Tannin-rich forage as a methane mitigation strategy for cattle and the implications for rumen microbiota

2021 ◽  
Vol 61 (1) ◽  
pp. 26 ◽  
Author(s):  
Gisele M. Fagundes ◽  
Gabriela Benetel ◽  
Mateus M. Carriero ◽  
Ricardo L. M. Sousa ◽  
James P. Muir ◽  
...  
Keyword(s):  
Production Systems ◽  
Condensed Tannin ◽  
Quantitative Polymerase Chain Reaction ◽  
Methanogenic Archaea ◽  
Polymerase Chain Reaction Analysis ◽  
Gliricidia Sepium ◽  
Desmodium Ovalifolium ◽  
Ruminal Bacteria ◽  
Rumen Microbiota

Context Methane from ruminant livestock systems contributes to the greenhouse effect on the environment, which justifies the adoption of novel feed strategies that mitigate enteric emissions. Aims We investigated the effects of the condensed tannin (CT)-rich legumes Flemingia macrophylla, Leucaena leucocephala, Stylosanthes guianensis, Gliricidia sepium, Cratylia argentea, Cajanus cajan, Desmodium ovalifolium, Macrotyloma axillare, Desmodium paniculatum and Lespedeza procumbens on in vitro methane emissions and rumen microbiota for beef cattle. Methods Four rumen-cannulated Nellore cattle grazing a tropical grass pasture were used as inoculum donors. Key results Real-time quantitative polymerase chain reaction analysis revealed that the abundance of Ruminococcus flavefaciens, methanogenic archaea and protozoa populations were reduced (P £ 0.05), whereas total ruminal bacteria were enhanced in the presence of CT. Our study also revealed a positive (P £ 0.05) relationship between CT and Fibrobacter succinogenes abundance. Reactive CT from L. leucocephala, D. paniculatum and L. procumbens resulted in decreased (P £ 0.05) isoacid content and methane production. Conclusions L. leucocephala, D. paniculatum and L. procumbens have the potential to suppress rumen methanogenesis. However, in vitro fermentation of L. leucocephala resulted in greater (P £ 0.05) degradability percentages than the other two species. Implications CT in legume species will have potential as part of an overall nutritional strategy to manipulate rumen microbiota and mitigate enteric methanogenesis in livestock production systems.

scholarly journals CircATRNL1 promotes epithelial–mesenchymal transition in endometriosis by upregulating Yes-associated protein 1 in vitro

2020 ◽  
Vol 11 (7) ◽  
Author(s):  
Dandan Wang ◽  
Yajuan Luo ◽  
Guangwei Wang ◽  
Qing Yang
Keyword(s):  
Epithelial Mesenchymal Transition ◽  
Quantitative Polymerase Chain Reaction ◽  
Polymerase Chain Reaction Analysis ◽  
In Vitro Assays ◽  
Migration And Invasion ◽  
Circular Rnas ◽  
Mesenchymal Transition ◽  
Inhibitory Function ◽  
Ishikawa Cells

Abstract Endometriosis is a common and benign gynecological disorder but exhibits malignant features. However, the underlying pathogenesis and pathophysiology of endometriosis remain unclear. Circular RNAs have been demonstrated to participate in the occurrence and progression of multiple diseases. This study was aimed to explore the roles of circATRNL1 in endometriosis in vitro. Based on the results of reverse transcription-quantitative polymerase chain reaction analysis, we found significant upregulation of circATRNL1 and Yes-associated protein 1 (YAP1), while downregulation of miR-141-3p and miR-200a-3p in ectopic tissues compared to eutopic tissues. The immunohistochemistry and western blot analysis showed differentially expressed epithelial–mesenchymal transition (EMT) markers between EuEM and EcEM tissues. The in vitro assays indicated that overexpression of circATRNL1 could promote the proliferation, migration, and invasion of Ishikawa cells, and induce EMT process, while circATRNL1 silencing showed the opposite effect. The mechanical investigation indicated that circATRNL1 upregulated YAP1 by sponging miR-141-3p and miR-200a-3p. Gain-of-function assays validated the inhibitory function of miR-141-3p and miR-200a-3p in endometriosis. The results of rescue assays confirmed the function of circATRNL1–miR-141-3p/miR-200a-3p–YAP1 axis on Ishikawa cells. Our findings demonstrate that abnormal upregulation of circATRNL1 regulates cell proliferation and motility and promotes EMT process via the miR-141-3p/miR-200a-3p–YAP1 axis in vitro, which could contribute to the progression of endometriosis.

