Culture bag systems for clinical applications of adult human neural crest-derived stem cells

Introduction Facing the challenging treatment of neurodegenerative diseases as well as complex craniofacial injuries such as those common after cancer therapy, the field of regenerative medicine increasingly relies on stem cell transplantation strategies. Here, neural crest-derived stem cells (NCSCs) offer many promising applications, although scale up of clinical-grade processes prior to potential transplantations is currently limiting. In this study, we aimed to establish a clinical-grade, cost-reducing cultivation system for NCSCs isolated from the adult human nose using cGMP-grade Afc-FEP bags. Methods We cultivated human neural crest-derived stem cells from inferior turbinate (ITSCs) in a cell culture bag system using Afc-FEP bags in human blood plasma-supplemented medium. Investigations of viability, proliferation and expression profile of bag-cultured ITSCs were followed by DNA-content and telomerase activity determination. Cultivated ITSCs were introduced to directed in vitro differentiation assays to assess their potential for mesodermal and ectodermal differentiation. Mesodermal differentiation was determined using an enzyme activity assay (alkaline phosphatase, ALP), respective stainings (Alizarin Red S, Von Kossa and Oil Red O), and RT-PCR, while immunocytochemistry and synaptic vesicle recycling were applied to assay neuroectodermal differentiation of ITSCs. Results When cultivated within Afc-FEP bags, ITSCs grew three-dimensionally in a human blood plasma-derived matrix, thereby showing unchanged morphology, proliferation capability, viability and expression profile in comparison to three dimensionally-cultured ITSCs growing in standard cell culture plastics. Genetic stability of bag-cultured ITSCs was further accompanied by unchanged telomerase activity. Importantly, ITSCs retained their potential to differentiate into mesodermal cell types, particularly including ALP-active, Alizarin Red S-, and Von Kossa-positive osteogenic cell types, as well as adipocytes positive in Oil Red O assays. Bag culture further did not affect the potential of ITSCs to undergo differentiation into neuroectodermal cell types coexpressing β-III-tubulin and MAP2 and exhibiting the capability for synaptic vesicle recycling. Conclusions Here, we report for the first time the successful cultivation of human NCSCs within cGMP-grade Afc-FEP bags using a human blood plasma-supplemented medium. Our findings particularly demonstrate the unchanged differentiation capability and genetic stability of the cultivated NCSCs, suggesting the great potential of this culture system for future medical applications in the field of regenerative medicine.

Total absolute horospherical curvature of submanifolds in hyperbolic space

We study the horospherical geometry of submanifolds in hyperbolic space. The main result is a formula for the total absolute horospherical curvature of M, which implies, for the horospherical geometry, the analogues of classical inequalities of the Euclidean Geometry. We prove the horospherical Chern-Lashof inequality for surfaces in 3-space and the horospherical Fenchel and Fary-Milnor`s theorems.

O modelo de avaliação de ativos: (Capital asset pricing model)

Examina o modelo de seleção de portfólios desenvolvido por Markowitz, principalmente no que concerne: as suas relações com a teoria da utilidade de Von Neumann-Morgenstern; aos algo ritmos de solução do problema de Programação Quadrática paramétrica dele decorrente; a simplificação proporcionada pelo Modelo Diagonal de Sharpe. Mostra que a existência de um título sem risco permite a especificação do Teorema da Separação e a simplificação do problema de seleção de portfólios. Analisa o modelo denominado por CAPM, de equilíbrio no Mercado de Capitais sob condições de incerteza, comparando os processos dedutivos empregados por Lintner e Mossin. Examina as implicações decorrentes do relaxamento dos pressupostos subjacentes ã esse modelo de equilíbrio geral, principalmente a teoria do portfólio Zero-Beta desenvolvida por Black.

