Caracterização morfológica e funcional de células aderentes de baço de camundongo

A separação, caracterização e ensaio funcional das células inflamatórias presentes no local de lesão têm se tornado imperiosos no estudo de diversas doenças. Através da utilização de métodos histoquímicos para esterase e fosfatase ácida, bem como do Teste de Fagocitose e da coloração pelo Giemsa, realizados nas células esplénicas de dez camundongos, foi possível se caracterizar bem os componentes do Sistema Fagocítico Mononuclear e distinguir os outros tipos de células presentes, além de permitir a quantificação diferencial das mesmas.

Três novas espécies de tripanosomatídeos de insetos isolados em Alfenas, Minas Gerais, Brasil

Três novas espécies de tripanosomatídeos foram isoladas em Alfenas, MG, Brasil: Herpetomonas anglusteri sp. n., do intestino posterior de Liopygia ruficorins (Diptera: Sarcophagidae); Crithidia roitmani sp. e.e Crithidia de souzai sp. n., do intestino médio e o posterior de Ornidia obesa (Diptera: Syrphidae). O isolamento foi feito em meio complexo de roitmanmas os três isolados cresceram bem no meio definido do mesmo Autor. Os clones foram obtidos em ágar-sangue de carneiro, desfibrinado, em placas de Petri, a 28ºC, por 2-7 dias. Um único clone de cada espécie foi utilizado neste trabalho. Dados morfológicos e morfométricos foram obtidos em câmara clara após coloração dos flagelados. H. anglusteri cresceu em meio complexo tanto a 28 como a 37ºC e, em meio definido, apenas a 28ºC. Não exige treonina e biotina para seu crescimento. C. roitmani apresenta tamanho médio maior que C. desouzai, não cresce em água de coco e seu crescimento é mais lento comparativamente a C. desouzai, apesar de terem sido isoladas critídias exige hemina e adenina para seu crescimento. Alguns ácidos aminados e vitaminas componentes do meio definido utilizado no ensaio, também não são exigidos, o que sugere serem estes tripanosomatídeos portadores de endossimbiontes.

Criptosporidiose em crianças de 1 a 2 anos de idade, com diarréia aguda em Belém, Pará, Brasil

Num período de 12 meses, 201 espécimes fecais de 61 crianças foram examiandos com vista a detecção de Cryptosporidium. Cento e quinze espécimes foram obtidos durante os episódios diarréicos e 86 de crianças sem diarréia (grupo controle). Todos os espécimes fecais foram examinados pelo método de coloração de Ziehl-Neelsen modificado. Cryptosporidum foi identificado em seis (5,2%) das 115 amostras das crianças com diarréia. Em nenhum dos controles foi obtida a presença desse parasito. O presente estudo sugere que o Cryptosporidium é um agente causal de diarréia auto-limitada em crianças imunocompetentes em Belém, Pará.

Interpretación y traducción de texto y matemáticas en Braille escrito a máquina

El trabajo expuesto en la presente memoria, forma parte de un proyecto de colaboración entre el Centro de Visión por Computador de la UAB y el Centro Joan Amades (ONCE), cuyo objetivo es la creación de recursos educativos que faciliten la integración de niños invidentes en las aulas. Se presenta el proceso de implementación de un intérprete y traductor de documentos escritos en Braille con contenido matemático y de texto, que permite a un profesor que no conozca el sistema Braille, la lectura de documentos creados por alumnos invidentes. Dicho intérprete forma parte de una herramienta que permite el reconocimiento de documentos escritos con una máquina Perkins.

ICR a partir de traços d'escriptura natural captats amb un bolígraf digital

Aquest projecte consisteix en la realització d’una anàlisi de diferents reconeixedors de caracters manuscrits, concretament de nombres, per a una possible implantació en la digitalització de formularis en la industria. Al llarg del document s’estudien dos reconeixedors diferents, concretament l’incorporat al paquet "Tablet PC and Recognition Pack" de Microsoft i el Heloise Hse, proporcionat per la Universitat de Berkeley a Califòrnia.

