1,720,984 research outputs found
Effect of Estradiol and Progesterone on ovine Amniotic Epithelial Cells
Full text linkThis study was designed to clarify Estradiol (E2) and Progesterone (P4) steroid effects on ovine Amniotic Epithelial
Cells (oAECs) that has a conserved plasticity and highly self-renewable capacity (Parolini et al., Stem Cells, 26(2),
300–311, 2008; Barboni et al., Stem Cell Rev Rep, 10:725–741, 2014). Based on their conserved
immunomodulatory properties, oAECs are suitable for allo and xeno-transplantation (Barboni et al., Cell Transplant,
21(11), 2377–2395, 2012; Muttini et al., Res Vet Sci, 94(1),158–169, 2013). To date, no information is present on
the effects of prolonged steroid exposition on AECs. oAECs were cultured as previously reported (Barboni et al.,
Cell Transplant. 21(11), 2377–2395, 2012) and treated with 12.5μM and 25μM of E2 or P4 (Sigma-Aldrich, Milan,
Italy), alone and in both combinations, for three passages. Untreated cells were marked control (CTR). At 70%
confluency, cells were detached for doubling time (DT) evaluation. Cells at fourth passage were differentiated for 21
days in osteogenic media (DM) (Mattioli et al., Cell Biol Int 36(1):7-19, 2012) without steroid. Alizarin Red and
Alcian-Blue (Sigma-Aldrich, Milano, Italy) stainings were performed. RNA and cDNA were obtained as previously
reported (Barboni et al., Cell Transplant. 21(11), 2377–2395, 2012). Real Time for NANOG, SOX2 ,OCT4 stemness
genes expression were performed by SensiFast SYBR (Bioline, Aurogene, Rome, Italy) using specific primers
(Mattioli et al., Cell Biol Int. 36(1):7-19, 2012).The protocol was: 5 min at 95°C, 30 cycles at 95°C for 15 sec, 60°C
for 30 sec, 72°C for 15 sec. Comparative Ct 2-ΔΔC(t) normalization to GAPDH was applied. IHC analyses were
carried out for Cytokeratin 8 and αSMA expression as previously reported (Barboni et al. PLoS ONE 7(2): e30974,
2012). Data expressed as mean (±SD), compared by one-way ANOVA followed by Tukey’s test (GraphPad Prism
5). Significant values for P < 0.05. Steroids treated ovine AECs proliferate with significant differences between
concentrations. While P4 treated cells showed cuboidal shape and Cytokeratin expression until third passage, CTR
and E2 treated cells showed a rapid downregulation of Cytokeratin and increased αSMA expression. oAECs with
E2+P4 showed both cell type morphology. Steroids modified stemness genes based on the concentration. 12.5 μM
E2, 25μM P4 and 25μM of both E2+P4 treatments maintained higher OCT4, NANOG and SOX2 expressions in
treated cells despite their progressive downregulation in the CTR. Moreover, compared to CTR, after Alizarin
staining, steroid pretreated cells suffered morphological changes under DM acquiring Alcian Blue-positive
chondrogenic-like morphology. AECs stemness properties and plasticity can be modified by prolonged steroidal
treatment. These data improve our knowledge, opening new prospective on oAEC use in stem cell-based therapy.
Acknowledgments. Research supported by H2020-MSCA ITN EJD-REP BIOTECH 675526
Going Beyond Counting First Authors in Author Co-citation Analysis
Full text linkThe present study examines one of the fundamental aspects of author co-citation analysis (ACA) - the way co-citation
counts are defined. Co-citation counting provides the data on which all subsequent statistical analyses and mappings
are based, and we compare ACA results based on two different types of co-citation counting - the traditional type that
only counts the first one among a cited work's authors on the one hand and a non-traditional type that takes into
account the first 5 authors of a cited work on the other hand. Results indicate that the picture produced through this non-traditional author co-citation counting contains more coherent author groups and is therefore considerably clearer. However, this picture represents fewer specialties in the research field being studied than that produced through the traditional first-author co-citation counting when the same number of top-ranked authors is selected and analyzed. Reasons for these effects are discussed
The Epithelial-Mesenchymal-Tendon Transition State of Amniotic Epithelial Stem Cells Influences Tendon Regeneration: Insights from an Experimental Sheep Achilles Tendon Injury Model
No full textVariations on the Author
Full text link“Variations on the Author” discusses two of Eduardo Coutinho’s recent films (Um Dia na Vida, from 2010, and Últimas Conversas, posthumously released in 2015) and their contribution to the general question of documentary authorship. The director’s filmography is characterized by a consistent yet self-effacing form of authorial self-inscription: Coutinho often features as an interviewer that rather than express opinions propels discourses; an interviewer that is good at listening. This mode of self-inscription characterizes him as an author who is not expressive but who is nonetheless markedly present on the screen. In Um Dia na Vida, however, Coutinho is completely absent form the image, while Últimas Conversas, on the contrary, includes a confessional prologue that moves the director from the margins to the center of his films. This article examines the ways in which these works stand out in the filmography of a director who offers new insights into the notion of cinematic authorship
Appropriate Similarity Measures for Author Cocitation Analysis