Álgebras de operadores, esperança condicional e a Entropia de Connes-Stormer

Neste trabalho fazemos um breve estudo de Álgebras de Operadores, mais especificamente Álgebras-C* e Álgebras de von Neumann. O objetivo é expor alguns resultados que seriam os análogos não-comutativos de teoremas em Teoria da Medida e Teoria Rrgódica. Inicialmente, enunciamos alguns resultados de Análise Funcional e Teoria Espectral, muitos destes sendo demonstrados, com ênfase especial aos que dizem respeito µas álgebras. Com isso, dispomos das ferramentas necessárias para falarmos de alguns tópicos da então chamada Teoria da Integração Não-Comutativa. Uma desigualdade tipo Jensen é provada e, com o teorema de Radon-Nikodym para funcionais normais positivos, construimos uma esperança condicional, provando que esta possui as mesmas propriedades da esperança condicional da Teoria das Probabilidades. Dada a Esperança Condicional, objeto este que faz parte do cenário atual de pesquisa na área de Álgebra de Operadores e que está relacionado com resultados fundamentais tal como o Índice de Jones, passamos à definição da Entropia de Connes-Stormer. Finalizamos o trabalho analisando esta entropia, que é a versão para as álgebras de von Neumann da entropia Kolmogorov-Sinai em Teoria Ergódica. Provamos algumas pro- priedades que são análogas às do conceito clássico de entropia e indicamos uma aplicação da mesma. O texto não possui resultados originais, trata-se apenas de uma releitura de artigos usando versões mais recentes de alguns teoremas.

Aplicação de modelos não-lineares para descrever a evolução de características de crescimento e carcaça em bovinos da raça Hereford

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Comparison of non-linear growth models to describe the growth curve in West African Dwarf sheep

The objectives of this study were to compare the goodness of fit of four non-linear growth models, i.e. Brody, Gompertz, Logistic and Von Bertalanffy, in West African Dwarf (WAD) sheep. A total of 5274 monthly weight records from birth up to 180 days of age from 889 lambs, collected during 2001 to 2004 in Betecoucou breeding farm in Benin were used. In the preliminary analysis, the General Linear Model Procedure of the Statistical Analysis Systems Institute was applied to the dataset to identify the significant effects of the sex of lamb (male and female), type of birth (single and twin), season of birth (rainy season and dry season), parity of dam (1, 2 and 3) and year of birth (2001, 2002, 2003 and 2004) on the observed birth weight and monthly weight up to 6 months of age. The models parameters (A, B and k), coefficient of determination (112), mean square error (MSE) were calculated using language of technical computing package Matlab(R), 2006. The mean values of A, B and k were substituted into each model to calculate the corresponding Akaike's Information Criterion (AIC). Among the four growth functions, the Brody model has been selected for its accuracy of fit according to the higher R(2), lower MSE and A/C Finally, the parameters A, B and k were adjusted in Matlab(R) 2006 for the sex of lamb, year of birth, season of birth, birth type and the parity of ewe, providing a specific slope of the Brody growth curve. The results of this study suggest that Brody model can be useful for WAD sheep breeding in Betecoucou farm conditions through growth monitoring.

Propagação de danos em sistemas cooperativos: propriedades termodinâmicas

High-precision calculations of the correlation functions and order parameters were performed in order to investigate the critical properties of several two-dimensional ferro- magnetic systems: (i) the q-state Potts model; (ii) the Ashkin-Teller isotropic model; (iii) the spin-1 Ising model. We deduced exact relations connecting specific damages (the difference between two microscopic configurations of a model) and the above mentioned thermodynamic quanti- ties which permit its numerical calculation, by computer simulation and using any ergodic dynamics. The results obtained (critical temperature and exponents) reproduced all the known values, with an agreement up to several significant figures; of particular relevance were the estimates along the Baxter critical line (Ashkin-Teller model) where the exponents have a continuous variation. We also showed that this approach is less sensitive to the finite-size effects than the standard Monte-Carlo method. This analysis shows that the present approach produces equal or more accurate results, as compared to the usual Monte Carlo simulation, and can be useful to investigate these models in circumstances for which their behavior is not yet fully understood