Anàlisi i resolució de puzzles

Aquest projecte és una anàlisi d'algunes tècniques de reconeixement d'objectes en 2D, la seva comparació i equivalència. La solució presentada en aquesta memòria ha estat implementada en C++ per a poder avaluar els resultats així com les diferents problemàtiques aparegudes durant tot el procés. La resolució de puzzles és tan sols un exemple dels múltiples casos d'objectes bidimensionals que ens podem trobar que necessiten tècniques de matching per obtenir un objecte global.

Extracció del mapa de profunditat a partir d'una seqüència d'imatges

Aquest projecte resol les fases inicials d'un altre projecte més gran que té com a objectiu la conversió automàtica de seqüències d'imatges a 3D. El projecte s'ha centrat en la reconstrucció calibrada de col·leccions d'imatges mitjançant la tècnica anomenada structure from motion. Aquesta tècnica forma part de l'àmbit de la visió per computador i s'utilitza per obtenir la posició i l'orientació de les diferents càmeres juntament amb una reconstrucció 3D de l'escena en forma de núvol de punts.

Las Bodegas españolas y su adaptación a los cambios del mercado a finales del siglo XX: análisis lingüístico de la publicidad

El sector vitivinícola español ha experimentado una serie de transformaciones en los últimos años, especialmente derivada de la caída del consumo interno y la creciente competencia en los mercados. Esto ha provocado que las bodegas españolas tuvieran que adaptarse a las nuevas circunstancias mediante la aplicación de modernas políticas de marketing. El objetivo principal del estudio es conocer el grado de aplicación de éstas, mediante la comparación y el análisis lingüístico de distintos tipos de incursiones publicitarias en medios impresos especializados, tratando aspectos como la frecuencia específica, la variación léxica, las categorías semánticas y las familias de palabras más utilizadas. La principal conclusión es que las bodegas españolas emprendieron un proceso de adaptación tardía, pasando de mensajes sencillos, sin imagen y relacionados con el sistema productivo, a otros mucho más elaborados, con imagen y nuevos términos que pretenden transmitir que estamos ante un producto de calidad.

Use of an immunodominant p17 antigenic fraction of Neospora caninum in detection of antibody response in cattle

A Neospora caninum 17 kDa protein fraction (p17) has been described as an immunodominant antigen (IDA) under reducing and non-reducing conditions. The aim of the present study was to investigate the diagnostic utility of p17 in cattle. In order to achieve this, p17 was purified by electroelution from whole N. caninum tachyzoite soluble extract and a p17-based Western blot (WB-p17) was developed. The p17 recognition was measured by densitometry and expressed as OD values to check the validity of the WB-p17. A total of 131 sera including sequential samples from naturally- and experimentally-infected calves and breeding cattle were analysed by WB-p17 and compared with IFAT using whole formalin-fixed tachyzoites as a reference test. The results obtained highlight the feasibility of using the N. caninum p17 in a diagnostic test in cattle. Firstly, the assay based on the p-17 antigen discriminated between known positive and negative sera from different cattle populations, breeding cattle and calves. Secondly, the p17 antigen detected fluctuations in the antibody levels and seroconversion in naturally- and experimentally-infected cattle. Significant differences in p-17 antigen recognition were observed between naturally infected aborting and non-aborting cattle, as well as significant antibody fluctuations over time in experimentally infected cattle, which varied between groups. Furthermore, the results obtained with WB-p17 are in accordance with the results obtained with the IFAT, as high agreement values were obtained when all bovine subpopulations were included (kappa = 0.86).

Functionalized nanostructures with application in regenerative medicine

In the last decade, both regenerative medicine and nanotechnology have been broadly developed leading important advances in biomedical research as well as in clinical practice. The manipulation on the molecular level and the use of several functionalized nanoscaled materials has application in various fields of regenerative medicine including tissue engineering, cell therapy, diagnosis and drug and gene delivery. The themes covered in this review include nanoparticle systems for tracking transplanted stem cells, self-assembling peptides, nanoparticles for gene delivery into stem cells and biomimetic scaffolds useful for 2D and 3D tissue cell cultures, transplantation and clinical application.

Cannabinoids reduce ErbB2-driven breast cancer progression through Akt inhibition.

