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1.
Minh H. Dinh Meegan R. Anderson Michael D. McRaven Gianguido C. Cianci Scott G. McCoombe Z. L. Kelley Casey J. Gioia Angela J. Fought Alfred W. Rademaker Ronald S. Veazey Thomas J. Hope 《PLoS pathogens》2015,11(3)
To gain insight into female-to-male HIV sexual transmission and how male circumcision protects against this mode of transmission, we visualized HIV-1 interactions with foreskin and penile tissues in ex vivo tissue culture and in vivo rhesus macaque models utilizing epifluorescent microscopy. 12 foreskin and 14 cadaveric penile specimens were cultured with R5-tropic photoactivatable (PA)-GFP HIV-1 for 4 or 24 hours. Tissue cryosections were immunofluorescently imaged for epithelial and immune cell markers. Images were analyzed for total virions, proportion of penetrators, depth of virion penetration, as well as immune cell counts and depths in the tissue. We visualized individual PA virions breaching penile epithelial surfaces in the explant and macaque model. Using kernel density estimated probabilities of localizing a virion or immune cell at certain tissue depths revealed that interactions between virions and cells were more likely to occur in the inner foreskin or glans penis (from local or cadaveric donors, respectively). Using statistical models to account for repeated measures and zero-inflated datasets, we found no difference in total virions visualized at 4 hours between inner and outer foreskins from local donors. At 24 hours, there were more virions in inner as compared to outer foreskin (0.0495 +/− 0.0154 and 0.0171 +/− 0.0038 virions/image, p = 0.001). In the cadaveric specimens, we observed more virions in inner foreskin (0.0507 +/− 0.0079 virions/image) than glans tissue (0.0167 +/− 0.0033 virions/image, p<0.001), but a greater proportion was seen penetrating uncircumcised glans tissue (0.0458 +/− 0.0188 vs. 0.0151 +/− 0.0100 virions/image, p = 0.099) and to significantly greater mean depths (29.162 +/− 3.908 vs. 12.466 +/− 2.985 μm). Our in vivo macaque model confirmed that virions can breach penile squamous epithelia in a living model. In summary, these results suggest that the inner foreskin and glans epithelia may be important sites for HIV transmission in uncircumcised men. 相似文献
2.
Cristiana Leite N. Tatiana Silva Sandrine Mendes Andreia Ribeiro Joana Paes de Faria Tania Louren?o Francisco dos Santos Pedro Z. Andrade Carla M. P. Cardoso Margarida Vieira Artur Paiva Cláudia L. da Silva Joaquim M. S. Cabral Jo?o B. Relvas Mário Gr?os 《PloS one》2014,9(10)
Mesenchymal stem cells (MSCs) are viewed as safe, readily available and promising adult stem cells, which are currently used in several clinical trials. Additionally, their soluble-factor secretion and multi-lineage differentiation capacities place MSCs in the forefront of stem cell types with expected near-future clinical applications. In the present work MSCs were isolated from the umbilical cord matrix (Wharton''s jelly) of human umbilical cord samples. The cells were thoroughly characterized and confirmed as bona-fide MSCs, presenting in vitro low generation time, high proliferative and colony-forming unit-fibroblast (CFU-F) capacity, typical MSC immunophenotype and osteogenic, chondrogenic and adipogenic differentiation capacity. The cells were additionally subjected to an oligodendroglial-oriented step-wise differentiation protocol in order to test their neural- and oligodendroglial-like differentiation capacity. The results confirmed the neural-like plasticity of MSCs, and suggested that the cells presented an oligodendroglial-like phenotype throughout the differentiation protocol, in several aspects sharing characteristics common to those of bona-fide oligodendrocyte precursor cells and differentiated oligodendrocytes. 相似文献
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Antonio Hernandes Chaves Neto Karla Cristiana Queiroz Renato Milani Edgar Julian Paredes‐Gamero Giselle Zenker Justo Maikel P. Peppelenbosch Carmen Veríssima Ferreira 《Journal of cellular biochemistry》2011,112(1):71-77
