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Orientador: Helion Vargas Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Fisica Gleb Wataghin Resumo: Não informado Abstract: Not informed Mestrado Física Mestre em Física

Agradecimentos: The equipment used for the development of this work was purchased with the help of Fapesp (the São Paulo Research Foundation; 08/57906-3) and CNPq (the National Council for Scientific and Technological Development; 573913/2008-0). We reviewed the content of the manuscript, followed by Ms Diane Ellis, B.A. in education. Biostatistician Cleide Aparecida Moreira Silva, Research Committee, Biostatistics Division, Medical Sciences School, Unicamp, provided statistical consultation Abstract: Solid tumors typically contain high levels of fibrillar collagen. The increased stromal collagen deposition usually promotes cancer progression since biochemical and biophysical cues from tumor-associated collagen fibers stimulate neoplastic cells. Few studies have investigated the relationship between Merkel cell carcinoma (MCC) and the extracellular matrix (ECM), but there are no works evaluating collagen. This is an observational, analytical, retrospective study including 11 patients with MCC. Primary tumor-stained sections were evaluated by second harmonic generation microscopy and texture analysis. Peritumoral texture features (area fraction, mean gray value, entropy, and contrast) showed much lower values than normal skin (P < .0001) revealing extensively altered structure of peritumoral collagen fibers. These differences were not significant between tumors with unfavorable and favorable known prognostic factors. Profound changes in collagen fibers present in the stroma accompanying primary MCC may contribute to the aggressive behavior of this tumor. Our results indicate that whatever MCC histological subtype, size or anatomical location, MCC promotes the same type of ECM for its development. As an outlook, therapies using ECM macromolecules or fibroblasts (the architects of ECM remodeling) as target could be useful in the treatment of MCC FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQ Aberto

Agradecimentos: AF acknowledges the National Institute of Photonics (INFo). CC acknowledges National Institute of Science and Technology in Photonics Applied to Cell Biology [INFABIC] CNPq (573913/2008-00/FAPESP (08/57906-3); CNPq grant (312049/2014-5); "Física do Petróleo em Meios Porosos", PETROBRAS-UFC (2016/00328-4) and F020/WIPPS II—Simulação numérica de invasão de água em poços produtores", PETROGAL-UFC T.G.E. acknowledges the support from the Winton Program for the Physics of Sustainability, the Isaac Newton Trust, and the Leverhulme Trust (RPG-2018-256) Abstract: We report analytical expressions for optical forces acting on particles inside waveguides. The analysis builds on our previously reported Fourier Transform method to obtain Beam Shape Coefficients for any beam. Here we develop analytical expressions for the Beam Shape Coefficients in cylindrical and rectangular metallic waveguides. The theory is valid for particle radius a ranging from the Rayleigh regime to large microparticles, such as aerosols like virus loaded droplets. The theory is used to investigate how optical forces within hollow waveguides can be used to sort particles in "optical chromatography" experiments in which particles are optically propelled along a hollow-core waveguide. For Rayleigh particles, the axial force is found to scale with a (6), while the radial force, which prevents particles from crashing into the waveguide walls, scales with a (3). For microparticles, narrow Mie resonances create a strong wavelength dependence of the optical force, enabling more selective sorting. Several beam parameters, such as power, wavelength, polarization state and waveguide modes can be tuned to optimize the sorting performance. The analysis focuses on cylindrical waveguides, where meter-long liquid waveguides in the form of hollow-core photonic crystal fibers are readily available. The modes of such fibers are well-approximated by the cylindrical waveguide modes considered in the theory CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQ FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP Aberto

Agradecimentos: The authors are thankful to the National Council for Scientific and Technological Development (CNPq), the Coordination for the Improvement of Higher Education Personnel (CAPES) and the Foundation for Science and Technology of Pernambuco (FACEPE). This work is also linked to the National Institute of Photonics (INFo) and National Institute of Science and Technology in Photonics Applied to Biology (INFABIC). C. L. Cesar is thankful to Física do Petróleo em Meios Porosos", PETROBRAS-UFC 2016/00328-4 and WIPPS II – Simulação numérica de invasão de água em poços produtores", PETROGAL-UFC Abstract: Biomechanical and electrical properties are important to the performance and survival of red blood cells (RBCs) in the microcirculation. This study proposed and explored methodologies based on optical tweezers and cationic quantum dots (QDs) as biophotonic tools to characterize, in a complementary way, viscoelastic properties and membrane electrical charges of RBCs. The methodologies were applied to normal (HbA) and ß-thalassemia intermedia (Hbß) RBCs. The ß-thalassemia intermedia disease is a hereditary hemoglobinopathy characterized by a reduction (or absence) of ß-globin chains, which leads to a-globin chains precipitation. The apparent elasticity (µ) and membrane viscosity (?m) of RBCs captured by optical tweezers were obtained in just a single experiment. Besides, the membrane electrical charges were evaluated by flow cytometry, exploring electrostatic interactions between cationic QDs, stabilized with cysteamine, with the negatively charged RBC surfaces. Results showed that Hbß RBCs are less elastic, have a higher ?m, and presented a reduction in membrane electrical charges, when compared to HbA RBCs. Moreover, the methodologies based on optical tweezers and QDs, here proposed, showed to be capable of providing a deeper and integrated comprehension on RBC rheological and electrical changes, resulting from diverse biological conditions, such as the ß-thalassemia intermedia hemoglobinopathy CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQ COORDENAÇÃO DE APERFEIÇOAMENTO DE PESSOAL DE NÍVEL SUPERIOR - CAPES FUNDAÇÃO DE AMPARO À CIÊNCIA E TECNOLOGIA DO ESTADO DE PERNAMBUCO - FACEPE FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP Fechado

Agradecimentos: We gratefully acknowledge the Brazilian agencies CNPq, CAPES and FUNCAP, and the National Institute of Science and Technology for Complex Systems in Brazil for financial support. L.M acknowledges David Quispe Aruquipa for the help provided with the data mining processing and Biviana Orbegozo for her valuable comments that improved the manuscript. C.L.C acknowledges CNPq grant (312049/2014-5); FAPESP (11/51959-0); "Física do Petróleo em Meios Porosos", PETROBRAS-UFC 2016/00328-4 and WIPPS II—Simulação numérica de invasão de água em poços produtores", PETROGAL-UFC Abstract: The follow up of Representative behavior after elections is imperative for a democratic Representative system, at the very least to punish betrayal with no re-election. Our goal was to show how to follow Representatives' and how to show behavior in real situations and observe trends in political crises including the onset of game changing political instabilities. We used correlation and correlation distance matrices of Brazilian Representative votes during four presidential terms. Re-ordering these matrices with Minimal Spanning Trees displays the dynamical formation of clusters for the sixteen year period, which includes one Presidential impeachment. The reordered matrices, colored by correlation strength and by the parties clearly show the origin of observed clusters and their evolution over time. When large clusters provide government support cluster breaks, political instability arises, which could lead to an impeachment, a trend we observed three years before the Brazilian President was impeached. We believe this method could be applied to foresee other political storms CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQ FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP COORDENAÇÃO DE APERFEIÇOAMENTO DE PESSOAL DE NÍVEL SUPERIOR - CAPES FUNDAÇÃO CEARENSE DE APOIO AO DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - FUNCAP Aberto

Agradecimentos: The authors thank the National Institute of Science and Technology on Photonics Applied to Cell Biology (INFABiC) at the State University of Campinas for access to equipment and assistance; INFABiC is co-funded by Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) (08/57906-3) and Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) (573913/2008-0). C. L. Cesar also acknowledges the grant by FAPESP (11/51959-0) and CNPq (312049/2014-5) Abstract: One of the promising tools to evaluate collagen in the extracellular matrix is the second-harmonic generation microscopy (SHG). This approach may shed light on the biological behavior of cancers and their taxonomy, but has not yet been applied to characterize collagen fibers in cases diagnosed as invasive breast carcinoma (BC) of histological special types (IBC-ST). Tissue sections from 99 patients with IBC-ST and 21 of invasive breast carcinoma of no special type (IBC-NST) were submitted to evaluation of collagen parameters by SHG. Tissue microarray was performed to evaluate immunohistochemical-based molecular subtype. In intratumoral areas, fSHG and bSHG (forward-SHG and backward-SHG) collagen parameters achieved their lowest values in mucinous, papillary and medullary carcinomas, whereas the highest values were found in classic invasive lobular and tubular carcinomas. Unsupervised hierarchical cluster analysis and minimal spanning tree using intratumoral collagen parameters allowed the identification of three main groups of breast cancer: group A (classic invasive lobular and tubular carcinomas); group B (IBC-NST, metaplastic, invasive apocrine and micropapillary carcinomas); and group C (medullary, mucinous and papillary carcinomas). Our findings provide further characterization of the tumor microenvironment of IBC-ST. This understanding may add information to build more consistent tumor categorization and to refine prognostication CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQ FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP Aberto

