19 documents found, page 1 of 2
PD-1 and ICOS are coexpressed in T follicular helper cells but define three sta...
Ribeiro, Filipa; Antunes, Diogo; Pires, Ana R.; Rino, José; Filipe, Beatriz; Jesus, Kátia; Correia, Ricardo; Fonseca, Válter R; Kumar, SaumyaHumoral responses to infection or vaccination require T cell-B cell interactions. T follicular helper (TFH) cells drive germinal center (GC) responses by providing help to B cells, whereas T follicular regulatory (TFR) cells regulate them. Both mature GC-located TFH and TFR cells are widely characterized by the expression of ICOS and PD-1. However, although human TFR cells share many phenotypic characteristics ...
A new protocol for whole-brain biodistribution analysis of AAVs by tissue clear...
Lopes, Miguel M.; Paysan, Jacques; Rino, José; Lopes, Sara M.; Pereira de Almeida, Luís; Cortes, Luísa; Nobre, Rui JorgeRecombinant adeno-associated virus (rAAV) has become one of the most promising gene delivery systems for both in vitro and in vivo applications. However, a key challenge is the lack of suitable imaging technologies to evaluate delivery, biodistribution and tropism of rAAVs and efficiently monitor disease amelioration promoted by AAV-based therapies at a whole-organ level with single-cell resolution. Therefore, ...
Low-dose ionizing radiation induces therapeutic neovascularization in a pre-cli...
Ministro, Augusto; Oliveira, Paula de; Nunes, Raquel J.; Rocha, André dos Santos; Correia, Adriana; Carvalho, Tânia; Rino, José; Faísca, PedroAims: We have previously shown that low-dose ionizing radiation (LDIR) induces angiogenesis but there is no evidence that it induces neovascularization in the setting of peripheral arterial disease. Here, we investigated the use of LDIR as an innovative and non-invasive strategy to stimulate therapeutic neovascularization using a model of experimentally induced hindlimb ischemia (HLI). Methods and results: Afte...
Quantification of cell cycle kinetics by EdU (5-ethynyl-2′-deoxyuridine)-couple...
Pereira, Pedro D.; Caetano, Ana Serra; Cabrita, Marisa; Bekman, Evguenia; Braga, José; Rino, José; Santus, Renè; Filipe, Paulo L.; Sousa, Ana E.We propose a novel single-deoxynucleoside-based assay that is easy to perform and provides accurate values for the absolute length (in units of time) of each of the cell cycle stages (G1, S and G2/M). This flow-cytometric assay takes advantage of the excellent stoichiometric properties of azide-fluorochrome detection of DNA substituted with 5-ethynyl-2'-deoxyuridine (EdU). We show that by pulsing cells with EdU...
Pharmacological inhibition of the spliceosome subunit SF3b triggers EJC-indepen...
Carvalho, Teresa; Martins, Sandra; Rino, José; Marinho, Sérgio; Carmo-Fonseca, MariaSpliceostatin A, meayamycin, and pladienolide B are small molecules that target the SF3b subunit of the spliceosomal U2 small nuclear ribonucleoprotein (snRNP). These compounds are attracting much attention as tools to manipulate splicing and for use as potential anti-cancer drugs. We investigated the effects of these inhibitors on mRNA transport and stability in human cells. Upon splicing inhibition, unspliced...
Decrease of CD68 Synovial Macrophages in Celastrol Treated Arthritic Rats
Cascão, Rita; Vidal, Bruno; Lopes, Inês P.; Paisana, Eunice; Rino, José; Moita, Luis F.; Fonseca, João E.Rheumatoid arthritis (RA) is a chronic immune-mediated inflammatory disease characterized by cellular infiltration into the joints, hyperproliferation of synovial cells and bone damage. Available treatments for RA only induce remission in around 30% of the patients, have important adverse effects and its use is limited by their high cost. Therefore, compounds that can control arthritis, with an acceptable safet...
Decrease of CD68 synovial macrophages in celastrol treated arthritic rats
Cascao, Rita; Vidal, Bruno; Lopes, Inês; Paisana, Eunice; Rino, José; Moita, Luis; Fonseca, João EuricoBackground: Rheumatoid arthritis (RA) is a chronic immune-mediated inflammatory disease characterized by cellular infiltration into the joints, hyperproliferation of synovial cells and bone damage. Available treatments for RA only induce remission in around 30% of the patients, have important adverse effects and its use is limited by their high cost. Therefore, compounds that can control arthritis, with an acce...
Adenosine A2A receptors modulate α-Synuclein aggregation and toxicity
Ferreira, Diana G.; Batalha, Vânia; Vicente Miranda, Hugo; Coelho, Joana E; Gomes, Rui; Gonçalves, Francisco Q.; Real, Joana I.; Rino, JoséAbnormal accumulation of aggregated α-synuclein (aSyn) is a hallmark of sporadic and familial Parkinson's disease (PD) and related synucleinopathies. Recent studies suggest a neuroprotective role of adenosine A2A receptor (A2AR) antagonists in PD. Nevertheless, the precise molecular mechanisms underlying this neuroprotection remain unclear. We assessed the impact of A2AR blockade or genetic deletion (A2AR KO) o...
Synthesis, characterization and biological evaluation of carboranylmethylbenzo[...
Da Silva, A. Filipa F.; Seixas, Raquel S. G. R.; Silva, Artur M. S.; Coimbra, Joana; Fernandes, Ana C.; Santos, Joana P.; Matos, António; Rino, JoséHerein we present the synthesis and characterization of benzo[b]acridin-12(7H)-ones bearing carboranyl moieties and test their biological effectiveness as boron neutron capture therapy (BNCT) agents in cancer treatment. The cellular uptake of these novel compounds into the U87 human glioblastoma cells was evaluated by boron analysis (ICP-MS) and by fluorescence imaging (confocal microscopy). The compounds enter...
Live-cell visualization of pre-mRNA splicing with single-molecule sensitivity
Martin, Robert M.; Rino, José; Carvalho, Célia; Kirchhausen, Tomas; Carmo-Fonseca, MariaRemoval of introns from pre-messenger RNAs (pre-mRNAs) via splicing provides a versatile means of genetic regulation that is often disrupted in human diseases. To decipher how splicing occurs in real time, we directly examined with single-molecule sensitivity the kinetics of intron excision from pre-mRNA in the nucleus of living human cells. By using two different RNA labeling methods, MS2 and λN, we show that ...