Despite numerous reports on the ability of ascorbic acid and β‐glycerophosphate (AA/β‐GP) to induce osteoblast differentiation, little is known about the molecular mechanisms involved in this phenomenon. In this work, we used a peptide array containing specific consensus sequences (potential substrates) for protein kinases and traditional biochemical techniques to examine the signaling pathways modulated during AA/β‐GP‐induced osteoblast differentiation. The kinomic profile obtained after 7 days of treatment with AA/β‐GP identified 18 kinase substrates with significantly enhanced or reduced phosphorylation. Peptide substrates for Akt, PI3K, PKC, BCR, ABL, PRKG1, PAK1, PAK2, ERK1, ERBB2, and SYK showed a considerable reduction in phosphorylation, whereas enhanced phosphorylation was observed in substrates for CHKB, CHKA, PKA, FAK, ATM, PKA, and VEGFR‐1. These findings confirm the potential usefulness of peptide microarrays for identifying kinases known to be involved in bone development in vivo and in vitro and show that this technique can be used to investigate kinases whose function in osteoblastic differentiation is poorly understood. J. Cell. Biochem. 112: 71–77, 2011. © 2010 Wiley‐Liss, Inc. 相似文献
6.
Laura Roesler Nery Natalia Silva Eltz Cristiana Hackman Raphaela Fonseca Stefani Altenhofen Heydi Noriega Guerra Vanessa Morais Freitas Carla Denise Bonan Monica Ryff Moreira Roca Vianna 《PloS one》2014,9(9)
Alzheimer’s disease (AD) is a devastating neurodegenerative disorder with no effective treatment and commonly diagnosed only on late stages. Amyloid-β (Aβ) accumulation and exacerbated tau phosphorylation are molecular hallmarks of AD implicated in cognitive deficits and synaptic and neuronal loss. The Aβ and tau connection is beginning to be elucidated and attributed to interaction with different components of common signaling pathways. Recent evidences suggest that non-fibrillary Aβ forms bind to membrane receptors and modulate GSK-3β activity, which in turn phosphorylates the microtubule-associated tau protein leading to axonal disruption and toxic accumulation. Available AD animal models, ranging from rodent to invertebrates, significantly contributed to our current knowledge, but complementary platforms for mechanistic and candidate drug screenings remain critical for the identification of early stage biomarkers and potential disease-modifying therapies. Here we show that Aβ1–42 injection in the hindbrain ventricle of 24 hpf zebrafish embryos results in specific cognitive deficits and increased tau phosphorylation in GSK-3β target residues at 5dpf larvae. These effects are reversed by lithium incubation and not accompanied by apoptotic markers. We believe this may represent a straightforward platform useful to identification of cellular and molecular mechanisms of early stage AD-like symptoms and the effects of neuroactive molecules in pharmacological screenings. 相似文献
7.
Cândido Barreto de Novais Cristiana Sbrana Orivaldo José Saggin Júnior José Oswaldo Siqueira Manuela Giovannetti 《Mycorrhiza》2013,23(4):325-331
Hyphal anastomoses which play a key role in the formation of interconnected mycorrhizal networks and in genetic exchange among compatible individuals have been studied in a limited number of species and isolates of arbuscular mycorrhizal fungi (AMF), mainly in symbiotic mycelium. In this work, the occurrence and frequency of anastomosis between hyphae of the same and different germlings were assessed in tropical isolates belonging to Acaulospora, Claroideoglomus, Gigaspora, Glomus, Rhizophagus and Scutellospora. Germlings belonging to Acaulospora, Claroideoglomus, Glomus and Rhizophagus formed perfect hyphal fusions, with frequencies ranging from 9.29?±?3.01 to 79.84?±?4.39 % within the same germling and from 14.02?±?7.36 to 91.41?±?3.92 % between different germlings. Rare fusions, occurring within the same hypha, were detected in Gigaspora species, and no anastomoses were observed in Scutellospora species. The consistent detection of nuclei in perfect fusions suggests that nuclear migration is active both within and between germlings. Present data on anastomosis formation, nuclear migration and germling viability in tropical isolates of AMF widen our knowledge on the extensive and consistent occurrence of successful hyphal fusions in this group of beneficial symbionts. The ability to anastomose and establish protoplasm flow, fundamental for the maintenance of physiological and genetic continuity, may produce important fitness consequences for the obligately biotrophic AMF. 相似文献
8.