Agradecimentos: The authors are members of Centros de Pesquisa, Inovação e Difusão Redoxoma-Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Grant 2013/07937-8 and received further support from Fundação Zerbini. L. Y. Tanaka, T. L. S. Araujo, and A. I. Rodriguez were supported by FAPESP Scholarship Grants 2013/17115-5, 2015/06210-2, and 2013/06241-0. M. C. C. Morais was supported by Programa de Pós-Graduação em Oncologia, Instituto do Câncer do Estado de São Paulo, by a Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) scholarship grant, Programa Nacional de Pós-Doutoramento. A. F. Ramos is thankful to CAPES for financial support Abstract: Although redox processes closely interplay with mechanoresponses to control vascular remodeling, redox pathways coupling mechanostimulation to cellular cytoskeletal organization remain unclear. The peri/epicellular pool of protein disulfide isomerase-A1 (pecPDIA1) supports postinjury vessel remodeling. Using distinct models, we investigated whether pecPDIA1 could work as a redox-dependent organizer of cytoskeletal mechanoresponses. In vascular smooth muscle cells (VSMCs), pecPDIA1 immunoneutralization impaired stress fiber assembly in response to equibiaxial stretch and, under uniaxial stretch, significantly perturbed cell repositioning perpendicularly to stretch orientation. During cyclic stretch, pecPDIA1 supported thiol oxidation of the known mechanosensor beta(1)-integrin and promoted polarized compartmentalization of suifenylated proteins. Using traction force microscopy, we showed that pecPDIA1 organizes intracellular force distribution. The net contractile moment ratio of platelet-derived growth factor-exposed to basal VSMCs decreased from 0.90 +/- 0.09 (IgG-exposed controls) to 0.70 +/- 0.08 after pecPDIA1 neutralization (P < 0.05), together with an enhanced coefficient of variation for distribution of force modules, suggesting increased noise. Moreover, in a single cell model, pecPDIA1 neutralization impaired migration persistence without affecting total distance or velocity, whereas siRNA-mediated total PDIA1 silencing disabled all such variables of VSMC migration. Neither expression nor total activity of the master mechanotransmitter/regulator RhoA was affected by pecPDIA1 neutralization. However, cyclic stretch-induced focal distribution of membrane-bound RhoA was disrupted by pecPDI inhibition, which promoted a nonpolarized pattern of RhoA/caveolin-3 cluster colocalization. Accordingly, FRET biosensors showed that pecPDIA1 supports localized RhoA activity at cell protrusions versus perinuclear regions. Thus. pecPDI acts as a thiol redox-dependent organizer and noise reducer mechanism of cytoskeletal repositioning, oxidant generation, and localized RhoA activation during a variety of VSMC mechanoresponses. NEW & NOTEWORTHY Effects of a peri/epicellular pool of protein disulfide isomerase-A1 (pecPDIA1) during mechanoregulation in vascular smooth muscle cells (VSMCs) were highlighted using approaches such as equibiaxial and uniaxial stretch, random single cell migration, and traction force microscopy. pecPDIA1 regulates organization of the cytoskeleton and minimizes the noise of cell alignment, migration directionality, and persistence. pecPDIA1 mechanisms involve redox control of beta(1)-integrin and localized RhoA activation. pecPDIA1 acts as a novel organizer of mechanoadaptation responses in VSMCs COORDENAÇÃO DE APERFEIÇOAMENTO DE PESSOAL DE NÍVEL SUPERIOR - CAPES FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP Fechado

Agradecimentos: Authors are grateful to FAPESP (projects 2014/03691-7 and 2014/25979-2), Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq—Brasil), and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES—Brasil) for financial support Abstract: Reduced graphene oxide (rGO) layers are known to be significantly conductive along the basal plane throughout delocalized sp(2) domains Defects present in rGO implies in disordered systems with numerous localized sites, resulting in a charge transport governed mainly by a 2D variable range hopping (VRH) mechanism These characteristics are observed even in multilayered rGO since the through-plane conduction is expected to be insubstantial. Here, we report on the multilayer assembly of functionalized rGO quantum dots (GQDs) presenting 3D VRH transport that endows elevated charge carrier mobility, ca similar to 236 cm(2) V(-1 )s(-1). Polyelectrolyte-wrapped GQDs were assembled by layer-by-layer technique (LbL), ensuring molecular level thickness control for the formed nanostructures, along with the adjustment of the film transparency (up to 92% in the visible region). The small size and the random distribution of GQDs in the LbL structure are believed to overcome the translational disorder in multilayered films, contributing to a 3D interlayer conduction that enhances the electronic properties. Such high-mobility, transparency-tunable films assembled by a cost-effective method possess interesting features and wide applicability in optoelectronics FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP COORDENAÇÃO DE APERFEIÇOAMENTO DE PESSOAL DE NÍVEL SUPERIOR - CAPES CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQ Fechado

Agradecimentos: We thank Dr. J. Xavier Neto for providing the DS-red mice and Dr.Mario J. Saad for the iNOS KO mice. This study was funded byFAPESP (grant 2009/016150-6) and CNPq. INFABiC is cofunded byFAPESP and CNPq Abstract: Prostate development and function are regulated by androgens. Epithelial cell apoptosis in response to androgen deprivation is caspase-9-dependent and peaks at Day 3 after castration. However, isolated epithelial cells survive in the absence of androgens. Znf142 showed an on-off expression pattern in intraepithelial CD68-positive macrophages, with the on-phase at Day 3 after castration. Rats treated with gadolinium chloride to deplete macrophages showed a significant drop in apoptosis, suggesting a causal relationship between macrophages and epithelial cell apoptosis. Intraepithelial M1-polarization was also limited to Day 3, and the inducible nitric oxide synthase (iNOS) knockout mice showed significantly less apoptosis than wild-type controls. The epithelial cells showed focal DNA double-strand breaks (DSB), 8-oxoguanine, and protein tyrosine-nitrosylation, fingerprints of exposure to peroxinitrite. Cultured epithelial cells induced M1-polarization and showed focal DSB and underwent apoptosis. The same phenomena were reproduced in LNCaP cells cocultured with Raw 264.7 macrophages. In conclusion, the M1 (142)-macrophage (named after Znf142) attack causes activation of the intrinsic apoptosis pathway in epithelial cells after castration FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQ Fechado

Agradecimentos: U.N. was recipient of a TWAS/CNPq fellowship. This work was funded by a grant from FAPESP (Grant 2009/16150-6) Abstract: The high incidence of prostatic diseases, including malignant tumors, makes the understanding of prostate biology very important. Androgen deprivation, blockade by orchiectomy, or chemical castration causes prostate and tumor shrinkage. The gene networks involved in a cell type-specific fashion are rather unknown. This work was undertaken to identify genes with annotated function in transcription regulation that might define transitions in gene expression. A total of 15 potential regulatory genes were identified. Validation by qRT-PCR showed that Zfp703 and Arid1a exhibit expression maxima at day 1; Ash2l, Nelf, Pbx3, Eya2 at day 4; Dmrt2 at day 5 and Lbh and Sox1 at day 7 after castration. Using immunohistochemistry, we further determined that PBX3 was found in both stromal and epithelial cells, whereas ARID1A and NELF were restricted to the epithelium, and DMRT2 and EYA2 were exclusively found in the stroma. Though the proteins ZFP703 and ASH2l were not found in any experimental condition, their mRNAs were located by in situ hybridization in both epithelium and stroma. In conclusion, androgen deprivation triggers the expression of temporally regulated gene sets in both epithelial and stromal cells. These gene subsets will help establish the regulatory gene expression programs orchestrating the castration-induced remodeling of the prostate gland, and represent putative targets to increase the efficacy of androgen-deprivation to induce epithelial (and cancer) cell death CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQ FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP Fechado