scholarly journals The Viability and Anti-Inflammatory Effects of Hyaluronic Acid-Chitlac-Tracimolone Acetonide-β-Cyclodextrin Complex on Human Chondrocytes

Cartilage ◽  
2020 ◽  
pp. 194760352090865 ◽  
Author(s):  
Elena Tarricone ◽  
Rossella Elia ◽  
Elena Mattiuzzo ◽  
Alessia Faggian ◽  
Assunta Pozzuoli ◽  
...  
Keyword(s):  
Gene Expression ◽  
Hyaluronic Acid ◽  
Articular Chondrocytes ◽  
Quantitative Polymerase Chain Reaction ◽  
Polymerase Chain Reaction Analysis ◽  
Human Chondrocytes ◽  
Cyclodextrin Complex ◽  
Modified Chitosan ◽  
Anti Inflammatory

Objective To compare the effects of the complex triamcinolone acetonide-hydroxypropyl-β-cyclodextrin (TA-CD) on in vitro inflamed primary human articular chondrocytes in the presence or absence of the mixture hyaluronic acid-Chitlac, a lactose-modified chitosan (HA-CTL). Design Changes in cell viability and pro-inflammatory cytokines gene expression were analyzed in human chondrocytes using an in vitro model of macrophage-mediated inflammation. Human monocytes U937 were differentiated to macrophages by phorbol 12-myristate 13-acetate (PMA) and lipopolysaccharides (LPS). The anti-inflammatory effects of the complex TA-CD and HA-CTL mixture were assessed on chondrocytes exposed for 24 hours to U937 conditioned medium (CM), by quantitative polymerase chain reaction analysis. Results The TA-CD viability was enhanced by the presence of the HA-CTL mixture in chondrocyte cultures. The exposure of cells to CM significantly increased interleukin-1β and interleukin-6 gene expression, and when the complex TA-CD was added to the inflamed cells, gene transcription of cytokines was restored to near baseline values, both in the presence or in the absence of HA-CTL mixture. Conclusion The addition of HA-CTL mixture significantly attenuated cytotoxicity induced by TA and preserved the anti-inflammatory effects, thus confirming the chondroprotective role of the HA-CTL mixture.

scholarly journals In Vivo Association of Ku with Mammalian Origins of DNA Replication

2001 ◽  
Vol 12 (11) ◽  
pp. 3386-3401 ◽  
Author(s):  
Olivia Novac ◽  
Diamanto Matheos ◽  
Felipe D. Araujo ◽  
Gerald B. Price ◽  
Maria Zannis-Hadjopoulos
Keyword(s):  
Dna Replication ◽  
Quantitative Polymerase Chain Reaction ◽  
Polymerase Chain Reaction Analysis ◽  
Origin Firing ◽  
Origin Binding Protein ◽  
M Phase ◽  
Polymerase Chain ◽  
In Cells

Ku is a heterodimeric (Ku70/86-kDa) nuclear protein with known functions in DNA repair, V(D)J recombination, and DNA replication. Here, the in vivo association of Ku with mammalian origins of DNA replication was analyzed by studying its association withors8 and ors12, as assayed by formaldehyde cross-linking, followed by immunoprecipitation and quantitative polymerase chain reaction analysis. The association of Ku with ors8 and ors12 was also analyzed as a function of the cell cycle. This association was found to be approximately fivefold higher in cells synchronized at the G1/S border, in comparison with cells at G0, and it decreased by approximately twofold upon entry of the cells into S phase, and to near background levels in cells at G2/M phase. In addition, in vitro DNA replication experiments were performed with the use of extracts from Ku80+/+ and Ku80−/− mouse embryonic fibroblasts. A decrease of ∼70% in in vitro DNA replication was observed when the Ku80−/− extracts were used, compared with the Ku80+/+ extracts. The results indicate a novel function for Ku as an origin binding-protein, which acts at the initiation step of DNA replication and dissociates after origin firing.