Marcella Barbarino Daniele Cesari Maria Bottaro Luca Luzzi Asadoor Namagerdi Franca Maria Bertolino Cristiana Bellan Fabrizio Proietti Pasquale Somma Mariacarolina Micheli Maria Margherita de Santi Raffaella Guazzo Luciano Mutti Luigi Pirtoli Piero Paladini Paola Indovina Antonio Giordano 《Journal of cellular and molecular medicine》2020,24(10):5565-5577
Malignant mesothelioma (MM) is an aggressive asbestos-related cancer of the serous membranes. Despite intensive treatment regimens, MM is still a fatal disease, mainly due to the intrinsic resistance to current therapies and the lack of predictive markers and new valuable molecular targets. Protein arginine methyltransferase 5 (PRMT5) inhibition has recently emerged as a potential therapy against methylthioadenosine phosphorylase (MTAP)-deficient cancers, in which the accumulation of the substrate 5'-methylthioadenosine (MTA) inhibits PRMT5 activity, thus sensitizing the cells to further PRMT5 inhibition. Considering that the MTAP gene is frequently codeleted with the adjacent cyclin-dependent kinase inhibitor 2A (CDKN2A) locus in MM, we assessed whether PRMT5 could represent a therapeutic target also for this cancer type. We evaluated PRMT5 expression, the MTAP status and MTA content in normal mesothelial and MM cell lines. We found that both administration of exogenous MTA and stable PRMT5 knock-down, by short hairpin RNAs (shRNAs), selectively reduced the growth of MTAP-deleted MM cells. We also observed that PRMT5 knock-down in MTAP-deficient MM cells reduced the expression of E2F1 target genes involved in cell cycle progression and of factors implicated in epithelial-to-mesenchymal transition. Therefore, PRMT5 targeting could represent a promising new therapeutic strategy against MTAP-deleted MMs. 相似文献
9.
Pinto José de Azevedo Cristiana Rodrigues Oliveira Rui von Stosch Moritz 《Bioprocess and biosystems engineering》2019,42(11):1853-1865
Bioprocess and Biosystems Engineering - Hybrid semi-parametric modeling, combining mechanistic and machine-learning methods, has proven to be a powerful method for process development. This paper... 相似文献
10.
Mutational and epigenetic driver events profoundly alter intercellular communication pathways in cancer. This effect includes deregulated release, molecular composition, and biological activity of extracellular vesicles (EVs), membranous cellular fragments ranging from a few microns to less than 100 nm in diameter and filled with bioactive molecular cargo (proteins, lipids, and nucleic acids). While EVs are usually classified on the basis of their physical properties and biogenetic mechanisms, recent analyses of their proteome suggest a larger than expected molecular diversity, a notion that is also supported by multicolour nano‐flow cytometry and other emerging technology platforms designed to analyze single EVs. Both protein composition and EV diversity are markedly altered by oncogenic transformation, epithelial to mesenchymal transition, and differentiation of cancer stem cells. Interestingly, only a subset of EVs released from mutant cells may carry oncogenic proteins (e.g., EGFRvIII), hence, these EVs are often referred to as “oncosomes”. Indeed, oncogenic transformation alters the repertoire of EV‐associated proteins, increases the presence of pro‐invasive cargo, and alters the composition of distinct EV populations. Molecular profiling of single EVs may reveal a more intricate effect of transforming events on the architecture of EV populations in cancer and shed new light on their biological role and diagnostic utility. 相似文献