Prova automática de satisfatibilidade módulo teoria aplicada ao método B

Este trabalho apresenta uma extensão do provador haRVey destinada à verificação de obrigações de prova originadas de acordo com o método B. O método B de desenvolvimento de software abrange as fases de especificação, projeto e implementação do ciclo de vida do software. No contexto da verificação, destacam-se as ferramentas de prova Prioni, Z/EVES e Atelier-B/Click n Prove. Elas descrevem formalismos com suporte à checagem satisfatibilidade de fórmulas da teoria axiomática dos conjuntos, ou seja, podem ser aplicadas ao método B. A checagem de SMT consiste na checagem de satisfatibilidade de fórmulas da lógica de primeira-ordem livre de quantificadores dada uma teoria decidível. A abordagem de checagem de SMT implementada pelo provador automático de teoremas haRVey é apresentada, adotando-se a teoria dos vetores que não permite expressar todas as construções necessárias às especificações baseadas em conjuntos. Assim, para estender a checagem de SMT para teorias dos conjuntos destacam-se as teorias dos conjuntos de Zermelo-Frankel (ZFC) e de von Neumann-Bernays-Gödel (NBG). Tendo em vista que a abordagem de checagem de SMT implementada no haRVey requer uma teoria finita e pode ser estendida para as teorias nãodecidíveis, a teoria NBG apresenta-se como uma opção adequada para a expansão da capacidade dedutiva do haRVey à teoria dos conjuntos. Assim, através do mapeamento dos operadores de conjunto fornecidos pela linguagem B a classes da teoria NBG, obtem-se uma abordagem alternativa para a checagem de SMT aplicada ao método B

Mechanical behavior of ceramic veneer in zirconia-based restorations: a 3-dimensional finite element analysis using microcomputed tomography data

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Influence of Implant Inclination Associated With Mandibular Class I Removable Partial Denture

The aim of this study was to use two-dimensional finite element method to evaluate the displacement and stress distribution transmitted by a distal extension removable partial denture (DERPD) associated with an implant placed at different inclinations (0, 5, 15, and 30 degrees) in the second molar region of the edentulous mandible ridge. Six hemimandibular models were created: model A, only with the presence of the natural tooth 33; model B, similar to model A, with the presence of a conventional DERPD replacing the missing teeth; model C, similar to the previous model, with a straight implant (0 degrees) in the distal region of the ridge, under the denture base; model D, similar to model C, with the implant angled at 5 degrees in the mesial direction; model E, similar to model C, with the implant angled at 15 degrees in the mesial direction; and model F, similar to ME, with the implant angled at 30 degrees in the mesial direction. The models were created with the use of the AutoCAD 2000 program (Autodesk, Inc, San Rafael, CA) and processed for finite element analysis by the ANSYS 8.0 program (Swanson Analysis Systems, Houston, PA). The force applied was vertical of 50 N on each cusp tip. The results showed that the introduction of the RPD overloaded the supporting structures of the RPD and that the introduction of the implant helped to relieve the stresses of the mucosa alveolar, cortical bone, and trabecular bone. The best stress distribution occurred in model D with the implant angled at 5 degrees. The use of an implant as a support decreased the displacement of alveolar mucosa for all inclinations simulated. The stress distribution transmitted by the DERPD to the supporting structures was improved by the use of straight or slightly inclined implants. According to the displacement analysis and von Mises stress, it could be expected that straight or slightly inclined implants do not represent biomechanical risks to use.

