New NCI-N87-derived human gastric epithelial line after human telomerase catalytic subunit over-expression

AIM: To establish a cellular model correctly mimicking the gastric epithelium to overcome the limitation in the study of Helicobacter pylori (H. pylori ) infection. METHODS: Aiming to overcome this limitation, clones of the heterogenic cancer-derived NCI-N87 cell line were isolated, by stably-transducing it with the human telomerase reverse-transcriptase (hTERT) catalytic subunit gene. The clones were first characterized regarding their cell growth pattern and phenotype. For that we measured the clones' adherence properties, expression of cell-cell junctions' markers (ZO-1 and E-cadherin) and ability to generate a sustained transepithelial electrical resistance. The gastric properties of the clones, concerning expression of mucins, zymogens and glycan contents, were then evaluated by haematoxylin and eosin staining, Periodic acid Schiff (PAS) and PAS/Alcian Blue-staining, immunocytochemistry and Western blot. In addition, we assessed the usefulness of the hTERT-expressing gastric cell line for H. pylori research, by performing co-culture assays and measuring the IL-8 secretion, by ELISA, upon infection with two H. pylori strains differing in virulence. RESULTS: Compared with the parental cell line, the most promising NCI-hTERT-derived clones (CL5 and CL6) were composed of cells with homogenous phenotype, presented higher relative telomerase activities, better adhesion properties, ability to be maintained in culture for longer periods after confluency, and were more efficient in PAS-reactive mucins secretion. Both clones were shown to produce high amounts of MUC1, MUC2 and MUC13. NCI-hTERT-CL5 mucins were shown to be decorated with blood group H type 2 (BG-H), Lewis-x (Lex), Ley and Lea and, in a less extent, with BG-A antigens, but the former two antigens were not detected in the NCI-hTERT-CL6. None of the clones exhibited detectable levels of MUC6 nor sialylated Lex and Lea glycans. Entailing good gastric properties, both NCIhTERT- clones were found to produce pepsinogen-5 and human gastric lipase. The progenitor-like phenotype of NCI-hTERT-CL6 cells was highlighted by large nuclei and by the apical vesicular-like distribution of mucin 5AC and Pg5, supporting the accumulation of mucus-secreting and zymogens-chief mature cells functions. CONCLUSION: These traits, in addition to resistance to microaerobic conditions and good responsiveness to H. pylori co-culture, in a strain virulence-dependent manner, make the NCI-hTERT-CL6 a promising model for future in vitro studies.info:eu-repo/semantics/publishedVersio

New NCI-N87-derived human gastric epithelial line after human telomerase catalytic subunit over-expression

Supported by Grants from the Fundacao para a Ciencia e a Tecnologia (FCT, Portugal), No. PPCDT/SAL-IMI/57297/2004 and No. PTDC/BIM-MEC/1051/2012; The Swedish Cancer foundation; The Swedish Research Council, No. K2010-79X-21372-01-3; Forska utan djurforsok, Animal Free Research; and by Research fellowship 2011 from the Sociedade Portuguesa de Gastrenterologia (Portugal).AIM: To establish a cellular model correctly mimicking the gastric epithelium to overcome the limitation in the study of Helicobacter pylori (H. pylori) infection. METHODS: Aiming to overcome this limitation, clones of the heterogenic cancer-derived NCI-N87 cell line were isolated, by stably-transducing it with the human telomerase reverse-transcriptase (hTERT) catalytic subunit gene. The clones were first characterized regarding their cell growth pattern and phenotype. For that we measured the clones' adherence properties, expression of cell-cell junctions' markers (ZO-1 and E-cadherin) and ability to generate a sustained transepithelial electrical resistance. The gastric properties of the clones, concerning expression of mucins, zymogens and glycan contents, were then evaluated by haematoxylin and eosin staining, Periodic acid Schiff (PAS) and PAS/Alcian Blue-staining, immunocytochemistry and Western blot. In addition, we assessed the usefulness of the hTERT-expressing gastric cell line for H. pylori research, by performing co-culture assays and measuring the IL-8 secretion, by ELISA, upon infection with two H. pylori strains differing in virulence. RESULTS: Compared with the parental cell line, the most promising NCI-hTERT-derived clones (CL5 and CL6) were composed of cells with homogenous phenotype, presented higher relative telomerase activities, better adhesion properties, ability to be maintained in culture for longer periods after confluency, and were more efficient in PAS-reactive mucins secretion. Both clones were shown to produce high amounts of MUC1, MUC2 and MUC13. NCI-hTERT-CL5 mucins were shown to be decorated with blood group H type 2 (BG-H), Lewis-x (Le(x)), Le(y) and Le(a) and, in a less extent, with BG-A antigens, but the former two antigens were not detected in the NCI-hTERT-CL6. None of the clones exhibited detectable levels of MUC6 nor sialylated Le(x) and Le(a) glycans. Entailing good gastric properties, both NCI-hTERT-clones were found to produce pepsinogen-5 and human gastric lipase. The progenitor-like phenotype of NCI-hTERT-CL6 cells was highlighted by large nuclei and by the apical vesicular-like distribution of mucin 5AC and Pg5, supporting the accumulation of mucus-secreting and zymogens-chief mature cells functions. CONCLUSION: These traits, in addition to resistance to microaerobic conditions and good responsiveness to H. pylori co-culture, in a strain virulence-dependent manner, make the NCI-hTERT-CL6 a promising model for future in vitro studies.publishersversionpublishe