Agradecimentos: The authors are thankful to Dr. Iscia Lopes-Cendes for providing facilities in her laboratory. The authors are grateful to FAPESP (Grants 2009/16150-6 and 2017/04377-2), CNPq and CAPES for the funding of this research. INFABiC is co-funded by FAPESP (Grant 2014/50938-8) and CNPq (Grant 465699/2014-6) Abstract: Environmental and nutritional factors, including fatty acids (FA), are associated with prostatitis, benign prostate hyperplasia and prostate cancer. We hypothesized that different FA in normolipidic diets (7%) affect prostate physiology, increasing the susceptibility to prostate disorders. Thus, we fed male C57/BL6 mice with normolipidic diets based on linseed oil, soybean oil or lard (varying saturated and unsaturated FA contents and omega-3/omega-6 ratios) for 12 or 32 weeks after weaning and examined structural and functional parameters of the ventral prostate (VP) in the systemic metabolic context. Mongolian gerbils were included because they present a metabolic detour for low water consumption (i.e., oxidize FA to produce metabolic water). A linseed oil-based diet (LO, 67.4% PUFAs, omega-3/omega-6 = 3.70) resulted in a thermogenic profile, while a soybean oil-based diet (SO, 52.7% PUFAs, omega-3/omega-6 = 0.11) increased body growth and adiposity. Mice fed lard (PF, 13.1% PUFA, omega-3/omega-6 = 0.07) depicted a biphasic growth, resulting in decreased adiposity in adulthood. SO and PF resulted in hepatic steatosis and steatohepatitis, respectively. PF and SO increased prostate epithelial volume, and lard resulted in epithelial hyperplasia. Animals in the LO group had smaller prostates with predominant atrophic epithelia and inflammatory loci. Inflammatory cells were frequent in the VP of PF mice (predominantly stromal) and LO mice (predominantly luminal). RNAseq after 12 weeks revealed good predictors of a later-onset inflammation. The transcriptome unveiled ontologies related to ER stress after 32 weeks on PF diets. In conclusion, different FA qualities result in different metabolic phenotypes and differentially impact prostate size, epithelial volume, inflammation and gene expression FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQ COORDENAÇÃO DE APERFEIÇOAMENTO DE PESSOAL DE NÍVEL SUPERIOR - CAPES Aberto

Agradecimentos: The authors acknowledge the Brazilian agencies: Coordenação de Pessoal de Nível Superior (CAPES), Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), and Fundação de Amparo a Ciência e a Tecnologia do Estado de Pernambuco (FACEPE), and Instituto Serrapilheira. This work is also linked to the National Institute of Photonics (INCT-INFo), the National Institute of Photonics Applied to Cell Biology (INCT-INFABIC), and LARnano/UFPE. The authors thank Mariana O. Baratti from INFABIC/UNICAMP/SP for confocal microscopy images and Professor Wilson Barros from the Physics Department of the Federal University of Pernambuco for both MR images acquisition and data treatments. CFGCG also acknowledges the portuguese agency FCT (Fundação para a Ciência e a Tecnologia) Abstract: Magnetic resonance imaging (MRI) is a powerful non-invasive diagnostic tool that enables distinguishing healthy from pathological tissues, with high anatomical detail. Nevertheless, MRI is quite limited in the investigation of molecular/cellular biochemical events, which can be reached by fluorescence-based techniques. Thus, we developed bimodal nanosystems consisting in hydrophilic quantum dots (QDs) directly conjugated to Gd(III)-DO3A monoamide chelates, a Gd(III)-DOTA derivative, allowing for the combination of the advantages of both MRI and fluorescence-based tools. These nanoparticulate systems can also improve MRI contrast, by increasing the local concentration of paramagnetic chelates. Transmetallation assays, optical characterization, and relaxometric analyses, showed that the developed bimodal nanoprobes have great chemical stability, bright fluorescence, and high relaxivities. Moreover, fluorescence correlation spectroscopy (FCS) analysis allowed us to distinguish nanosystems containing different amounts of chelates/QD. Also, inductively coupled plasma optical emission spectrometry (ICP - OES) indicated a conjugation yield higher than 75%. Our nanosystems showed effective longitudinal relaxivities per QD and per paramagnetic ion, at least 5 times [per Gd(III)] and 100 times (per QD) higher than the r(1) for Gd(III)-DOTA chelates, suitable for T-1-weighted imaging. Additionally, the bimodal nanoparticles presented negligible cytotoxicity, and efficiently labeled HeLa cells as shown by fluorescence. Thus, the developed nanosystems show potential as strategic probes for fluorescence analyses and MRI, being useful for investigating a variety of biological processes COORDENAÇÃO DE APERFEIÇOAMENTO DE PESSOAL DE NÍVEL SUPERIOR - CAPES CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQ FUNDAÇÃO DE AMPARO À CIÊNCIA E TECNOLOGIA DO ESTADO DE PERNAMBUCO - FACEPE Aberto

Agradecimentos: This work was supported by grant # 07/59319-5 Sao Paulo Research Foundation - Fapesp - http://www.fapesp.br/en/ - NBM; grant # 015/2014 - Fundo de Apoio a Dermatologia do Estado de Sao Paulo "Sebastiao Sampaio" - Funadersp - http://www.sbd-sp.org.br/funadersp.php - PENFV; grant #043/2016 - Fundo de Apoio a Dermatologia do Estado de Sao Paulo "Sebastiao Sampaio" - Funadersp - http://www.sbd-sp.org.br/funadersp.php - PENFV; grant # 01P-04520-2013 - Coordenacao de Aperfeicoamento de Pessoal de Nivel Superior - Capes - http://www.capes.gov.br/ - BGL; Fundo de Apoio ao Ensino, a Pesquisa e Extensao, Universidade Estadual de Campinas (0044/17). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript Abstract: The use of medicinal plants and their derivatives is increasing, and approximately one-third of all traditional herbal medicines are intended for wound treatment. Natural products used in these treatments include vegetable oils, which are rich in essential fatty acids. Once in contact with an ulcerative surface, the oil reaches the blood and lymphatic vessels, thus eliciting systemic effects. Objective This study evaluated the local and possible systemic effects of essential fatty acids (sunflower oil) applied topically to rat wounds. Methods Cutaneous punch wounds (6 mm) were produced on the dorsa of 30 rats. Saline (SS), mineral oil (MO) or essential fatty acid (EFA) solutions were applied topically. Healing was evaluated after 2, 4 and 10 days (n = 5 per group) by visual and histological/morphometric examination, second harmonic generation (SHG) microscopy, and cytokine and growth factor quantification in the scar tissue (real-time PCR) and in serum (ELISA). Results MO/EFA-treated animals had higher IGF-1, leptin, IL-6 and IFN-gamma mRNA expression and lower serum IL-6 levels than the control (SS/MO) animals. SHG analysis showed no difference in collagen density between the animals treated with MO and EFA. Conclusion EFA treatment induces topical (observed by local IGF-1, leptin, IL-6 and IFN-gamma production) and systemic effects, lowering IL-6 levels in the serum. As the oil is widely used to shorten ulcer healing time, studies are needed to evaluate the treatment safety and possible undesired effects FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP COORDENAÇÃO DE APERFEIÇOAMENTO DE PESSOAL DE NÍVEL SUPERIOR - CAPES Aberto