Full text linkWe provide a number of new insights into the methodological discussion about author cocitation analysis. We first argue that the use of the Pearson correlation for measuring the similarity between authors’ cocitation profiles is not very satisfactory. We then discuss what kind of similarity measures may be used as an alternative to the Pearson correlation. We consider three similarity measures in particular. One is the well-known cosine. The other two similarity measures have not been used before in the bibliometric literature. Finally, we show by means of an example that our findings have a high practical relevance.information science;Pearson correlation;cosine;similarity measure;author cocitation analysis
Dispelling the Myths Behind First-author Citation Counts
Full text linkWe conducted a full-scale evaluative citation analysis study of scholars in the XML research field to explore just how different from each other author rankings resulting from different citation counting methods actually are, and to demonstrate the capability of emerging data and tools on the Web in supporting more realistic citation counting methods. Our results contest some common arguments for the continued
use of first-author citation counts in the evaluation of scholars, such as high correlations between author rankings by first-author citation counts and other citation
counting methods, and high costs of using more realistic citation counting methods that are not well-supported by the ISI databases. It is argued that increasingly available digital full text research papers make it possible for citation analysis studies to go beyond what the ISI databases have directly supported and to employ more
sophisticated methods
Opposing Roles of KLHL14 and X1 Isoforms in the Regulation of E-Cadherin-mediated Epithelial-Mesenchymal Transition
No full textKLHL14 belongs to the Kelch-like (KLHL) family of E3 ubiquitin ligases. Proteins of this family share a conserved dimerization domain (BTB), a linker domain (BACK), and a C-terminal Kelch domain responsible for substrate recognition [1]. KLHL14 has recently emerged as a tumor suppressor in several cancers [2, 3]. Although it was originally implicated in the inhibition of epithelial-mesenchymal transition (EMT) in amniotic epithelial stem cells (AECs), its precise role and molecular mechanisms remain unclear [4]. Understanding KLHL14 function is highly relevant, as EMT is involved not only in cancer progression but also in stem cell development and tissue regeneration. Moreover, KLHL14 is expressed as two isoforms: the full-length form (KLHL14) and a short splice variant known as X1, which lacks the substrate-binding domain. However, the role of X1 has remained completely unexplored to date.
To elucidate the roles of KLHL14 and X1 in EMT, we employed two complementary models: AECs, a physiological EMT model, and HepG2, an epithelial hepatocarcinoma cell line with high endogenous KLHL14 expression. We investigated the contribution of both isoforms through gain- and loss-of-function approaches using c-Myc-tagged overexpression constructs and isoform-specific siRNA-mediated silencing.
Our results show that both isoforms contribute to the maintenance of epithelial identity through modulation of E-Cadherin, via two distinct mechanisms. Full-length KLHL14, predominantly localized at the plasma membrane in AECs and HepG2 cells, co-localized with E-Cadherin at adherens junctions. This result was further confirmed in situ in the whole amniotic membrane (AM). In contrast, X1 exhibited an endoplasmic reticulum (ER)-like pattern. Co-immunoprecipitation confirmed the specific interaction of both isoforms with E-Cadherin. This interaction was also observed in the AM, supporting the physiological relevance of our findings. Mechanistically, KLHL14 promotes E-Cadherin degradation via the ubiquitin-proteasome and lysosomal pathways, whereas X1 enhances its de novo synthesis. Functionally, silencing of both isoforms leads to overall E-Cadherin downregulation, increased cell proliferation, and EMT induction. Remarkably, these effects are conserved in the native AM, where silencing reduces E-Cadherin expression, disrupts adherens junctions, and increases Vimentin levels.
Overall, this study reveals the opposing functions of KLHL14 and X1 in balancing E-Cadherin-mediated EMT regulation. While KLHL14 promotes E-Cadherin degradation, X1 supports its expression, together modulating epithelial stability. These findings uncover a previously unrecognized regulatory axis and underscore KLHL14 as a crucial pivot for epithelial integrity and plasticity. Elucidating this mechanism opens new perspectives for targeting EMT in regenerative strategies and anti-cancer therapies, where reprogramming cell fate and preventing aberrant transitions remain major clinical challenges
koamabayili/VECTRON-author-checklist: VECTRON author checklist
No full textWe have done our best to complete the author checklist relating to the use of animals in the hut study. Note that the objective for the hut study was to evaluate the IRS treatment applications for residual efficacy against Anopheles mosquitoes, including the local An. coluzzii mosquito population. Cows were only used to attract mosquitoes into the huts and no tests were carried out directly on the cows. The author checklist is intended for use with studies where experiments are carried out on animals, which is why we have had such difficulty in completing this for the hut study, as many of the questions do not relate to how the cows were used
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