scholarly journals Improvement of Bacterial Vaginosis by Oral Lactobacillus Supplement: A Randomized, Double-Blinded Trial

2021 ◽  
Vol 11 (3) ◽  
pp. 902
Author(s):  
Ta-Chin Lin ◽  
I-Ling Hsu ◽  
Wan-Hua Tsai ◽  
Yi-Chih Chu ◽  
Lung-Ching Kuan ◽  
...  
Keyword(s):  
Bacterial Vaginosis ◽  
In Vitro Selection ◽  
Quantitative Polymerase Chain Reaction ◽  
Polymerase Chain Reaction Analysis ◽  
Beneficial Effects ◽  
Vaginal Microflora ◽  
Improvement Effect ◽  
Double Blinded ◽  
Blinded Trial

Bacterial vaginosis (BV) is the most common vaginal infection globally, with a high recurrent rate after antibiotic treatment. Probiotics consumption is known to improve BV with different efficacy among species or strains. After in vitro selection of Lactobacillus strains with growth inhibition and preventing adhesion to HeLa cervical epithelial cells, a randomized and double-blinded trial of two Lactobacillus formula, namely, VGA-1 and VGA-2, in BV patients with Nugent scores of 4–10 was conducted. Among 37 subjects who completed the trial, we observed significantly decreased Nugent scores in both VGA-1 (n = 18) and VGA-2 (n = 19) consumption groups. VGA-1 consumption significantly improved vaginal discharge odor/color and itching at both 2-week and 4-week-consumption, but those only observed after a 4-week-consumption in the VGA-2 group. We also observed a tendency to reduce recurrent rates among enrolled participants after VGA-1 or VGA-2 consumption. The improvement effect of VGA-1/VGA-2 was associated with the significant reduction of interleukin-6 expression after 4-week-consumption and the restoration of normal vaginal microflora by quantitative polymerase chain reaction analysis. In conclusion, VGA-1 or VGA-2 displayed beneficial effects in BV patients, but the VGA-1 formula showed a better efficacy, potentially used for BV intervention.

scholarly journals Lovastatin as a supplement to mitigate rumen methanogenesis: an overview

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Amaury Ábrego-Gacía ◽  
Héctor M. Poggi-Varaldo ◽  
Vania Robles-González ◽  
Teresa Ponce-Noyola ◽  
Graciano Calva-Calva ◽  
...  
Keyword(s):  
Volatile Fatty Acids ◽  
Methanogenic Archaea ◽  
Methane Emissions ◽  
Ruminal Fermentation ◽  
Enteric Fermentation ◽  
Feed Composition ◽  
Rumen Microbiota ◽  
Alpha And Beta Diversity