Agradecimentos: The authors acknowledge J. H. Clerici, V. B. Pelegati, and M. O. Baratti for technical assistance, and Prof. H. F. de Carvalho and Prof. A. A. de Thomaz, from the University of Campinas, for valuable discussions. A.C. and E.P.A.M.B. would like to acknowledge Rene Van Veldhoven for the exceptional work and care for the MOVPE systems, and the technical support from the NanoLab@TU/e cleanroom. This work was financially supported by the FAPESP (Fundacao de Amparo a Pesquisa do Estado de Sao Paulo, Brazil), through grants 2010/51748-7, 2013/02300-1, and 2013/10957-0 and the CNPq (Conselho Nacional de Desenvolvimento Cientifico e Tecnologico, Brazil), grant number 479486/2012-3. A.M.d.S. acknowledges scholarship from the CNPq; P.K.S. acknowledges the FAPESP for funding his scholarship. M.A.C., A.A.d.S., and C.L.C. acknowledge the CNPq research fellowships. The authors thank the access to confocal microscopy equipment provided by the National Institute of Science and Technology on Photonics Applied to Cell Biology (INFABIC) at the University of Campinas; INFABIC is cofunded by the FAPESP (08/57906-3) and the CNPq (573913/2008-0). The authors also acknowledge the National Nanotechnology Laboratory (LNNano) at the CNPEM, Brazil, for granting access to their electron microscopy facilities. A.C. and E.P.A.M.B. acknowledge the funding of the Dutch Technology Foundation STW (Project 11826), which is part of the Netherlands Organisation for Scientific Research (NWO), and which is partly funded by the Dutch Ministry of Economic Affairs Abstract: The design and application of indium phosphide (InP) nanowire arrays to acquire Xylella fastidiosa bacterial cell vector force maps are discussed. The nanowire deflections are measured with subdiffraction localization confocal laser scanning microscopy (CLSM). The nanowire mechanical stability in air and liquid media as well as methods to average out thermally induced oscillations are investigated. The accuracy of center determination of the CLSM reflected laser intensity profile at nanowire apex is studied using Gaussian fitting and localization microscopy techniques. These results show that the method is reliable for measuring nanowire displacements above approximate to 25 nm. Corresponding force ranges probed by this method can be customized depending on nanowire geometry and array configuration. The method is applied to explore X. fastidiosa cell adhesion forces on the InP nanowire surface, and in situ probes the effect of N-acetylcysteine on adhered cells. Future perspectives for application of this method in microbiology studies are also outlined FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQ Fechado

Agradecimentos: V.A.E. acknowledges FAPESP (grant 2013/26385-6). J.M.C.d.S.F. acknowledges CNPq (grant 165756/2014-4). E.J.-V. acknowledges FAPESP (grant PV-2017/05854-9). C.L.C. acknowledges the resources obtained from Instituto Nacional de Fotonica Aplicada a Biologia Celular-INFABIC (CNPq grant 573913/2008-0, FAPESP grant 08/57906-3); Biologia das Doencas Neoplasicas da Medula Ossea (FAPESP grant 11/51959-0), and Bolsa de Produtividade (CNPq grant 312049/2014-5) and Centro de Optica e Fotonica, CEPOF (FAPESP grant 05/51689-2). F.C.M. acknowledges FAPESP (grant 2012/10127-5), INES/INCT/CNPq (grant 465423/2014-0), CNPq (grant 407887/2013-0), CAPES, and; Lamult/IFGW/Unicamp Abstract: In the last two decades, many experiments were conducted in self-organization of nanocrystals into two- and three-dimensional (3D) superlattices and the superlattices were synthesized and characterized by different techniques, revealing their unusual properties. Among all characterization techniques, X-ray diffraction (XRD) is the one that has allowed the confirmation of the 3D superlattice formation due to the presence of sharp and intense diffraction peaks. In this work, we study self-organized superlattices of quantum dots of PbS prepared by dropping a monodispersed colloidal solution on a glass substrate at different temperatures. We showed that the intensity of the low-angle XRD peaks depends strongly on the drying time (substrate temperature). We claim that the peaks are originated from the 3D superlattice. Scanning electron microscopy images show that this 3D superlattice (PbS quantum dots) is formed in flake's shape, parallel to the substrate surface and randomly oriented in the perpendicular planes FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQ COORDENAÇÃO DE APERFEIÇOAMENTO DE PESSOAL DE NÍVEL SUPERIOR - CAPES Aberto

Agradecimentos: FAPESP (2013/16911-2); Instituto Nacional de Fotônica Aplicada à Biologia Celular-INFABIC (CNPq grant 465699/2014-6, FAPESP grant 2014/50938-8); Biologia das Doenças Neoplásicas da Medula Óssea (FAPESP grant 11/51959-0); CLC - CNPq produtividade grant (312049/2014-5); and Centro de Óptica e Fotônica - CEPOF (FAPESP grant 05/51689-2) Abstract: Acquiring images of biological tissues and cells without the assistance of exogenous labels with a fast repetition rate and chemical specificity is what coherent anti-Stokes Raman Scattering (CARS) imaging offers. Nonresonant background (NRB) is one of the main drawbacks of the CARS microscopy technique because it limits the detection of weak Raman lines and the detection of low-concentration molecules. We show that a six-wave mixing process with two beams, which is a cascade effect of CARS, show better signal/NRB ratio and can be utilized for biological tissues imaging. The cascade CARS (CCARS) depends on chi-3 to the fourth power, instead of chi-3 squared as in the usual CARS signal||therefore, the contrast ratio with NRB is higher for CCARS than for CARS. We present analytic calculations showing that CCARS have better contrast over CARS in any situation. Comparison of the signals of both techniques generated on water-ethanol solutions confirm these results. Finally, we acquired CCARS images of fresh biological tissues, attesting that it is a useful tool for biological studies FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQ Aberto

Agradecimentos: The author(s) disclosed receipt of the following financial support for the research, authorship, and/or publication of this article: The authors thank the National Institute of Science and Technology on Photonics Applied to Cell Biology (INFABiC) at the State University of Campinas for access to equipment and assistance; INFABiC is co-funded by Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) (08/57906-3) and Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) (573913/2008-0). Rodrigo A Natal and José Vassallo contributed equally to this article Abstract: Second-harmonic generation microscopy represents an important tool to evaluate extracellular matrix collagen structure, which undergoes changes during cancer progression. Thus, it is potentially relevant to assess breast cancer development. We propose the use of second-harmonic generation images of tumor stroma selected on hematoxylin and eosin–stained slides to evaluate the prognostic value of collagen fibers analyses in peri and intratumoral areas in patients diagnosed with invasive ductal breast carcinoma. Quantitative analyses of collagen parameters were performed using ImageJ software. These parameters presented significantly higher values in peri than in intratumoral areas. Higher intratumoral collagen uniformity was associated with high pathological stages and with the presence of axillary lymph node metastasis. In patients with immunohistochemistry-based luminal subtype, higher intratumoral collagen uniformity and quantity were independently associated with poorer relapse-free and overall survival, respectively. A multivariate response recursive partitioning model determined 12.857 and 11.894 as the best cut-offs for intratumoral collagen quantity and uniformity, respectively. These values have shown high sensitivity and specificity to differentiate distinct outcomes. Values of intratumoral collagen quantity and uniformity exceeding the cut-offs were strongly associated with poorer relapse-free and overall survival. Our findings support a promising prognostic value of quantitative evaluation of intratumoral collagen by second-harmonic generation imaging mainly in the luminal subtype breast cancer CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQ FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP Aberto

Agradecimentos: R.J., P.K.S. and D.S.O. acknowledge scholarships from FAPESP and A.M.S. and D.B.A. from CAPES/CNPq. Access to confocal microscopy was granted by INFABIC/UNICAMP (FAPESP 08/57906-3, CNPq 573913/2008-0). We acknowledge access to electron microscopy (LME) and microfabrication (LMF) facilities at the National Nanotechnology Laboratory (LNNano, Brazil) and clean room facilities at both Semiconductor Component Center (CCSNano, UNICAMP) and Device Research Laboratory (IFGW, UNICAMP). Xf.XadA2 antibodies were provided by Dr. A. A. de Souza (IAC, Cordeirópolis, São Paulo, Brazil). The authors are greatly indebted to A. Gobbi and M.H. Piazzetta from LMF/LNNano for PDMS microfluidics processing, Dr. A. Kisner from IQ/UNICAMP and P .C. Galvão, from IQ/UNESP, for technical assistance during this work Abstract: Electrically active field-effect transistors (FET) based biosensors are of paramount importance in life science applications, as they offer direct, fast, and highly sensitive label-free detection capabilities of several biomolecules of specific interest. In this work, we report a detailed investigation on surface functionalization and covalent immobilization of biomarkers using biocompatible ethanolamine and poly(ethylene glycol) derivate coatings, as compared to the conventional approaches using silica monoliths, in order to substantially increase both the sensitivity and molecular selectivity of nanowire-based FET biosensor platforms. Quantitative fluorescence, atomic and Kelvin probe force microscopy allowed detailed investigation of the homogeneity and density of immobilized biomarkers on different biofunctionalized surfaces. Significantly enhanced binding specificity, biomarker density, and target biomolecule capture efficiency were thus achieved for DNA as well as for proteins from pathogens. This optimized functionalization methodology was applied to InP nanowires that due to their low surface recombination rates were used as new active transducers for biosensors. The developed devices provide ultrahigh label-free detection sensitivities similar to 1 fM for specific DNA sequences, measured via the net change in device electrical resistance. Similar levels of ultrasensitive detection of similar to 6 fM were achieved for a Chagas Disease protein marker (IBMP8-1). The developed InP nanowire biosensor provides thus a qualified tool for detection of the chronic infection stage of this disease, leading to improved diagnosis and control of spread. These methodological developments are expected to substantially enhance the chemical robustness, diagnostic reliability, detection sensitivity, and biomarker selectivity for current and future biosensing devices FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQ COORDENAÇÃO DE APERFEIÇOAMENTO DE PESSOAL DE NÍVEL SUPERIOR - CAPES Fechado

