1,721,097 research outputs found
A blend of botanicals reduces LPS-induced inflammation and disruptive effects on apical-out porcine enteroids
No full textTo better study host-microbial interactions or stressful conditions that affect the porcine intestinal epithelium in-vitro it is possible to use porcine apical-out enteroids, that better represent the intestinal epithelium compared to enterocytes monolayers or basal-out organoids. Botanicals are an interesting class of bioactive compounds with antioxidant and anti-inflammatory properties that help to support intestinal health.
The aim of this study was to investigate the ability of a thymol-based blend of botanicals (BOT) to reduce the disruptive effects of LPS-induced inflammation on apical-out porcine enteroids.
Intestinal crypts were obtained from 35 days-old pigs’ jejunum and they were used to generate apical-out enteroids. Those were then challenged with lipopolysaccharide (LPS) for 6h, in the absence or presence of BOT. FD4 paracellular permeability and gene expression were then evaluated.
Apical-out porcine enteroids responded to the LPS challenge by showing a 70% increase in FD4 paracellular-permeability, a 2-fold increase in IL1β, IL8, and mucin 4 (MUC4) levels (p<0.01), and a 3-fold increase in IL6 mRNA expression (p<0.01). Moreover, a decrease in zonula occludens 1 (ZO1) and occludin (OCCL) levels was observed. BOT showed beneficial properties in reducing all the effects connected to the LPS challenge. FD4 permeability was decreased by 35% (p<0.05), and IL1β, IL6, IL8, and MUC4 expression were significantly reduced compared to the challenged control. Moreover, BOT increased the ZO1 and OCCL levels by 50% (p<0.05), acting as a barrier-reinforcing agent.
BOT showed interesting anti-inflammatory properties, being able to maintain epithelial integrity thus reducing LPS-induced damage in-vitro. This thymol-based blend has the potential to be further investigated as a feed additive to improve the pigs’ ability to overcome stressful phases during their lifecycle
A blend of botanicals can reduce LPS-induced inflammation and disruptive effects on apical-out chicken and porcine enteroids
No full textIn-vitro studies about the efficacy of feed additives to fight intestinal inflammation or pathogens are usually performed on 2D primary enterocyte monolayers or immortalized cells (1). Enterocyte monolayers are useful but they lack the different epithelial components, present in the intestine, which are essential to better study the various properties of botanicals. For this purpose, intestinal organoids could be a valid alternative. Nash and colleagues (2021) published a study about the deep characterization of chicken apical-out enteroids (2) and recently, Joo and colleagues (2022) demonstrated that by sub-culturing basal-out pig enteroids on ultra-low attachment plates it is possible to obtain porcine apical-out enteroids (3), which are both ideal for in-vitro functional studies.
The aim of this study was to investigate the ability of a thymol-based blend of botanicals (BOT) to reduce the disruptive effects of LPS-induced inflammation on apical-out chicken and porcine enteroids.
Methods
To obtain chicken enteroids, intestines of 18day-old chicken embryos were recovered and digested with collagenase type I. The digested tissue was filtered through cell-strainers to recover the >40 μm fraction. The obtained cell aggregates were then cultured in suspension for 4 days with a proper organoid floating medium to generate mature chicken enteroids. To obtain pig enteroids, intestinal crypts were obtained from 35 days-old pigs’ jejunum using EDTA. Recovered intestinal crypts were cultured embedded in extracellular matrix for 2 days to allow the formation of basal-out organoids. Then, on day 3 basal-out enteroids were recovered and cultured for 24h in suspension on ultra-low attachment plates to generate apical-out organoids.
Mature enteroids of both species were then challenged with lipopolysaccharide (LPS) for 6h, in the absence or presence of BOT. FD4 paracellular permeability (PCP) and gene expression for selected markers were then evaluated.
Results
Chicken enteroids responded to the LPS challenge by showing a 60% increase in FD4 PCP, a near 60-fold increase in IL1β expression, a 4-fold increase in IL6, and IL8 levels (p<0.01), a 7-fold increase in tumor necrosis factor-α (TNFα) mRNA expression (p<0.01) and a 30-fold increase in interferon-γ (INFγ - p<0.01) expression. Moreover, a decrease in zonula occludens 1 (ZO1 - p<0.05), occludin (OCCL - p<0.05), and toll-like receptor 4 mRNA levels were observed (p<0.01). BOT showed anti-inflammatory properties in reducing all the effects connected to the LPS challenge. FD4 PCP was decreased by 40% (p<0.05), and the expression of all pro-inflammatory markers was significantly reduced compared to the challenged control. BOT restored the ZO1 and OCCL levels near the control levels (p<0.05). Moreover, BOT significantly increased the defensin beta 4a (DEFB4A) expression over the negative control (p<0.05). This could be an interesting effect to improve the defense ability of the host against pathogens.
Furthermore, apical-out porcine enteroids also well responded to the inflammatory challenge by showing a 70% increase in FD4 PCP, a 2-fold increase in IL1β, IL8, and mucin 4 (MUC4) levels (p<0.01), and a 3-fold increase in IL6 mRNA expression (p<0.01). Moreover, a decrease in ZO1 and occludin OCCL levels was observed. BOT showed beneficial properties in reducing all the effects connected to the LPS challenge. FD4 PCP was decreased by 35% (p<0.05), and IL1β, IL6, IL8, and MUC4 expression were significantly reduced compared to the challenged control. Moreover, BOT increased the ZO1 and OCCL levels by 50% (p<0.05), acting as a barrier-reinforcing agent.
