scholarly journals Condensed Tannins in White Clover (Trifolium repens) Foliar Tissues Expressing the Transcription Factor TaMYB14-1 Bind to Forage Protein and Reduce Ammonia and Methane Emissions in vitro

2022 ◽  
Vol 12 ◽  
Author(s):  
Marissa B. Roldan ◽  
Greig Cousins ◽  
Stefan Muetzel ◽  
Wayne E. Zeller ◽  
Karl Fraser ◽  
...  
Keyword(s):  
Nitrous Oxide ◽  
Transcription Factor ◽  
White Clover ◽  
Trifolium Repens ◽  
Condensed Tannins ◽  
Animal Health ◽  
Methane Emissions ◽  
Nitrous Oxide Emissions

Grazing ruminants contribute to global climate change through enteric methane and nitrous oxide emissions. However, animal consumption of the plant polyphenolics, proanthocyanidins, or condensed tannins (CTs) can decrease both methane emissions and urine nitrogen levels, leading to reduced nitrous oxide emissions, and concomitantly increase animal health and production. CTs are largely absent in the foliage of important temperate pasture legumes, such as white clover (Trifolium repens), but found in flowers and seed coats. Attempts at enhancing levels of CT expression in white clover leaves by mutagenesis and breeding have not been successful. However, the transformation of white clover with the TaMYB14-1 transcription factor from Trifolium arvense has resulted in the production of CTs in leaves up to 1.2% of dry matter (DM). In this study, two generations of breeding elevated foliar CTs to >2% of DM. The CTs consisted predominantly of prodelphinidins (PD, 75–93%) and procyanidins (PC, 17–25%) and had a mean degree of polymerization (mDP) of approximately 10 flavan-3-ol subunits. In vitro studies showed that foliar CTs were bound to bovine serum albumin and white clover proteins at pH 6.5 and were released at pH 2.-2.5. Using rumen in vitro assays, white clover leaves containing soluble CTs of 1.6–2.4% of DM significantly reduced methane production by 19% (p ≤0.01) and ammonia production by 60% (p ≤ 0.01) relative to non-transformed wild type (WT) controls after 6 h of incubation. These results provide valuable information for further studies using CT expressing white clover leaves for bloat prevention and reduced greenhouse gas emissions in vivo.

scholarly journals Use of Tannin-Containing Plants as Antimicrobials Influencing the Animal Health

2022 ◽  
Vol 45 (2) ◽  
pp. 33-40
Author(s):  
Mohammed M Dakheel ◽  
Afnan A Al-Mnaser ◽  
Jessica Quijada ◽  
Martin J Woodward ◽  
Caroline Rymer
Keyword(s):  
Nutritional Quality ◽  
Condensed Tannins ◽  
Animal Health ◽  
Degree Of Polymerization ◽  
Gram Negative Bacteria ◽  
Animal Feeding ◽  
The Subject

The antimicrobial effects of diverse tannin-containing plants, particularly condensed tannins (CTs) produced from various plants, are the subject of this study. CT components can be determined using CT-specific procedures such the HCl-Butanol Acetone assay, Thiolysis reaction, and HPLC/MS analysis. These methods indicate CT contents, including mean degree of polymerization, the procyanidins and prodelphinidins ratio (PC/PD%), the isomers of trans- and cis-, and CT concentration. Tannin-containing plants possess antibacterial action, which can be attributed to their protein linkage technique, and tannin-type variations, particularly CTs extract and their PC/PD%. The effects of CT components on the development of Gram-positive and Gram-negative bacteria have been documented for their relative PC/PD%; this is regarded to be a key predictor of tannin characteristics in terms of antimicrobials. In conclusion, tannins, more specific CT compositions, have significant impacts on in vivo trials of animal productions and utilization of metabolites and fermentation in vitro experiments. These findings need further investigations to fully understand how CT-types act on animal feeding in terms of enhanced nutritional quality of animal diets, which may have implications for human and animal health.

scholarly journals Methane production from in vitro incubation of kikuyu grass, lucerne and forages containing condensed tannins

2005 ◽  
pp. 147-153 ◽  
Author(s):  
M.H. Tavendale ◽  
L.P. Meagher ◽  
Z.A. Park-Ng ◽  
G.C. Waghorn ◽  
G.T. Attwood
Keyword(s):  
Fatty Acid ◽  
New Zealand ◽  
Methane Production ◽  
Condensed Tannins ◽  
Volatile Fatty Acid ◽  
Dry Matter ◽  
Methane Emissions ◽  
Kikuyu Grass