Agradecimentos: The authors would like to thank the funding agencies for financial aid for research: FAPESP/CEPID (05/51689-2) and (11/51959-0); CNPq/INCT (573.916/2008-0) and (573913/2008-00); CNPq/PQ (312397/2013-5); CAPES/PROCAD (8881.068505/2014-01) and FAPEAM Programa RH-POSGRAD Abstract: Thousands of people die every year from burn injuries. The aim of this study is to evaluate the feasibility of high intensity femtosecond lasers as an auxiliary treatment of skin burns. We used an in vivo animal model and monitored the healing process using 4 different imaging modalities: histology, Optical Coherence Tomography (OCT), Second Harmonic Generation (SHG), and Fourier Transform Infrared (FTIR) spectroscopy. 3 dorsal areas of 20 anesthetized Wistar rats were burned by water vapor exposure and subsequently treated either by classical surgical debridement, by laser ablation, or left without treatment. Skin burn tissues were non-invasively characterized by OCT images and biopsied for further histopathology analysis, SHG imaging and FTIR spectroscopy at 3, 5, 7 and 14 days after burn. The laser protocol was found as efficient as the classical treatment for promoting the healing process. The study concludes to the validation of femtosecond ultra-short pulses laser treatment for skinburns, with the advantage of minimizing operatory trauma FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQ COORDENAÇÃO DE APERFEIÇOAMENTO DE PESSOAL DE NÍVEL SUPERIOR - CAPES FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DO AMAZONAS - FAPEAM Aberto

Agradecimentos: The authors would like to thank Prof. Benedicto de Campos Vidal from University of Campinas for providing collagen type II, and Tereza Salles and Paulo Latuf Filho for their valuable technical assistance. We also thank Raquel Foglio for English review and Michel Moraes for figure formatting and design. This work was supported by Conselho Nacional de Desenvolvimento Cientifico e Tecnologico (CNPq/MCT), Fundavao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP) and Fundacao de Coordenacao de Aperfeicoamento de Pessoal de Nivel Superior (CAPES) Abstract: Objective. Articular cartilage is an avascular tissue with limited ability of self-regeneration and the current clinical treatments have restricted capacity to restore damages induced by trauma or diseases. Therefore, new techniques are being tested for cartilage repair, using scaffolds and/or stem cells. Although type II collagen hydrogel, fibrin sealant, and adipose-derived stem cells (ASCs) represent suitable alternatives for cartilage formation, their combination has not yet been investigated in vivo for focal articular cartilage defects. We performed a simple experimental procedure using the combination of these 3 compounds on cartilage lesions of rabbit knees. Design. The hydrogel was developed in house and was first tested in vitro for chondrogenic differentiation. Next, implants were performed in chondral defects with or without ASCs and the degree of regeneration was macroscopically and microscopically evaluated. Results. Production of proteoglycans and the increased expression of collagen type II (COL2 alpha 1), aggrecan (ACAN), and sex-determining region Y-box 9 (SOX9) confirmed the chondrogenic character of ASCs in the hydrogel in vitro. Importantly, the addition of ASC induced a higher overall repair of the chondral lesions and a better cellular organization and collagen fiber alignment compared with the same treatment without ASCs. This regenerating tissue also presented the expression of cartilage glycosaminoglycan and type II collagen. Conclusions. Our results indicate that the combination of the 3 compounds is effective for articular cartilage repair and may be of future clinical interest CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQ FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP COORDENAÇÃO DE APERFEIÇOAMENTO DE PESSOAL DE NÍVEL SUPERIOR - CAPES Fechado

Agradecimentos: V.E. acknowledges FAPESP (grant 2013/26385-6). E.J.V. acknowledges FACEPE (grant 0116-1.06/13, APQ0071-1.06/14). C.L.C. acknowledges the resources obtained from Institute Nacional de Fotonica Aplicada a Biologia Celular - INFABIC (CNPq grant 573913/2008-0, FAPESP grant 08/57906-3); Biologia das Doencas Neoplasicas da Medula Ossea (FAPESP grant 11/51959-0); Bolsa de Produtividade (CNPq grant 312049/2014-5); and Centro de Optica e Fotonica CEPOF (FAPESP grant 05/51689-2). F.C.M. acknowledges FAPESP (grant 2012/10127-5), INES/INCT/CNPq (grant 465423/2014-0), CNPq (grant 407887/2013-0), CAPES, and Lamult/IFGW/Unicamp Abstract: Aqueous colloidal silver nanoparticles have substantial potential in biological application as markers and antibacterial agents and in surface-enhanced Raman spectroscopy applications. A simple method of fabrication and encapsulation into an inert shell is of great importance today to make their use ubiquitous. Here we show that colloids of silver-core/silica-shell nanoparticles can be easily fabricated by a laser-ablation-assisted chemical reduction method and their sizes can be tuned in the range of 2.5 to 6.3 nm by simply choosing a proper water ethanol proportion. The produced silver nanoparticles possess a porous amorphous silica shell that increases the inertness and stability of colloids, which decreases their toxicity compared with those without silica. The presence of a thin 2 to 3 nm silica shell was proved by EDX mapping. The small sizes of nanoparticles achieved by this method were analyzed using optical techniques, and they show typical photoluminescence in the UV-vis range that shifts toward higher energies with decreasing size FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP FUNDAÇÃO DE AMPARO À CIÊNCIA E TECNOLOGIA DO ESTADO DE PERNAMBUCO - FACEPE CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQ COORDENAÇÃO DE APERFEIÇOAMENTO DE PESSOAL DE NÍVEL SUPERIOR - CAPES Fechado

Agradecimentos: This study was funded by the São Paulo Research Foundation (08/57906–3), the National Council for Scientific and Technological Development (573913/2008–0), and the Biologia das Doenças Neoplásicas da Medula Óssea (FAPESP grant 11/51959-0) project with fellowship support from CNPq (310518/2015-6) Abstract: Background: The diagnosis of idiopathic atrophoderma of Pasini and Pierini (IAPP) relies on typical clinical features, particularly distinctive pigmented ovular/round depressed plaques. Histologic examination often reveals no obvious changes, but patterns of collagen distribution, using multiphoton imaging and second harmonic generation can help track hidden details of tissue organization contributing to atrophy. Objective: To identify histologic features that distinguish IAPP from unaffected skin. Methods: Eleven patients were included for conventional analyses. Masson trichromee and Unna-Tanzer orcein-stained sections were evaluated using automated morphometry. Hematoxylin-eosinestained sections were analyzed by multiphoton imaging using 2-photon excited fluorescence and second harmonic generation. Results: No abnormalities were found under light microscopy or by automated quantification. Multiphoton imaging revealed no difference in optical density of either collagen or elastic fibers in lesioned and unaffected skin; however, horizontal collagen fiber organization in lesion specimens increased toward the lower dermis, whereas elastic fibers featured greater disorganization within the upper dermis. Limitations: The low number of patients evaluated. Conclusion: The atrophic appearance of IAPP lesions reflects changes in organization, but not in collagen and elastic tissue content. Minute organizational differences that are imperceptible to the experienced pathologist and undetectable by automated analyses were revealed by multiphoton analyses, particularly second harmonic generation, in association with texture analyses FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQ Fechado

