Glycoengineered cell models for the characterization of cancer O-glycoproteome: an innovative strategy for biomarker discovery

Glycosylation is one of the most abundant forms of protein posttranslational modification. O-glycosylation is a major type of protein glycosylation, comprising different types and structures expressed in several physiologic and pathologic conditions. The understanding of protein attachment site and glycan structure is of the utmost importance for the clarification of the role glycosylation plays in normal cells and in pathological conditions. Neoplastic transformation frequently shows the expression of immature truncated O-glycans. These aberrantly expressed O-glycans have been shown to induce oncogenic properties and can be detected in premalignant lesions, meaning that they are an important source of biomarkers. This article addresses the recent application of genetically engineered cancer cell models to produce simplified homogenous O-glycans allowing the characterization of cancer cells O-glycoproteomes, using advanced mass spectrometry methods and the identification of potential cancer-specific O-glycosylation sites. This article will also discuss possible applications of these biomarkers in the cancer field.IPATIMUP integrates the i3S Research Unit, which is partially supported by FCT, the Portuguese Foundation for Science and Technology. The authors were supported by FEDER funds through the Operational Programme for Competitiveness Factors-COMPETE and National Funds through the FCT-Foundation for Science and Technology, under the projects: PEst-C/SAU/LA0003/2013 and PTDC/BBB-EBI/0786/2012. D Campos has received a grant from the FCT (grant number: SFRH/BD/73717/2010). D Freitas has received a grant from the FCT (grant number: PD/BI/105913/2014)

Revision of Recreation and Leisure Practicum

In a 1-ha plot divided into 100 subplots of 10 x 10 m, all trees with at least 15 cm of perimeter at breast height (DBH = 4.8 cm) were marked and had their heights estimated and perimeter taken. The rock cover (rocks over 50 cm diameter) was estimated in five classes of frequency, and records were made for individuals growing directly on rocks. We found 1,274 trees matching the sampling criteria, which belong to 41 botanical families (highlighting Myrtaceae, Rubiaceae and Fabaceae) and 142 species or morphotypes. The most important (Importance Value Index) species are: Euterpe edulis, Mollinedia schottiana, Bathysa mendoncaei, Coussarea accedens, Rustia formosa and Guapira opposita. Shannon's diversity index was 4.05 nats/ind and Pielou's equability was 0.82. The average tree height is of 9 m and the canopy is at around 18 m. The trees' average diameter is 13.9 cm, and 29 individuals surpass 50 cm DBH. The basal area for the I-ha plot (live trees only) is 30.27 m(2). A direct relation was found between rock cover and lesser species richness and number of individuals per subplot. No relationship was found between rock cover and the mean height or mean diameter of stems in the subplots. 34 tree species in this area are able to grow on rocks; 11 of which do not grow roots to the soil, particularly Euterpe edulis and Guapira opposita. The height and diameter of the individuals that grow on rocks is not statistically different from the remaining in the plot

Impact of Truncated O-glycans in Gastric-Cancer-Associated CD44v9 Detection

CD44 variant isoforms are often upregulated in cancer and associated with increased aggressive tumor phenotypes. The CD44v9 is one of the major protein splice variant isoforms expressed in human gastrointestinal cancer cells. Immunodetection of CD44 isoforms like CD44v9 in tumor tissue is almost exclusively performed by using specific monoclonal antibodies. However, the structural variability conferred by both the alternative splicing and CD44 protein glycosylation is disregarded. In the present work, we have evaluated the role of O-glycosylation using glycoengineered gastric cancer models in the detection of CD44v9 by monoclonal antibodies. We demonstrated, using different technical approaches, that the presence of immature O-glycan structures, such as Tn and STn, enhance CD44v9 protein detection. These findings can have significant implications in clinical applications mainly at the detection and targeting of this cancer-related CD44v9 isoform and highlight the utmost importance of considering glycan structures in cancer biomarker detection and in therapy targeting.This work was funded by FEDER funds through the Operational Programme for Competitiveness Factors-COMPETE, grant numbers POCI-01-0145-FEDER-016585; POCI-01-0145-FEDER-007274; OCI-01-0145-FEDER-031028; and national funds through the Foundation for Science and Technology (FCT), grant numbers PTDC/BBB-EBI/0567/2014 (to CAR), UID/BIM/04293/2013, and PTDC/MED-QUI/29780/2017; and the project NORTE-01-0145-FEDER-000029, supported by Norte Portugal Regional Programme (NORTE 2020), under the PORTUGAL 2020 Partnership Agreement, through the European Regional Development Fund (ERDF). F. Pinto received a fellowship from FCT (SFRH/BPD/115730/2016)

Synthesis of Carbonated Hydroxyapatite Nanorods in Liquid Crystals

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Syntheses of calcium phosphate nanoparticles, carried out in systems formed from surfactant, oil and water, have resulted in materials with promising possibilities for application. The calcium phosphate particles were synthesized using two different liquid crystals, formed from Renex (TM), cyclohexane and a salts solution. The morphology of the nanoparticles synthesized in the liquid crystals is similar to that of hydroxyapatite particles that form bone mineral, where collagen fibers connect these particles so as to form a composite. Therefore, the synthesis of calcium phosphate nanoparticles in the systems used in this work can advance current understanding of mineralization processes that result in the formation of bone mineral.123265268Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq

Different isolation approaches lead to diverse glycosylated extracellular vesicle populations