AbstractMethane from enteric fermentation is the gas with the greatest environmental impact emitted by ruminants. Lovastatin (Lv) addition to feedstocks could be a strategy to mitigate rumen methane emissions via decreasing the population of methanogenic archaea (MA). Thus, this paper provides the first overview of the effects of Lv supplementation, focusing on the inhibition of methane production, rumen microbiota, and ruminal fermentation. Results indicated that Lv treatment had a strong anti-methanogenic effect on pure strains of MA. However, there are uncertainties from in vitro rumen fermentation trials with complex substrates and rumen inoculum.Solid-state fermentation (SSF) has emerged as a cost-effective option to produce Lv. In this way, SSF of agricultural residues as an Lv-carrier supplement in sheep and goats demonstrated a consistent decrease in ruminal methane emissions. The experimental evidence for in vitro conditions showed that Lv did not affect the volatile fatty acids (VFA). However, in vivo experiments demonstrated that the production of VFA was decreased. Lv did not negatively affect the digestibility of dry matter during in vitro and in vivo methods, and there is even evidence that it can induce an increase in digestibility. Regarding the rumen microbiota, populations of MA were reduced, and no differences were detected in alpha and beta diversity associated with Lv treatment. However, some changes in the relative abundance of the microbiota were induced. Further studies are recommended on: (i) Lv biodegradation products and stability, as well as its adsorption onto the solid matter in the rumen, to gain more insight on the “available” or effective Lv concentration; and (ii) to determine whether the effect of Lv on ruminal fermentation also depends on the feed composition and different ruminants.

Density and morphological characterization of bacteria in integrated and exclusive production systems

2020 ◽  
Vol 13 (11) ◽  
pp. 1
Author(s):  
A. R. B. Zanco ◽  
A. Ferreira ◽  
G. C. M. Berber ◽  
E. N. Gonzaga ◽  
D. C. C. Sabino
Keyword(s):  
Bacterial Community ◽  
Rainy Season ◽  
Production Systems ◽  
Cropping System ◽  
Morphological Characterization ◽  
Native Forest ◽  
Exclusive Production ◽  
Colony Forming Units

The different integrated production systems can directly interfere with its bacterial community. The present study aimed to assess density, bacterial diversity and the influence of dry and rainy season in different integrated and an exclusive production system. The fallow and a native forest area was assessed to. Samples were collected in 2012 March and September. The isolation were carried out into Petri dishes containing DYGS medium. The number of colony forming units (CFU) was counted after 48 hours and. The bacterial density ranged between 106 and 107 CFU g-1 soil. The crop system affected the dynamics of the bacterial community only in the rainy season. The rainy season showed greater density of total bacteria when compared to the dry period regardless of the cropping system. The dendrograms with 80 % similarity showed thirteen and fourteen groups in the rainy and dry seasons. Isolates with the capacity to solubilize phosphate in vitro were obtained from all areas in the two seasons, but this feature has been prevalent in bacteria isolated during the rainy season

scholarly journals The Differential Metabolomes in Cumulus and Mural Granulosa Cells from Human Preovulatory Follicles

2021 ◽  
Author(s):  
Er-Meng Gao ◽  
Bongkoch Turathum ◽  
Ling Wang ◽  
Di Zhang ◽  
Yu-Bing Liu ◽  
...  
Keyword(s):  
Oocyte Maturation ◽  
Granulosa Cells ◽  
Quantitative Polymerase Chain Reaction ◽  
Cumulus Cells ◽  
Cholesterol Transport ◽  
Vitro Fertilization ◽  
Expression Of Genes ◽  
Preovulatory Follicles ◽  
Morphological Differences

AbstractThis study evaluated the differences in metabolites between cumulus cells (CCs) and mural granulosa cells (MGCs) from human preovulatory follicles to understand the mechanism of oocyte maturation involving CCs and MGCs. CCs and MGCs were collected from women who were undergoing in vitro fertilization (IVF)/intracytoplasmic sperm injection (ICSI) treatment. The differences in morphology were determined by immunofluorescence. The metabolomics of CCs and MGCs was measured by liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) followed by quantitative polymerase chain reaction (qPCR) and western blot analysis to further confirm the genes and proteins involved in oocyte maturation. CCs and MGCs were cultured for 48 h in vitro, and the medium was collected for detection of hormone levels. There were minor morphological differences between CCs and MGCs. LC-MS/MS analysis showed that there were differences in 101 metabolites between CCs and MGCs: 7 metabolites were upregulated in CCs, and 94 metabolites were upregulated in MGCs. The metabolites related to cholesterol transport and estradiol production were enriched in CCs, while metabolites related to antiapoptosis were enriched in MGCs. The expression of genes and proteins involved in cholesterol transport (ABCA1, LDLR, and SCARB1) and estradiol production (SULT2B1 and CYP19A1) was significantly higher in CCs, and the expression of genes and proteins involved in antiapoptosis (CRLS1, LPCAT3, and PLA2G4A) was significantly higher in MGCs. The level of estrogen in CCs was significantly higher than that in MGCs, while the progesterone level showed no significant differences. There are differences between the metabolomes of CCs and MGCs. These differences may be involved in the regulation of oocyte maturation.