A series of in vitro incubations with kikuyu grass (Pennisetum clandestinum), lucerne and six legumes containing condensed tannins (CT) were undertaken to evaluate this technique against in vivo trials in New Zealand, measuring methane emissions. Published trials have demonstrated a reduction in methane emissions associated with CT and in one instance from kikuyu. The incubations used fresh minced forage (equivalent to 0.5 g dry matter (DM)) and were carried out in 50 ml sealed bottles containing buffer and rumen inoculum. Gas was sampled through a septum to monitor volume and composition throughout the 24h incubation. Incubation for 24 h resulted in 2.4-6.6 % conversion of DM to methane, and suggested CT concentrations below about 8% of the DM can reduce methane production without inhibiting fermentation rate. Higher concentrations of CT (> 8%) were associated with a lower rate of digestion. In common with in vivo trials, CT concentration in forage DM was inversely related to methane (adjusted R2 = 0.49; P = 0.01) and volatile fatty acid (adjusted R2=0.86; P

Solubilization and degradation of ribulose-1,5-bisphosphate carboxylase/oxygenase (EC 4.1.1.39; Rubisco) protein from white clover (Trifolium repens) and Lotus corniculatus by rumen microorganisms and the effect of condensed tannins on these processes

2000 ◽  
Vol 134 (3) ◽  
pp. 305-317 ◽  
Author(s):  
B. R. MIN ◽  
W. C. MCNABB ◽  
T. N. BARRY ◽  
J. S. PETERS
Keyword(s):  
White Clover ◽  
Trifolium Repens ◽  
Condensed Tannins ◽  
Rumen Fluid ◽  
Lotus Corniculatus ◽  
Plant Protein ◽  
Bisphosphate Carboxylase ◽  
Fraction I

In situ and in vitro rumen incubations were used to determine the effect of condensed tannins (CT) on the solubilization and degradation of the plant protein from white clover (Trifolium repens) and Lotus corniculatus. These forages contained, respectively 0·3 and 22·1 g CT/kg dry matter (DM). The sheep used for the experiments were also fed either white clover or L. corniculatus. Effects of CT were determined by making measurements in the presence and absence of polyethylene glycol (PEG; molecular weight 3500), which binds and inactivates CT. The loss of DM, neutral detergent fibre (NDF), total nitrogen (N) and Rubisco (ribulose-1,5-bisphosphate carboxylase/oxygenase; EC 4.1.1.39; fraction I leaf protein) from polyester bags suspended in the rumen of sheep was measured. The loss of these constituents from polyester bags suspended in the rumen was used as a measurement of their solubilization. Degradation was defined as the disappearance of Rubisco from white clover and L. corniculatus added to in vitro incubations with rumen fluid obtained from the same fistulated sheep fed either white clover or L. corniculatus.In the absence of PEG, the in situ loss of Rubisco from L. corniculatus was less rapid than the loss of this protein from white clover when each forage was incubated in the rumen of sheep fed the same diet. Addition of PEG tended to increase the loss of Rubisco from L. corniculatus, suggesting that CT slowed the rates of solubilization of Rubisco from this forage. Effects of rumen fluid were small, but there was some evidence that the rumen fluid in sheep fed L. corniculatus reduced the solubilization of Rubisco from white clover. The action of CT did not inhibit the in situ loss of NDF from either white clover or L. corniculatus.In the absence of PEG, the in vitro degradation of Rubisco from L. corniculatus was slower when compared to the degradation of this protein from white clover; PEG addition increased the degradation of Rubisco from L. corniculatus, but not from white clover, showing that CT was the causal agent. The addition of CT extracted from L. corniculatus markedly depressed the degradation of Rubisco from white clover, with the effect being completely reversible by PEG. The large subunit (LSU) of Rubisco was consistently degraded at a faster rate than the small subunit (SSU) and added CT had a greater effect in slowing the degradation of the LSU compared to the SSU. There was little difference in the degradation of Rubisco when rumen fluid from sheep fed either white clover or L. corniculatus was used for in vitro incubations.It was concluded that the action of CT from L. corniculatus reduces the digestion of protein in the rumen of sheep. This effect is predominantly due to the action of CT reducing the degradation of plant protein, although CT also reduced the solubilization of plant protein. The main effects of CT on protein solubilization and degradation seemed to be produced locally by CT present in plant tissue; transfer of these effects through rumen fluid was small in magnitude.

scholarly journals The digestion of perennial ryegrass (Lolium perenne cv. Melle) and white clover (Trifolium repens cv. Blanca) by grazing cattle