Estrategias utilizadas en la prevención de la neumonía asociada a la ventilación mecánica

La primera causa de mortalidad en cuanto a infecciones nosocomiales (IN) es la neumonía asociada a la ventilación mecánica (NAV); reto muy grande al que se deben enfrentar el personal médico y el paciente.Es importante que en las unidades de cuidado intensivo se realicen actividades que permitan prevenir esta patología, ya que genera múltiples complicaciones. El objetivo de esta revisión es identificar las diferentes estrategias utilizadas en la prevención de la neumonía asociada a la ventilación mecánica, mediante una revisión de la literatura reciente. Se realizó una revisión bibliográfica de diferentes bases de datos, como archivos de bronconeumología, SciELO, pubmed, entre otros. Se reunió suficiente información para realizar el desarrollo del tema y cumplir los objetivos. Se encontraron múltiples estrategias que han demostrado ser efectivas en la prevención de la NAVM por múltiples investigaciones realizadas. Como terapeutas respiratorios, es necesario que seamos exigentes en la implementación y cumplimiento de estas medidas

Homoclinic standing waves in focussing DNLS equations --Variational approach via constrained optimization

We study focussing discrete nonlinear Schr\"{o}dinger equations and present a new variational existence proof for homoclinic standing waves (bright solitons). Our approach relies on the constrained maximization of an energy functional and provides the existence of two one-parameter families of waves with unimodal and even profile function for a wide class of nonlinearities. Finally, we illustrate our results by numerical simulations.Comment: new version with revised introduction and improved condition (A3); 16 pages, several figure

Calidad del grano de arroz

La calidad de un producto es definida por el conjunto de propiedades y características del mismo que satisfacen las necesidades particulares de los usuarios finales. En este contexto, la calidad del grano de arroz constituye el conjunto de todas las propiedades y características del arroz que satisfagan las necesidades de los consumidores (Bao, 2014). Actualmente, existe una gran cantidad de información genética sobre las diferentes características que definen la calidad del arroz y que pueden ayudar a mejorar el producto producido y consumido por la población

Co-periodic stability of periodic waves in some Hamiltonian PDEs

International audienceThe stability theory of periodic traveling waves is much less advanced than for solitary waves, which were first studied by Boussinesq and have received a lot of attention in the last decades. In particular, despite recent breakthroughs regarding periodic waves in reaction-diffusion equations and viscous systems of conservation laws [Johnson–Noble–Rodrigues–Zumbrun, Invent math (2014)], the stability of periodic traveling wave solutions to dispersive PDEs with respect to 'arbitrary' perturbations is still widely open in the absence of a dissipation mechanism. The focus is put here on co-periodic stability of periodic waves, that is, stability with respect to perturbations of the same period as the wave, for KdV-like systems of one-dimensional Hamiltonian PDEs. Fairly general nonlinearities are allowed in these systems, so as to include various models of mathematical physics, and this precludes complete integrability techniques. Stability criteria are derived and investigated first in a general abstract framework, and then applied to three basic examples that are very closely related, and ubiquitous in mathematical physics, namely, a quasilinear version of the generalized Korteweg–de Vries equation (qKdV), and the Euler–Korteweg system in both Eulerian coordinates (EKE) and in mass Lagrangian coordinates (EKL). Those criteria consist of a necessary condition for spectral stability , and of a sufficient condition for orbital stability. Both are expressed in terms of a single function, the abbreviated action integral along the orbits of waves in the phase plane, which is the counterpart of the solitary waves moment of instability introduced by Boussinesq. However, the resulting criteria are more complicated for periodic waves because they have more degrees of freedom than solitary waves, so that the action is a function of N + 2 variables for a system of N PDEs, while the moment of instability is a function of the wave speed only once the endstate of the 1 solitary wave is fixed. Regarding solitary waves, the celebrated Grillakis–Shatah– Strauss stability criteria amount to looking for the sign of the second derivative of the moment of instability with respect to the wave speed. For periodic waves, stability criteria involve all the second order, partial derivatives of the action. This had already been pointed out by various authors for some specific equations, in particular the generalized Korteweg–de Vries equation — which is special case of (qKdV) — but not from a general point of view, up to the authors' knowledge. The most striking results obtained here can be summarized as: an odd value for the difference between N and the negative signature of the Hessian of the action implies spectral instability, whereas a negative signature of the same Hessian being equal to N implies orbital stability. Furthermore, it is shown that, when applied to the Euler–Korteweg system, this approach yields several interesting connexions between (EKE), (EKL), and (qKdV). More precisely, (EKE) and (EKL) share the same abbreviated action integral, which is related to that of (qKdV) in a simple way. This basically proves simultaneous stability in both formulations (EKE) and (EKL) — as one may reasonably expect from the physical point view —, which is interesting to know when these models are used for different phenomena — e.g. shallow water waves or nonlinear optics. In addition, stability in (EKE) and (EKL) is found to be linked to stability in the scalar equation (qKdV). Since the relevant stability criteria are merely encoded by the negative signature of (N + 2) × (N + 2) matrices, they can at least be checked numerically. In practice, when N = 1 or 2, this can be done without even requiring an ODE solver. Various numerical experiments are presented, which clearly discriminate between stable cases and unstable cases for (qKdV), (EKE) and (EKL), thus confirming some known results for the generalized KdV equation and the Nonlinear Schrödinger equation, and pointing out some new results for more general (systems of) PDEs