scholarly journals Anti-Metastatic and Anti-Angiogenic Effects of Curcumin Analog DK1 on Human Osteosarcoma Cells In Vitro

2021 ◽  
Vol 14 (6) ◽  
pp. 532
Author(s):  
Muhammad Nazirul Mubin Aziz ◽  
Nurul Fattin Che Rahim ◽  
Yazmin Hussin ◽  
Swee Keong Yeap ◽  
Mas Jaffri Masarudin ◽  
...  
Keyword(s):  
Cell Lines ◽  
Safety Profile ◽  
Quantitative Polymerase Chain Reaction ◽  
Tube Formation ◽  
Osteosarcoma Cell ◽  
Curcumin Analog ◽  
Human Osteosarcoma ◽  
Human Osteosarcoma Cells ◽  
Human Osteosarcoma Cell

Osteosarcoma (OS) is a life-threatening malignant bone tumor associated with poor prognosis among children. The survival rate of the patient is still arguably low even with intensive treatment provided, plus with the inherent side effects from the chemotherapy, which gives more unfavorable outcomes. Hence, the search for potent anti-osteosarcoma agent with promising safety profile is still on going. Natural occurring substance like curcumin has gained a lot of attention due to its splendid safety profile as well as it pharmacological advantages such as anti-metastasis and anti-angiogenesis. However, natural curcumin was widely known for its poor cellular uptake, which undermines all potential that it possesses. This prompted the development of synthetically synthesized curcuminoid analog, known as (Z)-3-hydroxy-1-(2-hydroxyphenyl)-3-phenylprop-2- en-1-one (DK1). In this present study, in vitro scratch assay, transwell migration/invasion assay, HUVEC tube formation assay, and ex vivo rat aortic ring assays were performed in order to investigate the anti-metastatic and anti-angiogenic potential of DK1. For further comprehension of DK1 mechanism on human osteosarcoma cell lines, microarray gene expression analysis, quantitative polymerase chain reaction (qPCR), and proteome profiler were adopted, providing valuable forecast from the expression of important genes and proteins related to metastasis and angiogenesis. Based on the data gathered from the bioassays, DK1 was able to inhibit the metastasis and angiogenesis of human osteosarcoma cell lines by significantly reducing the cell motility, number of migrated and invaded cells as well as the tube formation and micro-vessels sprouting. Additionally, DK1 also has significantly regulated several cancer pathways involved in OS proliferation, metastasis, and angiogenesis such as PI3K/Akt and NF-κB in both U-2 OS and MG-63. Regulation of PI3K/Akt caused up-regulation of genes related to metastasis inhibition, namely, PTEN, FOXO, PLK3, and GADD45A. Meanwhile, NF-κB pathway was regulated by mitigating the expression of NF-κB activator such as IKBKB and IKBKE in MG-63, whilst up-regulating the expression of NF-κB inhibitors such as NFKBIA and NFKBIE in U-2 OS. Finally, DK1 also has successfully hindered the metastatic and angiogenic capability of OS cell lines by down-regulating the expression of pro-metastatic genes and proteins like MMP3, COL11A1, FGF1, Endoglin, uPA, and IGFBP2 in U-2 OS. Whilst for MG-63, the significantly down-regulated oncogenes were Serpin E1, AKT2, VEGF, uPA, PD-ECGF, and Endoglin. These results suggest that curcumin analog DK1 may serve as a potential new anti-osteosarcoma agent due to its anti-metastatic and anti-angiogenic attributes.

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