1988 ◽  
Vol 60 (1) ◽  
pp. 137-149 ◽  
Author(s):  
M. J. Ulyatt ◽  
D. J. Thomson ◽  
D. E. Beever ◽  
R. T. Evans ◽  
M. J. Haines
Keyword(s):  
Small Intestine ◽  
Perennial Ryegrass ◽  
White Clover ◽  
Trifolium Repens ◽  
Water Soluble ◽  
Soluble Carbohydrate ◽  
Derived Relations ◽  
Primary Growth

1. Pure swards of perennial ryegrass (Loliwn perenne cv. Melle) as a primary growth (May), trimmed primary growth (early June) and regrowth (late June), and white clover (Trifolium repens cv. Blanca) as a mature primary growth (July) and vegetative regrowth (August), were grazed by twelve Friesian steers (mean body-weights throughout experiment 152–231 kg) at daily allocation rates of forage which provided dry matter (DM) intakes ranging from 28 to 36 g/kg body-weight).2. Total nitrogen contents of the three ryegrasses declined with season (37–20 g/kg DM), but in vitro organic matter (OM) digestibilities were relatively constant (0.76–0.79). The clovers had higher N (average 366 g/kg DM) and lower fibre and water-soluble carbohydrate contents than the grasses, and in vitro OM digestibilities of 0.70 and 076 respectively.3. Duodenal digesta samples were obtained, using a portable sampling apparatus from the animals grazing the pasture, and estimates of the flow of nutrients into the small intestine were derived using two indigestible markers which were continuously infused into the rumen using a portable infusion apparatus. Forage intakes were calculated from estimates of faecal output of indigestible OM and the predicted in vivo OM digestibilities of the forages consumed. Coefficients of variation for OM flow to the small intestine and OM intake were 11.8 and 10.9 % respectively.4. The apparent digestion of OM in the rumen ranged from 722 to 741 g/kg digestible OM intake and from 681 to 711 g/kg digestible OM intake for the grass and clover diets respectively. Substantial losses of ingested N before the small intestine were measured on all diets except the regrowth ryegrass. Losses were 0.30 and 040 g/g N intake on the primary growth ryegrass and the regrowth clover respectively; N contents were 37 and 39 g/kg DM respectively.5. Comparison of the values obtained at pasture with that obtained when similar diets were offered to housed cattle (Beever et al. 1985) indicated that combined relations relating duodenal OM and non-ammonia-N (NAN) flows to OM and N intakes respectively could be established for each diet. For two diets (primary-growth grass and regrowth clover) the relations were curvilinear; for the remaining diets, the derived relations were linear.6. The effects of forage species, stage of maturity and variations in the stem: leaf ratio in the grasses, on nutrient supply in grazing animals are discussed, and a combined relation for all grazed and harvested forages examined in this and in a previous study reported by Beever et al. (1985) is established. The relation Y = 1.430 - 0.0169X (r072, residual sd 0.140). where Y is NAN flow/N intake (g/g) and X is N content in the crop (gfkg OM), indicates that net losses of N across the rumen will occur on diets which contain more than 25.5 g N/kg OM.7. Possible limitations in the techniques available to measure nutrient digestion and supply in grazing ruminants are discussed.

Subnuclear compartmentalization of sequence-specific transcription factors and regulation of eukaryotic gene expression

2005 ◽  
Vol 83 (4) ◽  
pp. 535-547 ◽  
Author(s):  
Gareth N Corry ◽  
D Alan Underhill
Keyword(s):  
Gene Expression ◽  
Transcription Factor ◽  
Transcription Factors ◽  
Transcriptional Activation ◽  
Current Knowledge ◽  
Nuclear Architecture ◽  
Subnuclear Localization ◽  
Modular Structures

To date, the majority of the research regarding eukaryotic transcription factors has focused on characterizing their function primarily through in vitro methods. These studies have revealed that transcription factors are essentially modular structures, containing separate regions that participate in such activities as DNA binding, protein–protein interaction, and transcriptional activation or repression. To fully comprehend the behavior of a given transcription factor, however, these domains must be analyzed in the context of the entire protein, and in certain cases the context of a multiprotein complex. Furthermore, it must be appreciated that transcription factors function in the nucleus, where they must contend with a variety of factors, including the nuclear architecture, chromatin domains, chromosome territories, and cell-cycle-associated processes. Recent examinations of transcription factors in the nucleus have clarified the behavior of these proteins in vivo and have increased our understanding of how gene expression is regulated in eukaryotes. Here, we review the current knowledge regarding sequence-specific transcription factor compartmentalization within the nucleus and discuss its impact on the regulation of such processes as activation or repression of gene expression and interaction with coregulatory factors.Key words: transcription, subnuclear localization, chromatin, gene expression, nuclear architecture.