Agradecimentos: The authors acknowledge technical assistance from Vitor Pelegatti, from INFABIC/UNICAMP, Maria H. Piazetta and A.Gobbi, from LNNano/CNPEM. The authors are also greatly indebted to Dr. Richard Janissen, from Kavli Institute of Nanoscience, Delft University of Technology, and M.Sc. Duber Murillo from Applied Physics Department at the State University of Campinas, for fruitful discussions during this work Abstract: The pathogenicity of Xylella fastidiosa is associated with its systematic colonization of the plant xylem, forming bacterial biofilms. Mechanisms of bacterial transport among different xylem vessels, however, are not completely understood yet, but are strongly influenced by the presence of extracellular polymeric substances (EPS), which surrounds the assembly of cells forming the biofilm. In this work, we show quantitative measurements on the elastic properties of the system composed by EPS and bacterial cell. In order to investigate the mechanical properties of this system, force spectroscopy and confocal Raman measurements were carried out during Xylella fastidiosa subsp. pauca initial stages of adhesion and cluster formation. We show that stiffness progressively decreases with increasing culture growth time, from two to five days. For early adhesion samples, stiffness values are quite different at the bacterial polar and body regions. Lower stiffness values at the cell pole suggest a flexible mechanical, response at this region, associated with first cell adhesion to a surface. These results correlate very well with our observations of cell motion within microchannels, under conditions simulating xylem flow. Both the oscillatory movement of vertically attached single cells, as well as the transport of cell clusters within the biofilm matrix can be explained by the presence of softer materials at the cell pole and EPS matrix. Our results may thus add to a more detailed understanding of mechanisms used by cells to migrate among vessels in plant xylem FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQ COORDENAÇÃO DE APERFEIÇOAMENTO DE PESSOAL DE NÍVEL SUPERIOR - CAPES Fechado

Agradecimentos: This work was supported by the Mexican founding agency Consejo Nacional de Ciencia y Tecnologia (CONACyT) under project 183728. Authors also acknowledge the financial support from Instituto Politecnico Nacional under projects SIP20151327 and SIP20160171. E. Jimenez-Villar acknowledges FACEPE and CNPq (Grant 0116-1.06/13, APQ-0071-1.06/14), Brazil. E.J-V also acknowledges the FAPESP for financial supports (Grant PV-2017/05854-9) Abstract: The synthesis processes of metallic nanoparticles have seen a growing interest in recent years, mainly by the potential applications of the phenomenon of localized surface plasmon resonance associated with metallic nanoparticles. This paper shows a fast method to synthesize silver, gold and silver/gold alloy nanoparticles coated with a porous silica shell by the assisted laser ablation method in three steps. The method involves a redox chemical reaction where the reducing agent is supplied in nanometric form by laser ablation. In the first step, a silicon target immersed in water is ablated for several minutes. Later, AgNO3 and HAuCl4 aliquots are added to the solution. The redox reaction between the silver and gold ions and products resulting from ablation process can produce silver, gold or silver/gold alloy nanoparticles coated with a porous silica shell. The influence of the laser pulse energy, ablation time, Ag+ and Au3+ concentration, as well as the Ag+/Au3+ ratio, on optical and structural properties of the nanostructures was investigated. This work represents a step forward in the study of reaction mechanisms that take place during the synthesis of nanoscale materials by the assisted laser ablation technique FUNDAÇÃO DE AMPARO À CIÊNCIA E TECNOLOGIA DO ESTADO DE PERNAMBUCO - FACEPE CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQ FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP Fechado

Bartonella henselae is a causative agent of anemia, cat scratch disease, bacillary angiomatosis, recurrent fever, hepatitis, endocarditis, chronic lymphadenopathy, joint and neurological disorders. B. henselae are intra-erythrocytic bacteria. The goal of this study was to visualize the B. henselae invasion into enucleated human red blood cells in real time using bacterium endogenous fluorescence. We took advantage of the unique fluorescence emission spectral profile of the bacteria. We used a linear unmixing approach to separate the fluorescence emission spectra of human erythrocytes from native B. henselae when excited at 488nm. Human blood samples were inoculated with B. henselae and incubated for 60 hours. 3-D live images were captured at select intervals using multi-photon laser scanning microscopy. Uninfected blood samples were also analyzed. This study revealed bacteria entering mature erythrocytes over a 60 hour time period Aberto

Agradecimentos: The authors acknowledge J. H. Clerici, V. B. Pelegati, and M. O. Baratti for technical assistance and C. B. Santos and Prof. A. P. de Souza, from CBMEG/UNICAMP for providing the XadA1 protein. A.C. and E.P.A.M.B. would like to acknowledge Rene Van Veldhoven for the exceptional work and care for the MOVPE systems and the technical support from the Nano-Lab@TU/e cleanroom. This work was financially supported by FAPESP (Fundacao de Amparo a Pesquisa do Estado de Sao Paulo, Brazil), through grants 2010/51748-7 and 2013/02300-1, and CNPq (Conselho Nacional de Desenvolvimento Cientifico e Tecnologico, Brazil), grant number 479486/2012-3. D.M.M. and M.P.M. acknowledge scholarships from CAPES (Coordenacao de Aperfeicoamento de Pessoal de Nivel Superior, Brazil); P.K.S. and R.J. acknowledge FAPESP for funding their scholarships. M.A.C., A.A.S., and H.F.C. acknowledge CNPq fellowships. The authors thank the access to confocal microscopy equipment provided by the National Institute of Science and Technology on Photonics Applied to Cell Biology (INFABIC) at the State University of Campinas; INFABIC is cofunded by FAPESP (08/57906-3) and CNPq (573913/2008-0). We also acknowledge the National Nano technology Laboratory (LNNano) at CNPEM, Brazil, for granting access to their electron microscopy facilities. A.C. and E.P.A.M.B. acknowledge the funding of the Dutch Technology Foundation STW (Project 11826), which is part of The Netherlands Organisation for Scientific Research (NWO), and which is partly funded by the Dutch Ministry of Economic Affairs Abstract: Surface attachment of a planktonic bacteria, mediated by adhesins and extracellular polymeric substances (EPS), is a crucial step for biofilm formation. Some pathogens can modulate cell adhesiveness, impacting host colonization and virulence. A framework able to quantify cell-surface interaction forces and their dependence on chemical surface composition may unveil adhesiveness control mechanisms as new targets for intervention and disease control. Here we employed InP nanowire arrays to dissect factors involved in the early stage biofilm formation of the phytopathogen Xylella fastidiosa. Ex vivo experiments demonstrate single-cell adhesion forces up to 45 nN, depending on the cell orientation with respect to the surface. Larger adhesion forces occur at the cell poles; secreted EPS layers and filaments provide additional mechanical support. Significant adhesion force enhancements were observed for single cells anchoring a biofilm and particularly on XadA1 adhesincoated surfaces, evidencing molecular mechanisms developed by bacterial pathogens to create a stronger holdfast to specific host tissues FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQ COORDENAÇÃO DE APERFEIÇOAMENTO DE PESSOAL DE NÍVEL SUPERIOR - CAPES Aberto

Agradecimentos: Contract grant sponsor: FAPESP; Contract grant number: 2009/16150-6 (to H.F.C.); Contract grant sponsor: FAPESP; Contract grant number: 2011/51591-3; Contract grant sponsors: CEPOF (Optics and Photonics Research Center, FAPESP), INFABIC (National Institute of Photonics Applied to Cell Biology, FAPESP and CNPq), PICTO UNER-INTA-CAFEGS; Contract grant number: 2009-209; Contract grant sponsor: PIO CONICET-UNER Res: 4337/15; Contract grant number: No 14620140100004 CO) Abstract: A plethora of optical techniques is currently available to obtain non-destructive, contactless, real time information with subcellular spatial resolution to observe cell processes. Each technique has its own unique features for imaging and for obtaining certain biological information. However none of the available techniques can be of universal use. For a comprehensive investigation of biological specimens and events, one needs to use a combination of bioimaging methods, often at the same time. Some modern confocal/multiphoton microscopes provide simultaneous fluorescence, fluorescence lifetime imaging, and four-dimensional imaging. Some of them can also easily be adapted for harmonic generation imaging, and to permit cell manipulation technique. In this work we present a multimodal optical workstation that extends a commercially available confocal microscope to include nonlinear/multiphoton microscopy and optical manipulation/stimulation tools. The nonlinear microscopy capabilities were added to the commercial confocal microscope by exploiting all the flexibility offered by the manufacturer. The various capabilities of this workstation as applied directly to reproductive biology are discussed FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQ Fechado

