Use of fluorescence lifetime imaging microscopy (FLIM) as a timer of cell cycle S phase

Incorporation of thymidine analogues in replicating DNA, coupled with antibody and fluorophore staining, allows analysis of cell proliferation, but is currently limited to monolayer cultures, fixed cells and end-point assays. We describe a simple microscopy imaging method for live real-time analysis of cell proliferation, S phase progression over several division cycles, effects of anti-proliferative drugs and other applications. It is based on the prominent (~ 1.7-fold) quenching of fluorescence lifetime of a common cell-permeable nuclear stain, Hoechst 33342 upon the incorporation of 5-bromo-2’-deoxyuridine (BrdU) in genomic DNA and detection by fluorescence lifetime imaging microscopy (FLIM). We show that quantitative and accurate FLIM technique allows high-content, multi-parametric dynamic analyses, far superior to the intensity-based imaging. We demonstrate its uses with monolayer cell cultures, complex 3D tissue models of tumor cell spheroids and intestinal organoids, and in physiological study with metformin treatment.

‘Crows’ vs. ‘Avatar’, or: 3D vs. total-dimensional immersion

3D film’s explicit new space depth arguably provides both an enhanced realistic quality to the image and a wealth of more acute visual and haptic sensations (a ‘montage of attractions’) to the increasingly involved spectator. But David Cronenberg’s related ironic remark that ‘cinema as such is from the outset a «special effect»’ should warn us against the geometrical naiveté of such assumptions, within a Cartesian ocularcentric tradition for long overcome by Merleau-Ponty’s embodiment of perception and Deleuze’s notion of the self-consistency of the artistic sensation and space. Indeed, ‘2D’ traditional cinema already provides the accomplished «fourth wall effect», enclosing the beholder behind his back within a space that no longer belongs to the screen (nor to ‘reality’) as such, and therefore is no longer ‘illusorily’ two-dimensional. This kind of totally absorbing, ‘dream-like’ space, metaphorical for both painting and cinema, is illustrated by the episode ‘Crows’ in Kurosawa’s Dreams. Such a space requires the actual effacement of the empirical status of spectator, screen and film as separate dimensions, and it is precisely the 3D caracteristic unfolding of merely frontal space layers (and film events) out of the screen towards us (and sometimes above the heads of the spectators before us) that reinstalls at the core of the film-viewing phenomenon a regressive struggle with reality and with different degrees of realism, originally overcome by film since the Lumière’s Arrival of a Train at Ciotat seminal demonstration. Through an analysis of crucial aspects in Avatar and the recent Cave of Forgotten Dreams, both dealing with historical and ontological deepening processes of ‘going inside’, we shall try to show how the formal and technically advanced component of those 3D-depth films impairs, on the contrary, their apparent conceptual purpose on the level of contents, and we will assume, drawing on Merleau-Ponty and Deleuze, that this technological mistake is due to a lack of recognition of the nature of perception and sensation in relation to space and human experience.

6-(Methylsulfonyl) hexyl isothiocyanate as potential chemopreventive agent: molecular and cellular profile in leukaemia cell lines