scholarly journals Integrin α3β1 Promotes Invasive and Metastatic Properties of Breast Cancer Cells through Induction of the Brn-2 Transcription Factor

Cancers ◽  
2021 ◽  
Vol 13 (3) ◽  
pp. 480
Author(s):  
Rakshitha Pandulal Miskin ◽  
Janine S. A. Warren ◽  
Abibatou Ndoye ◽  
Lei Wu ◽  
John M. Lamar ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Transcription Factor ◽  
Cancer Cells ◽  
Cell Invasion ◽  
Breast Cancer Cells ◽  
Gene Promoter ◽  
Akt Inhibitor ◽  
Integrin Α3β1

In the current study, we demonstrate that integrin α3β1 promotes invasive and metastatic traits of triple-negative breast cancer (TNBC) cells through induction of the transcription factor, Brain-2 (Brn-2). We show that RNAi-mediated suppression of α3β1 in MDA-MB-231 cells caused reduced expression of Brn-2 mRNA and protein and reduced activity of the BRN2 gene promoter. In addition, RNAi-targeting of Brn-2 in MDA-MB-231 cells decreased invasion in vitro and lung colonization in vivo, and exogenous Brn-2 expression partially restored invasion to cells in which α3β1 was suppressed. α3β1 promoted phosphorylation of Akt in MDA-MB-231 cells, and treatment of these cells with a pharmacological Akt inhibitor (MK-2206) reduced both Brn-2 expression and cell invasion, indicating that α3β1-Akt signaling contributes to Brn-2 induction. Analysis of RNAseq data from patients with invasive breast carcinoma revealed that high BRN2 expression correlates with poor survival. Moreover, high BRN2 expression positively correlates with high ITGA3 expression in basal-like breast cancer, which is consistent with our experimental findings that α3β1 induces Brn-2 in TNBC cells. Together, our study demonstrates a pro-invasive/pro-metastatic role for Brn-2 in breast cancer cells and identifies a role for integrin α3β1 in regulating Brn-2 expression, thereby revealing a novel mechanism of integrin-dependent breast cancer cell invasion.

scholarly journals Thymoquinone, a Dietary Bioactive Compound, Exerts Anti-Inflammatory Effects in Colitis by Stimulating Expression of the Colonic Epithelial PPAR-γ Transcription Factor

Nutrients ◽  
2021 ◽  
Vol 13 (4) ◽  
pp. 1343
Author(s):  
Balaji Venkataraman ◽  
Saeeda Almarzooqi ◽  
Vishnu Raj ◽  
Abdullah T. Alhassani ◽  
Ahmad S. Alhassani ◽  
...  
Keyword(s):  
Transcription Factor ◽  
Activity Index ◽  
Nigella Sativa ◽  
Mapk Signaling ◽  
Mitogen Activated Protein Kinase ◽  
Ppar Γ ◽  
Anti Inflammatory ◽  
Ht 29

Inflammatory bowel diseases (IBD) are chronic inflammatory disorders with increasing incidence and prevalence worldwide. Here, we investigated thymoquinone (TQ), a naturally occurring phytochemical present in Nigella sativa, for anti-inflammatory effects in colonic inflammation. To address this, we used in vivo (mice) and in vitro (HT-29 cells) models in this investigation. Our results showed that TQ treatment significantly reduced the disease activity index (DAI), myeloperoxidase (MPO) activity, and protected colon microscopic architecture. In addition, TQ also reduced the expression of proinflammatory cytokines and mediators at both the mRNA and protein levels. Further, TQ decreased phosphorylation of the activated mitogen-activated protein kinase (MAPK) signaling pathway and nuclear factor kappa B (NF-κB) proteins and enhanced colon epithelial PPAR-γ transcription factor expression. TQ significantly decreased proinflammatory chemokines (CXCL-1 and IL-8), and mediator (COX-2) mRNA expression in HT-29 cells treated with TNF-α. TQ also increased HT-29 PPAR-γ mRNA, PPAR-γ protein expression, and PPAR-γ promoter activity. These results indicate that TQ inhibits MAPK and NF-κB signaling pathways and transcriptionally regulates PPAR-γ expression to induce potent anti-inflammatory activity in vivo and in vitro models of colon inflammation.