BACKGROUND ErbB2-positive breast cancer is characterized by highly aggressive phenotypes and reduced responsiveness to standard therapies. Although specific ErbB2-targeted therapies have been designed, only a small percentage of patients respond to these treatments and most of them eventually relapse. The existence of this population of particularly aggressive and non-responding or relapsing patients urges the search for novel therapies. The purpose of this study was to determine whether cannabinoids might constitute a new therapeutic tool for the treatment of ErbB2-positive breast tumors. We analyzed their antitumor potential in a well established and clinically relevant model of ErbB2-driven metastatic breast cancer: the MMTV-neu mouse. We also analyzed the expression of cannabinoid targets in a series of 87 human breast tumors. RESULTS Our results show that both Delta9-tetrahydrocannabinol, the most abundant and potent cannabinoid in marijuana, and JWH-133, a non-psychotropic CB2 receptor-selective agonist, reduce tumor growth, tumor number, and the amount/severity of lung metastases in MMTV-neu mice. Histological analyses of the tumors revealed that cannabinoids inhibit cancer cell proliferation, induce cancer cell apoptosis, and impair tumor angiogenesis. Cannabinoid antitumoral action relies, at least partially, on the inhibition of the pro-tumorigenic Akt pathway. We also found that 91% of ErbB2-positive tumors express the non-psychotropic cannabinoid receptor CB2. CONCLUSIONS Taken together, these results provide a strong preclinical evidence for the use of cannabinoid-based therapies for the management of ErbB2-positive breast cancer.

Functional characterization of E- and P-cadherin in invasive breast cancer cells

BACKGROUND Alterations in the cadherin-catenin adhesion complexes are involved in tumor initiation, progression and metastasis. However, the functional implication of distinct cadherin types in breast cancer biology is still poorly understood. METHODS To compare the functional role of E-cadherin and P-cadherin in invasive breast cancer, we stably transfected these molecules into the MDA-MB-231 cell line, and investigated their effects on motility, invasion and gene expression regulation. RESULTS Expression of either E- and P-cadherin significantly increased cell aggregation and induced a switch from fibroblastic to epithelial morphology. Although expression of these cadherins did not completely reverse the mesenchymal phenotype of MDA-MB-231 cells, both E- and P-cadherin decreased fibroblast-like migration and invasion through extracellular matrix in a similar way. Moreover, microarray gene expression analysis of MDA-MB-231 cells after expression of E- and P-cadherins revealed that these molecules can activate signaling pathways leading to significant changes in gene expression. Although the expression patterns induced by E- and P-cadherin showed more similarities than differences, 40 genes were differentially modified by the expression of either cadherin type. CONCLUSION E- and P-cadherin have similar functional consequences on the phenotype and invasive behavior of MDA-MB-231 cells. Moreover, we demonstrate for the first time that these cadherins can induce both common and specific gene expression programs on invasive breast cancer cells. Importantly, these identified genes are potential targets for future studies on the functional consequences of altered cadherin expression in human breast cancer.

Breast cancer 1 (BrCa1) may be behind decreased lipogenesis in adipose tissue from obese subjects.

CONTEXT Expression and activity of the main lipogenic enzymes is paradoxically decreased in obesity, but the mechanisms behind these findings are poorly known. Breast Cancer 1 (BrCa1) interacts with acetyl-CoA carboxylase (ACC) reducing the rate of fatty acid biosynthesis. In this study, we aimed to evaluate BrCa1 in human adipose tissue according to obesity and insulin resistance, and in vitro cultured adipocytes. RESEARCH DESIGN AND METHODS BrCa1 gene expression, total and phosphorylated (P-) BrCa1, and ACC were analyzed in adipose tissue samples obtained from a total sample of 133 subjects. BrCa1 expression was also evaluated during in vitro differentiation of human adipocytes and 3T3-L1 cells. RESULTS BrCa1 gene expression was significantly up-regulated in both omental (OM; 1.36-fold, p = 0.002) and subcutaneous (SC; 1.49-fold, p = 0.001) adipose tissue from obese subjects. In parallel with increased BrCa1 mRNA, P-ACC was also up-regulated in SC (p = 0.007) as well as in OM (p = 0.010) fat from obese subjects. Consistent with its role limiting fatty acid biosynthesis, both BrCa1 mRNA (3.5-fold, p<0.0001) and protein (1.2-fold, p = 0.001) were increased in pre-adipocytes, and decreased during in vitro adipogenesis, while P-ACC decreased during differentiation of human adipocytes (p = 0.005) allowing lipid biosynthesis. Interestingly, BrCa1 gene expression in mature adipocytes was restored by inflammatory stimuli (macrophage conditioned medium), whereas lipogenic genes significantly decreased. CONCLUSIONS The specular findings of BrCa1 and lipogenic enzymes in adipose tissue and adipocytes reported here suggest that BrCa1 might help to control fatty acid biosynthesis in adipocytes and adipose tissue from obese subjects.