Despite numerous therapeutic interventions cancer is still today the second leading cause of death. A growing interest has been addressed to isothiocytanates and more recently, the 6- (methylsulfonyl) hexyl isothiocyanate (6-MITC), the main constituent of the rhizome of Wasabia Japonica, has stimulated the interest of researchers. Aim of the research was to study if 6-MITC is able to modulate the main mechanisms underlying chemopreventive process in leukemic cells lines, verify the selectivity of action and the safety of use in terms of mutagenicity. The study was conducted on different cell types. In particular, Jurkat and HL-60 cells were treated with increasing concentrations of 6-MITC and cell viability, induction of apoptosis, cell cycle analysis, autophagy modulation and stimulation of differentiation were evaluated by flow cytometry. PBL, the non-transformed counterparty of leukemia cells, was used to analyse the selectivity of action by studying the same mechanisms previously indicated. Finally, safety of use and antimutagenicity were studied in TK6 cells adopting an automated protocol in flow cytometry. The achieved results have demonstrated that isothiocyanate modulates many signaling pathways involved in chemopreventive mechanism. In fact, 6-MITC induces apoptosis of both transformed cells, limits tumor growth by slowing down the cell cycle of Jurkat cells and blocks HL-60 cell cycle, increases the autophagic flux and induces cytodifferentiation of promyelocytic HL-60 into macrophage and granulocytic phenotypes. Furthermore, the results obtained with 6-MITC on PBL from healthy donors suggest that the isothiocyante is a good selective cytotoxic agent. Essential feature of a good chemopreventive agent is selectivity toward cancer cells and low toxicity towards non-transformed cells. Finally, the analysis of the micronuclei revealed that 6-MITC is not mutagenic, ensuring safe use, and that instead, it is able to counteract the mutagenic activity of the aneuploidogen Vinblastine, demonstrating another important and interesting chemopreventive activity.

Implementazione del Sistema Informativo 3D: sviluppo di un lessico codificato per una funzionale e produttiva comparazione dei dati

Il progetto del dottorato di ricerca ha permesso di estendere il lavoro svolto sul Sistema Informativo 3D del Cantiere della Fontana del Nettuno al fine di definire la struttura concettuale e i relativi contenuti tematici di una piattaforma open source in grado di sviluppare la documentazione di restauro, sia per opere complesse caratterizzate dalla presenza di molti materiali costitutivi, sia per interventi più semplici nel quale preservare memoria e fornire libero accesso ai dati. Il confronto tra il SI del Cantiere della Fontana del Nettuno con le attuali metodologie utilizzate in campo nazionale ed internazionale ha permesso di ampliare i lessici necessari per la documentazione grafica e testuale da effettuare su diverse classi di materiali, creando delle cartelle integrate da combinare in base ai materiali costitutivi delle opere da restaurare. Il lavoro ha permesso la redazione di un lessico specifico per i diversi materiali costitutivi fornendo una banca dati informatizzata di facile consultazione.

Mineralogy and Geochemistry of Phosporite Deposits from Tropic Seamount (NE Tropical Atlantic Ocean)

Marine phosphorite deposits can form in a variety of environments, and despite of similar P contents, their mineralogy can change substantially. Seamount phosphorites are increasingly recognized for their P resources and are known to concentrate rare earth elements (REEs) and yttrium (Y) during early diagenetic formation, much more than continental-margin phosphorite deposits. Their importance is increasing in terms of economic potential but have always been studied for paleoenvironmental reconstruction purposes. The Canary Island Seamount Province (CISP) has been extensively studied for its Fe-Mn crusts and nodules deposits, but to date there has not been any systematic study on the phosphorite substrate rocks. This study aims at characterizing the mineralogy and geochemistry of the Tropic seamount phosphorites and offer insights into their mechanisms of formation. The Tropic seamount is a guyot that presents a variety of depositional environments. Two types of phosphorite slabs were identified: (1) a massive facies with oxic enrichments of Mn, Cr, Co, Ni and Cu located on the summit edges and flanks, and (2) a complex facies with suboxic-to-anoxic enrichments of U and V observed on the phosphorites located on the summit of the guyot. Both phosphorite types experienced advanced phosphatization (P2O5 between 24 and 31 wt.%, 3-4 wt.% of F). Differences in uptake of rare earth elements + yttrium (REY) and variations in mineralogy (e.g., presence of foraminifera vs. rounded glauconite grains, carbonate fragments or bioclasts) between the two types, allow phosphorites that formed in upwelling, nutrient-rich and oxic-suboxic environments to be distinguished from those which formed in suboxic-anoxic organic-poor environments. A potential combined ore deposit (Fe-Mn crusts + phosphorites) with high REY contents, like the seamount phosphorites analyzed in this study (ΣREY=870 μg/g on average), could help supply the resources needed for green-tech and high-tech applications.