Agradecimentos: A.A.d.T. was a recipient of a fellowship of the State of Sao Paulo Funding Agency (FAPESP). FAPESP and National Research Council (CNPq) grants to H.F.C. and to C.L.C. (FAPESP Grant Nos. 08/57906-3 and 09/54164-9) are acknowledged. INFABIC is cofounded by CNPq and FAPESP Abstract: One of the most important properties of quantum dots (QDs) is their size. Their size will determine optical properties and in a colloidal medium their range of interaction. The most common techniques used to measure QD size are transmission electron microscopy (TEM) and X-ray diffraction. However, these techniques demand the sample to be dried and under a vacuum. This way any hydrodynamic information is excluded and the preparation process may alter even the size of the QDs. Fluorescence correlation spectroscopy (FCS) is an optical technique with single molecule sensitivity capable of extracting the hydrodynamic radius (HR) of the QDs. The main drawback of FCS is the blinking phenomenon that alters the correlation function implicating in a QD apparent size smaller than it really is. In this work, we developed a method to exclude blinking of the FCS and measured the HR of colloidal QDs. We compared our results with TEM images, and the HR obtained by FCS is higher than the radius measured by TEM. We attribute this difference to the cap layer of the QD that cannot be seen in the TEM images FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQ Fechado

Androgens regulate prostate physiology, and exert their effects through the androgen receptor. We hypothesized that androgen deprivation needs additional transcription factors to orchestrate the changes taking place in the gland after castration and for the adaptation of the epithelial cells to the androgen-deprived environment, ultimately contributing to the origin of castration-resistant prostate cancer. This study was undertaken to identify transcription factors that regulate gene expression after androgen deprivation by castration (Cas). For the sake of comparison, we extended the analysis to the effects of administration of a high dose of 17 beta-estradiol (E2) and a combination of both (Cas+ E2). We approached this by (i) identifying gene expression profiles and enrichment terms, and by searching for transcription factors in the derived regulatory pathways; and (ii) by determining the density of putative transcription factor binding sites in the proximal promoter of the 10 most up- or down-regulated genes in each experimental group in comparison to the controls Gapdh and Tbp7. Filtering and validation confirmed the expression and localized EVI1 (Mecom), NFY, ELK1, GATA2, MYBL1, MYBL2, and NFkB family members (NFkB1, NFkB2, REL, RELA and RELB) in the epithelial and/or stromal cells. These transcription factors represent major regulators of epithelial cell survival and immaturity as well as an adaptation of the gland as an immune barrier in the absence of functional stimulation by androgens. Elk1 was expressed in smooth muscle cells and was up-regulated after day 4. Evi1 and Nfy genes are expressed in both epithelium and stroma, but were apparently not affected by androgen deprivation FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQ Aberto

Agradecimentos: This work was funded by Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP; Grants 2006/54878-3, 2007/55930-1, 2007/59442-1, 2008/53519-5, 2008/57805-2, 2010/02628-9) and Conselho Nacional de Pesquisa (CNPq: Grants 304366/2009-9, 475158/2010-5). We thank the access to equipment and assistance provided by the National Institute of Science and Technology on Photonics Applied to Cell Biology (INFABIC) at the State University of Campinas Abstract: Focal adhesion kinase (FAK) contributes to cellular homeostasis under stress conditions. Here we show that alpha B-crystallin interacts with and confers protection to FAK against calpain-mediated proteolysis in cardiomyocytes. A hydrophobic patch mapped between helices 1 and 4 of the FAK FAT domain was found to bind to the beta 4-beta 8 groove of alpha B-crystallin. Such an interaction requires FAK tyrosine 925 and is enhanced following its phosphorylation by Src, which occurs upon FAK stimulation. alpha B-crystallin silencing results in calpain-dependent FAK depletion and in the increased apoptosis of cardiomyocytes in response to mechanical stress. FAK overexpression protects cardiomyocytes depleted of alpha B-crystallin against the stretch-induced apoptosis. Consistently, load-induced apoptosis is blunted in the hearts from cardiac-specific FAK transgenic mice transiently depleted of alpha B-crystallin by RNA interference. These studies define a role for alpha B-crystallin in controlling FAK function and cardiomyocyte survival through the prevention of calpain-mediated degradation of FAK FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP Aberto

Agradecimentos: We would like to express our thanks for the access to equip¬ment and assistance provided by the National Institute of Sci¬ence and Technology on Photonics Applied to Cell Biology (INFABIC) at the State University of Campinas; INFABIC is co-funded by Fundação de Amparo a Pesquisa do Estado de São Paulo (FAPESP) (08/57906–3) and Conselho Nacio¬nal de Desenvolvimento Cientifico e Tecnológico (CNPq) (573913/2008–0). We thank the FI-UNER teachers: Mrs Menis and Mrs Monzón for her English review. This paper was supported by the PID-UNER6116: "Estudios Moleculares y Celulares del Sistema de Endotelinas en cáncer colo-rectal" Abstract: This work reviews the most relevant present-day processing methods used to improve the accuracy of multimodal nonlinear images in the detection of epithelial cancer and the supporting stroma. Special emphasis has been placed on methods of non linear optical (NLO) microscopy image processing such as: second harmonic to autofluorescence ageing index of dermis (SAAID), tumor-associated collagen signatures (TACS), fast Fourier trans-form (FFT) analysis, and gray level co-occurrence matrix (GLCM)-based methods. These strategies are presented as a set of potential valuable diagnostic tools for early cancer detection. It may be proposed that the combination of NLO microscopy and informatics based image analysis approaches described in this review (all carried out on free software) may represent a powerful tool to investigate collagen organization and remodeling of extracellular matrix in carcinogenesis processes FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQ Fechado

Agradecimentos: The authors thank FAPESP, CNPq and CAPES for the finantial support and scholarship. Andre A. de Thomaz, Hernandes F. Carvalho for the collaborating in multiphoton confocal microscopy techniques and FLIM Abstract: A gastro-resistant system of acryl-EZE (R) MP coated alginate/chitosan microparticles was developed to improve the controlled release of oxytetracycline (OTC). Microparticles were obtained by complex coacervation and, thereafter, were coated using fluidized polymer dispersion with acryl-EZE (R) MP solution. OTC distribution inside the microparticles was determined by multiphoton confocal microscopy, demonstrating the efficiency of encapsulation process. In vitro OTC release kinetic was performed in order to obtain the release profile in gastric and intestinal simulated fluids. A fast initial release, or burst effect, was observed with uncoated microparticles loaded with OTC in gastric conditions. When a 50% mass increase in acryl-EZE (R) MP coating was achieved, OTC release in acidic medium was greatly reduced, resulting in the expected gastro-resistant effect. Different mathematical models were applied to describe the drug diffusion across the polymer matrix. The Logistic model was the best tool to interpret the experimental data in most of the systems studied FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQ COORDENAÇÃO DE APERFEIÇOAMENTO DE PESSOAL DE NÍVEL SUPERIOR - CAPES Fechado