scholarly journals Progression and Differentiation of Alveolar Rhabdomyosarcoma Is Regulated by PAX7 Transcription Factor—Significance of Tumor Subclones

Cells ◽  
2021 ◽  
Vol 10 (8) ◽  
pp. 1870
Author(s):  
Klaudia Skrzypek ◽  
Grażyna Adamek ◽  
Marta Kot ◽  
Bogna Badyra ◽  
Marcin Majka
Keyword(s):  
Transcription Factor ◽  
Cell Lines ◽  
Migration Activity ◽  
Id Proteins ◽  
Rh30 Cell ◽  
Str Profile ◽  
And Migration

Rhabdomyosarcoma (RMS), is the most frequent soft tissue tumor in children that originates from disturbances in differentiation process. Mechanisms leading to the development of RMS are still poorly understood. Therefore, by analysis of two RMS RH30 cell line subclones, one subclone PAX7 negative, while the second one PAX7 positive, and comparison with other RMS cell lines we aimed at identifying new mechanisms crucial for RMS progression. RH30 subclones were characterized by the same STR profile, but different morphology, rate of proliferation, migration activity and chemotactic abilities in vitro, as well as differences in tumor morphology and growth in vivo. Our analysis indicated a different level of expression of adhesion molecules (e.g., from VLA and ICAM families), myogenic microRNAs, such as miR-206 and transcription factors, such as MYOD, MYOG, SIX1, and ID. Silencing of PAX7 transcription factor with siRNA confirmed the crucial role of PAX7 transcription factor in proliferation, differentiation and migration of RMS cells. To conclude, our results suggest that tumor cell lines with the same STR profile can produce subclones that differ in many features and indicate crucial roles of PAX7 and ID proteins in the development of RMS.

scholarly journals RIP3 impedes transcription factor EB to suppress autophagic degradation in septic acute kidney injury

2021 ◽  
Vol 12 (6) ◽  
Author(s):  
Ruizhao Li ◽  
Xingchen Zhao ◽  
Shu Zhang ◽  
Wei Dong ◽  
Li Zhang ◽  
...  
Keyword(s):  
Acute Kidney Injury ◽  
Transcription Factor ◽  
Nuclear Translocation ◽  
Kidney Injury ◽  
Septic Acute Kidney Injury ◽  
Transcription Factor Eb ◽  
Autophagic Degradation ◽  
Lps Treatment

AbstractAutophagy is an important renal-protective mechanism in septic acute kidney injury (AKI). Receptor interacting protein kinase 3 (RIP3) has been implicated in the renal tubular injury and renal dysfunction during septic AKI. Here we investigated the role and mechanism of RIP3 on autophagy in septic AKI. We showed an activation of RIP3, accompanied by an accumulation of the autophagosome marker LC3II and the autophagic substrate p62, in the kidneys of lipopolysaccharide (LPS)-induced septic AKI mice and LPS-treated cultured renal proximal tubular epithelial cells (PTECs). The lysosome inhibitor did not further increase the levels of LCII or p62 in LPS-treated PTECs. Moreover, inhibition of RIP3 attenuated the aberrant accumulation of LC3II and p62 under LPS treatment in vivo and in vitro. By utilizing mCherry-GFP-LC3 autophagy reporter mice in vivo and PTECs overexpression mRFP-GFP-LC3 in vitro, we observed that inhibition of RIP3 restored the formation of autolysosomes and eliminated the accumulated autophagosomes under LPS treatment. These results indicated that RIP3 impaired autophagic degradation, contributing to the accumulation of autophagosomes. Mechanistically, the nuclear translocation of transcription factor EB (TFEB), a master regulator of the lysosome and autophagy pathway, was inhibited in LPS-induced mice and LPS-treated PTECs. Inhibition of RIP3 restored the nuclear translocation of TFEB in vivo and in vitro. Co-immunoprecipitation further showed an interaction of RIP3 and TFEB in LPS-treated PTECs. Also, the expression of LAMP1 and cathepsin B, two potential target genes of TFEB involved in lysosome function, were decreased under LPS treatment in vivo and in vitro, and this decrease was rescued by inhibiting RIP3. Finally, overexpression of TFEB restored the autophagic degradation in LPS-treated PTECs. Together, the present study has identified a pivotal role of RIP3 in suppressing autophagic degradation through impeding the TFEB-lysosome pathway in septic AKI, providing potential therapeutic targets for the prevention and treatment of septic AKI.

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