Evaluation of the influence of location of osseointegrated implants associated with mandibular removable partial dentures

Purpose: The aim of this research was to assess, by means of, the bi-dimensional finite element method, the best implant location in the alveolar edge, through stress distribution and support structure displacement of a distal extension removable partial denture associated with an osseointegrated implant of 10.0 x .75 mm, acting as abutment for the denture base.Methods and Materials: Five models in sagittal cut were used to represent: model A-hemi arch containing natural tooth 33 and the distal alveolar edge; model B-similar to model A, but with a conventional removable partial denture to replace the absent teeth; model C (MC)-similar to the previous one, with an implant in the distal region of the edge under the denture base; model D-similar to MC, with the implant in the central region of the edge; model E-similar to MC, with an implant in the mesial region of the edge. With the aid of the finite element program ANSYS 8.0, the models were loaded with strictly vertical forces of 50 N on each cusp tip. Displacement and von Mises Maps were plotted for visualization of results.Results: The introduction of implant diminished the tendency of intrusion of the removable partial denture in all situations. The maximum stress was observed on implant in all situations. Approximating implant in direction of support teeth was benefit for stress distribution.Conclusion: Model D presented the lowest value for maximum tendency to displacement when compared with those found in the other models; model E demonstrated better relief with regard to demand from the abutment tooth; locating the implant near of the abutment tooth influenced positively the distribution of stresses on the analyzed structures.

Study of stress distribution with finite element method in a root with prefabricated threaded split shaft post - Part I

A finite element analysis was carried out to study the role of prefabricated threaded split shaft post (Flexi-Post) on dentinal stress in pulpless tooth. Three dimensional plane strain model of mesio-distal section of a human maxillary central incisor without restoration was analysed with the MSC/NASTRAN (MacNeal/ Schwendler) general purpose finite analysis program was executed on a microcomputer. The model as discretized into 48.954 axisymmetric finite elements defined by 10.355 nodes. Each element was assigned unique elastic properties to represent the materials modeled. Homogeneity, isotropy and linear elasticity were assume for all material. A simulation of static load of 100N was applied to the incisal edge of the post; vertical. Maximal principal stresses and von Mises equivalent stress were calculated. Using the element analysis model employed in this study, the following can be concluded concerning threaded split shaft post (Flexi-Post): Maximum principal stresses in dentin were located at cervical place and at the post apex. The apical threads of the post not redirecting stresses away from the root.

Partial hyperbolicity for symplectic diffeomorphisms

We prove that every robustly transitive and every stably ergodic symplectic diffeomorphism on a compact manifold admits a dominated splitting. In fact, these diffeomorphisms are partially hyperbolic. (c) 2005 Elsevier SAS. All rights reserved.

Reaction of rat connective tissue to implanted dentin tubes filled with mineral trioxide aggregate or calcium hydroxide

This study was conducted to observe the rat subcutaneous connective tissue reaction to the implanted dentin tubes filled with calcium hydroxide or mineral trioxide aggregate. The animals were sacrificed after 7 and 30 days, and the specimens were prepared for morphological study. Some undecalcified specimens were prepared for histological analysis with polarized light and Von Kossa technique for calcium. The results were similar for both studied materials. At the tube openings, there were Von Kossa-positive granules that were birefringent to polarized light. Next to these granulations, there was an irregular tissue like a bridge that was Von Kossa-positive. The dentin walls of the tubes exhibited in the tubules a structure highly birefringent to polarized light, usually like a layer and at different depths. It is possible that the mechanism of action of both materials has some similarity. Copyright © 1999 by The American Association of Endodontists.

Reaction of rat connective tissue to implanted dentin tube filled with mineral trioxide aggregate, Portland cement or calcium hydroxide.

The subject of this study was to observe the rat subcutaneous connective tissue reaction to implanted dentin tubes filled with mineral trioxide aggregate, Portland cement or calcium hydroxide. The animals were sacrificed after 7 or 30 days and the undecalcified specimens were prepared for histological analysis with polarized light and Von Kossa technique for mineralized tissues. The results were similar for the studied materials. At the tube openings, there were Von Kossa-positive granules that were birefringent to polarized light. Next to these granulations, there was an irregular tissue like a bridge that was Von Kossa-positive. The dentin walls of the tubes exhibited in the tubules a structure highly birefringent to polarized light, usually like a layer and at different depths. The mechanism of action of the studied materials has some similarity.