Extracellular vesicles (EVs) are a heterogeneous group of small secreted particles involved in inter-cellular communication and mediating a broad spectrum of biological functions. EVs cargo iscomposed of a large repertoire of molecules, including glycoconjugates. Herein, we report the firststudy on the impact of the isolation strategy on the EV populations’glycosylation profile. The use ofdifferent state-of-the-art protocols, namely differential ultracentrifugation (UC), total exosome isola-tion (TEI), OptiPrepTMdensity gradient (ODG) and size exclusion chromatography (SEC) resulted in EVpopulations displaying different sets of glycoconjugates. The EV populations obtained by UC, ODGand SEC methods displayed similar protein and glycan profiles, whereas TEI methodology isolated themost distinct EV population. In addition, ODG and SEC isolation protocols provided an enhanced EVglycoproteins detection. Remarkably, proteins displaying the tumour-associated glycan sialyl-Tn(STn) were identified as packaged cargo into EVs independently of the isolation methodology. STncarrying EV samples isolated by UC, ODG and SEC presented a considerable set of cancer-relatedproteins that were not detected in EVs isolated by TEI. Our work demonstrates the impact of usingdifferent isolation methodologies in the populations of EVs that are obtained, with consequences inthe glycosylation profile of the isolated population. Furthermore, our results highlight the importanceof selecting adequate EV isolation protocols and cell culture conditions to determine the structuraland functional complexity of the EV glycoconjugates.This work was funded by FEDER funds through theOperational Programme for Competitiveness Factors-COMPETE (POCI-01-0145-FEDER-016585; POCI-01-0145-FEDER-007274; POCI-01-0145-FEDER-028489) and NationalFunds through the Foundation for Science and Technology(FCT), under the projects: PTDC/BBB-EBI/0567/2014 (toCAR), PTDC/MED-ONC/28489/2017 (to AM) and UID/BIM/04293/2013; and the project NORTE-01-0145-FEDER-000029, supported by Norte Portugal Regional Programme(NORTE 2020), under the PORTUGAL 2020 PartnershipAgreement, through the European Regional DevelopmentFund (ERDF). DF acknowledges the FCT PhD Programmesand Programa Operacional Potencial Humano (POPH), speci-fically the Biotech Health Programme (Doctoral Programme onCellular and Molecular Biotechnology Applied to HealthSciences), with the reference PD/0016/2012 funded by FCTand the grant SFRH/BD/110636/2015 from FCT, POPH andFSE (Fundo Social Europeu); MB acknowledges the EuropeanUnion’s Horizon 2020 research and innovation programmeunder the Marie Sklodowska-Curie grant agreement No.748880; and JP acknowledges FCT (SFRH/BD/137319/2018).The authors acknowledge Rede Nacional de Espectrometria deMassa, ROTEIRO/0028/2013, ref. LISBOA-01-0145-FEDER-022125, supported by COMPETE and North PortugalRegional Operational Programme (Norte2020), under thePORTUGAL 2020 Partnership Agreement, through theEuropean Regional Development Fund (ERDF). SV acknowl-edges the Danish National Research Foundation (DNRF107)

Author Correction: iLoF: An intelligent Lab on Fiber Approach for Human Cancer Single-Cell Type Identification

An amendment to this paper has been published and can be accessed via a link at the top of the paper.This work was partially funded by the projects NanoSTIMA and NORTE-01-0145-FEDER-000029, both supported by the North Portugal Regional Operational Program (NORTE 2020), under the PORTUGAL 2020 Partnership Agreement, and through the European Regional Development Fund (ERDF); and by the Portuguese Foundation for Science and Technology, within the scope of the PhD grant PD/BD/135023/2017 and the projects: PTDC/BBB-EBI/0567/2014 (to CAR) and UID/BIM/04293/2013. It was also funded by FEDER funds through the Operational Programme for Competitiveness Factors-COMPETE (POCI-01-0145-FEDER-016585; POCI-01-0145-FEDER-007274; PPBI-POCI-01-0145-FEDER-022122). MB acknowledges the Marie Sklodowska-Curie grant agreement No. 748880

Upper tropospheric ozone production from lightning NOx-impacted convection: Smoke ingestion case study from the DC3 campaign

©2015. American Geophysical Union. All Rights Reserved. As part of the Deep Convective Cloud and Chemistry (DC3) experiment, the National Science Foundation/National Center for Atmospheric Research (NCAR) Gulfstream-V (GV) and NASA DC-8 research aircraft probed the chemical composition of the inflow and outflow of two convective storms (north storm, NS, south storm, SS) originating in the Colorado region on 22 June 2012, a time when the High Park wildfire was active in the area. A wide range of trace species were measured on board both aircraft including biomass burning (BB) tracers hydrogen cyanide (HCN) and acetonitrile (ACN). Acrolein, a much shorter lived tracer for BB, was also quantified on the GV. The data demonstrated that the NS had ingested fresh smoke from the High Park fire and as a consequence had a higher VOC OH reactivity than the SS. The SS lofted aged fire tracers along with other boundary layer ozone precursors and was more impacted by lightning NOx (LNOx) than the NS. The NCAR master mechanism box model was initialized with measurements made in the outflow of the two storms. The NS and SS were predicted to produce 11 and 14ppbv of O3, respectively, downwind of the storm over 2days. Sensitivity tests revealed that the ozone production potential of the SS was highly dependent on LNOx. Normalized excess mixing ratios, ΔX/ΔCO, for HCN and ACN were determined in both the fire plume and the storm outflow and found to be 7.0±0.5 and 2.3±0.5pptvppbv-1, respectively, and 1.4±0.3pptvppbv-1 for acrolein in the outflow only

Super-resolving phase measurements with a multi-photon entangled state

Using a linear optical elements and post-selection, we construct an entangled polarization state of three photons in the same spatial mode. This state is analogous to a ``photon-number path entangled state'' and can be used for super-resolving interferometry. Measuring a birefringent phase shift, we demonstrate two- and three-fold improvements in phase resolution.Comment: 4 pages, 3 figure