Própolis do sudeste e nordeste do Brasil: influência da sazonalidade na atividade antibacteriana e composição fenólica

The composition and biological activities of propolis, a resinous hive product collected by honeybees from various plant sources, depends on various factors such as season and vegetation of the area. The aim of this study was to evaluate the influence of the seasonal effect on the ethanolic extracts of Brazilian propolis (EEP) type 6 and type 12, collected during 6 months in terms of antibacterial activity and phenolic composition. The antimicrobial properties were evaluated by MIC and MBC on S. mutans Ingbritt 1600 and the profile of chemical composition by UV-visible spectrophotometry, HPLC-RF and GC-MS. The results demonstrated that the season in which propolis is collected influences its chemical composition, resulting in modifications in its antibacterial activity.

Condutividade e difusividade térmica do figo (Ficus carica L.) Roxo de Valinhos

The post harvest cooling and/or freezing processes for horticultural products have been carried out with the objective of removing the heat from these products, allowing them a bigger period of conservation. Therefore, the knowledge of the physical properties that involve heat transference in the fig fruit Roxo de Valinhos is useful for calculating projects and systems of food engineering in general, as well as, for using in equations of thermodynamic mathematical models. The values of conductivity and thermal diffusivity of the whole fig fruit-rami index were determined, and from these values it was determined the value of the specific heat. For these determination it was used the transient method of the Line Heat Source. The results shown that the fig fruit has a thermal conductivity of 0.52 W m-1°C, thermal diffusivity of 1.56 x 10-7 m² s-1, pulp density of 815.6 kg m-3 and specific heat of 4.07 kJ kg-1 °C.

Influência da temperatura do ar de secagem no calor latente de vaporização de água em feijão macassar (Vigna unguiculata (L.) Walp.), variedade sempre-verde

In order to determine the energy needed to artificially dry an agricultural product the latent heat of vaporization of moisture in the product, H, must be known. Generally, the expressions for H reported in the literature are of the form H = h(T)f(M), where h(T) is the latent heat of vaporization of free water, and f(M) is a function of the equilibrium moisture content, M, which is a simplification. In this article, a more general expression for the latent heat of vaporization, namely H = g(M,T), is used to determine H for cowpea, always-green variety. For this purpose, a computer program was developed which automatically fits about 500 functions, with one or two independent variables, imbedded in its library to experimental data. The program uses nonlinear regression, and classifies the best functions according to the least reduced chi-squared. A set of executed statistical tests shows that the generalized expression for H used in this work produces better results of H for cowpea than other equations found in literature.

História do lazer na natureza no Rio de Janeiro entre 1779 e 1838 : um estudo de caso

Universidade Estadual de Campinas . Faculdade de Educação Física

Analise de volumes parciais do tronco durante a respiração por videogrametria

Universidade Estadual de Campinas . Faculdade de Educação Física

Desvelando os significados da vivencia da ginastica geral para adolescentes de uma instituição salesiana de proteção a criança e ao adolescente

Universidade Estadual de Campinas. Faculdade de Educação Física

Adaptações das curvas da coluna vertebral na marcha em função da altura dos saltos dos calçados

Universidade Estadual de Campinas. Faculdade de Educação Física

Influencia da atividade fisica na imbagem corporal e percepção de dor de pessoas idosas com dores cronicas

Universidade Estadual de Campinas . Faculdade de Educação Física

Proposição, validação e aplicação de um novo metodo para analise cinematica tridimensional da movimentação da caixa toracica durante a respiração

Universidade Estadual de Campinas . Faculdade de Educação Física

Analise da disciplina de educação fisica especial nas Institutições de Ensino Superior publicas do estado do Parana

Universidade Estadual de Campinas . Faculdade de Educação Física