On the ill-posedness result for the BBM equation

We prove that the initial value problem (IVP) for the BBM equation is ill-posed for data in Hs(R), s < 0 in the sense that the ow-map u0 7! u(t) that associates to initial data u0 the solution u cannot be continuous at the origin from Hs(R) to even D0(R) at any _xed t > 0 small enough. This result is sharp.Fundação para a Ciência e a Tecnologia (FCT

Sustainability disclosure and reputation: a comparative study

“This is a post-peer-review, pre-copyedit version of an article published in Corporate Reputation Review. The definitive publisher-authenticated version Corporate Reputation Review 14(2), pp.79-96 is available online at: http://www.palgrave-journals.com/crr/index.html”Drawing on legitimacy theory, we discuss that a company’s reputation is a determinant of sustainability disclosure. Specifically, we consider the concept of reputation into three dimensions for analysis: stakeholders’ commitment, financial performance and media exposure. This paper differs from previous social and environmental reporting studies in that it investigates both internal and external contextual factors that influence disclosure practice. We claim that companies with a good financial performance, that are adopting an active strategic position towards stakeholders and that are exposed to significant public pressure are more likely to use sustainability disclosure in order to communicate their legitimacy to operate to stakeholders. Moreover the paper analyses a wide range of corporate reports for their social and environmental content using an international sample that allows for a comparison of disclosure practices among Continental European, UK and USA companies. Our results show that stakeholder commitment and media exposure are positively associated with sustainability disclosure. Moreover, we find evidence that the drivers of disclosure vary by information type

Transcription and Expression of Plasmodium falciparum Histidine-Rich Proteins in Different Stages and Strains: Implications for Rapid Diagnostic Tests

Background: Although rapid diagnostic tests (RDTs) for Plasmodium falciparum infection that target histidine rich protein 2 (PfHRP2) are generally sensitive, their performance has been reported to be variable. One possible explanation for variable test performance is differences in expression level of PfHRP in different parasite isolates. Methods: Total RNA and protein were extracted from synchronised cultures of 7 P. falciparum lines over 5 time points of the life cycle, and from synchronised ring stages of 10 falciparum lines. Using quantitative real-time polymerase chain reaction, Western blot analysis and ELISA we investigated variations in the transcription and protein levels of pfhrp2, pfhrp3 and PfHRP respectively in the different parasite lines, over the parasite intraerythrocytic life cycle. Results: Transcription of pfhrp2 and pfhrp3 in different parasite lines over the parasite life cycle was observed to vary relative to the control parasite K1. In some parasite lines very low transcription of these genes was observed. The peak transcription was observed in ring-stage parasites. Pfhrp2 transcription was observed to be consistently higher than pfhrp3 transcription within parasite lines. The intraerythrocytic lifecycle stage at which the peak level of protein was present varied across strains. Total protein levels were more constant relative to total mRNA transcription, however a maximum 24 fold difference in expression at ring-stage parasites relative to the K1 strain was observed. Conclusions: The levels of transcription of pfhrp2 and pfhrp3, and protein expression of PfHRP varied between different P. falciparum strains. This variation may impact on the detection sensitivity of PfHRP2-detecting RDTs