cells

A single-cell diagnostic technique for epithelial cancers is developed by utilizing laser trapping and Raman spectroscopy to differentiate cancerous and normal epithelial cells. Single-cell suspensions were prepared from surgically removed human colorectal tissues following standard primary culture protocols and examined in a near-infrared laser-trapping Raman spectroscopy system, where living epithelial cells were investigated one by one. A diagnostic model was built on the spectral data obtained from 8 patients and validated by the data from 2 new patients. Our technique has potential applications from epithelial cancer diagnosis to the study of cell dynamics of carcinogenesis. (c) 2006 Optical Society of America.

From molecules to organs : Microscopy and multi-scale nature of development

Morphogenesis is a phenomenon of intricate balance and dynamic interplay between processes occurring at a wide range of scales (spatial, temporal and energetic). During development, a variety of physical mechanisms are employed by tissues to simultaneously pattern, move, and differentiate based on information exchange between constituent cells, perhaps more than at any other time during an organism's life. To fully understand such events, a combined theoretical and experimental framework is required to assist in deciphering the correlations at both structural and functional levels at scales that include the intracellular and tissue levels as well as organs and organ systems. Microscopy, especially diffraction-limited light microscopy, has emerged as a central tool to capture the spatio-temporal context of life processes. Imaging has the unique advantage of watching biological events as they unfold over time at single-cell resolution in the intact animal. In this work I present a range of problems in morphogenesis, each unique in its requirements for novel quantitative imaging both in terms of the technique and analysis. Understanding the molecular basis for a developmental process involves investigating how genes and their products- mRNA and proteins-function in the context of a cell. Structural information holds the key to insights into mechanisms and imaging fixed specimens paves the first step towards deciphering gene function. The work presented in this thesis starts with the demonstration that the fluorescent signal from the challenging environment of whole-mount imaging, obtained by in situ hybridization chain reaction (HCR), scales linearly with the number of copies of target mRNA to provide quantitative sub-cellular mapping of mRNA expression within intact vertebrate embryos. The work then progresses to address aspects of imaging live embryonic development in a number of species. While processes such as avian cartilage growth require high spatial resolution and lower time resolution, dynamic events during zebrafish somitogenesis require higher time resolution to capture the protein localization as the somites mature. The requirements on imaging are even more stringent in case of the embryonic zebrafish heart that beats with a frequency of ~ 2-2.5 Hz, thereby requiring very fast imaging techniques based on two-photon light sheet microscope to capture its dynamics. In each of the hitherto-mentioned cases, ranging from the level of molecules to organs, an imaging framework is developed, both in terms of technique and analysis to allow quantitative assessment of the process in vivo. Overall the work presented in this thesis combines new quantitative tools with novel microscopy for the precise understanding of processes in embryonic development.