Agradecimentos: This study was supported by the Fundação do Amparo à Pesquisa do Estado de São Paulo - FAPESP (www.fapesp.br). FAPESP Process No 2010/12565-4. FAPESP-INCT-INFABIC Process No 2008/57906-3. The funding institution had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript Abstract: Nonlinear optical (NLO) microscopy techniques have potential to improve the early detection of epithelial ovarian cancer. In this study we showed that multimodal NLO microscopies, including two-photon excitation fluorescence (TPEF), second-harmonic generation (SHG), third-harmonic generation (THG) and fluorescence lifetime imaging microscopy (FLIM) can detect morphological and metabolic changes associated with ovarian cancer progression. We obtained strong TPEF + SHG + THG signals from fixed samples stained with Hematoxylin & Eosin (H&E) and robust FLIM signal from fixed unstained samples. Particularly, we imaged 34 ovarian biopsies from different patients (median age, 49 years) including 5 normal ovarian tissue, 18 serous tumors and 11 mucinous tumors with the multimodal NLO platform developed in our laboratory. We have been able to distinguish adenomas, borderline, and adenocarcinomas specimens. Using a complete set of scoring methods we found significant differences in the content, distribution and organization of collagen fibrils in the stroma as well as in the morphology and fluorescence lifetime from epithelial ovarian cells. NLO microscopes provide complementary information about tissue microstructure, showing distinctive patterns for serous and mucinous ovarian tumors. The results provide a basis to interpret future NLO images of ovarian tissue and lay the foundation for future in vivo optical evaluation of premature ovarian lesions FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP Aberto

Agradecimentos: The authors are grateful to Coordenação de Aperfeiçoamento de Pessoal de Nível Supeior (CAPES), Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), Fundação de Amparo à Ciência e Tecnologia do Estado de Pernambuco (FACEPE), Fundação de Hematologia e Hemoterapia de Pernambuco (HEMOPE), L’óreal, Academia Brasileira de Ciências, Organização das Nações Unidas para a Educação, a Ciência e a Cultura (UNESCO) and Aggeu Magalhães Institute (FIOCRUZ - Fundação Oswaldo Cruz). This work is also linked to Optics and Photonics Research Center (CEPOF) from Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP), National Institute of Photonics Applied to Cell Biology, National Institute of Photonics and National Institute of Science and Blood Technology Abstract: During storage, red blood cells (RBCs) for transfusion purposes suffer progressive deterioration. Sialylated glycoproteins of the RBC membrane are responsible for a negatively charged surface which creates a repulsive electrical zeta potential. These charges help prevent the interaction between RBCs and other cells, and especially among each RBCs. Reports in the literature have stated that RBCs sialylated glycoproteins can be sensitive to enzymes released by leukocyte degranulation. Thus, the aim of this study was, by using an optical tweezers as a biomedical tool, to measure the zeta potential in standard RBCs units and in leukocyte reduced RBC units (collected in CPD-SAGM) during storage. Optical tweezers is a sensitive tool that uses light for measuring cell biophysical properties which are important for clinical and research purposes. This is the first study to analyze RBCs membrane charges during storage. In addition, we herein also measured the elasticity of RBCs also collected in CPD-SAGM. In conclusion, the zeta potential decreased 42% and cells were 134% less deformable at the end of storage. The zeta potential from leukodepleted units had a similar profile when compared to units stored without leukoreduction, indicating that leukocyte lyses were not responsible for the zeta potential decay. Flow cytometry measurements of reactive oxygen species suggested that this decay is due to membrane oxidative damages. These results show that measurements of zeta potentials provide new insights about RBCs storage lesion for transfusion purposes COORDENAÇÃO DE APERFEIÇOAMENTO DE PESSOAL DE NÍVEL SUPERIOR - CAPES CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQ FUNDAÇÃO DE AMPARO À CIÊNCIA E TECNOLOGIA DO ESTADO DE PERNAMBUCO - FACEPE FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP Aberto

Agradecimentos: The authors are grateful to FAPESP. This work is also linked with the CEPOF (Optics and Photonics Research Center, FAPESP) and National Institute of Photonics Applied to Cell Biology (INFABIC) Abstract: We show that combined multimodal nonlinear optical (NLO) microscopies, including two-photon excitation fluorescence, second-harmonic generation (SHG), third harmonic generation, and fluorescence lifetime imaging microscopy (FLIM) can be used to detect morphological and metabolic changes associated with stroma and epithelial transformation during the progression of cancer and osteogenesis imperfecta (01) disease. NLO microscopes provide complementary information about tissue microstructure, showing distinctive patterns for different types of human breast cancer, mucinous ovarian tumors, and skin dermis of patients with OI. Using a set of scoring methods (anisotropy, correlation, uniformity, entropy, and lifetime components), we found significant differences in the content, distribution and organization of collagen fibrils in the stroma of breast and ovary as well as in the dermis of skin. We suggest that our results provide a framework for using NLO techniques as a clinical diagnostic tool for human cancer and OI. We further suggest that the SHG and FLIM metrics described could be applied to other connective or epithelial tissue disorders that are characterized by abnormal cells proliferation and collagen assembly FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP Aberto

Agradecimentos: The authors would like to thank Brazilian agencies CNPq (CTEnergy project) and FAPESP for financial support, as well as CePOF-Unicamp Abstract: We have been able to produce soft glass conventional core-clad and micro-structured fibers using rod-and-tube and stack-and-draw method respectively. The stack-and-draw technique shows several difficulties when used with soft glasses, that we managed to avoid using two different lead and alkaline glasses. Non commercial glasses and fibers were thermo-mechanically and optically characterized CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQ FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP Fechado

Agradecimentos: This work was supported by the FIRB project "Laboratorio nazionale sulle nanotecnologie per la genomica e postgenomica" (NG-Laboratory) Abstract: Axial trapping through a dielectric interface is investigated in the framework of the angular spectrum representation and of the generalized Lorenz-Mie theory. We determine the optical force for an arbitrarily polarized non-paraxial, strongly aberrated, axially symmetric focusing beam and apply this description to the case of an arbitrarily positioned dielectric microsphere, commonly employed in optical tweezers, not taking into account the contribution of evanescent waves at the interface. We derive the analytical expression of the force profile, finding that the incident polarization does not affect the axial optical force. In addition, we derive an approximated expression for the axial force as a function of beam displacement just outside the microsphere and we show how the information provided by the ripple structure of the optical trapping efficiency versus sphere displacement curve, due to the aberration effect, could be exploited to calibrate the bead axial position versus the experimental beam positioning controls Fechado

Agradecimentos: This research was supported by FAPESP, PRONEX and CEPOF Abstract: Mode-mismatched Thermal Lens (TL) measurements were performed in 70TeO(2)-19WO(3)-7Na(2)O-4Nb(2)O(5) (% mol) tellurite glasses doped with either Er3+ or Tm3+ and co-doped with Er3+/Tm3+ ions. Thermo-optical parameters (D, K, ds/dQ and ds/dT) were obtained in function of thulium concentrations (0.39-1.6) x 10 20 ions/cm(3). For Er3+/Tm3+ co-doped tellurite glasses, D and K values are practically independent of the Tm3+ concentrations used in this study. The average values of D and ds/dT obtained for tellurite glasses are: (3.1 +/- 0.2) x 10(-3) supercript stopcm(2)/s and (16 +/- 3) x 10(-6) stopK(-1), respectively FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP Fechado

Agradecimentos: The authors acknowledge the financial support from CNPq (Brazilian National Council for Research) and the CePOF-Unicamp (Center for Optics and Photonics) supported by FAPESP (São Paulo State Foundation for Research) Abstract: A new polymeric open optical fiber with a star cross section from extruded PMMA grains was developed. We have painted the external surface of this fiber with the Europium chelate or Antracene. We have observed the typical luminescence of these compounds when excited with nanosecond 355 nm pulsed laser. These results show that the developed fiber presents potentialities for optical sensing CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQ FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP Fechado

Agradecimentos: The authors are grateful for the financial support of the following Brazilian agencies: CNPq, FAPESP (Cepof), FACEPE, RENAMI and IMMC. We thank Prof. F. Galembeck and Dr. C.P. Leite for the TEM images Abstract: Fluorescent semiconductor nanocrystals in quantum confinement regime (quantum dots) present several well-known features which make them very useful tools for biological labeling purposes. Low photobleaching rates, high chemical stability and active surface allowing conjugation to living cells explain the success of this labeling procedure over the commonly used fluorescent dyes. In this paper we report the results obtained with highly fluorescent core-shell CdTe-CdS (diameter=3-7 nm) colloidal nanocrystals synthesized in aqueous medium and conjugated to glucose molecules. The conjugated nanocrystals were incubated with living yeast cells, in order to investigate their glucose up-take activity in real time, by confocal microscopy analysis CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQ FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP FUNDAÇÃO DE AMPARO À CIÊNCIA E TECNOLOGIA DO ESTADO DE PERNAMBUCO - FACEPE Fechado