Executive functions profile in extreme eating/weight conditions: from anorexia nervosa to obesity.

BACKGROUND Extreme weight conditions (EWC) groups along a continuum may share some biological risk factors and intermediate neurocognitive phenotypes. A core cognitive trait in EWC appears to be executive dysfunction, with a focus on decision making, response inhibition and cognitive flexibility. Differences between individuals in these areas are likely to contribute to the differences in vulnerability to EWC. The aim of the study was to investigate whether there is a common pattern of executive dysfunction in EWC while comparing anorexia nervosa patients (AN), obese subjects (OB) and healthy eating/weight controls (HC). METHODS Thirty five AN patients, fifty two OB and one hundred thirty seven HC were compared using the Wisconsin Card Sorting Test (WCST); Stroop Color and Word Test (SCWT); and Iowa Gambling Task (IGT). All participants were female, aged between 18 and 60 years. RESULTS There was a significant difference in IGT score (F(1.79); p<.001), with AN and OB groups showing the poorest performance compared to HC. On the WCST, AN and OB made significantly more errors than controls (F(25.73); p<.001), and had significantly fewer correct responses (F(2.71); p<.001). Post hoc analysis revealed that the two clinical groups were not significantly different from each other. Finally, OB showed a significant reduced performance in the inhibition response measured with the Stroop test (F(5.11); p<.001) compared with both AN and HC. CONCLUSIONS These findings suggest that EWC subjects (namely AN and OB) have similar dysfunctional executive profile that may play a role in the development and maintenance of such disorders.

"The 3/3 strategy": a successful multifaceted hospital wide hand hygiene intervention based on WHO and continuous quality improvement methodology.

BACKGROUND Only multifaceted hospital wide interventions have been successful in achieving sustained improvements in hand hygiene (HH) compliance. METHODOLOGY/PRINCIPAL FINDINGS Pre-post intervention study of HH performance at baseline (October 2007-December 2009) and during intervention, which included two phases. Phase 1 (2010) included multimodal WHO approach. Phase 2 (2011) added Continuous Quality Improvement (CQI) tools and was based on: a) Increase of alcohol hand rub (AHR) solution placement (from 0.57 dispensers/bed to 1.56); b) Increase in frequency of audits (three days every three weeks: "3/3 strategy"); c) Implementation of a standardized register form of HH corrective actions; d) Statistical Process Control (SPC) as time series analysis methodology through appropriate control charts. During the intervention period we performed 819 scheduled direct observation audits which provided data from 11,714 HH opportunities. The most remarkable findings were: a) significant improvements in HH compliance with respect to baseline (25% mean increase); b) sustained high level (82%) of HH compliance during intervention; c) significant increase in AHRs consumption over time; c) significant decrease in the rate of healthcare-acquired MRSA; d) small but significant improvements in HH compliance when comparing phase 2 to phase 1 [79.5% (95% CI: 78.2-80.7) vs 84.6% (95% CI:83.8-85.4), p<0.05]; e) successful use of control charts to identify significant negative and positive deviations (special causes) related to the HH compliance process over time ("positive": 90.1% as highest HH compliance coinciding with the "World hygiene day"; and "negative":73.7% as lowest HH compliance coinciding with a statutory lay-off proceeding). CONCLUSIONS/SIGNIFICANCE CQI tools may be a key addition to WHO strategy to maintain a good HH performance over time. In addition, SPC has shown to be a powerful methodology to detect special causes in HH performance (positive and negative) and to help establishing adequate feedback to healthcare workers.