Conclusion
BOT showed interesting anti-inflammatory properties both on chicken and pig enteroids, being able to maintain epithelial integrity thus reducing LPS-induced damage in-vitro. This thymol-based blend has the potential to be further investigated as a feed additive to improve chickens and pigs ability to overcome stressful phases during their lifecycle
In-ovo injections of Glutamine and Sodium Butyrate Regulate Intestinal Epithelium Development in a Broiler Intestinal Organoid Model
No full textEarly life events play significant roles in the tissue development and animal’s health in their later life.
Early nutrition, through in ovo delivery, has shown beneficial effects on improving intestinal health in broiler chickens.
However, the underlying mechanism is not fully investigated. The proliferation and differentiation of intestinal epithelial
stem cells (IESCs) and progenitor cells govern the rapid development and functions of intestinal epithelium. A recently
developed 3D intestinal organoids culture technique allows investigations on intestinal epithelial functions that are
close to the physiological conditions. In this study, we evaluated the effects of in ovo feeding of two well-known gut
beneficial nutrients, glutamine (Gln) or sodium butyrate (SB), on intestinal epithelial development and functions by
using intestinal enteroids culture. A hundred eggs of commercial Cobb500 broilers were injected with 0.2 ml of either
PBS, 3% Gln, or 1%SB on day E18. Chicks were euthanized at the day of hatch. The small intestines were sampled
for epithelial isolation and enteroids culture. The enteroids formation and growth were monitored by microscope. After
4 days of culture, the enteroids were harvested for FITC-dextran (FD4) permeability analysis, and gene expression
analysis of epithelial cell markers and functional proteins. One-way ANOVA and Student’s T test were used for
statistical analysis. Our results indicated that in ovo injection of both Gln and SB significantly (P < 0.05) increased the
enteroids formation rate compared with PBS control. In ovo SB injection significantly (P < 0.05)
increased the gene expressions of nutrient transporters (B0AT1, SGLT1, EAAT3) and tight junction proteins (TJP1,
ZO2) compared with the PBS control group. Moreover, both Gln and SB treatments significantly (P < 0.01)
reduced the FD4 permeability of the cultured enteroids. The gene expressions of epithelial cell markers were
analyzed. In comparison with PBS control, ChgA (enteroendocrine cell) was upregulated (P < 0.1) in Gln group,
MUC2 (goblet cells) was upregulated (P < 0.05) in SB group, while LYZ (Paneth cells) was down regulated in
SB group, Lgr5 (IESCs) were numerically downregulated in both of Gln and SB groups. Together, our results
suggested that the in ovo injection of Gln or SB simulated intestinal epithelium proliferation, and programmed
the epithelial cell differentiation towards absorptive cells, which may improve the early post-hatch feed adaptation.
Understanding the programming effects of early nutrition would shed lights on developing novel feeding strategies for
improving gut health
Thymol and thyme essential oil can reduce LPS-induced inflammation and disruptive effects on apical-out chicken enteroids
No full textIn vitro studies on efficacy of feed additives in combating intestinal inflammation or pathogens are typically conducted on 2D enterocyte monolayers. While these monolayers are useful, they lack the different cell types present in the intestinal mucosa, which are essential for studying the diverse properties of botanicals. To address this issue, intestinal apical-out organoids could be a viable alternative. The objective of this study was to investigate the ability of thymol (THY) and thyme essential oil (TEO) to mitigate the disruptive effects of LPS-induced inflammation on apical-out chicken enteroids.
Enteroids were obtained from the intestines of 18-day-old chicken embryos. The tissues were recovered, digested with collagenase and filtered through cell strainers to recover the >40 μm fraction. The obtained cell aggregates were then cultured in suspension for 4 days with an organoid floating medium to generate mature enteroids. They were then challenged with lipopolysaccharide (LPS – n=6) for 6 h, in the absence or presence of THY 10 ppm (n=6) and TEO 20 ppm (n=6). A negative control (CTR- - n=6) group, which was neither challenged nor treated, was also included. Fluorescein isothiocyanate-4 kDa dextran (FD4) paracellular permeability (PCP), reported as relative fluorescence units (RFU), and gene expression for selected markers were then evaluated. Data were analyzed using one-way ANOVA with Tukey’s multiple comparisons (p<0.05).
The enteroids responded to the LPS challenge by exhibiting a 2.46-fold increase in FD4 PCP compared to the CTR- (CTR- 1680 ± 330 vs LPS 4121 ± 721 – p<0.0001) RFU, as well as an increase in mRNA levels of interleukin (IL)1β (60-fold), IL6, and IL8 (both 4-fold), tumor necrosis factor-α (7-fold), and interferon-γ (30-fold) (p<0.01). Moreover, a significant (p<0.05) decrease in zonula occludens 1 (ZO1) and occludin (OCCL) expression was observed.
THY and TEO reduced the effects associated with the LPS challenge. FD4 PCP was significantly decreased by 30% with THY (LPS 4121 ± 721 vs THY 2896 ± 667 – p=0.0002) RFU and by 40% with TEO (LPS 4121 ± 721 vs TEO 2511 ± 371 – p<0.0001) RFU. Furthermore, the gene expression of all the pro-inflammatory markers was significantly reduced compared to LPS (p<0.05). THY and TEO restored ZO1 and OCCL mRNA levels to CTR- values (p<0.05). Finally, THY significantly increased the expression of defensin beta 4a, avian beta-defensin 3, and cathelicidin-2 mRNA expression closer to the CTR- (p<0.05).
In conclusion, THY and TEO exhibited intriguing properties by reducing LPS-induced inflammation and damage in vitro. These two botanicals have the potential to be used as feed additives to help chickens overcome stressful phases during their lifecycle
Going Beyond Counting First Authors in Author Co-citation Analysis
Get PDFThe 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
Variations on the Author
Get PDF“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
Get PDFWe 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
Get PDFWe 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
- …
